(Advances in Agronomy Volume 143) Donald L. Sparks (Eds.) - Academic Press (2017) PDF

ADVANCES IN AGRONOMY
Advisory Board
MARY BETH KIRKHAM RONALD L. PHILLIPS
Kansas State University University of Minnesota
KATE M. SCOW ALFRED E. HARTEMINK

University of California, Davis University of Wisconsin - Madison
Emeritus Advisory Board Members

JOHN S. BOYER LARRY P. WILDING
University of Delaware Texas A&M University
EUGENE J. KAMPRATH PAUL M. BERTSCH

North Carolina State University University of Kentucky
MARTIN ALEXANDER
Cornell University
Academic Press is an imprint of Elsevier
50 Hampshire Street, 5th Floor, Cambridge, MA 02139, United States
525 B Street, Suite 1800, San Diego, CA 92101–4495, United States
The Boulevard, Langford Lane, Kidlington, Oxford OX5 1GB, United Kingdom
125 London Wall, London EC2Y 5AS, United Kingdom
First edition 2017
Copyright © 2017 Elsevier Inc. All rights reserved.
No part of this publication may be reproduced or transmitted in any form or by any means,
electronic or mechanical, including photocopying, recording, or any information storage and
retrieval system, without permission in writing from the publisher. Details on how to seek
permission, further information about the Publisher’s permissions policies and our
arrangements with organizations such as the Copyright Clearance Center and the Copyright
Licensing Agency, can be found at our website: www.elsevier.com/permissions.
This book and the individual contributions contained in it are protected under copyright by
the Publisher (other than as may be noted herein).
Notices
Knowledge and best practice in this field are constantly changing. As new research and
experience broaden our understanding, changes in research methods, professional practices,
or medical treatment may become necessary.
Practitioners and researchers must always rely on their own experience and knowledge in
evaluating and using any information, methods, compounds, or experiments described
herein. In using such information or methods they should be mindful of their own safety and
the safety of others, including parties for whom they have a professional responsibility.
To the fullest extent of the law, neither the Publisher nor the authors, contributors, or editors,
assume any liability for any injury and/or damage to persons or property as a matter of
products liability, negligence or otherwise, or from any use or operation of any methods,
products, instructions, or ideas contained in the material herein.
ISBN: 978-0-12-812421-5
ISSN: 0065-2113
For information on all Academic Press publications

visit our website at https://www.elsevier.com/books-and-journals
Publisher: Zoe Kruze

Acquisition Editor: Alex White
Editorial Project Manager: Helene Kabes
Production Project Manager: Magesh Kumar Mahalingam
Cover Designer: Miles Hitchen
Typeset by SPi Global, India
CONTRIBUTORS
Ronald Corstanje
Cranfield Soil and AgriFood Institute, School of Environment, Energy and AgriFood,
Cranfield University, Bedfordshire, United Kingdom
Richard M. Cruse
Iowa State University, Ames, IA, United States
Graham Halcro
Jerry L. Hatfield
USDA-ARS, National Laboratory for Agriculture and the Environment, Ames, IA,
United States
Michael H.B. Hayes
Carbolea Research Group, Chemical Sciences, University of Limerick, Limerick, Ireland
Feng-Min Li
State Key Laboratory of Grassland Agroecosystems, School of Life Sciences, Institute of Arid
Agroecology, Lanzhou University, Lanzhou, China
Abdul M. Mouazen
Cranfield University, Bedfordshire, United Kingdom; Precision Soil and Crop Engineering
Group, Faculty of Bioscience Engineering, Ghent University, Ghent, Belgium
David Mulla
University of Minnesota, Saint Paul, MN, United States
Rosaleen Mylotte
Chemical Sciences, University of Limerick, Limerick, Ireland
Said Nawar
Thomas J. Sauer
USDA-ARS, National Laboratory for Agriculture and the Environment, Ames, IA,
United States
Kadambot H.M. Siddique
UWA Institute of Agriculture, The University of Western Australia, Perth, WA, Australia
Roger S. Swift
Queensland Alliance for Agriculture and Food Innovation, University of Queensland,
Brisbane, QLD, Australia
vii
viii Contributors
Neil C. Turner
Jian-Yong Wang
Agroecology, Lanzhou University, Lanzhou, China; UWA Institute of Agriculture,
The University of Western Australia, Perth, WA, Australia
You-Cai Xiong
Agroecology, Lanzhou University, Lanzhou, China
PREFACE
Volume 143 contains four outstanding reviews on topics in the crop and soil
sciences. Chapter 1 is a thought-provoking review on soils and the too often
lack of recognition of their critical role in the water, food, and energy nexus.
Chapter 2 presents advances in our understanding of soil organic matter, par-
ticularly the humin fraction. Chapter 3 presents a meta-analysis of impacts of
drought stress on morphophysiological traits, biochemical characteristics,
and yield in wheat. Chapter 4 is a review on delineation of soil management
zones for variable rate fertilization.
I am grateful to the authors for their first-rate contributions.
Donald L. Sparks
Newark, DE, United States
ix
CHAPTER ONE
Soil: The Forgotten Piece of the

Water, Food, Energy Nexus
Jerry L. Hatfield*,1, Thomas J. Sauer*, Richard M. Cruse†
*USDA-ARS, National Laboratory for Agriculture and the Environment, Ames, IA, United States
†
Iowa State University, Ames, IA, United States
1
Corresponding author: e-mail address: jerry.hatfield@ars.usda.gov
Contents
1. Introduction 2
2. Soil—A Missing Nexus Component 5
2.1 Functionality of Soil 5
2.2 Current State of Soils 8
2.3 Soil Degradation 10
2.4 Soil Erosion 12
3. Scale of the Interface Between Soils and Ecosystem Services 28
3.1 Field 28
3.2 Landscape 30
3.3 Watershed 30
4. Implications of Soil as a Component of the Food, Energy, Water Nexus 32
5. Challenges 34
5.1 Enhancing the Soil Resource to Reduce Soil Degradation 34
5.2 Soil as a Component in the Food, Energy, Water Nexus 35
References 36
Abstract
The water, food, energy nexus has prompted sustainability concerns as interactions
between these interdependent human needs are degrading natural resources required
for a secure future world. Discussions about the future needs for food, water, and energy
to support the increasing world population have ignored our soil resource that is the
cornerstone or our capacity to produce food, capture water, and generate energy from
biological systems. Soil scientists often recognize soils as a critical component of food,
energy, or water security; however, the translation of that awareness into action strat-
egies to either enhance public recognition of soil resource importance or improve soil
management is lacking. Food, water, and energy security represents the current and
future challenge of sustaining humankind while protecting the environment. These
interactions are recognized by scientists, but the linkage to policy decisions or imple-
mentation of strategies to create positive outcomes for food, energy, or water enhance-
ment is lacking. If we consider that soil is responsible for 99% of the world’s food
production, then the importance of soil in the food, energy, water nexus becomes
Advances in Agronomy, Volume 143 # 2017 Elsevier Inc. 1

ISSN 0065-2113 All rights reserved.
http://dx.doi.org/10.1016/bs.agron.2017.02.001
2 Jerry L. Hatfield et al.
apparent. If we further consider that soil erosion is the major factor, affecting soil deg-
radation and declines in productivity are directly related to degradation of the soil
resource, then the implications of soil in the context of increasing food, energy, and
water security becomes more evident. However, if the attitude is one that technology
will provide answers to these problems, then the soil degradation rate will continue to
increase and we will reach a tipping point in which technological advances will not be
able to overcome the impacts of a reduced topsoil depth coupled with a more variable
climate. Soil is the forgotten piece of the food, energy, water nexus; however, the over-
sight extends beyond this nexus to include many of the ecological services required by
humankind.
1. INTRODUCTION
The water, food, energy nexus has prompted sustainability concerns as
interactions between these interdependent human needs is degrading natural
resources required for a secure future world. Discussions about the future
needs for food, water, and energy to support the increasing world population
have ignored our soil resource that is the cornerstone or our capacity to pro-
duce food, capture water, and generate energy from biological systems. Soil
scientists often recognize soils as a critical component of food, energy, or
water security; however, the translation of that awareness into action strat-
egies to either enhance public recognition of soil resource importance or to
improve soil management is lacking. Food, water, and energy security rep-
resents the current and future challenge of sustaining humankind while
protecting the environment. These interactions are recognized by scientists,
but the linkage to policy decisions or implementation of strategies to create
positive outcomes for food, energy, or water enhancement is lacking
(Bouma and McBratney, 2013). Banwart (2011) recently identified and cau-
tioned against the potential impact of soil degradation on our ability to
achieve sustainability, further supported by Hatfield and Walthall (2015)
articulating that soil degradation and loss of soil resources would impact
our ability to produce sufficient quantity and quality food to meet world
demands. As a framework for enhancing our understanding of soil’s role
in the food–energy–water nexus, we propose the following conceptual dia-
gram (Fig. 1). Hewitt et al. (2015) provided a bold statement that soil is the
most overlooked component in ecosystem services and policy level deci-
sions. Recent food, energy, water nexus evaluations have highlighted the
nexus component interactions (Endo et al., 2015). In the assessment by
Endo et al. (2015) soils are identified as a critical factor only in their
Soil: The Forgotten Piece of the Water, Food, Energy Nexus 3
Fig. 1 Conceptual diagram of the role of soil in the food, energy, water nexus.
supporting role for crop production; this is particularly concerning as

Adhikari and Hartemink’s (2016) review support the importance of soils
in ecosystem functions of which provisioning of food, water, and energy
are central themes and that the recognition of soil’s critical role is lacking.
We have apparently lost sight of the important role of soil and assume that
the soils capability to supply ecosystem functions will forever exist. The
future food, water, and energy challenges facing us require that we under-
stand the value of our soil and that the condition of the soil resource dictates
functions within the food, energy, water nexus.
Critical to this discussion is defining what is encompassed in food,
energy, and water security to quantitatively determine the demands for food,
water, and energy projected for the reminder of the century (Gulati et al,
2013; Smajgl et al., 2016; Taniguchi et al., 2015). The projections for food
requirements are often targeted toward the population of 9 + billion people
by 2050; however, there is a need to sustain that population beyond 2050
and meet their water and energy needs as well. Feeding 9 billion people pre-
sents a major challenge because of multiple factors (Fabiola and Dalila, 2016).
Increased production is not possible without new lands under cultivation
and/or increased crop yields (Gregory and George, 2011). Recent estimates
by Smith et al. (2010) show per capita land area will continue to decrease,
having already declined from 0.415 ha in 1961 to 0.214 ha in 2007, and they
estimated that average cereal yields will need to increase by 25% from the
average of 3.23 t ha1 in 2005–07 to 4.34 t ha1 in 2030. Gregory and
George (2011) found that to meet production requirements for food supply
only 20% would come from new land and 80% from intensification
(increased yields and greater cropping intensity). Increasing land area for
production will lead to increased carbon losses, and for the tropics, the efforts
must be directed toward increasing crop yields rather than clearing more
land (West et al., 2010). Increasing productivity will require increased man-
agement intensity and improved agronomic techniques, and, since the land
resource will become a premium, this option will remain as the most viable
solution.
Water availability becomes a larger issue within regions and distribution
among regions. A recent UNESCO report (WWAP, 2015) summarized
that current agricultural water demand to meet food demands is
unsustainable and will require greater emphasis on increasing water use effi-
ciency (WUE) and reducing water losses (leaching and runoff ). They esti-
mated that water demands for energy production would increase, and
directing resources toward improving energy production efficiency should
be linked with agricultural production systems. Linking efficiency gains in
agriculture with energy production requires examining the connection
between agriculture and energy water use patterns. The water demand
for manufacturing was estimated to increase by 400% by 2050, and this
report suggested that greater emphasis be placed on evaluation of the water
footprint of all sectors (WWAP, 2015). They also suggested that the negative
impacts of climate change on water supply and disruptions in the stability of
the water supply caused by more extreme events would add to the stress in
the food production and water for irrigation. This report proposed that for a
sustainable future a path toward water security be developed in coordination
with food security. Precipitation is the primary source of fresh water, and soil
serves as the reservoir of available water for agriculture. Thus, soil becomes a
central component of ensuring a sustainable path toward food and water
security. However, there will have to be combination of practices integrated
together to achieve all of the nexus-related goals over the next 30+ years.
If we assume that the food, energy, water nexus represents an integration
of ecosystem services, then a framework can be developed to facilitate infor-
mation exchange among different services. Adhikari and Hartemink (2016)
proposed four ecosystem services: provisioning services (providing food,
energy, or water), regulatory services (climate and greenhouse gas regula-
tion, carbon sequestration, water regulation, water purification, soil erosion
and flood control, pest and disease regulation, and pollination and seed dis-
persal), cultural services (recreation/ecotourism, esthetics of the landscape,
cultural heritage, knowledge, and education), and supporting services (hab-
itat, nutrient cycling, and soil formation). Soils are closely linked to provi-
sioning, regulatory, and supporting services and are closely tied with our
ability to develop and preserve a sustainable future. Soils will become the
foundation for building our future, and it will become necessary to deter-
mine how we interject soils into the mainstream discussion about how
we achieve the goals for abundant and clean water, food security, and ade-
quate energy.
2. SOIL—A MISSING NEXUS COMPONENT

2.1 Functionality of Soil
Biophysical soil functions include nutrient cycling, water dynamics, filtering
and buffering, physical stability and support of plant systems and human
structures, and promotion of biodiversity and habitat. There is no one single
definition of soil functions; however, the summary from the World Soil
Information network (2016; www.iSRIC.org) as “Soil is our life support
system. Soils provide anchorage for roots, hold water and nutrients. Soils
are home to myriad microorganisms that fix nitrogen and decompose
organic matter, and armies of microscopic animals as well as earthworms
and termites. We build on soil as well as with it and in it” provides a very
useful summary of the importance of soils in humankind. These broad cat-
egories of soil functions require some expansion to fully understand the vital
nature or soils in the food, energy, water nexus. There are a number of soil
properties related to soil functionality as shown in Fig. 2; however, the abil-
ity of the soil to provide these functions is more complex than merely listing
the soil properties. Our understanding of the linkage between soil properties
and soil functions and the resultant ecosystem services is incomplete
(Adhikari and Hartemink, 2016; Daily et al., 1997; Swinton et al., 2006).
The ability of the soil to provide these functions will, however, depend upon
the state of the soil properties. For example, soil water holding capacity is a
soil property related to water dynamics and filtering and buffering; however,
a sandy soil with a low water holding capacity will not provide those func-
tions as well as a clay loam soil with a high water holding capacity.
For the purpose of incorporating soil into the food, energy, water nexus,
we have to expand our concept of soil functions to link with ecosystem
functions (Adhikari and Hartemink, 2016). Soil is a foundational component
of environmental sustainability problems that link climate change, decline in
biodiversity, water security, energy security, and food security (Bouma and
McBratney, 2013; Hartemink and McBratney, 2008). The concept of soil
functionality may need expansion to include the concept of soil security that
in turn provides the linkage between soil properties, soil functionality, and
ecosystem services (Bouma, 2015; Bouma and McBratney, 2013;
Food, energy, water security
Ecosystem services
Provisioning, regulating, cultural, supporting
Soil functions
Food production
Biological diversity
Carbon sequenstration
Source or raw materials
Availability of nutrients and water
Support for plants and infrastructure
Soil Properties
Soil organic carbon

Soil pH Bulk density Soil biology
Cation exchange capacity
Available water capacity Soil texture
Electrical conductivity Soil porosity
Soil aggregate stability Hydroulic conductivity
Fig. 2 Interface of soil properties relative to soil functions and ecosystem services.
McBratney et al., 2014). Bouma (2015) stated that soil security must result
from enhanced global soil resource and requires the elimination of soil deg-
radation. He suggested four areas in which soil science should be linked to
foster increased awareness in environmental and societal issues. These are
“(i) demonstrating the importance of soils in inter- and transdisciplinary pro-
grams focusing on food, water, climate, biodiversity, and energy problems,
which are environmental issues that are widely acknowledged to be impor-
tant; (ii) focusing research on the seven soil functions (soil water availability,
nutrient cycling, carbon cycling, soil structure, aggregate stability, adequate
rooting depth, and gas exchange) to demonstrate the importance of soil for
widely recognized Ecosystem Services and Sustainable Development Goals;
(iii) reframing reporting of soil studies by not only including technical data
but embedding this in human-interest storylines, building on the deep emo-
tional links between soil and man, and (iv) educating and involving knowl-
edge brokers that can link science with societal partners not only during a
given project but also in the preparatory and implementation phase.”
A key part of this awareness is understanding the linkage among soil and eco-
system services and sustainable development and the well-being of humans
with these latter points being the goals of understanding the food, energy,
water nexus (Bouma, 2014). In their review, Adhikari and Hartemink
(2016) concluded that future evaluations of ecosystem services should focus
on soil functions as the basis for sustainability and recognize that soils are very
complex in order to fully understand the interface of soils with ecosystem
services in the food, energy, water nexus. This will require a more holistic
approach than has been considered in previous studies.
Soil organic carbon is a key contributor to soil function. Lal (2004,
2009a,b, 2011) stated that increasing soil organic carbon by 1 t SOC ha1
year1 would have the potential to increase food grain production in devel-
oping countries by 24–32 million tons annually and would help improve
food security. The cycling of carbon and nutrients, only two of many soil
functions, is dependent upon soil biological diversity (Brussaard, 1997),
and soil degradation threatens soil’s ability to perform all of the functions
(Bender et al., 2016; Hatfield, 2014). Cycling carbon found in plant residues,
roots, and organic manures involves actions of a diverse biological system.
The soil biological system plays a crucial role in soil property modification,
conceptually shown in Fig. 3; preserving and/or enhancing soil biology is a
key to soil biological processes. In a recent review, Madhu and Hatfield
(2013) found that the combination of changes in atmospheric CO2 concen-
tration coupled with changing soil management will affect the soil physical,
chemical, and biological properties. A changing climate will create changes
in the soil biological system and although soils possess the most diverse bio-
logical systems (Bender et al., 2016), soil degradation affects more than one
quarter of the world’s soils, and the increasing intensity of land use will fur-
ther increase degradation and reduce soil biological diversity (Bai et al.,
2008; Lal and Stavi, 2015; Tsiafouli et al., 2015). To improve soil quality
will require a diversity of soil biology but intensively managed systems show
a reduction in soil biodiversity (Tsiafouli et al., 2015). Another component
of promoting soil biological diversity is the creation of a soil microclimate
Soil aggradation climb
es
m
tco
ou
l e
sib Improved water
Vi availability
Improved soil
structure
Improved nutrient
cycling
ss
oce
Organic matter pr
ic
turnover m
na
dy
Biological d
an
e
activity i bl
vis
In
Fig. 3 Process of soil aggradation to restore soil functions.
that provides food, water, oxygen, and shelter from extremes. Soil functions
depicted in Fig. 2 are dependent upon the interaction among soil properties
and represent a spatial and temporal representation of the interaction
between how we manage our soils and link this with the ecosystem
processes.
2.2 Current State of Soils

Mollisols and Alfisols are typically considered the world’s most productive
soils (Eswaran et al., 2012). These soils are derived under native prairie and
forest vegetation, respectively, have inherently fertile parent materials, and
have had favorable climates that supports rich growth of the respective veg-
etation types. The topography is such that the surface soil is suitably stable;
that is, the soils with prairie or forest vegetation cover were not sufficiently
erosive to preclude soil profile development with relatively high levels of soil
organic matter accumulations near the soil surface. Mollisols, considered the
world’s breadbasket soils, are typically found in the northern and southern
hemispheres in mid-latitudes and constitute about 7% of the world’s soil
resource base (Eswaran et al., 2012). They typically support well-developed
and relatively prosperous cultures of the United States, South America,
Europe, and Asia and most experience intensive agricultural production
practices. Climatic conditions that have favored rich prairie plant growth
for centuries fostering Mollisol development are also generally favorable for
domesticated crop production. This poses a soil degradation challenge—
while past climates, especially rainfall, coupled with typically rolling topog-
raphy and heavy vegetation favored sustaining the soil resource, modern
production approaches with intensive tillage leaving the soil surface bare
much of the year accelerates degradation (Liu et al., 2012; Montgomery,
2007). Alfisols occupy approximately 10% of the world’s soil resource
base, are typically found in cooler regions than are Mollisols, and favorably
support agricultural production but to a lesser degree than Mollisols
(Eswaran et al., 2012). Climate supporting forest growth also typically sup-
ports agricultural crops, but when left exposed or are aggressively disturbed
through activities such as tillage are easily degraded. These two soil orders
occupy approximately 17% of the world’s landscape, yet only 10%–12%
of the world’s lands have no natural limitations for agricultural production
(Purakayastha et al., 2012). That is, even our best soils for crop production
often have production limitations.
Not all cultures have the rich soil resources coupled with favorable cli-
mates such as occurs on many of the Mollisols and Alfisols. Soils in the tro-
pics or subtropics, warmer areas with shorter or no winters, tend to be much
older, more weathered, and less productive than Mollisols and Alfisols even
with favorable precipitation. These soils are susceptible to and have experi-
enced substantial degradation, dominantly caused by marginal land manage-
ment practices and water-induced soil erosion. None of the world’s prime
farmland is found in the tropics (Eswaran et al., 2001); the tropics cover
about 40% of the world’s land surface and are home to about 40% of the
world’s population (Edelman et al., 2014).
Agricultural soils can be subdivided into dryland and irrigated soils. Irri-
gated land area constitutes approximately 20% of the world’s agricultural
soils. However, they are the source of approximately 40% of the world’s
agricultural production (FAO, 2011). In contrast, dryland soils (soils for
which crops rely exclusively on rainfall for water) occupy a much greater
agricultural area, but production per unit area averages much lower than that
for irrigated soils. This strongly suggests a critical factor limiting crop yield
on dryland soils is water—as does arguments by others: for example, see
Hatfield and Walthall (2015). That is, soil functionality most critical for food
production and security revolves around soil–plant–water relations; soil
alterations negatively impacting soil water storage and crop water availability
are highly important soil degradation considerations. Fertility adjustments
can be made to soils through nutrient additions such as with commercial
fertilizers or manures if physically and economically available, with the

caveat that many soils in economically deprived areas have been mined of
their fertility with no currently feasible opportunity for nutrient replacement
(FAO, 2011).
2.3 Soil Degradation

Soil degradation is a nebulous term suggesting that the capacity of a soil to
perform selected specified service(s), such as growing crops, has been dimin-
ished. The concept seems rather simple, but quantifying degradation has
been very challenging and creates uncertainty associated with quantifying
land degradation efforts (Bai et al., 2008; Safriel, 2007). A global evaluation
of soil or land degradation requires sampling and/or evaluation methodol-
ogy and a land degradation metric that meets needs and interests for multiple
different groups. Sampling intensity required for a quantitatively defendable
evaluation varies between different landscapes and soil characteristics under
investigation. For example, within a given field, hill tops or side slopes may
be degraded from soil erosion, while level areas may be unchanged or even
improved from eroded topsoil deposition. A single value to describe change
across multiple widely differing spatial conditions in small areas has multiple
interpretation challenges. The temporal sampling period required to defen-
sively characterize rate of soil change, or intensity of change, may be mul-
tiple decades. In spite of multiple challenges, various efforts to address and
quantify soil degradation at the global scale exist in the literature. Safriel
(2007) identifies five different global land degradation assessments con-
ducted in roughly the last 35 years: Generalized Map of the Status of Desert-
ification in Arid Lands (Dregne, 1977); Desertification of Arid Lands
(Dregne, 1983); Global Desertification Dimensions and Costs (Dregne
and Chou, 1992); Global Assessment of Human-induced Land Degradation
(Oldeman, 1994; Oldeman et al., 1990, 1991); and Synthesis on the Main
Areas of Land Cover and Land Use Change (Lepers, 2003). All assessments
indicate soil degradation is occurring, but degrees and most impacted loca-
tions vary with methodology and interpretation.
The approach to considering soil or land degradation has recently incor-
porated land function as opposed to earlier approaches that considered pri-
marily soil properties such as soil organic matter or carbon content, structural
condition, degree of salinization, compaction or bulk density, or degree of
erosion. Utilizing available data from a variety of sources and a process that
included change in land function, the FAO (2011) suggests the relatively
small area of the earth’s surface devoted to agriculture (about 11%), is 25%
highly degraded—see Fig. 4 (FAO, 2011). As one would expect, land deg-
radation was spatially variable. About 10% is improving. This evaluation is in
contrast to the Global Assessment of Human-Induced Soil Degradation
(GLASOD) project that identified 15% of the agricultural land area as being
degraded (Bai et al., 2008). The FAO report relied heavily on changes in
precipitation use efficiency in contrast to the GLASOD project that relied
Typology of degradation Intervention options

of ecosystem benefits
Type 1—High degradation trend Rehabilitate if economically feasible; mitigate

or highly degraded lands where degrading trends are high
Type 2—Moderate degradation trend Introduce measure to mitigate degradation

in slightly or moderately degraded land
Type 3—Stable land, slightly Preventive interventions

or moderately degraded
Type 4—Improving lands Reinforcement of enabling

conditions which foster SLM
Type 4:
Improving lands Bare areas
10%
18%
Water
Type 3: 2%
Stable land,
slightly or
moderately
36%
degraded
25%
Type 1:
High degradation
8%
or highly degraded lands
Type 2:
Moderate degradation in slightly
or moderately degraded land
Fig. 4 Types and extent of soil degradation. Food and Agriculture Organization of the
United Nations, 2011, The State of the World’s Land and Water Resources for Food and
Agriculture (SOLAW)—Managing Systems at Risk, http://www.fao.org/nr/water/docs/
SOLAW_EX_SUMM_WEB_EN.pdf, reproduced with permission.
heavily on human judgement. GLASOD estimates soil erosion is responsible

for 83% of global land degradation.
2.4 Soil Erosion

The surface layer of virtually all soils is the most important layer of the soil
profile, and the soil layer most impacted by soil erosion. The surface soil is
the entrance point of rainwater and air exchange; the surface layer controls
infiltration rates, which is especially important as intense rain storm fre-
quency is increasing in many parts of the globe (Hansen et al., 2012) and
water that does not infiltrate is not only unavailable for plant use, runoff
water carries with it soil materials contributing to the soil degradation pro-
cess and offsite damage. Soil erosion increases by a factor of about 1.7 times as
precipitation intensity increases (Nearing et al., 2004). Soil organic matter
content typically is highest at the soil surface and decreases with depth; soil
organic matter favors nutrient storage and release during decomposition and
supports stable aggregate structure formation and maintenance (Stevenson,
1972). Soil containing the highest soil organic matter content is the first to be
eroded. As soil erosion proceeds, the soil profile component with the highest
soil organic matter is lost and profile soil organic matter content decreases
(Fenton et al., 2005).
Soil erosion is conceptually simple; it involves detachment of soil parti-
cles from the soil surface, transport of those particles, followed by deposition.
Quantifying soil erosion is not so conceptually simple, however. For exam-
ple, no clear, consistent defining rules exist identifying travel distance
required before transported soil particles are considered eroded materials.
Soil erosion is too often interpreted by the public as soil lost, but in reality,
it is soil movement on (or from) a defined slope with typically a portion lost
from the defined area. The quantity of lost soil depends on a myriad of fac-
tors including topography, size of the area considered, soil texture, and water
runoff rate. Nonetheless, literature contains many statements regarding soil
erosion rates, too often without clearly defining the basis for quantification.
Even with this caveat, evidence strongly suggests soil movement down slope
is at an alarming rate, a rate that dominates the ongoing soil degradation
process.
The Status of the Worlds Soil Resources Report suggests globally,
approximately 20–30 Gt of soil are eroded annually by water with about
5 Gt eroded by tillage (FAO and ITPS, 2015) from approximately 1.6 billion
hectares of cultivated land (FAO, 2011). Using these published erosion and
land area estimates, soil erosion (using 25 Gt of annual soil erosion for the
calculation) is occurring at an average rate of 15.6 Mg/ha/year (on agricul-
tural lands) or an annual surface soil depth reduction of approximately
1.2 mm (assumes the soil bulk density is 1.3 g cm3 for this calculation).
Other sources indicate global soil loss is approximately 30 Mg/ha or
2.3 mm (Pimentel, 2006; Pimentel et al., 1995).
Soil degradation through erosion is partially offset by soil renewal
through weathering of soil parent material. Estimated rates of soil renewal
vary widely and are dependent on study approaches used. Typical global soil
renewal rate estimates fall within 0.002–0.09 mm/year (Alexander, 1988;
Wakatuski and Rasyidin, 1992). Using the most generous soil renewal rate
above, the estimated average global erosion rate exceeds renewal rates by an
order of magnitude, and using the slower renewal rate, losses are more than
two orders of magnitude in excess of renewal rates. Others have also con-
cluded soil erosion rates are from one to two orders of magnitude greater
than soil renewal rates (Cruse et al., 2013; Montgomery, 2007). While these
statements should bring alarm to all, the short-term productivity loss associ-
ated with average soil erosion rates is not particularly alarming. Evidence
suggests we are losing only about 4% production potential for each 0.1 m
of soil depth reduction (Bakker et al., 2007; den Biggelaar et al., 2001;
Fenton et al., 2005); a soil depth loss of 1 mm would impact crop production
the subsequent year less than 0.5%, a loss that could not be detected in most
research plots or farm fields. However, over time, the “yield drag” imposed
by annual accumulation of soil losses have become noticeable and are having
major impacts.
Understanding soil erosion is highly variable in space and time; average
values across the globe or even across a field can mask much higher soil ero-
sion rates occurring at smaller scales. Cox et al. (2011) illustrated that sheet
and rill erosion estimates for selected townships in Iowa, USA, for 2007 var-
ied more than an order of magnitude from the long-term mean sheet and rill
erosion estimates for Iowa (11.6 Mg/ha) which itself is an order of magni-
tude greater than estimated soil renewal rates. Some of the more highly
erodible areas globally have annual average soil loss rates in the hundreds
of Mg/ha (Pimentel et al., 1995). Using average soil erosion rates over rel-
atively large areas camouflages the extensive production damage being done
over major agricultural land areas. Pimentel (2006) estimates global agricul-
ture loses 10 million hectares annually due to soil erosion. Soil loss and deg-
radation is excessive and occurring are rates that compromise our capacity to
meet rising global food demands (Delong et al., 2015).
2.4.1 Climate Change, Especially Extreme Rainfall and Heat

Climate change is rapidly altering our concept of normal weather relative to
agricultural production with the expectation that agricultural production
will become more susceptible to the changing climate over the next decades
(Hatfield et al., 2014). Projections revealed in the last National Climate
Assessment (Melillo et al., 2014) showed that by mid-century there would
be a continued increase in annual precipitation across the temperate regions
with a shifting seasonality of more spring precipitation and more variable
precipitation in the summer period. In fact, these changes are already occur-
ring across the globe and especially in the areas of the world with intensive
agricultural production. A changing climate increases the probability of
changes in the frequency, intensity, spatial extent, duration, and timing of
extreme weather and climate events. Extreme events in temperature and
precipitation could have a major impact on global ecosystems and the soil
resource because of the increased potential for erosion and accelerated loss
of soil carbon or organic matter (Handmer et al., 2012). With climate
change, deviations occur in the mean temperature or precipitation, and
Hansen et al. (2012) showed that shifts in the distribution of temperature
or precipitation will lead to increased occurrence in extreme events.
Changes in the climate and increased demand for food, energy, and water
require a better understanding of the linkages between climate variability
and the intersection between ecosystem services and soils. The continued
change in temperature and patterns of precipitation will contribute to
increased problems of water availability required to support agricultural pro-
duction to achieve food security and of a variable water supply on the phys-
ical, chemical, or biological processes within the soil profile (Calanca, 2007;
Fuhrer et al., 2006; Torriani et al., 2007). Climate change will increase the
potential negative environmental impacts, such as higher erosion rates
(Nearing et al., 2004), or faster decomposition of soil organic matter and
increased nitrogen (N) leaching (Bindi and Olesen, 2010). Development
of adaptation strategies to offset the negative impacts of climate change
on agriculture will require a more comprehensive view of agriculture
and ecosystem services (Hatfield and Walthall, 2015; Hatfield et al.,
2014). As a result, there will be increased pressure to develop strategies to
adapt agricultural land management practices to offset climate change and
reduce the competition for land and water resources (Lotze-Campen and
Schellnhuber, 2009). Given the different responses to climate variation,
Klein et al. (2013) suggested that adaptation to climate change for agricul-
tural lands would require a multiobjective optimization approach to link
climate, management, and soils information together with simulation

models to evaluate a range of solutions for management practices. This
approach developed an optimum set of solutions that could be spatially dis-
tributed across the landscape and provided a framework for how we need to
be evaluating the linkages among the components of agricultural systems.
Increased intensity and extreme precipitation events increase the poten-
tial for accelerated soil erosion (Garbrecht et al., 2015, Nearing et al., 2004).
This increase in potential erosive events will require greater adoption of
available conservation practices to protect the soil resource (Garbrecht
et al., 2015; Hatfield et al., 2014). Changes in precipitation across the globe
are not expected to be uniform (Fig. 5) with increases in the low and high
Fig. 5 Projected changes in average annual precipitation over the 2071–99 period for
the globe (compared to the 1970–99 period) under a low emission scenario that
assumes rapid reductions in emissions and concentrations of heat-trapping gases
(RCP 2.6) and a higher emission scenario that assumes continued increase in emissions
(RCP 8.5). Hatched areas indicate confidence that the projected changes are significant
and consistent among models. White areas indicate the changes are not projected to be
larger than expected from natural variability (Walsh et al., 2014. Source: NOAA NCDC/
CICS-NC).
latitudes and much of the food producing regions becoming drier under a
high emission scenario. Increasing the intensity of precipitation could
potentially lead to more runoff from soils and could reduce the amount
of effective precipitation available to the crop-inducing conditions where
water limitation affects plant productivity. More critical in many areas is
the shift toward reduced and more variable precipitation during the summer
when crop water use is the highest and where any water deficit would
reduce productivity. Changing precipitation patterns and amounts will
affect the water and food resources of the world showing that soil manage-
ment practices focused on water capture, storage, and availability will
become even more critical to preserve our production capability. The
combination of extreme temperature and precipitation events will have impli-
cations for crop productivity and food security. Evaluations of ground-based-
meteorological stations coupled with a suite of climate models
were conducted to determine the patterns of temperature and precipitation
occurrences over the world (Hao et al., 2013). They evaluated four combina-
tions of temperature and precipitation: warm/wet (high temperature/high
precipitation), warm/dry (high temperature/low precipitation), cold/wet
(cold temperatures/high precipitation), and cold/dry (cold temperatures/
low precipitation) and compared the 1978–2004 period with the 1951–77
period on a global scale. They found warm/wet extremes increased in the high
latitudes and tropics, while the warm/dry extremes increased in many areas, e.-
g., central Africa, eastern Australia, northern China, parts of Russia, and the
Middle East (Hao et al., 2013). Conversely, the extremes in the cold/wet and
cold/dry combinations decreased over most of the earth. The increase in the
warm/wet and warm/dry extremes over many of the agriculture areas will
have a negative impact on agricultural productivity and change the
distribution of viable crop production.
Two aspects often overlooked about climate change are the effect of
increasing temperature on evaporative demand and rate of crop water use
and the effect on soil temperature and the resultant rate of chemical and bio-
logical reactions in the soil profile. It is often assumed that the effect of the
projected increase in air temperature of 1.5°C under the low emission sce-
nario and 4–5°C under the high emission scenario will reduce crop produc-
tivity because of the direct impact on the metabolic processes in plants
(Hatfield and Walthall, 2015; Hatfield et al., 2011, 2014; Izaurralde et al.,
2011). This effect is enhanced because minimum temperatures are increasing
at a faster rate than maximum temperatures, and the effect of increased
minimum temperatures on the physiology of organisms reveals a disruption

in the efficiency of metabolic processes (Hatfield et al., 2011). One of
impacts on plant productivity is water deficits that prevent the plant from
functioning at optimum levels. Increased air temperature increases atmo-
spheric demand for water, leading to the potential for increased water def-
icits especially in degraded soils (Hatfield and Prueger, 2015). Development
of frameworks for integrating these pieces together will be critical for under-
standing the role of soil in the food, energy, water nexus. There are emerg-
ing frameworks as indicators for crop suitability as proposed by Holzk€amper
et al. (2013) where they incorporated six factors into a suitability index.
These indicators included average daily minimum temperatures below 0°
C for frost impacts, daily mean temperature to determine plant growth,
average daily maximum temperature above 35°C for heat stress, average
daily soil water availability (precipitation–reference evapotranspiration),
and length of the phenological period (days) to account for the effects of
changing phenological development on biomass accumulation and crop
yield. They related their suitability index to maize yields for a number of
locations around the world with a positive relationship between productiv-
ity and their suitability index. Estimation of the climate impacts on food pro-
duction and security and the interface between climate (temperature) and
soils (soil water availability) were easily identified in this indicator.
Soil temperature is expected to increase in proportion to air temperature.
A rise in the average annual soil temperatures between 1.5°C and 5°C will
have major impacts on the rate of nutrient cycling and biological activity.
This aspect is rarely discussed and may have significant consequences on
all of the below ground processes. The increased rates of biological and
chemical reactions in the soil will exacerbate the negative impacts of soil
degradation because the effectiveness of soil functions will be reduced. Cli-
mate change threatens to further degrade soils through increased erosion
caused by exposure to more extreme events and reduced capability of cur-
rent conservation practices to protect the soil resource (Garbrecht et al.,
2015; Nearing et al., 2004). Management of soil water through conservation
practices (Hatfield et al., 2001) and preservation of the soil resource to max-
imize the capability to provide water to the crop are foundational to food
security.
Future efforts to incorporate soils into enhancing our ability to under-
stand the food, energy, water nexus will require assessment frameworks sim-
ilar to those developed and used for agroclimatic assessments. These indices
provide an assessment of where crops could be grown and have been based
on temperature and soil water availability. There are many examples in the
literature of agroclimatic indices (Araya et al., 2010; Daccache et al., 2012;
Falasca et al., 2012; Moeletsi and Walker, 2012; Neild and Richman, 1981;
Simane and Struik, 1993; van Wart et al., 2013). The current forms have
shown that assessment of crop suitability is dependent upon soil water avail-
ability, and recent agroclimatic assessment tools have incorporated soils into
the framework because of the importance of soil water on crop productivity.
As we continue to experience climate change, the future climate effects on
crop productivity reveal the sensitivity to temperature and precipitation
with precipitation being the dominant factor affecting productivity in the
short-term (Hao et al., 2013; Maracchi et al., 2005; Motha and Baier,
2005; Sivakumar et al., 2005; Tao et al., 2009; Zabel et al., 2014). Soil man-
agement practices showing a positive benefit on soil water storage and soil
water availability will have a significant impact on crop productivity and
biomass production and subsequent improvement of soil functions and eco-
system services. These changes in the soil and resultant changes in manage-
ment practices will be more significant given the projections that extreme
precipitation events, including drought, are expected to increase with cli-
mate change (Calanca, 2007; Collins et al., 2013; Hansen et al., 2012;
Walsh et al., 2014).
Climate change will impact the capability of soils to provide the ecosys-
tem services necessary for food, energy, and water to meet the needs of
humans. Our challenge will be how to increase the capacity of our soils
to function at the highest level necessary.
2.4.2 Urban Expansion—Covering of Productive Soils

The increasing world population and the associated increase in infrastructure
(e.g., roads and buildings) will increase the competition for land used for
food production (Smith et al., 2010). Ajmone-Marsan et al. (2015) suggested
that urban soils were increasing around the world, and management of these
soils will be a challenge because of the classic definitions of soil functions are
not applicable to urban soils. Urban soils lose their productive and filtering
capabilities because of sealing and are valued for their support functions
more than other ecosystem services (Ajmone-Marsan et al., 2015). This is
not a new revelation about the changing landscape with evidence that from
1960 to 1970, 5.5 million hectares were urbanized at the rate of 0.056 ha per
person increase in the population during that period (Ramsey and Corty,
1982). The changes in the landscape as suggested by Ajmone-Marsan et al.

(2015) would indicate that the overall ecosystem services of urban soils
would be diminished and that these soils are valued more for structural sup-
port than other functions. Fazal (2000) used geographic information systems
to quantify the changes in the land use in Saharanpur City, India, from 1988
to 1998 and found urbanization was decreasing the amount of high quality
agricultural lands. In Europe, over 970,000 ha of agricultural soil was lost
between 1990 and 2000 with an estimation of the same amount to be lost
in the next decade (EEA, 2010). In their analysis, the threats to agricultural
productivity were not due to the loss of land but to soil degradation; how-
ever, the sealing of the soil caused by urbanization was 4% and increased to
6% from 1990 to 2000 and productive agricultural soils continue to be lost to
urbanization and transportation infrastructures (EEA, 2010).
We will not be able to slow the rate of conversion of agricultural lands to
nonagricultural uses. Therefore, we need to direct our attention to increas-
ing the functionality of the soil resource. The continued degradation of the
soil resource must be reversed to enhance our production capacity to
achieve food, energy, and water security.
2.4.3 Growing Food Demand Placing Increased Pressure on the Soil

Resource
The increased demand for food and concern for food security places an ever-
increasing pressure on the soil resource. If we take the estimates for increased
cereal production of 25%–50% by 2050 (Smith et al., 2010) then the soil
resource will be required to supply more water, nutrients, and increase
the support functions for a viable crop. These estimates often focus on cereal
production; however, crops directly consumed by humans, e.g., vegetables
are largely ignored in these assessments. Food security requires a balanced
diet along with nutritious food supply and concerns should be directed
toward the capacity of the soil resource to supply food security from all food
production systems. In a recent analysis by Wise (2013), there is a summary
of the potential for increasing food supply and the land area needed to sup-
port this increase. Tilman et al. (2011) found that meeting global food
demands would require an additional billion hectares of land being brought
into production by 2050 and would require high levels of nitrogen inputs
and increased greenhouse gas emissions under intensive agricultural prac-
tices. In contrast, applying moderate intensification in low productivity
regions could reduce land demands by 80%, reduce greenhouse gas
emissions, and reduce nitrogen use and produce sufficient amounts to meet
food demands (Tilman et al, 2011). The effects of climate change on future
crop productivity have been summarized in many reports; however, these
studies fail to integrate the potential impact of a degrading soil resource
and the increased demand on the soil from all agricultural systems that
demand higher levels of production. The concept of sustainable intensifica-
tion provides a framework for the integration of production systems with
ecological principles (Campbell et al, 2014).
It will be important to realize that sustainable production will require a
soil resource to support high levels of production. Conceptually, we can
view this as the integration of practices that increase productivity and soil
health (Fig. 6). Vanlauwe et al. (2014) showed that there may be multiple
paths to achieve sustainable agriculture systems. Lal (2015b) stated that there
must be a systems approach to understanding and quantifying the dynamics
of conservation agriculture. In his analysis he found there were four basic
components: management of residue mulch, cover crops, complex crop
rotations, and integrated nutrient management necessary to transform
no-till agriculture into conservation agriculture. Earlier, he had summarized
that the source of yield reductions in no-till systems were due to a range of
factors from reduced early seedling growth, N and P availability, residue
removal and poor residue management, and greater disease pressure (Lal,
2015a). The development and implementation of conservation or climate
smart agriculture systems cannot be considered as a single factor approach
but will require an integration of a number of components to achieve sus-
tainable intensification. The foundation for any changes will be the soil
e
bl n
na
Productivity
ta
i
a tio
s
Su ns ific
te
In
Soil health
Fig. 6 Conceptual diagram for the interaction of increased productivity and sustainable
intensification relative to soil health.
resources ability to provide the soil functions, especially, water, nutrients,

and support for the growing plant.
2.4.4 Linkage of Soils to Food

Throughout history there have been repeated linkages between the vitality
and strength of a culture and its ability to produce food (Hillel, 1991).
A reliable source of food was prerequisite in order for a stable society to
develop, sustain itself, and grow in population and technology. Regardless
of their military strength or other human and natural resources, no culture
was immune to decline if there were significant disruptions in their food sup-
ply. Whether the cause was salinity, erosion, drought, or soil exhaustion, food
insecurity was often caused by poor soil management and contributed to the
decline of some of the most powerful cultures in history. Long-term sustain-
able food production was only achieved through judicious recycling of
organic nutrients, as described by King (1911) regarding agriculture in China,
through natural regeneration via deposition of nutrient-laden sediment in
annual floods of the Nile and other rivers (Hillel, 1994), or via shifting crops
to new areas as in slash and burn systems. Shortfalls in food supply due to over-
population or soil degradation were frequent drivers of human exploration
and migration as populations shifted to exploit fertile new lands.
Agricultural practices in the New World often included clearing forest
land, burning the unused wood and understory, and then sowing crops into
the soil that was enriched by the nutrients released from the forest biomass.
After several growing seasons crop yields would decline and another part of
the forest would be cleared. Westward expansion of agricultural settlement
in the eastern United States in the 18th and 19th centuries was driven by the
need to constantly find new, uncultivated soils (Stoll, 2002). Although
important discoveries of the role of soil properties and management in crop
production had already been made (Dokuchaev, 1967; King, 1906; Tull,
1762), it was not until the mid-20th century that the available knowledge
of soil properties and processes were broadly applied to systematic soil
management.
Soils provide three vital functions to enable vigorous plant growth: (1)
physical support for roots to anchor plants to the soil, (2) a pore structure
that provides conduits for water and facilitates the exchange of gases between
the roots and the atmosphere, and (3) a habitat for soil fauna to facilitate
decomposition and recycling or organic materials and provide a reservoir
for nutrients. Understanding the intricacies of root growth, distribution,
and function across plant species and soil properties has been a challenging
area of research in spite of the recognized importance of roots on crop pro-
duction (Arkin and Taylor, 1981; Dexter, 2004; Skaggs and Shouse, 2008).
Bishopp and Lynch (2015) suggest that the tremendous increases in crop
yields in the 20th century can be attributed to fertilizer use and management
of the aboveground parts of the plant. However, they conclude that to meet
food production demands in the 21st century, greater attention must be
given to managing belowground plant parts, i.e., roots. Rooting patterns
result from complex interactions between crop genetics, agronomic prac-
tices (fertility and pest management), and soil physical and chemical prop-
erties. Greater attention and management need to be devoted to
optimizing root growth and function, while avoiding practices like soil
compaction that are known to restrict root growth (Batey, 2009; Unger
and Kaspar, 1994).
Soil particles of varying sizes, shapes, and mineralogy are gathered into
aggregates, and the arrangement of these particles and aggregates (soil struc-
ture) determines the characteristics of the soil pore space. Soil structure is a
result of physical processes such as shrink–swell forces due to wetting/drying
and freeze/thaw cycles and biological processes associated with root growth
and exudates and the activities of soil fauna (Angers and Caron, 1998; Horn
and Smucker, 2005). It is through the soil pore space that water and gases are
transported to and from plant roots. Soils with strong, stable structure are
best suited for maintaining the balance between draining excess water and
retaining plant-available water while still maintaining well-aerated void
space for healthy root growth (Gli nski and Ste˛pniewski, 1985; Kirkham,
2014). Healthy soils with stable or increasing soil organic matter content
are also more likely to have optimal soil structure and able to store a greater
amount of plant-available water (Hudson, 1994).
As water and air have very different thermal properties, soil thermal
properties are strongly influenced by soil water content. Water content
and heat transfer in soils are therefore closely coupled with important impli-
cations for biological activity and chemical reactions (Parlange et al., 1998).
Soil structure and porosity influence the soil thermal and water regimes that
affect the biological processes associated with nutrient transformations and
organic matter recycling. The biochemical characteristics of plant litter,
roots, and animal manures including lignin content and C-nutrient stoichi-
ometry will affect the mode and rate of decomposition, which affects the
rates of nutrient cycling and carbon sequestration (Himes, 1997; Kirkby
et al., 2014; Sardans et al., 2012). Biological soil processes are often highly
spatially and temporally heterogeneous, involve a large diversity of organ-

isms, and include an abundance of interacting processes (Andren et al.,
2008). The complexity of processes involved in the decomposition of
organic nutrient sources like compost and animal manures results in a greater
degree of uncertainty in the amount and timing of nutrient availability than
is common for inorganic fertilizers.
2.4.5 Linkage of Soils to Water

A key linkage between soil and food is the use of water by plants. Water
provides many functions in plant physiology, but some of the most impor-
tant functions are in the transmission of nutrients and photosynthates and in
providing evaporative cooling for the plant. The relationship between plant
water use and plant growth is a fundamental facet of agriculture and food
production and has thus received considerable attention (Ali and
Talukder, 2008; Hatfield et al., 2001; Stewart and Steiner, 1990; Taylor
et al, 1983). Arkley (1963) provides a comprehensive review of early liter-
ature for many crops and illustrates the remarkably consistent and often lin-
ear relationship between yield and transpiration for a particular crop and
location. The yield-water use relationship is often expressed as WUE
WUE ¼ Y=ET
where Y is the crop yield (grain yield or dry matter production) and ET is
evapotranspiration. There are many ways of expressing WUE based on units
of measure of yield and whether ET, transpiration, or the amount of irriga-
tion are used as the measure of water use. Agriculture competes for fresh-
water with industry and domestic users and currently consumes about
70% of the global groundwater withdrawals (FAO, 2011), which represents
43% of the total irrigation water use on over 300 million hectares of irrigated
lands (Siebert et al., 2010). Pimentel and Pimentel (2008) estimated that
pumping of irrigation water alone consumed 15% of all energy used in global
crop production. Thus, pumping of irrigation water is a major energy input
into irrigated agriculture so improving WUE has a significant impact on
both water and energy consumption.
There are multiple approaches to improving WUE and reducing energy
consumption in irrigated agriculture that include improvements in water
capture, storage, conveyance, and application (Ali and Talukder, 2008;
Levidow et al., 2014; Pereira et al., 2012). Changing irrigation practices
can have a large impact on WUE, however, may involve trade-offs between
energy and water consumption. Converting from flood to sprinkler or drip
irrigation may save considerable quantities of water but may also require
greater energy input for manufacturing or operation of the more efficient
water delivery systems. Deficit irrigation, where less than the full irriga-
tion water requirement is applied, can be used to optimize limited water
resources but still enhance crop yields (Ali and Talukder, 2008). Choice
of crop also has a large impact on water use as crop water requirements
vary from over 1500 L kg1 for soybean and rice to less than 700 L kg1
for corn, potatoes, and millet (Pimentel et al., 2004). In rainfed systems,
climate change is altering traditional crop production zones and will
lead to both expansion of the extent of some crops while limiting the
economic range of production of others (Fischer et al., 2005; van
Wart et al., 2013). Thus, climate change and resource constraints are both
likely to affect the amount and distribution of crops throughout the 21st
century.
Irrigated crop production is concentrated in arid and semiarid regions,
where ET is high so reducing evaporation of water from after it has been
delivered to the field but prior to plant uptake is a key element of increasing
WUE at the field scale. Soil management practices offer great potential to
improve WUE in both irrigated and rainfed agriculture (Hatfield et al.,
2001; Passioura, 2006; Unger and Stewart, 1983; Viets, 1966). Practices that
increase infiltration and reduce soil water evaporation and percolation losses
will improve WUE by increasing the proportion of precipitation or irriga-
tion that transpires from the growing plants. No-till crop production with
retention of surface crop residues and/or cover crops often increases plant-
available water by reducing runoff and evaporation from the soil (Klocke
et al., 2009; Sauer and Daniel, 1987; Sauer et al., 1998; Swella et al.,
2015). In addition to optimizing water retention and availability, Hatfield
et al. (2001), in a review of WUE research, concluded that modifying nutri-
ent management practices could increase WUE 15%–25%. It is likely that a
systems approach, optimizing many aspects of the crop production system,
will produce the greatest improvement in WUE and reduction in energy
consumption.
2.4.6 Linkage of Soils to Energy

One of the unique characteristics of soil with regard to the water–food–
energy nexus is that soil is the only component of the nexus that is not
mobile. While transportation of water, food, and energy are all common fea-
tures of modern food production systems, the land resource is fixed in space
and therefore faces greater threats from climate change, urbanization, and
soil degradation. Optimizing crop production on a land resource base will

require optimizing water and energy inputs. Between 2002 and 2012 the
correlation between food and oil price indices had a correlation coefficient
of 0.94 indicating a very strong link between energy usage and the price of
food (Ringler et al., 2013). This relationship illustrates the degree of energy
intensity in modern production agriculture and, especially for the United
States, competition for resources for biofuel production (Ringler
et al., 2013).
Agricultural energy use can be divided into direct (fuels and electricity)
and indirect uses, which are energy-intensive inputs like fertilizer and pes-
ticides. The greatest energy input into US agriculture is fuel use for field
operations and the largest consumption of liquid fuels in the food system
is for transportation (Beckman et al., 2013; Dale et al., 2011). For US agri-
culture, direct energy inputs are approximately twice that of the indirect
inputs and of the indirect inputs, fertilizer manufacture accounts for one-half
of the energy inputs (Beckman et al., 2013). Soil management decisions
regarding tillage and fertilizer use therefore have important implications
for energy use by agricultural systems, where the greatest gains in energy
use efficiency are likely to occur with increased adoption of no-till or
reduced tillage practices and improved nutrient use efficiency.
Miranowski (2005) concluded energy conservation should be a goal of
US agriculture as a way to reduce vulnerability to energy price shocks
and long-term price increases while reducing environmental impacts of fossil
fuel consumption. Improved production efficiencies are also likely to have
the greatest potential to reduce productions costs and improve system
profitability.
Biofuel, especially corn grain ethanol in the United States, and other
renewable energy production over recent years has had a dramatic effect
on the agricultural sector as it became a significant energy producer
(Beckman et al., 2013; Wallander et al., 2011). US corn production
increased dramatically during the decade of 2000–09 through increased crop
yield but also conversion of acreage from other crops, especially soybean and
cotton, and expansion into marginal lands and former Conservation Reserve
Program (CRP) acreage (Wallander et al., 2011). This sudden change in
corn production had significant impacts on grain and farmland prices and
precipitated intense policy debates regarding converting land use from food
to fuel production (Gamborg et al., 2012). There is also concern regarding
the sustainability of intensified production practices in response to high
commodity prices, especially on marginal lands susceptible to erosion and
drought (Wright and Wimberly, 2012), and on the environmental impacts

of these intensified cropping systems.
The Energy Independence and Security Act (EISA) of 2007 established a
Renewable Fuel Standard (RFS) mandating 36 billion gallons of biofuels be
produced annually in the US by 2022. Of this amount, 44.4% of the RFS is
to be based on cellulosic feedstocks. In addition to ethical concerns regarding
diverting grain from food to energy production, grain-based bioenergy pro-
duction has potential negative environmental consequences including SOC
depletion, GHG production, nitrate leaching, and loss of biodiversity
(Gelfand et al., 2013). For these reasons, there is heightened interest in lig-
nocellulosic bioenergy production utilizing perennial herbaceous or woody
feedstock grown primarily on lands marginal for crop production (Cai et al.,
2011; Fischer and Shcrattenholzer, 2001; Gelfand et al., 2013; Rosenberg
and Smith, 2009). A comprehensive study of lignocellulosic feedstock sup-
ply, the so-called Billion Ton Report was completed by the US Department
of Energy (DOE) in 2005 and updated in 2011 (U.S. Department of Energy,
2011). This report considers various scenarios for estimation of feedstock
supply and bioenergy production including constraints imposed to reduce
or prevent environmental degradation such as SOC depletion through crop
residue removal from cultivated cropland.
2.4.7 Improving Soils

The ability of soils to grow crops and additionally provide multiple ecosys-
tem services is often unquestioned in developing countries. However,
assessments of soil degradation provide what can be considered a sobering
perspective on actual soil conditions in both developed and developing
countries. Oldeman et al. (1991) provided a global assessment of human-
induced soil degradation that summarized physical, chemical, and climate
constraints on agricultural production. Major constraints include loss of
organic matter and nutrients and poor structure or too much or insufficient
moisture for plant growth. A more recent assessment summarizing changes
over 1981–2003 concluded that 23.5% of the global land area was degrading
over this time interval, an area of 35 million km2 affecting 1.5 billion people
(Bai et al., 2008). Climate change introduces serious additional threats to
crop growth and food security, and these threats, due to increasing fre-
quency and duration of floods and droughts, will likely have greater impact
on soils that are already degraded.
Reducing limitations on food production due to impaired soil function-
ing is therefore a key component of any strategy for food security and climate
change adaptation. Practices to maintain productive soils are generally well

known (Lal, 2009a,b; Syers, 1997), yet conventional soil management is
often reduced to providing sufficient nutrients, water, and pest control
for a single growing season with little planning or provision for subsequent
years. As soil degradation typically occurs gradually over many years or
decades as with wind and water erosion, salinization, desertification, or
nutrient depletion, practices needed to restore full functionality may also
need to be applied over long periods, especially when resources are limited.
This long-term commitment to soil restoration is especially challenging for
already food insecure developing countries. Soil rehabilitation generally
involves multiple components including cessation of damaging practices
and initiation of remediation practices. Many of these techniques were
developed for mineland reclamation applications where overburden mate-
rial was managed to enhance development of functioning growth media
(Barnhisel and Hower, 1997; Bendfeldt et al., 2001). Common remediation
practices include targeted tillage to alleviate compaction and enhance soil
structural development, planting of deep-rooting plants to facilitate nutrient
cycling and pore network development, and addition of organic amend-
ments to increase SOC content and facilitate soil aggregation (Frye
et al., 1985).
Recently, there has been an increased focus on biological aspects of soil
health, where traditionally more focus has been placed on physical and
chemical properties. In reality, a more holistic approach recognizing the
important linkages among physical, chemical, and biological processes is
likely to enhance soil management and rehabilitation efforts. Ecological per-
spectives on soil science recognize these linkages (Coleman, 2008; Lavelle,
2000) and the value of managing or enhancing components of the soil bio-
logic communities and processes to sustain or improve biological function-
ing (Curry and Good, 1992). One example is the important linkage between
soil organic matter and plant-available water. Hudson (1994) studied this
relationship and found significant enhancement of plant-available water
with increasing soil organic matter content for sand, silt loam, and silty clay
loam textures. Managing soils to increase their organic matter content not
only improves the ability of the soil to retain water for plant growth but also
enhances the internal carbon and nutrient cycling, important processes for
development of resilient soils. The more holistic approach to soil function-
ing has also led to investigations of soil properties and their relationships with
human health (Brevik and Sauer, 2015; Wall et al., 2015). These analyses
provide new perspectives on soils and how their properties and management
can impact human health directly and indirectly through interactions with
food and water.
3. SCALE OF THE INTERFACE BETWEEN SOILS AND

ECOSYSTEM SERVICES
Incorporating soil into the food, energy, water nexus at different scales
can best be visualized through an evaluation of the ecosystem services and
soil functions at a range of spatial scales (Grêt-Regamey et al., 2015). The
direct impact of soil functions and degradation of these functions are realized
at the field, landscape, and watershed scale, and the linkage of soil properties
evident at the field scale become even more evident at the watershed and
landscape scale because of the integrative effect of the interactions. Ecosys-
tem services show the value of soil, and Table 1 illustrates the linkage
between ecosystem services and the spatial scales at which they become
dominant. The ability of soil properties to effectively provide soil functions
has an impact at all scales; however, soil degradation effects that prevent the
fulfillment of soil functions will impact ecosystem functions.
3.1 Field
A field often consists of a complex set of soil each with its own values of soil
properties creating a mixture of the magnitude of soil functions. For exam-
ple, there is a wide range in soils in their soil water holding capacity as
depicted from a central Iowa field (Fig. 7). Water availability is a direct func-
tion of soil organic matter content (Hudson, 1994), and as we degrade soils,
there are two changes that affect water availability: the loss of soil organic
matter and water holding capacity and the loss of infiltration capacity that
prevents water from entering into the soil. Soil water availability differences
across a field directly affect crop production (Baskan et al., 2013; Hatfield,
Table 1 Overview of Ecosystem Services at Different Spatial Scales of Agricultural

Systems
Ecosystem Services
Spatial Scale Provisioning Regulating Cultural Supporting
Field X
Watershed X X X X
Landscape X X X X
Fig. 7 Representations of soil types within a production field in central Iowa and the
available soil water holding capacity to a rooting depth of the crop.
2012). The outcome of soil degradation is to reduce organic matter and infil-
tration rates and is the primary contribution to the spatial variation of crop
response across a field. The changes in crop productivity within a field are
related to soil factors (Perez-Quezada et al., 2003). Of these factors, water
availability is the major factor affecting crop production (Nazarifar et al.,

2012). The interrelationships among soil carbon, soil biology, nutrient
cycling, and soil water availability to ecosystems have been documented
at the field scale (Barrios, 2007; Ghaley et al., 2014; Krishnaswamy et al.,
2013; Marks et al., 2009; van Eekeren et al., 2010). At this scale, there have
been direct measurements related to soil functions and to ecosystem services
oriented to provisional aspects and are more closely linked to food and water
dynamics.
3.2 Landscape
If we consider a landscape as an area of land with a specific quality or attribute
then ecosystem services and the relationship of the ability of the soil to pro-
vide these functions relative to ecosystem functions. The primary ecosystem
function at the field scale is provisioning and when we expand to the land-
scape scale then regulating, cultural, and supporting functions become crit-
ical factors linking the ability of the soil function. These services are a direct
result of the ability to the soil to provide its functions as evidenced by
methods to spatially place different practices on the landscape (Delgado
and Berry, 2008; Tomer et al., 2015a,b). These methods are based on the
integration of processes that link water dynamics to practices that regulate
the impact of erosion or water quality. One of the major factors affecting
how ecosystem services are affected at the landscape scale is the land use
and cover across the landscape (Grêt-Regamey et al., 2015; Metzger
et al., 2006; Nelson et al., 2009; Sutton and Costanza, 2002).
3.3 Watershed
All four ecosystem services are evident at the watershed scale and the effect of
the ability of the soil to provide the functions. There is an intersection
among water availability, nutrient cycling, and soil biology and regulating
services related to water quality or erosion. If we examine the dynamics
within the field, then the linkage between within field processes related
to water availability to the plant and offsite movement of water and nutrients
become evident (Hatfield et al, 2009; Schilling and Zhang, 2004). The major
factor impacting the watershed scale is water movement and this is basis of
the planning tool developed by Tomer et al. (2015a,b) to determine where
different watershed scale practices could be placed within a watershed. The
nexus of food, energy, water systems is most evident at the watershed scale,
and one of the challenges is to quantify the role that changing soil properties
have on the ability of a watershed has to provide ecosystem functions.
Patterns of soil degradation within a field and across a landscape begin to
magnify at the watershed scale. The differences in water holding capacity
create the potential for runoff and erosion under situations with extreme
rainfall events. The projected increase in extreme precipitation events will
interact with soil water holding capacity and soil degradation to create a sit-
uation in which the infiltration capacity is exceeded. This will create scenar-
ios in which different portions of the watershed may experience runoff and
even flooding while other parts will retain their ability to store water within
the profile. An example of this integration is the Revised Universal Soil Loss
Equation (RUSLE2, Dabney et al., 2012). This model accounts for the
effect of climate, soils, topography, and land management and their interac-
tions and reveals that changing the land cover or water infiltration rate will
have a large impact on erosion. The interactions of soil, land management,
position in the watershed, and climate will have a large impact on the food,
energy, water nexus. In a review, den Biggelaar et al. (2004) evaluated the
impact of soil erosion at the global scale among different soil orders and crops
and concluded that soil erosion limits our ability to obtain food security and
increases the potential for offsite impacts of production and increases the
inputs necessary to maintain production levels. Variation of soil and topog-
raphy across a watershed will lead to variations in the rate of soil degradation
and productivity. Salley et al. (2016) and Littleboy et al. (1992b) showed that
across a watershed the rates of soil erosion and yield decline were greatest in
the shallow soils. They introduced the concept of soil productivity half-life
and found for combinations of soil depth, climate, slope, and management
that combinations of these effects reduced the half-life of soil to less than 100
years. These conclusions were based on a soil erosion model that allowed for
the incorporation of soils, slope, management, and climate at the watershed
scale (Littleboy et al., 1992a). If we consider how long many of the world
soils have been under cultivation and the need for increased productivity,
then understanding the spatial and temporal dynamics across the field, land-
scape, and watershed will become increasingly important in order to opti-
mize ecosystem services and be able to apply the appropriate land
management practices required to enhance the ability of the soil to function.
This metric applied to the watershed scale provides a framework for linking
soil degradation to ecosystem services for an assessment of the potential
impacts of changing soil quality and land management practices on food
and water security.
4. IMPLICATIONS OF SOIL AS A COMPONENT OF THE

FOOD, ENERGY, WATER NEXUS
If we consider that soil is responsible for 99% of the world’s food pro-
duction, then the importance of soil in the food, energy, water nexus
becomes apparent (Pimentel and Pimentel, 2000). If we further consider that
soil erosion is the major factor, affecting soil degradation and declines in pro-
ductivity are directly related to degradation of the soil resource, then the
implications of soil in the context of increasing food, energy, and water secu-
rity become more evident. However, if the attitude is one that technology
will provide answers to these problems then the soil degradation rate will
continue to increase and we will reach a tipping point in which technolog-
ical advances will not be able to overcome the impacts of a reduced topsoil
depth coupled with a more variable climate.
The capacity of soil to provide soil functions begin with the incorpora-
tion of carbon into the soil as mediated by soil biodiversity and soil ecology
(Bender et al., 2016; Blankinship et al., 2016). All of the soil properties
(Fig. 2) are directly related to the ability of the soil to sequester carbon from
the atmosphere and result in a change of soil properties. However, the pro-
cess by which this occurs is not chemical but biological with soil biotic
diversity and stability having the largest role (Fig. 3). If soil erosion is the
largest degradation process then land management practices that contribute
to erosion, e.g., lack of soil cover, or tillage, coupled with the variable cli-
mate (Hatfield, 2014) will have to be reversed (Lal, 2015a,b) if we are to
ensure water and food security. If we are to optimize soil functions related
to food production (nutrient cycling, water dynamics, physical stability, bio-
diversity, and support of plant functions), then maximizing the rate of car-
bon accumulation in the soil will become imperative to achieving these
goals. Murphy (2015) points out that change in soil organic matter affects
a number of soil properties. The soil properties modified by changing soil
organic matter content include aggregate stability, water holding capacity,
soil erodibility, soil texture, cation exchange capacity, nutrient cycling, soil
acidity, and biological diversity. These changes were not uniform in the soil
but isolated near the surface mainly in the upper 10 cm of the soil profile
(Murphy, 2015). This is one of the more critical zones of the soil because
it is the interface between the soil and the atmosphere and controls the rate
of water infiltration and gas exchange. If we extend our consideration of soil
erosion to focus on the ability of precipitation to enter the soil profile, then
the stability and protection of the upper 10 cm become even more impor-
tant. Food production requires water transpired by the plant and soil man-
agement practices linked with improving WUE (production per unit of
water transpired) demonstrate the value of increasing the available water
supply (Hatfield et al., 2001). Food security should be considered as a func-
tion of effective precipitation (amount of precipitation that infiltrates into
the soil and is available to the plant). The concept of soil organic matter
and the relationship to water holding capacity as shown by Hudson
(1994) are valid, but we need to consider the fact that the upper soil surface
may be the controlling layer for water dynamics in the soil and if we limit
infiltration we limit the availability of soil water to the plant.
Changes in soil organic matter content affect a number of soil properties
that are ultimately related to food production and the water dynamics in the
soil (Lal, 2004; Pan et al., 2009). One of the key factors in this change in the
soil aggregate stability and understanding aggregate stability is key to inter-
preting soil responses to management and biological activity (Blankinship
et al., 2016; Keil and Mayer, 2014; Six and Paustian, 2014). Aggregate sta-
bility is affected by how biological systems incorporate organic material into
material capable of promoting stability and this change results in improve-
ments in soil properties (Fig. 8). Soil degradation is a result of tillage and
Food
and
water
Food security
production Water
availability
Cation
exchange Infiltration
capacity
Nutrient Aggregate
cycling stability
Increasing
soil organic
matter
Fig. 8 Conceptual diagram of the linkage between increasing soil organic matter and
soil properties leading to food and water security.
residue removal and enhancement of soil will require a reduction in the

amount of tillage and maintenance of residue cover on the soil surface. These
processes are at work in all soils and Blankinship et al. (2016) found that in
dry soils the combination of dead plants and live microbes were sufficient to
maintain soil structure. Processes responsible for soil aggregation provide for
more efficient food production and increased WUE. In the content of
cropping system there is a combination of cropping and livestock systems
that utilize either annual or perennial pastures, and these systems offer the
potential to improve the soil properties, e.g., soil organic matter, carbon
sequestration, nutrient cycling, and water conservation, through the main-
tenance and increase in soil fungal populations (Davinic et al., 2013). All soil
properties contribute to the efficiency and stability of food and energy pro-
duction and the water cycle. Soil is the reservoir of water, oxygen, and nutri-
ents necessary for plants to grow and any limitation to those components will
limit productivity.
5. CHALLENGES
5.1 Enhancing the Soil Resource to Reduce Soil
Degradation
Soil security is a concept from which we can begin to expand the under-
standing of the value of soil as a critical component of the food, energy,
water nexus, and the need to counteract the negative impacts of soil degra-
dation (Bouma, 2015). The challenge is increasing the soil carbon content of
soil to offset carbon emisisons but also to increase soil functions (Fig. 8). The
first step in the process of restoring soil function is to reduce soil erosion
which will be a challenge given the changing precipitation regime with
more intense precipitation events and a shift toward more spring precipita-
tion when there is a lack of ground cover to protect the soil surface. Chappell
et al. (2016) point out that soil erosion needs to be incorporated into assess-
ment efforts on modeling the changes in soil organic carbon because this
reduces the uncertainty in estimates of the magnitude of the changes. If
we consider the changes required in soil to reverse soil degradation then
reducing erosion and the rate of soil carbon loss become the critical factors.
Soil degradation is driven by tillage and residue removal (Hatfield, 2014) and
as Lal (2015a,b) demonstrates that the benefits of conservation agriculture
extend beyond reducing erosion to overall improvement in the soil resource
capable of ensuring greater production and reduced degradation. Some of
the challenges we face in terms of reversing soil degradation include:
• Understanding the response of different soils to different management

practices that cause degradation; if we can map more susceptible soils,
then we can determine fragile regions requiring attention.
• Quantify the intersection between soil texture classes and the rate of soil
aggradation to determine the effectiveness of different soil management
practices
• Separate the rate of change in the surface layer of soil (0–5 cm) from the
rate of change in the 0–15 cm depth
• Determine the impact of reduced tillage methods on the rate of soil
change and determine the offsets between tillage and addition of organic
material on the soil surface
• Understand the linkage between soil biological activity and nutrient
cycling/availability in order to evaluate the effect of organic production
systems
• Provide a quantitative linkage between climate and soil management
practices to determine the optimum combination of management prac-
tices for different agroclimatic zones
• Increase the amount of water available to the crop from the root zone to
be able to offset the impact of the increasing variability in precipitation
patterns due to climate change
• Determine the increase required in soil management to offset the impact
of the increasing intensity of precipitation events causing erosion
• Evaluate the effectiveness of more diverse crop rotations and the role of
continuous cover in increasing soil aggradation
5.2 Soil as a Component in the Food, Energy, Water Nexus

Soil is the forgotten piece of the food, energy, water nexus; however, the
oversight extends beyond this nexus to include many of the ecological ser-
vices required by humankind. The linkage among soil properties, soil func-
tions, and ecosystem services (Fig. 2) provides a framework for how we
should be considering these interactions. Soil scientists need to become
more involved in the policy discussions about food production, water
resources, and energy derived from agricultural systems. There has been a
renewed call for attention of the role of soil in ecosystem services
(Adhikari and Hartemink; 2016; Bouma, 2014, 2015; Hartemink and
McBratney, 2008; Lal, 2016). The increased need for food security requires
we begin to assemble a different picture of soil functions in the context of
provisioning services and begin to develop a more ecological approach to
agricultural problems rather than an agronomic approach (Lal, 2016). That

does not mean we abandon agronomic aspects but demands that we place
agriculture into an ecological context. The food, energy, water nexus
revolves around soil and the value it brings to all of these endpoints
(Fig. 1). To achieve food security, water security, and energy security will
require a new paradigm of how we approach these problems. For example,
Ostergard et al. (2009) proposed that achievement of food security would
require a radical shift in crop production practices to address the problem
of soil degradation, loss of biodiversity, restricted and polluted water sup-
plies, limitations in fossil fuels, and climate change leading to more variation
in our weather during growing seasons. To achieve these solutions we are
going to have to understand and value the importance of soil in food pro-
duction and communicate that message to policymakers and the public.
There are some challenges we need to face in bringing an increased aware-
ness of soil in this context and we present these to increase the discussion
around this topic.
• Increase the awareness of the linkage between the soil and the capability
of producing food both in quality and quantity
• Increase the awareness of the role of soil in the water cycle and the need
to improve soil management practices to eliminate erosion and increase
the capability of the soil to store water
• Increase the awareness of the linkage between soil properties, soil func-
tions, and ecosystem services
• Increase the awareness that our soils are a limited resource and enhance-
ment of the soil resource will pay dividends in our ability to ensure food
and water security
REFERENCES
Adhikari, K., Hartemink, A.E., 2016. Linking soils to ecosystem services—a global review.
Geoderma 262, 101–111.
Ajmone-Marsan, F., Certini, G., Scalenghe, R., 2015. Describing urban soils through a fac-
eted system ensures more informed decision-making. Land Use Policy 51, 109–119.
Alexander, E.B., 1988. Rates of soil formation: implications for soil-loss tolerance. Soil Sci.
145, 37–45. Available at: http://coweeta.uga.edu/publications/2112.pdf.
Ali, M.H., Talukder, M.S.U., 2008. Increasing water productivity in crop production—a
synthesis. Agr. Water Manage. 95, 1201–1213. http://dx.doi.org/10.1016/j.
agwat.2008.06008.
Andren, O., Kirchmann, H., K€atterer, T., Magid, J., Paul, E.A., Coleman, D.C., 2008.
Visions of a more precise soil biology. Eur. J. Soil Sci. 59, 380–390.
Angers, D.A., Caron, J., 1998. Plant-induced changes in soil structure: processes and feed-
backs. Biogeochemistry 42, 55–72.
Araya, A., Keesstra, S.D., Stroosnijder, L., 2010. A new agro-climatic classification for crop
suitability zoning in northern semi-arid Ethiopia. Agr. Forest. Meteorol.
150, 1057–1064.
Arkin, G.G., Taylor, H.M., 1981. Modifying the Root Environment to Reduce Crop Stress.
American Society of Agricultural Engineers, St. Joseph, Michigan. 407 p.
Arkley, R.J., 1963. Relationships between plant growth transpiration. Hilgardia
34, 559–577.
Bai, Z.G., Dent, K.L., Olsson, L., Schaepman, M.E., 2008. Proxy global assessment of land
degradation. Soil Use and Manag. 24, 223–234. Available at: http://onlinelibrary.wiley.
com/doi/10.1111/j.1475-2743.2008.00169.x/full.
Bakker, M.M., Govers, G., Jones, R.A., Rounsevell, D.D.A., 2007. The effect of soil erosion
on Europe’s crop yields. Ecosystems 10 (7), 1209–1219. Available at: http://link.springer.
com/article/10.1007/s10021-007-9090-3.
Banwart, S., 2011. Save our soils. Nature 474, 151–152.
Barnhisel, R.I., Hower, J.M., 1997. Coal surface mine reclamation in the eastern United
States: the revegetation of disturbed lands to hayland/pasture or cropland. Adv. Agron.
61, 233–275.
Barrios, E., 2007. Soil biota, ecosystem services and land productivity. Ecol. Econ.
64, 269–285.
Baskan, O., Kosker, Y., Erpul, G., 2013. Spatial and temporal variation of moisture content
in the soil profiles of two different agricultural fields of semi-arid region. Environ. Monit.
Assess. 185, 10441–10458. http://dx.doi.org/10.1007/s10661-013-3343-8.
Batey, T., 2009. Soil compaction and soil management—a review. Soil Use Manag.
25, 335–345. http://dx.doi.org/10.1111/j.1475-2743.2009.00326.x.
Beckman, J., Borchers, A., Jones, C.A., 2013. Agriculture’s Supply and Demand for Energy
and Energy Products. EIB-112, U.S. Department of Agriculture, Economic Research
Service, Washington, DC. 29 p.
Bender, S.F., Wagg, C., van der Heijden, M.G.A., 2016. An underground revolution: bio-
diversity and soil ecological engineering for agricultural sustainability. Trends Ecol. Evol.
31, 440–452. http://dx.doi.org/10.1016/j.tree.2016.02.016.
Bendfeldt, E.S., Burger, J.A., Daniels, W.L., 2001. Quality of amended mine soils after six-
teen years. Soil Sci. Soc. Am. J. 65, 1736–1744.
Bindi, M., Olesen, J., 2010. The responses of agriculture in Europe to climate change. Reg.
Environ. Change 11, 151–158.
Bishopp, A., Lynch, J.P., 2015. The hidden half of crop yields. Nat. Plants 1, 1–2. http://dx.
doi.org/10.1038/NPLANTS.2015.117.
Blankinship, J.C., Fonte, S.J., Six, J., Schimel, J.P., 2016. Plant versus microbial controls on
soil aggregate stability in a seasonally dry ecosystem. Geoderma 272, 39–50.
Bouma, J., 2014. Soil science contributions towards sustainable development goals and their
implementation: linking soil functions with ecosystem services. J. Plant Nutr. Soil Sci.
177 (2), 111–120.
Bouma, J., 2015. Reaching out from the soil-box in pursuit of soil security. Soil Sci. Plant
Nutr. 61, 556–565. http://dx.doi.org/10.1080/00380768.2015.1045403.
Bouma, J., McBratney, A., 2013. Framing soils as an actor when dealing with wicked envi-
ronmental problems. Geoderma 200–201, 130–139.
Brevik, E.C., Sauer, T.J., 2015. The past, present, and future of soils and human health stud-
ies. Soil 1, 35–46.
Brussaard, L., 1997. Biodiversity and ecosystem functioning in soil. Ambio 26, 563–570.
Cai, X., Zhang, X., Wang, D., 2011. Land availability for biofuel production. Environ. Sci.
Technol. 45, 334–339.
Calanca, P., 2007. Climate change and drought occurrence in the Alpine region: how severe
are becoming the extremes? Glob. Planet. Change 57, 151–160.
Campbell, B.M., Thornton, P., Zoougmore, R., van Asten, P., Lipper, L., 2014. Sustainable
intensification: what is its role in climate smart agriculture? Curr. Opin. Environ. Sustain.
8, 39–43.
Chappell, A., Baldock, J., Sanderman, J., 2016. The global significance of omitting soil ero-
sion from soil organic carbon cycling scheme. Nat. Clim. Change 6, 187–191. http://dx.
doi.org/10.1038/NCCLIMATE2829.
Coleman, D.C., 2008. From peds to paradoxes: linkages between soil biota and their influ-
ences on ecological processes. Soil Biol. Biochem. 40, 271–289.
Collins, M., Knutti, R., Arblaster, J., Dufresne, J.-L., Fichefet, T., Friedlingstein, P.,
Gao, X., Gutowski, W.J., Johns, T., Krinner, G., Shongwe, M., Tebaldi, C.,
Weaver, C.J., Wehner, M., 2013. Long-term climate change: projections, commitments
and irreversibility. In: Stocker, T.F., Qin, D., Plattner, G.-K., Tignor, M., Allen, S.K.,
Boschung, J., Nauels, A., Xia, Y., Bex, V., Midgley, P.M. (Eds.), Climate Change 2013:
The Physical Science Basis. Contribution of Working Group I to the Fifth Assessment
Report of the Intergovernmental Panel on Climate Change. Cambridge University
Press, Cambridge and New York. 108 p.
Cox, C., Hug, A., Bruzelius, N., 2011. Losing Ground. Environmental Working Group,
Ames, IA. Available at: http://static.ewg.org/reports/2010/losingground/pdf/
losingground_report.pdf.
Cruse, R.M., Lee, S., Fenton, T.E., Wang, E., Laflen, J.L., 2013. Soil renewal and sustain-
ability. In: Lal, R., Stewart, B.A. (Eds.), Principles of Sustainable Soil Management in
Agroecosystems. CRC Press, Boca Raton, FL, pp. 477–500.
Curry, J.P., Good, J.A., 1992. Soil faunal degradation and restoration. Adv. Soil Sci.
17, 171–215.
Dabney, S.M., Yoder, D.C., Vieira, D.A.N., 2012. The application of the revised universal
soil loss equation, version 2, to evaluate the impacts of alternative climate change scenar-
ios on runoff and sediment yield. J. Soil Water Conserv. 67, 343–353.
Daccache, A., Keay, C., Jones, R.J.A., Weatherhead, E.K., Stalham, M.A., Knox, J.W.,
2012. Climate change and land suitability for potato production in England and Wales:
impacts and adaptation. J. Agric. Sci. 150, 161–177.
Daily, G.C., Matson, P.A., Vitousek, P.M., 1997. Ecosystem services supplied by soil.
In: Daily, G.C. (Ed.), Nature Services: Societal Dependence on Natural Ecosystems.
Island Press, Washington DC, pp. 113–132.
Dale, V.H., Efroymson, R.A., Kline, K.L., 2011. The land use-climate change-energy nexus.
Landsc. Ecol. 26, 755–773. http://dx.doi.org/10.1007/s10980-011-9606-2.
Davinic, M., Moore-Kucera, J., Acosta-Martı́nez, V., Zak, J., Allen, V., 2013. Soil fungal
distribution and functionality as affected by grazing and vegetation components of inte-
grated crop–livestock agroecosystems. Appl. Soil Ecol. 66, 61–70.
Delgado, J.A., Berry, J.K., 2008. Advances in precision conservation. Adv. Agron. 98, 1–44.
http://dx.doi.org/10.1016/S0065-2113(08)00201-0.
DeLong, C., Cruse, R.M., Wiener, J., 2015. The soil degradation paradox: compromising
our resources when we need them most. Sustainability 7, 866–879. Available at: http://
www.mdpi.com/2071-1050/7/1/866.
den Biggelaar, C., Lal, R., Wiebe, K., Breneman, V., 2001. Impact of soil erosion on crop
yields in North America. Adv. Agron. 72, 1–52. Available at: http://www.sciencedirect.
com/science/article/pii/S006521130172010X.
den Biggelaar, C., Lal, R., Wiebe, K., Eswaran, H., Breneman, V., Reich, P., 2004. The
global impact of soil erosion on productivity II: effects on crop yields and production
over time. Adv. Agron. 81, 49–95. http://dx.doi.org/10.1016/S0065-2113(03)
81002-7.
Dexter, A.R., 2004. Soil physical quality—part I. Theory, effects of soil texture, density, and
organic matter, and effects on root growth. Geoderma 120, 201–214.
Dokuchaev, V.V., 1967. Russian Chernozem (1883). (Translated from Russian into
English by N. Kaner). Israel Program for Scientific Translations Ltd. S. Monson,
Jerusalem. 419 p.
Dregne, H.E., 1977. Generalized Map of the Status of Desertification in Arid Lands.
A/CONF 74/31, In: United Nations Conference on Desertification, Nairobi, Kenya.
Dregne, H.E., 1983. Desertification of Arid Lands. Harwood Academic Publishers, London.
Dregne, H.E., Chou, N., 1992. Global desertification and costs. In: Dregne, H.E. (Ed.),
Degradation and Restoration of Arid Lands. Texas Tech University, Lubbock,
pp. 249–282.
Edelman, A., Gedling, A., Konovalov, E., McComiskie, R., Penny, A., Roberts, N.,
Templeman, S., Trewin, D., Ziembicki, M., 2014. State of the Tropics: 2014 Report.
James Cook University, Australia. Available at: https://www.jcu.edu.au/__data/assets/
pdf_file/0003/246621/SOTT_Final-Report_Low-Res_11-June-2014.pdf.
EEA, 2010. The European Environment: State and Outlook 2010: Soil. European
Environment Agency, Copenhagen, Denmark. 48 p. www.eea.europe.eu/soer/
europe/soil. Accessed 21-May-2016.
Endo, A., Tsurita, I., Burnett, K., Orencio, P.M., 2015. A review of the current state of
research on the water, energy, and food nexus. J. Hydrol.: Reg. Stud. http://dx.doi.
org/10.1016/j.ejrh.2015.11.010.
Eswaran, H., Lal, R., Reich, P.F., 2001. Land degradation: an overview. In: Bridges, E.M.,
Hannam, I.D., Oldeman, L.R., Pening de Vries, F.W.T., Scherr, S.J., Sompatpanit, S.
(Eds.), Responses to Land Degradation. Proceedings of the 2nd International Confer-
ence on Land Degradation and Desertification, Khon Kaen, Thailand. Oxford Press,
New Delhi, India.
Eswaran, H., Reich, P.F., Padmanabhan, E., 2012. World soil resources: opportunities and
challenges. In: Lal, R., Stewart, B.A. (Eds.), World Soil Resources and Food Security.
CRC Press, Boca Raton, FL, pp. 29–52.
Fabiola, R., Dalila, D.R., 2016. How the nexus of water/food/energy can be seen with the
perspective of people well being and the Italian BES framework. Agric. Agric. Sci.
Procedia 8, 732–740.
Falasca, S.L., Ulberich, A.C., Ulberich, E., 2012. Developing an agro-climatic zoning model
to determine potential production areas for castor bean (Ricinus communis L.). Ind. Crop
Prod. 40, 185–191.
FAO, 2011. The State of the World’s Land and Water Resources for Food and Agriculture
(SOLAW)—Managing Systems at Risk. Food and Agriculture Organization of the
United Nations and London, Earthscan, Rome.
FAO, ITPS, 2015. Status of the World’s Soil Resources (SWSR)—Main Report. Food and
Agriculture Organization of the United Nations and Intergovernmental Technical Panel
on Soils, Rome, Italy. Available at: http://www.fao.org/3/a-i5199e.pdf.
Fazal, S., 2000. Urban expansion and loss of agricultural land—a GIS based study of Saha-
ranpur City, India. Environ. Urban. 12, 133–149.
Fenton, T.E., Kazemi, M., Lauterbach-Barrett, M.A., 2005. Erosional impact on organic
matter content and productivity of selected Iowa soils. Soil Tillage Res. 81, 163–171.
Available at: http://www.sciencedirect.com/science/article/pii/S0167198704001928.
Fischer, G., Shcrattenholzer, L., 2001. Global bioenergy potentials through 2050. Biomass
Bioenergy 20, 151–159.
Fischer, G., Shah, M., Tubiello, F.N., van Velhuizen, H., 2005. Socio-economic and climate
change impacts on agriculture: an integrated assessment, 1990–2080. Philos. Trans. R.
Soc. B 360, 2067–2083. http://dx.doi.org/10.1098/rstb.20051744.
Frye, W.W., Bennett, O.L., Buntley, G.J., 1985. Restoration of crop productivity on eroded
or degraded soils. In: Follett, R.F., Stewart, B.A. (Eds.), Soil Erosion and Crop Produc-
tivity. ASA-CSSA-SSSA, Madison, WI, pp. 335–356.
Fuhrer, J., Beniston, M., Fischlin, A., Frei, C., Goyette, S., Jasper, K., Pfister, C., 2006. Climate
risks and their impact on agriculture and forests in Switzerland. Clim. Change 79, 79–102.
Gamborg, C., Millar, K., Shortall, O., Sandøe, P., 2012. Bioenergy and land use: framing the
ethical debate. J. Agric. Environ. Ethics 25, 909–925.
Garbrecht, J.D., Nearing, M.A., Steiner, J.L., Zhang, X.J., Nichols, M.H., 2015. Can con-
servation trump impacts of climate change on soil erosion? An assessment from winter
wheat cropland in the Southern Great Plains of the United States. Weather and Climate
Extremes 10, 32–39.
Gelfand, I., Sahajpal, R., Zhang, X., Izaurralde, R.C., Gross, K.L., Robertson, G.P., 2013.
Sustainable bioenergy production from marginal lands in the US Midwest. Nature
493, 514–517. http://dx.doi.org/10.1038/nature11811.
Ghaley, B.B., Porter, J.R., Sandhu, H.S., 2014. Soil-based ecosystem services: a synthesis of
nutrient cycling and carbon sequestration assessment methods. Int. J. Biodivers. Sci.
Ecosyst. Serv. Manag. 10, 177–186.
Gli
nski, J., Ste˛pniewski, W., 1985. Soil Aeration and its Role for Plants. CRC Press, Boca
Raton, FL. 229 p.
Gregory, P.J., George, T.S., 2011. Feeding nine billion: the challenge to sustainable crop
production. J. Exp. Bot. 62, 5233–5239.
Grêt-Regamey, A., Weibel, B., Kienast, F., Rabe, S.-E., Zulian, G., 2015. A tiered approach
for mapping ecosystem services. Ecosyst. Serv. 13, 16–27.
Gulati, M., Jacobs, I., Jooste, A., Naidoo, D., Fakir, S., 2013. The water-energy-food secu-
rity nexus: challenges and opportunities for food security in South Africa. Aquat.
Procedia 1, 150–164.
Handmer, J., Honda, Y., Kundzewicz, Z.W., Arnell, N., Benito, G., Hatfield, J.,
Mohamed, I.F., Peduzzi, P., Wu, S., Sherstyukov, B., Takahashi, K., Yan, Z., 2012.
Changes in impacts of climate extremes: human systems and ecosystems. In: Field, C.B.,
Barros, V., Stocker, T.F., Qin, D., Dokken, D.J., Ebi, K.L., Mastrandrea, M.D.,
Mach, K.J., Plattner, G.K., Allen, S.K., Tignor, M., Midgley, P.M. (Eds.), Managing
the Risks of Extreme Events and Disasters to Advance Climate Change Adaptation.
A Special Report of Working Groups I and II of the Intergovernmental Panel on Climate
Change (IPCC). Cambridge University Press, Cambridge, UK, and New York, NY,
pp. 231–290.
Hansen, J., Sato, M., Ruedy, R., 2012. Perception of climate change. Proc. Natl. Acad. Sci.
U.S.A. 109, 14726–14727.
Hao, Z., AghaKouchak, A., Phillips, T.J., 2013. Changes in concurrent monthly precipita-
tion and temperature extremes. Environ. Res. Lett. 8, 034014. http://dx.doi.org/
10.1088/1748-9326/8/3/034014. 7 p.
Hartemink, A.E., McBratney, A., 2008. A soil science renaissance. Geoderma 148, 123–129.
Hatfield, J.L., 2012. Spatial patterns of water and nitrogen response within corn production
fields. In: Aflakpui, G. (Ed.), Agricultural Science, Intech Publishers, Rijeka, Croatia, pp.
73–96. ISBN: 978-953-51-0567-1.
Hatfield, J.L., 2014. Soil degradation, land use, and sustainability. In: Songstad, D.D.,
Hatfield, J.L., Tomes, D.T. (Eds.), Convergence of Food Security, Energy
Security and Sustainable Agriculture. In: Biotechnology in Agriculture and Forestry,
vol. 67. Springer, New York, pp. 61–74. http://dx.doi.org/10.1007/978-3-642-
55262-5_4.
Hatfield, J.L., Prueger, J.H., 2015. Temperature extremes: effect on plant growth and devel-
opment. Weather and Climate Extremes 10, 4–10.
Hatfield, J.L., Walthall, C.L., 2015. Meeting global food needs: realizing the potential via
genetics x environment x management interactions. Agron. J. 107, 1215–1226.
Hatfield, J.L., Sauer, T.J., Prueger, J.H., 2001. Managing soils for greater water use efficiency:
a review. Agron. J. 93, 271–280.
Hatfield, J.L., McMullen, L.D., Jones, C.S., 2009. Nitrate-nitrogen patterns in the Raccoon
River Basin related to agricultural practices. J. Soil Water Conserv. 64, 190–199. http://
dx.doi.org/10.2489/jswc.64.3.190.
Hatfield, J.L., Boote, K.J., Kimball, B.A., Ziska, L.H., Izaurralde, R.C., Ort, D.,
Thomson, A.M., Wolfe, D.W., 2011. Climate impacts on agriculture: implications
for crop production. Agron. J. 10, 351–370.
Hatfield, J., Takle, G., Grotjahn, R., Holden, P., Izaurralde, R.C., Mader, T., Marshall, E.,
Liverman, D., 2014. Agriculture. In: Melillo, J.M., Richmond, T.C., Yohe, G.W.
(Eds.), Climate Change Impacts in the United States: The Third National Climate
Assessment. U.S. Global Change Research Program, Washington, DC, pp. 150–174.
http://dx.doi.org/10.7930/J02Z13FR. http://nca2014.globalchange.gov/report/sectors/
agriculture. Chapter 6.
Hewitt, A., Dominati, E., Webb, T., Cuthill, T., 2015. Soil natural capital quantification by
the stock adequacy method. Geoderma 241–242, 107–114.
Hillel, D., 1991. Out of the Earth—Civilization and the Life of the Soil. University of
California Press, Berkeley. 321 p.
Hillel, D., 1994. Rivers of Eden—The Struggle for Water and the Quest for Peace in the
Middle East. Oxford University Press, New York. 355 p.
Himes, F.L., 1997. Nitrogen, sulfur, and phosphorus and the sequestering of carbon. In:
Lal, R. et al., (Ed.), Soil Processes and the Carbon Cycle. CRC Press, Boca Raton,
Florida, pp. 315–319. 609 p.
Holzk€amper, A., Calanca, P., Fuhrer, J., 2013. Identifying climatic limitations to grain maize
yield potentials using a suitability evaluation approach. Agr. Forest. Meteorol.
168, 149–159.
Horn, R., Smucker, A., 2005. Structure formation and its consequences for gas and water
transport in unsaturated arable and forest soils. Soil Tillage Res. 82, 5–14.
Hudson, B.D., 1994. Soil organic matter and available water capacity. J. Soil Water Conserv.
49, 189–194.
Izaurralde, R.C., Thomson, A.M., Morgan, J.A., Fay, P.A., Polley, H.W., Hatfield, J.L.,
2011. Climate impacts on agriculture: implications for forage and rangeland production.
Agron. J. 103, 371–380.
Keil, R.G., Mayer, L.M., 2014. Mineralmatrices and organic matter. Treatise Geochem.
Org. Geochem. 12, 337–359.
King, F.H., 1906. The Soil. The Macmillan Company, London. 303 p.
King, F.H., 1911. Farmers for Forty Centuries. Rodale Press, Inc., Emmaus, PA. 441 p.
Kirkby, C.A., Richardson, A.E., Wade, L.J., Passioura, J.B., Batten, G.D., Blanchard, C.,
Kirkegaard, J.A., 2014. Nutrient availability limits carbon sequestration in arable soils.
Soil Biol. Biochem. 68, 402–409. http://dx.doi.org/10.1016/j.soilbio.2013.09.032.
Kirkham, M.B., 2014. Principles of Soil and Plant Water Relations, second ed. Elsevier
Academic Press, Burlington, MA. 598 p.
Klein, T., Holzk€amper, A., Calanca, P., Seppelt, R., Fuhrer, J., 2013. Adapting agricultural
land management to climate change: a regional multi-objective optimization approach.
Landsc. Ecol. 28, 2029–2047.
Klocke, N.L., Currie, R.S., Aiken, R.M., 2009. Soil water evaporation and crop residues.
Trans. ASABE 52, 103–110.
Krishnaswamy, J., Bonell, M., Venkatesh, B., Purandara, B.K., Rakesh, K.N., Lele, S.,
Kiran, M.C., Reddy, V., Badiger, S., 2013. The groundwater recharge response and
hydrologic services of tropical humid forest ecosystems to use and reforestation: support
for the “infiltration–evapotranspiration trade-off hypothesis” J. Hydrol. 498, 191–209.
Lal, R., 2004. Soil carbon sequestration impacts on global climate change and food security.
Science 304, 1623–1627.
Lal, R., 2009a. Soils and world food security. Soil Tillage Res. 102, 1–4.
Lal, R., 2009b. Ten tenets of sustainable soil management. J. Soil Water Conserv.
64, 20A–21A.
Lal, R., 2011. Sequestering carbon in soils of agro-ecosystems. Food Policy 36 (Suppl. 1),
S33–S39.
Lal, R., 2015a. Sequestering carbon and increasing productivity by conservation agriculture.
J. Soil Water Conserv. 70, 55a–62a.
Lal, R., 2015b. A system approach to conservation agriculture. J. Soil Water Conserv.
70, 82a–88a.
Lal, R., 2016. Global food security and nexus thinking. J. Soil Water Conserv. 71, 85a–90a.
Lal, R., Stavi, I., 2015. Achieving zero net land degradation: challenges and opportunities.
J. Arid Environ. 112 (Pt. A), 44–51.
Lavelle, P., 2000. Ecological challenges for soil science. Soil Sci. 165, 73–86.
Lepers, E., 2003. Synthesis of the Main Areas of Land-Cover and Land-Use Change:
Millennium Ecosystem Assessment, Final Report.
Levidow, L., Zaccaria, D., Mala, R., Vivas, E., Todorovic, M., Scardigno, A., 2014.
Improving water-efficient irrigation: prospects and difficulties of innovative practices.
Agric. Water Manage. 146, 84–94.
Littleboy, M., Silburn, D.M., Freebairn, D.M., Woodruff, D.R., Hammer, G.L.,
Leslie, D.L., 1992a. Impact of soil erosion on production in cropping systems I.
Development and validation of a simulation model. Aust. J. Soil. Res. 30, 757–774.
Littleboy, M., Freebairn, D.M., Hammer, G.L., Silburn, D.M., 1992b. Impact of soil erosion
on production in cropping systems II. Simulation of production and erosion risks for a
wheat cropping system. Aust. J. Soil. Res. 30, 775–788.
Liu, X., Burras, C.L., Kravchenko, Y.S., Duran, A., Huffman, T., Morras, H., Studdert, G.,
Zhang, X., Cruse, R.M., Yuan, X., 2012. Overview of Mollisols in the world: distri-
bution, land use and management. Can. J. Soil Sci. 92, 383–402. http://lib.dr.iastate.
edu/cgi/viewcontent.cgi?article¼1030&context¼agron_pubs.
Lotze-Campen, H., Schellnhuber, H.-J., 2009. Climate impacts and adaptation options in
agriculture: what we know and what we don’t know. J. Verbrauch. Lebensm. 4 (2),
145–150.
Madhu, M., Hatfield, J.L., 2013. Dynamics of plant root growth under increased atmospheric
carbon dioxide. Agron. J. 105, 657–669.
Maracchi, G., Sirotenko, O., Bindi, M., 2005. Impacts of present and future climate variabil-
ity on agriculture and forestry in the temperate regions: Europe. Clim. Change
70, 117–135.
Marks, E., Aflakpui, G.K.S., Nkem, J., Poch, R.M., Khouma, M., Kokou, K., Sagoe, R.,
Sebastia, M.T., 2009. Conservation of soil organic carbon, biodiversity and the provision
of other ecosystem services along climatic gradients in West Africa. Biogeosciences
6, 1825–1838.
McBratney, A., Field, D.J., Koch, A., 2014. The dimensions of soil security. Geoderma
213, 203–213.
Melillo, J.M., Richmond, T.C., Yohe, G.W. (Eds.), 2014. Climate Change Impacts in the
United States: The Third National Climate Assessment. U.S. Global Change Research
Program, Washington, DC, pp. 150–174. http://dx.doi.org/10.7930/J02Z13FR.
Metzger, M., Rounsevell, M., Acosta-Michlik, L., Leemans, R., Schr€ oter, D., 2006. The vul-
nerability of ecosystem services to land use change. Agric. Ecosyst. Environ. 114, 69–85.
Miranowski, J.A., 2005. Energy consumption in U.S. agriculture. In: Outlaw, J., Collins, K.,
Duffield, J. (Eds.), Agriculture as a Producer and Consumer of Energy. CAB
International, Cambridge, MA, pp. 68–108.
Moeletsi, M.E., Walker, S., 2012. Simple agroclimatic index to delineate suitable growing
areas for rainfed maize production in the Free State Province of South Africa. Agric.
For. Meteorol. 162–163, 63–70.
Montgomery, D., 2007. Soil erosion and agricultural stability. Proc. Natl. Acad. Sci. U.S.A.
104, 13268–13272.
Motha, R.P., Baier, W., 2005. Impacts of present and future climate change and climate var-
iability on agriculture in the temperate regions: North America. Clim. Change
70, 137–164.
Murphy, B.W., 2015. Impact of soil organic matter on soil properties—a review with
emphasis on Australian soils. Soil Res. 53, 605–635.
Nazarifar, M.H., Kanani, M.H., Momeni, R., 2012. Analysis of spatial and temporal
variations in crop water productivity of the rainfed wheat for a regional scale analysis.
Agriculture (Poľnohospodárstvo) 58, 65–73. http://dx.doi.org/10.2478/v10207-012-
0008-5.
Nearing, M., Pruski, F., O’Neal, M., 2004. Expected climate change impacts on soil erosion
rates: a review. J. Soil Water Conserv. 59, 43–50.
Neild, R.E., Richman, N.H., 1981. Agroclimatic normals for maize. Agric. Meteorol.
24, 83–95.
Nelson, E., Mendoza, G., Regetz, J., Polasky, S., Tallis, H., Cameron, D., Chan, K.M.,
Daily, G.C., Goldstein, J., Kareiva, P.M., 2009. Modeling multiple ecosystem services,
biodiversity conservation, commodity production, and tradeoffs at landscape scales.
Front. Ecol. Environ. 7, 4–11.
Oldeman, L.R., 1994. The global extent of soil degradation. In: Greenland, D.J., Szabolcs, I.
(Eds.), Soil Resilience and Sustainable Land Use. CAB International, Wallingford,
pp. 99–118.
Oldeman, L.R., Van Engelen, V.W.P., Pulles, J.H.M., 1990. The extent of human-induced
soil degradation. Annex 5. In: Oldeman, L.R., Hakkeling, R.T.A., Sombroek, W.G.
(Eds.), World Map of the Status of Human-Induced Soil Erosion: An Explanatory Note,
second ed. International Soil Reference and Information Centre, Wageningen,
The Netherlands.
Oldeman, L.R., Hakkeling, R.T.A., Sombroek, E.G., 1991. World Map of the Status of
Human-Induced Land Degradation. An Explanatory Note. International Soil
Reference and Information Center, and the United Nations Environment Program,
Nairobi, Kenya. http://www.isric.org/isric/webdocs/docs//ExplanNote.pdf.
Ostergard, H., Finckh, M.R., Fontaine, L., Goldringer, I., Hoad, S.P., Kristensen, K.,
Lammerts van Bueren, E.T., Mascher, F., Munck, L., Wolfe, M.S., 2009. Time for a
shift in crop production: embracing complexity through diversity at all levels. J. Sci. Food
Agric. 89, 1439–1445.
Pan, G.X., Smith, P., Pan, W.N., 2009. The role of soil organic matter in maintaining the
productivity and yield stability of cereals in China. Agric. Ecosyst. Environ.
129, 344–348.
Parlange, M.B., Cahill, A.T., Nielsen, D.R., Hopmans, J.W., Wendroth, O., 1998. Review
of heat and water movement in field soils. Soil Tillage Res. 47, 5–10.
Passioura, J., 2006. Increasing crop productivity when water is scarce—from breeding to field
management. Agr. Water Manage. 80, 176–196. http://dx.doi.org/10.1016/j.
agwat.2005.07.012.
Pereira, L.S., Cordery, I., Iocavides, I., 2012. Improved indicators of water use performance
and productivity for sustainable water conservation and saving. Agr. Water Manage.
108, 39–51. http://dx.doi.org/10.1016/j.agwat.2011.08.022.
Perez-Quezada, J.F., Pettygrove, G.S., Plant, R.E., 2003. Spatial-temporal analysis of yield
and soil factors in two four-crop- rotation fields in the Sacramento Valley, California.
Agron. J. 95, 676–687.
Pimentel, D., 2006. Soil erosion: a food and environmental threat. Environ. Dev. Sustain.
8, 119–137. Available at: http://link.springer.com/article/10.1007%2Fs10668-005-
1262-8.
Pimentel, D., Pimentel, M., 2000. Feeding the world’s population. Bioscience 50, 387.
Pimentel, D., Pimentel, M., 2008. Food, Energy, and Society, third ed. CRC Press, Boca
Raton, Florida.
Pimentel, D., Harvey, C., Resosudarmo, P., Sinclair, K., Kurz, D., McNair, M., Crist, S.,
Sphpritz, L., Fitton, L., Saffouri, R., Blair, R., 1995a. Environmental and economic costs
of soil erosion and conservation benefits. Science 267, 1117–1123. Available at: http://
science.sciencemag.org/content/267/5201/1117.
Pimentel, D., Berger, B., Filiberto, D., Newton, M., Wolfe, B., Karabinakis, E., Clark, S.,
Poon, E., Abbett, E., Nandagopal, S., 2004. Water resources: agricultural and environ-
mental issues. BioScience 54, 909–918.
Purakayastha, T.J., Singh, B.R., Narwal, R.P., Chhonkar, P.K., 2012. Soil resources affect-
ing food security and safety in South Asia. In: Lal, R., Stewart, B.A. (Eds.), World Soil
Resources and Food Security. CRC Press, Boca Raton, FL, pp. 29–52.
Ramsey, A.F., Corty, F.L., 1982. Conservation of prime agricultural land to nonagricultural
uses in one area of the Sunbelt. South. J. Agric. Econ, 23–29.
Ringler, C., Bhaduri, A., Lawford, R., 2013. The nexus across water, energy, land and food
(WELF): potential for improved resource use efficiency? Curr. Opin. Environ. Sustain.
5, 617–624. http://dx.doi.org/10.1016/j.cosust.2013.11.002.
Rosenberg, N.J., Smith, S.J., 2009. A sustainable biomass industry for the North American
Great Plains. Curr. Opin. Environ. Sustain. 1, 121–132.
Safriel, U.N., 2007. The assessment of global trends in land degradation. In: Sivakumar, M.V.K.,
Ndiang’ui, N. (Eds.), Climate and Land Degradation. Springer-Verlag, Berlin Heidelberg,
pp. 1–38.
Salley, S.W., Sleezer, R.O., Bergstrom, R.M., Martin, P.H., Kelly, E.F., 2016. A long-term
analysis of the historical dry boundary for the Great Plains of North America: implica-
tions of climatic variability and climatic change on temporal and spatial patterns in soil
moisture. Geoderma 274, 104–113.
Sardans, J., Rivas-Ubach, A., Peñuelas, J., 2012. The C:N:P stoichiometry of organisms and
ecosystems in a changing world: a review and perspectives. Perspect. Plant Ecol.
Evol. Syst. 14, 33–47. http://dx.doi.org/10.1016/j.ppees.2011.08.002.
Sauer, T.J., Daniel, T.C., 1987. Effect of tillage system on runoff losses of surface-applied
pesticides. Soil Sci. Soc. Am. J. 51, 410–415.
Sauer, T.J., Hatfield, J.L., Prueger, J.H., Norman, J.M., 1998. Surface energy balance of a
corn residue-covered field. Agr. Forest. Meteorol. 89, 155–168.
Schilling, K., Zhang, Y., 2004. Baseflow contribution to nitrate-nitrogen export from a large
agricultural watershed, USA. J. Hydrol. 295, 305–316.
Siebert, S., Burke, J., Faures, J.M., Frenken, K., Hoogeveen, J., D€ oll, P., Portmann, F.R.,
2010. Groundwater use for irrigation—a global inventory. Hydrol. Earth Syst. Sci.
14, 1863–1880. http://dx.doi.org/10.5194/hess-14-1863-2010.
Simane, B., Struik, P.C., 1993. Agroclimatic analysis: a tool for planning sustainable durum wheat
(Triticum turgidum var. durum) production in Ethiopia. Agr. Ecosyst. Environ. 47, 31–46.
Sivakumar, M.V.K., Das, H.P., Brunini, O., 2005. Impacts of present and future climate var-
iability and change on agriculture and forestry in the arid and semi-arid tropics. Clim.
Change 70, 31–72.
Six, J., Paustian, K., 2014. Aggregate-associated soil organic matter as an ecosystem property
and measurement tool. Soil Biol. Biochem. 68, A4–A9.
Skaggs, T.H., Shouse, P.J., 2008. Roots and root function: introduction. Vadose Zone J.
7, 1008–1009. http://dx.doi.org/10.2136/vzj2008.oo76.
Smajgl, A., Ward, J., Pluschke, L., 2016. The water-food-energy Nexus—Realising a new
paradigm. J. Hydrol. 533, 533–540.
Smith, P., Gregory, P.J., van Vuuren, D., Obersteiner, M., Havlik, P., Rounsevell, M.,
Woods, J., Stehfest, E., Bellarby, J., 2010. Competition for land. Philos. Trans. R.
Soc. B 365, 2941–2957.
Stevenson, F.J., 1972. Humus Chemistry: Genesis, Composition, Reactions. Wiley Press,
New York.
Stewart, B.A., Steiner, J.L., 1990. Water-use efficiency. In: Singh, R.P. et al., (Ed.), Dryland
Agriculture: Strategies for Sustainability. Advances in Soil Science, vol. 13. Springer-
Verlag, New York, pp. 151–173.
Stoll, S., 2002. Larding the Lean Earth—Soil and Society in Nineteenth-Century America.
Hill and Wang, New York. 287 p.
Sutton, P.C., Costanza, R., 2002. Global estimates of market and non-market values derived
from nighttime satellite imagery, land cover, and ecosystem service valuation. Ecol.
Econ. 41, 509–527.
Swella, G.B., Ward, P.R., Siddique, K.H.M., Flower, K.C., 2015. Combinations of tall stand-
ing and horizontal residue affect soil water dynamics in rainfed conservation agriculture
systems. Soil Tillage Res. 147, 30–38. http://dx.doi.org/10.1016/j.still.2014.11.004.
Swinton, S.M., Lupi, F., Robertson, G.P., Landis, D.A., 2006. Ecosystem services from agri-
culture: looking beyond the usual suspects. Am. J. Agric. Econ. 88, 1160–1166.
Syers, J.K., 1997. Managing soils for long-term productivity. Philos. Trans. R. Soc. Lond.
B Biol. Sci. 352, 1011–1021.
Taniguchi, M., Masuhara, N., Burnett, K., 2015. Water, energy, and food security in the Asia
Pacific region. J. Hydrol.: Reg. Stud. http://dx.doi.org/10.1016/j.ejrh.2015.11.005
Tao, F., Yokozawa, M., Liu, J., Zhang, Z., 2009. Climate change, land use change, and
China’s food security in the twenty-first century: an integrated perspective. Clim.
Change 93, 433–445.
Taylor, H.W., Jordan, W.R., Sinclair, T.S. (Eds.), 1983. Limitations to Efficient Water Use
in Crop Production. American Society of Agronomy, Madison, Wisconsin. 538 p.
Tilman, D., Balzer, C., Hill, J., Befort, B.L., 2011. Global food demand and the sustainable
intensification of agriculture. Proc. Natl. Acad. Sci. U.S.A. 108, 20260–20264.
Tomer, M.D., Porter, S.A., Boomer, K.M.B., James, D.E., Kostel, J.A., Helmers, M.J.,
Isenhart, T.M., McLellan, E., 2015a. Agricultural conservation planning framework:
1 developing multipractice watershed planning scenarios and assessing nutrient
reduction potential. J. Environ. Qual. 44, 754–767. http://dx.doi.org/10.2134/
jeq2014.09.0386.
Tomer, M.D., Boomer, K.M.B., Porter, S.A., Gelder, B.K., James, D.E., McLellan, E.,
2015b. Agricultural conservation planning framework: 2. Classification of riparian buffer
design types with application to assess and map stream corridors. J. Environ. Qual.
44, 768–779. http://dx.doi.org/10.2134/jeq2014.09.0387.
Torriani, D., Calanca, P., Schmid, S., Beniston, M., Fuhrer, J., 2007. Potential effects of
changes in mean climate and climate variability on the yield of winter and spring crops
in Switzerland. Climate Res. 34, 59–69.
Tsiafouli, M.A., Thebault, E., Sgardelis, S.P., De Ruiter, P.C., van der Putten, W.H.,
Birkhofer, K., Hemerik, L., De Vries, F.A., Bardgett, R.D., Brady, M.V.,
Bjornlund, L., Jorgensen, H.B., Christensen, S., Hertefeldt, T.D., Stary, S.,
Wolters, V., Hedlund, K., 2015. Intensive agriculture reduces soil biodiversity across
Europe. Glob. Chang. Biol. 21, 973–985.
Tull, J., 1762. Horse-Hoeing Husbandry or An Essay on the Principles of Vegetation and
Tillage, fourth ed. A. Millar, London. 432 p.
U.S. Department of Energy. 2011. U.S. Billion-Ton Update: Biomass Supply for a Bio-
energy and Bioproducts Industry. In: Perlack, R.D., Stokes, B.J. (Leads), ORNL/
TM-2011/224. Oak Ridge National Laboratory, Oak Ridge, TN. 227 p.
Unger, P.W., Kaspar, T.C., 1994. Soil compaction and root growth: a review. Agron. J.
86, 759–766.
Unger, P.W., Stewart, B.A., 1983. Soil management for efficient water use: an overview.
In: Taylor, H.M. et al., (Ed.), Limitation to Efficient Water Use in Crop Production.
American Society of Agronomy, Madison, Wisconsin, pp. 419–460. 538 p.
van Eekeren, N., de Boer, H., Hanegraaf, M., Bokhorst, J., Nierop, D., Bloem, J.,
Schouten, T., de Goede, R., Brussaard, L., 2010. Ecosystem services in grassland asso-
ciated with biotic and abiotic soil parameters. Soil Biol. Biochem. 42, 1491–1504.
van Wart, J., van Bussel, L.G.J., Wolf, J., Licker, R., Grassini, P., Nelson, A., Boogaard, H.,
Gerber, J., Mueller, N.I.D., Claessens, L., van Ittersum, M.K., Cassman, K.G., 2013.
Use of agro-climatic zones to upscale simulated crop yield potential. Field Crops Res.
143, 44–55.
Vanlauwe, B., Coyne, D., Gockowski, J., Hauser, S., Hussing, J., Masso, C., Nziguheba, G.,
Schut, M., Van Asten, P., 2014. Sustainable intensification and the African smallholder
former. Curr. Opin. Environ. Sustain. 8, 15–22.
Viets Jr., F.G., 1966. Increasing water use efficiency by soil management. In: Pierre, W.H.
et al., (Ed.), Plant Environment and Efficient Water Use. American Society of
Agronomy, Madison, Wisconsin, pp. 259–274. 295 p.
Wakatuski, T., Rasyidin, A., 1992. Rates of weathering and soil formation. Geoderma
52, 251–262. Available at: http://www.sciencedirect.com/science/article/pii/
001670619290040E.
Wall, D.H., Nielsen, U.N., Six, J., 2015. Soil biodiversity and human health. Nature
528, 69–76.
Wallander, S., Claassen, R., Nickerson, C., 2011. The Ethanol Decade: An Expansion of U.S.
Corn Production, 2000–09. EIB-79. U.S. Department of Agriculture, Economic
Research Service, Washington, DC. 16 p.
Walsh, J., Wuebbles, D., Hayhoe, K., Kossin, J., Kunkel, K., Stephens, G., Thorne, P.,
Vose, R., Wehner, M., Willis, J., Anderson, D., Doney, S., Feely, R., Hennon, P.,
Kharin, V., Knutson, T., Landerer, F., Lenton, T., Kennedy, J., Somerville, R.,
2014. Our changing climate. In: Melillo, J.M., Richmond, T.C., Yohe, G.W. (Eds.),
Climate Change Impacts in the United States: The Third National Climate Assessment.
U.S. Global Change Research Program, Washington, DC, pp. 19–67. http://dx.doi.
org/10.7930/J0KW5CXT. Chapter 2.
West, P.C., Gibbs, H.K., Monfreda, C., Wagner, J., Barford, C.C., Carpenter, S.R.,
Foley, J.A., 2010. Trading carbon for food: global comparison of carbon stocks vs. crop
yields on agricultural land. Proc. Natl. Acad. Sci. U.S.A. 46, 19645–19648.
Wise, T.A., 2013. Can we feed the world in 2050? A scoping paper to assess the evidence.
Global Development and Environment Institute. Working Paper No. 13-04. Tufts
University. 38 p. http://ase.tufts.edu/gdae. Accessed 21-May-2016.
Wright, C.K., Wimberly, M.C., 2012. Recent land use change in the Western Corn Belt
threatens grasslands and wetlands. Proc. Natl. Acad. Sci. U.S.A. 110, 4134–4139.
http://dx.doi.org/10.1073/pnas.1215404110.
WWAP (United Nations World Water Assessment Programme), 2015. The United Nations
World Water Development Report 2015: Water for a Sustainable World. UNESCO,
Paris. 139 p.
Zabel, F., Putzenlechner, B., Mauser, W., 2014. Global agricultural land resources—a high
resolution suitability evaluation and its perspectives until 2100 under climate change
conditions. PLoS One 9, e107522. www.plosone.org.
CHAPTER TWO
Humin: Its Composition and

Importance in Soil Organic Matter
Michael H.B. Hayes*, Rosaleen Mylotte†, Roger S. Swift{,1
*Carbolea Research Group, Chemical Sciences, University of Limerick, Limerick, Ireland
†
Chemical Sciences, University of Limerick, Limerick, Ireland
{
Queensland Alliance for Agriculture and Food Innovation, University of Queensland, Brisbane, QLD,
Australia
1
Corresponding author: e-mail address: r.swift@uq.edu.au
Contents
1. Introduction 48
2. Historical Perspectives of Humic Substances and Humin 51
2.1 Selected Theories for the Genesis of Humic Substances 52
2.2 Humin in the Historical Context 58
3. Modern Approaches to Studies of Humin 61
3.1 Isolation Processes for Humin 61
3.2 Applications of NMR Spectroscopy for Studies of the Compositions
of Humins 65
3.3 Two Dimensional NMR Spectroscopy 71
4. Information About the Isolation and Compositions of Humin From Applications
of Different NMR Procedures 74
4.1 Effects of Different Extraction and Fractionation Procedures 74
4.2 Information From DE 1H NMR Spectra 81
4.3 Information From CMP-NMR for Studies of Humin 85
4.4 Applications of 2D NMR Spectroscopy to Studies of Humin 87
5. Possible Sources of Humin Components Derived From Plant and Microbial
Sources 94
5.1 Cellulose 94
5.2 Hemicelluloses 95
5.3 Lignin 96
5.4 Tannins 98
5.5 Lipids 100
5.6 Cutin and Cutan 102
5.7 Suberin and Suberan 105
5.8 Latex Materials 108
5.9 Algaenan 108
5.10 Bacteran 110
5.11 Glomalin 110
5.12 Melanins 111
5.13 Black Carbon, Char/Biochar 111

48 Michael H.B. Hayes et al.
5.14 Summary of Section 5 113

6. Summation, Significance, and Future Directions 113
6.1 Summation 113
6.2 Significance of Humin Composition 117
6.3 Future Directions 119
References 122
Abstract
Humin is the insoluble component of soil organic matter (SOM) that remains after
extraction of the other components of SOM that are soluble in aqueous base. Humin
usually makes up a substantial component of SOM, but its lack of solubility and intrac-
table nature have made it difficult to study. To put this contemporary review of humin
into perspective, a summary of the historical development of SOM studies and the ter-
minology used to describe the various components of SOM is presented.
Two major recent advances, namely, sophisticated nuclear magnetic resonance
(NMR) techniques and the introduction of a solvent system (acidified dimethylsulfoxide)
capable of dissolving humin, have facilitated studies of its composition. The NMR tech-
niques used are briefly explained and the results obtained from their application are
presented. Based on this information, the possible contributions to humin of various
plant/soil components are assessed.
The major components of the humin are predominantly aliphatic hydrocarbon
functionalities, especially those found in lipids, waxes, cuticular materials, cutin/cutan,
and suberin/suberan, which are relatively minor components of plants. There is also evi-
dence for small amounts of carbohydrate (possibly highly ordered cellulose), peptide,
and peptidoglycans; but there is little evidence for lignin-derived structures. The com-
position of humin differs considerably from the base-soluble components of the SOM.
All components of SOM eventually decompose but at different rates, but the accumu-
lation of humin constituents relative to their levels in plants indicates that they are
relatively resistant to decomposition. The environmental consequences of humin
composition and the opportunities presented are discussed and future possibilities
outlined.
1. INTRODUCTION
Humin is operationally defined as being insoluble in aqueous alkali
(Hayes and Swift, 1978; Kononova, 1966), and in most cases, it is by far
the major component of soil organic matter (SOM). For many years, humin
was thought to be similar in basic structural features and chemical compo-
sition to the humic substances (HSs) such as the more readily extracted fulvic
and humic components (Hayes and Swift, 1978; Kononova, 1975). The dif-
ferences between humin and the more soluble humic substances were
Humin in Soil Organic Matter 49
thought to lie mainly in properties such as higher molecular weight and

lower levels of functional groups (particularly carboxyl and hydroxyl) that
give rise to lower charge density and decreased polarity resulting in
decreased solubility in alkaline aqueous extractants.
The senior authors of this review were, for many years, both proponents
and adherents of the above concept of “humin” as a more recalcitrant form
of HSs (Hayes and Swift, 1978, 1990), but they were, nonetheless, troubled
about some aspects of this concept. Over the last two decades, many pieces
of evidence have emerged from a number of workers which have led us (and
them) to question the prevailing view of humin composition. What began as
a trickle of information supporting a different view has swelled to a steady
stream of consistent data pointing to a different concept of the composition
of humin.
Accordingly, we believe that it is timely to carefully review the emerging
evidence and to express our current opinions on the composition of humin
based on a sound and growing body of evidence. In doing so, it is important
to set these recent advances against a backdrop of the history and the devel-
opment of concepts of SOM that have guided us to our collective under-
standing in this complex, often frustrating, but ultimately rewarding area
of study.
It is well recognized that the organic matter contents of soils in different
ecosystems, such as in long-term grassland, cultivation, or forest soils, reach a
steady state that depends on the natural ecosystem or the management sys-
tem employed. Under these conditions, the annual inputs of plant residues
into the soil are equal to the annual losses from the soil to the atmosphere,
principally as carbon dioxide from biological decomposition processes.
When the nature of the ecosystem or the management practice is chan-
ged, for example, from grassland or forest to intermittent or long-term cul-
tivation, there is a decline in the input of plant residues and an increase in soil
disturbance. As a consequence, there is a net decrease in the SOM content as
the result of the activity of soil microorganisms. The losses are most rapid in
the early stages of the changes because the readily labile materials are more
easily metabolized by microorganisms. Thus, materials such as starch, pep-
tides, and nucleic acids will be readily metabolized by the microorganisms,
unless these are protected by one or more of several mechanisms. Subse-
quently, as the more labile components are depleted, the rates of carbon
losses slow with time, and eventually a new steady state is reached in which
the SOM content is lower than at the start (Johnson, 1995). Studies of the
quantitative changes that take place when management systems are altered
have been summarized by Clapp et al. (2005).
It is well known that some of the components of SOM have a degree of
resistance to biological degradation. However, all will degrade eventually
because, as Jenkinson (1981) has stated, “in the long run, no fraction of
organic matter in plants and animals can withstand decomposition to carbon
dioxide and water. If this were not so, any completely resistant fraction
would by now cover the surface of the earth.”
An earlier review by the authors (Hayes and Swift, 1978) provided a clas-
sification of HSs based on solubility criteria built upon the proposals of
Kononova (1966, 1975) and utilized the best relevant information available
at the time. In that classification, SOM is separated into unaltered organic
materials that include fresh debris and nontransformed components of older
nonliving debris, and into transformed products (i.e., humus) that bear no
morphological resemblances to the structures from which they were
derived. These transformed products are referred to as humified materials
that can be divided into amorphous, brown-colored HSs and into com-
pounds that belong to recognizable classes such as saccharides, peptides,
nucleic acids, and altered lignins. These compounds can be fully synthesized
by microorganisms and/or formed from alterations of similar compounds in
the original debris. The latter set of materials, though not HSs per se, are
nevertheless components of the more broadly defined humus fraction.
Essentially, the thinking at that time was that humified, nonliving SOM
could be described based on solubility criteria in aqueous acidic or alkaline
solutions. The components that dissolved in alkali contained the humic acids
(HAs, precipitated at pH 1 from the alkaline solution) and the fulvic acids
(FAs), the fraction that remained in solution following acidification, and
humin, a term introduced by Berzelius (1839) to identify the alkali insoluble
fraction of SOM (Kononova, 1966). This relatively crude system of charac-
terization is still valuable and remains in common use.
The major research focus in the last 200 years (and particularly so in the
last 60 years) has been on the HAs and FAs, mainly because these could be
dissolved and isolated using aqueous media. The consensus was that these
HAs and FAs had the greatest significance of the defined humic fractions
insofar as their reactivities and beneficial functions in soils were concerned.
Now that procedures are available to isolate and to study compositional
aspects of humin, it is becoming evident that this fraction plays important
roles in soils and in agriculture. Also, the significance of humin in the seques-
tration of atmospheric carbon is being increasingly recognized. It is
estimated that there are approximately 2344 giga tons (Gt) of organic carbon
(OC) in the top 3 m of the world’s soils, and that is more than four times the
living pool of OC on the earth’s surface (Guo and Gifford, 2002; Jobbagy
and Jackson, 2000; Stockmann et al., 2013).
Humin is considered to make up 50% or more of the OC in mineral soils
and more than 70% of that in lithified sediments (Hedges and Keil, 1995).
Thus, it can be assumed that humin is the most abundant operationally
defined class of organic substances in the terrestrial environment, and the
major contributor to the sequestration of OC in soil. In recent years the term
“humin” is used to describe the dark precipitates formed during the hydro-
lysis of lignocellulosic biomass in second-generation biorefining operations.
The term used in that context has no relation to the concept introduced by
Berzelius (1839) or to the composition of humin as used in the context of
components of SOM. It is therefore important to have a better awareness of
the composition of humin in the context of SOM, of its associations with
other soil components, and of its role in carbon sequestration.
Kleber and Johnson (2010) have reviewed advances in our understanding
of the structural features of SOM and its role in the environment. We will
outline aspects of the evolutionary processes that have led to our current
understanding of the composition of HSs as a whole, but our major emphasis
will be on the humin fraction. Our rationale for this is based on its quanti-
tative importance, and because of the recent development of chemical pro-
cedures and instrumental techniques that give a greater understanding of its
origins and composition.
2. HISTORICAL PERSPECTIVES OF HUMIC SUBSTANCES

AND HUMIN
In the early part of the 19th century, it was thought that the uptake of
humus from soil was the source of the organic materials in plants. Sprengel
(1826) disproved this concept and showed that it was the mineral salts asso-
ciated with the soil and the HAs that provided the plant nutrients. Later,
Berzelius (1839) described what we now regard as HAs, FAs, and humin
based on their solubilities in aqueous basic and acidic media. Although it
may not be the direct source of some of the major plant nutrients, the impor-
tance of SOM for soil fertility has, nonetheless, long been recognized.
Around 400 BC, in a wide-ranging treatise on agriculture and soils, the
Greek historian and philosopher Xenophon wrote “whatever weeds are
grown upon the ground, being turned into the earth enrich the soil as much
as dung” (translation by Dakyns, 1897). In De Re Rustica, Lucius Columella

discusses in Book II the importance of organic matter amendments for
maintaining soil fertility (translated by Ash, 1941). The term humus, intro-
duced by Wallerius (1761) (Hayes, 2009; Russell, 1950, 1973; Waksman,
1936), refers to decomposed organic matter, and the term humus is widely
used in conversation to describe transformed OM in soils and composts.
Though aqueous base is still the solvent of choice for HSs, the emergence
of new isolation and fractionation procedures and above all else the availabil-
ity of advanced analytical instrumentation, in particular nuclear magnetic
resonance (NMR) techniques, have facilitated studies of the compositions
of the fractions of organic matter in isolates from soil. Recent research sug-
gests that the physical and chemical composition of the humin fraction dif-
fers substantially from the HSs. Thus, it is time to critically evaluate our
concepts of the nature of humic components and of humin in particular.
2.1 Selected Theories for the Genesis of Humic Substances

Until the microbiological sciences began to emerge, emphasis was placed in
chemical synthesis as origins for HSs (Hayes, 2009). Recognition of the key
involvement of microorganisms in decomposition processes, inspired by the
work of Pasteur, focussed interest on the roles that microorganisms play in
the transformations of organic debris in the soil environment. Studies, such
as those by von Post (1862), Darwin (1881), M€ uller (1887), and Ramann
(1888), indicated that the genesis of humus is essentially a biological, and
not a chemical, process. Their findings led to biological and chemical studies
based on the release of component molecules from the degradations of plant
components. Dokuchaev (1883) had a definitive influence when he defined
soil as a natural body formed through the combined action of natural factors,
and in particular to the biological factors contributing to soil formation, such
as vegetation cover and the activities of living organisms. Thus, humus was
seen to have an important role in soil formation and soil fertility. As a result, a
new era of soil humus studies was introduced at the end of the nineteenth
century, and there arose a general acceptance that HSs are complex com-
pounds formed as the result of decomposition of plant materials.
Schreiner and Shorey (1910) also regarded soil humus as a complex mix-
ture of organic substances arising from the decomposition of materials of
plant and animal origins. They considered that artifacts are formed during
the isolation of HSs in alkaline media and so they focussed on the isolation
of organic substances using the techniques and solvents of organic chemistry.
They used the “acid filtrates” after the precipitation of HAs, and the ethanol
extracts of the precipitated materials. In this way, they isolated and identified
more than 40 compounds that included hydrocarbons, sterols, fats, organic
acids, carbaldehydes, organophosphorus, and N-containing compounds.
Although this work was outstanding for its time, Shmuk (1924), in a review
of the work of Schreiner and Shorey, considered that their approach tended
to divide the humus concept into small groups of peripheral units and
overlooked the major reserve of organic substances in the soil. That is
true, and it would not be the last time that considerable time and effort
has been expended in detecting and identifying very minor components
of SOM that have shed little light on the composition of the major active
components.
Trusov (1914) carried out a systematic study of humus formation. Ini-
tially he subjected plant components, proteins, cellulose, plant oils, and tan-
nins to treatment with strong acids. Later, recognizing the importance of
biological processes, he (Trusov, 1915) studied the humification of plant
components under normal biological conditions and then (Trusov,
1916a,b) studied plant residues, leaves, grass, and woody species under sim-
ilar conditions. He concluded that the plant components most readily uti-
lized by microorganisms are first converted to microbial plasma and this
then participates in the formation of humus. Those plant residues not uti-
lized by microorganisms (such as lignin and tannins) were considered to
be direct sources of HSs. These concepts are highly relevant to modern
views. Trusov’s contemporary Shmuk (1924) was first to establish that soil
HAs contained benzenoid structures, although Hoppe-Seyler (1889) had
already shown that peats and coals had aromatic units in their compositions.
By esterification procedures, Shmuk showed that HSs had hydroxyl (of phe-
nolic origins) as well as carboxylic functional groups. He proposed that two
components were contained in the HA molecule—one of these was an
organic N-containing compound of microbial origin, and the second the
benzene ring. He regarded these components to be linked and not present
as a mixture.
Based on a series of studies commencing in 1902, Williams (1939) pro-
duced a set of perceptive conclusions (as listed by Kononova, 1966):
(1) that humus substances exist in soil as a natural body;
(2) that various plant materials that undergo complex biochemical transfor-
mations serve as sources of humus substances;
(3) that plant materials decompose to more simple products from which the
complex humic substances are synthesized; and
(4) that microbial enzymatic processes are involved in the decomposition

and in the synthesis processes.
Thus, it was accepted in the early decades of the 20th century that micro-
organisms had an important role in the synthesis of humus. Some scientists
considered the synthesis to be wholly biological, but there was a growing
concept that the compounds released in the microbial breakdown of organic
substrates could condense and give rise to chemical synthesis processes.
Deherain (1902) considered HSs to be products of interactions between
proteins and “encrusting substances” (mainly lignin). This concept preceded
the ligno-protein theory attributed to Waksman (1936) and Waksman and
Iyer (1932, 1933) that subsequently dominated concepts of the origins and
compositions of HSs for a generation. Their work indicated that oxidized
lignin when reacted with protein gave a product similar to HAs, and based
on their work the concept took hold of HAs as complexes of oxidized lignin
with protein.
Hayes (1960) repeated and extended the experimentation used by
Waksman and Iyer (1932, 1933) and isolated the product that had been reg-
arded as a ligno-casein complex. Although a product was formed that had
similar elemental compositions and electrophoretic mobility to those from
a Sapric Histosol HA, it was clear from the differential thermal analysis
(DTA) thermograms that the so-called complex was a mixture of oxidized
lignin and of casein, and was very different from the thermogram of the HA
(Hayes, 2009; Hayes et al, 1984). These results together with the general
observation that the amounts of amino acids present are insufficient to form
a ligno-protein complex led to the decline in the popularity of this theory.
Maillard (1912, 1916, 1917) showed that browning products from the
reaction of glucose with glycine had some resemblance to natural humic
materials from soil. Consequently, considerable emphasis was placed in
the middle of the 20th century on what is called the Browning reaction
(Ellis, 1959; Hodge, 1953) and on its possible contributions to the genesis
of HSs.
Ellis (1959) has defined the Maillard reaction as the “reaction of the
amino group of amino acids, peptides, or proteins with the glycosidic
hydroxyl group of sugars,” and he and Hodge (1953) have presented excel-
lent reviews of the chemistry of the Maillard–Browning reaction. The initial
reaction of glucose with glycine is followed by other more complex changes
that result eventually in the formation of brown pigments and macromole-
cules. Maillard (1917) showed that the products of the glucose–glycine reac-
tion resembled in many respects those of natural humic materials from soil.
That stimulated interest in the abiotic synthesis of humic substances from

sugars and amino acids liberated from the hydrolysis of polysaccharide
and of peptide materials.
Subsequently, Enders and Fries (1936) observed what they considered to
be a relationship between melanoidins (or browning products) and HAs, and
Enders and Marquardt (1941) showed that methylglyoxal [CH3C(O)C(O)
H] could give rise to melanoidins and “caramel” products. Then, based on
his experiments to establish mechanisms of formation of methylglyoxal from
hexose sugars, Enders (1942, 1943a) showed that glucose in aqueous solu-
tion was in equilibrium with the triose glyceraldehyde, and this triose could
give rise to methylglyoxal.
Enders and Sigurdsson (1943) postulated a pathway from methylglyoxal
to acetaldehyde (via pyruvic acid), and acetaldehyde could polymerize via
the aldol condensation mechanism. In order to show how HAs could be
formed under physiological conditions, Enders (1943b) showed that
methylglyoxal could be released by soil microorganisms under conditions
unfavorable for microbial growth (lack of substrate, low or high tempera-
ture, etc.) and then polymerize rapidly in the presence of amino acids. Later,
Enders and Sigurdsson (1947) showed the presence of methylglyoxal in 10 of
the 16 soils tested, and Enders et al. (1948) showed that the products of this
reaction had many properties similar to those of soil HSs.
Schuffelen and Bolt (1950) prepared methylglyoxal glycine products that
had C/N and exchange capacity values similar to those for Dalgrund peat,
and Hayes (1960, 2009) prepared a product from the same reagents that had
elemental analysis data, electrophoretic mobility, and DTA data similar to
those for a HA from a Sapric Histosol. Also there was a greater similarity
with the Histosol HA in the case of the methylglyoxal/glycine macromo-
lecular material than there was for the glucose/glycine reaction product
(Hayes, 1960, 2009).
Burdon (2001) has raised questions that would suggest that the Maillard
reaction is not a significant contributor to the synthesis of soil HSs. He has
pointed out that the concentrations of reducing sugars or of amino acids in
the soil solution are not sufficient to allow the reaction to take place to any
great extent. Since the Maillard reaction proceeds best under alkaline con-
ditions (Ellis, 1959), there should be more HSs in alkaline soils, and there are
not. Burdon referred to the spectroscopic data that show that lignin is a
major contributor to the aromaticity of humic materials, whereas the aro-
matic components in Maillard reaction products are largely composed of
heterocyclic N-containing substances.
However, based on the reasoning of Burdon (2001), acceptable evidence

about the extents to which methylglyoxal polymerization products, and of
the methylglyoxal/glycine reaction products, will be revealed by composi-
tional evidence from NMR data. Nevertheless, it should be stated here that
it is extremely unlikely that browning reaction products, regardless of the
reactants, can be major contributors to the compositions of soil HSs, but
we cannot rule out the possibility that these can make a minor contribution
to the mixture at least until the potential reactions are studied under condi-
tions that could prevail in soils. For example, in situations where soil tem-
perature is appropriate and sufficient moisture is maintained in the SOM for
microbial activity to prevail for a suitable period of time, then it would be
possible for condensation reactions to take place to give humic type
products.
Haworth and his colleagues (Cheshire et al., 1967; Haworth, 1971)
identified a variety of fused aromatic structures in the zinc dust distillation
and fusion reduction digests of “acid-boiled” HAs. They proposed that
“humic” molecules have a polycyclic aromatic core to which polysaccha-
rides, simple phenols, proteins or peptides, and metals are attached by
chemical or physical bonding processes. However, the zinc dust distillation
and fusion procedures involve very harsh reaction conditions and the prod-
uct yields are always very low. It has become accepted that such procedures
can lead to excessive bond breaking, and the recombination of fragments
can give fused aromatic structures (see Clapp et al., 2005; Hayes and
Swift, 1978). The formation of artifacts under these conditions is inevitable
(Burdon, 2001). Polyhydroxyaromatic compounds, quinones, and furfu-
rals, for example, would give rise to fused aromatic structures under
the conditions applied. Long-chain hydrocarbons were identified in the
digests, and these can be considered to have survived the distillation process.
Once again, the general consensus is that polycyclic aromatic compounds
do not make a significant contribution to the structure of naturally
occurring HSs.
A more likely source of polycyclic aromatic structures in soil has emerged
as a result of increased awareness of char, black carbon, or charcoal products
that have been produced by the burning of vegetation and fossil fuels over
time (Skjemstad et al., 2002). These char materials will be composed mainly
of fused aromatic structures, and the peripheral oxidation of the aromatic
structures will give rise to acidic carboxyl groups that could lead to the clas-
sification of these products such as HAs and FAs. However, in most soils, the
char/oxidized char contents will be small. Hence, even in this circumstance,
there is no convincing evidence to support Haworth’s structural hypothesis

for natural humic substances.
Based on the abundance of lignin in biomass, it is logical to consider that
lignin degradation products, or lignols, would contribute to the composi-
tions of HSs. Trusov (1915, 1916a,b; see Kononova, 1966) introduced
the concept that polyphenols and quinones contribute to the synthesis of
HSs. Subsequently, it was suggested that as a result of microbial enzymatic
oxidation, the phenols could oxidize to quinones, and these, through further
condensation, would be converted into dark-colored HSs. The contribution
of quinones to the compositions of HSs has been investigated extensively by
Flaig et al. (1975). Martin et al. (1967) took a different approach in which
they cultured the fungus Epicoccum nigrum on a glucose/asparagine medium
containing yeast extract and inorganic salts. After incubation, they isolated,
at pH 2, mycelium-free “HA” substances. Some of these were similar to
leonardite HAs in terms of elementary composition, total acidity, cation-
exchange capacity, carboxyl, phenolic, and hydroxyl contents, and in
molecular weight distributions. Haider and Martin (1967) illustrated a plau-
sible scheme for the genesis from nonaromatic precursors of phenols iden-
tified from the E. nigrum culture. There is strong evidence from that era to
indicate that fungi contribute significantly to the genesis of soil humus.
Intracellular enzymatic synthesis would seem to be likely, but extracellular
enzymatic catalytic synthesis cannot be ruled out (Hayes and Swift, 1978).
Burdon (2001) has posed a very relevant question when he asked “why
should a micro-organism expend energy and resources making a material
that it has no use for? Any organism that did this would become extinct
because of competition by organisms that did not waste energy and resources
in this way.” His succeeding arguments leave room to consider that the
“humic-type” materials are formed by enzymatic catalysis that render waste
products innocuous to the organism and are stored inside the fungal myce-
lium. A case can be made, using modern NMR spectroscopy techniques, for
further studies of the compositions of the fungal mycelia products along the
lines introduced by Haider and Martin.
The emergence of modern isolation, fractionation, and analytical proce-
dures has enabled us to get a good insight into the types of molecules that
compose HAs and FAs. The vast array of data by Schnitzer and his colleagues
(Schnitzer, 1978, 2000) from the gas chromatography–mass spectrometry
(GC–MS) analyses of the derivatized degradation products, aided by mech-
anistic awareness of the degradation reactions, allowed deductions to be
made of the types of molecules that compose the humic structures
(summarized by Clapp et al., 2005; Hayes and Swift, 1978, 1990). Identifi-
cations of the degradation products, coupled with NMR data, have made it
clear that lignin alteration products are significant contributors to these
humic fractions. There has been evidence for significant amounts of carbo-
hydrate and peptide degradation products, and for long-chain fatty acids and
hydrocarbon structures in degradation digests, but there is no convincing
evidence to indicate that these are part of the humic core structures. It is clear
that HAs and FAs isolated by the International Humic Substances Society
(IHSS) procedures (Swift, 1996) are derived to significant extents from lig-
nin and from altered carbohydrate and peptide components of plant and
microbial debris in the soil environment. The array of products identified
by Schnitzer and his colleagues were largely from the Bh horizon of a podzol,
but the same types of compounds have also been identified in the HAs and
FAs from a variety of surface soils.
Taking account of all of the foregoing information about the origins and
formation of HSs, it is our considered view that these are formed by a com-
bination of microbial transformations or significant alterations of plant mac-
romolecules, especially lignin and complex carbohydrates, together with the
resynthesis of some liberated compounds into the humic products. The evi-
dence for transformed lignin as a key component in the formation of HSs is
particularly strong (Clapp et al., 2005).
Applying many of the advanced techniques used for the studies of the
HAs and FAs, it is now possible to identify humin components from soils,
and the evidence that we will give in Section 4 will show that the compo-
sitions of the humin materials are very different from those of HAs and FAs.
2.2 Humin in the Historical Context

In 1806 the Swedish scientist Berzelius described as humin the “inert form”
of HSs, i.e., the fraction that was not alkali soluble (Kononova, 1961, p. 15).
Kononova (1961, pp. 69–74) has also reviewed approaches to the isolation
and characterization of soil humin materials by Russian scientists who
worked on coals, soils, and peats. In particular, Kononova agreed with
Shmuk (1930) that the humin of peat soils was derived from incompletely
humified plant residues,
Fuchs (1931) considered that humins arose from the denaturation, by
dehydration, condensation, polymerization, and the diminution in the
number of functional groups in the HAs associated with brown coals and
mineral coals. Williams (referred to in Kononova, 1961, 1966) attributed
the stabilization of soil crumbs to the conversion of HAs to humin, which
he viewed as irreversible. These concepts were reasonable assumptions based

on the information that was available at that time, and later Stevenson (1982,
1994) also considered that humin materials have intimate associations with
soil inorganic colloids.
The work of Tyurin and Gutkina (1940) is relevant to modern
approaches to the isolation of humin. HAs were first isolated from an H+-
exchanged chernozem soil using 0.1 M NaOH. The residual soil was acid-
ified (5 M HNO3) and then treated with HF to release the humin from its
associations with silicates. They concluded that the complex substances dis-
solved in alkali in the course of these operations were similar to the HAs and
FAs isolated in base from the chernozem. Similar results were obtained for a
podzol soil by Khan (1945; see Kononova, 1961, p. 71). In that work, Khan
first removed in alkali the HAs from the H+-exchanged soil, then subjected
the residual soil to from 7% to 25% H2SO4 (with slight heating), and isolated
HAs in 0.1 M NaOH. In this way, 88% of the so-called humin materials were
brought into solution. The HAs isolated by Khan and by Tyurin and Gutkina
had lower percentages of carbon and higher percentages of hydrogen and
oxygen than the HAs isolated from the whole H+-exchanged soils. In
our view, this indicates that the humin materials had undergone oxidation
during the treatment, and hence the products recovered could be regarded
as artifacts produced by the extraction processes.
Kononova (1961, pp. 73–74) considered that the failure of some HAs to
dissolve in alkali was attributable to their strong linkages to the mineral parts
of the soil. She considered that the stabilization of soil crumbs was caused by
these linkages. These associations had caused the HAs to lose their ability to
dissolve in aqueous base. That concept was strengthened by the gain in sol-
ubility of the materials in base after the associated silicates were removed by
treatment with HF.
In more recent times, humin has been variously described in the litera-
ture as HAs and/or FAs complexed to inorganic colloids or clay minerals
(Cloos et al., 1981; Theng, 1979), as plant and fungal residues at various
stages of transformation (Anderson et al., 1974), and as melanins (Russell
et al., 1983). Certainly, the associations between HSs and soil mineral col-
loids have relevance to soil aggregate stabilization, but extensive studies have
provided little evidence for significant adsorption by clay minerals of humic
preparations. Hydroxides of iron and aluminum, below their points of zero
charge, with or without associated clay particles could be expected to sorb
HAs and FAs. Chaney and Swift (1984) did find that aggregate stabilities
were improved in soils incubated with amendments of HAs and glucose.
Based on the studies of: Preston and Ripmeester (1982), Saiz-Jimenez

and De Leeuw (1986), Almendros and González-Vila (1987), Preston
et al. (1989), Rice and MacCarthy (1990); Preston and Newman (1992)
stated that “humins contain organic structural features similar to those found
in intact soils and HAs and that the lower solubility of humin may be due to
several factors including a higher proportion of poorly decomposed plant
material, higher molecular weight of the humified material, and strongly
associated organo-mineral complexes. Thus the structure of humin would
be expected to have a high level of spatial heterogeneity.” Similarly, Rice
and MacCarthy (1989, 1990, 1992) considered the humin fraction to be
composed of HA-like materials made up of HAs, bound lipids, bitumen,
and other insoluble residues. The HAs were regarded as being bound to clay
minerals, and/or with high C contents of large molecular sizes and insoluble
in the basic extraction media.
On the basis of what is now known, we consider that some of what were
regarded as the humins of mineral soils were actually HAs in strong associ-
ations where they were no longer extractable with dilute basic solutions.
The procedures used by Khan and by Tyurin and Gutkina will have altered
the compositions of what were immobilized HAs and the materials released
satisfied the operational definitions of HAs. We will show in Section 3.1 that
effective mild solvent systems will isolate lesser oxidized HAs leaving behind
true humin materials that are compositionally very different from those
extracted.
Toward the end of the 20th century, significant advances were made
when early NMR studies (Almendros et al., 1991; Hatcher et al., 1980,
1985; Preston et al., 1989) highlighted the substantial contributions of ali-
phatic hydrocarbon structures to the composition of humin, suggesting that
these complex, largely paraffinic structures had their origins in plant, algal, or
microbial sources. The major issue that slowed further progress at this stage
was the difficulty in making definitive interpretations of data originating
from the complex mixture of organic components in the soil. This was prin-
cipally due to the lack of a suitable procedure for the isolation of humin
required to enable more detailed and definitive analysis of its composition
and structure. This is not surprising since the insolubility of humin in aque-
ous media is one of its defining characteristics.
In recent years, new isolation procedures enabling the recovery of
unaltered humin materials, separately or in associations with the soil clays,
have facilitated studies of humin composition. In general, these studies
indicate that humin is composed largely of the aliphatic hydrocarbon and
wax-derived substances, of structures from microbial cell walls, and of some

carbohydrate- and peptide-derived materials (likely to be trapped in the
humin matrix) (Hayes et al., 2010; Simpson and Johnson, 2006; Simpson
et al., 2007c; Song et al., 2008, 2011). Evidence to substantiate this statement
will be presented in Section 5 following a consideration of the possible ori-
gins or sources of humin (Section 3) and of the NMR techniques (Section 4)
that have been used to study humin composition.
3. MODERN APPROACHES TO STUDIES OF HUMINa

In recent times, major advances have been made in the techniques
available for the isolation of humin, and over the same period, new
NMR techniques have been developed and applied for the characterization
of humin. These techniques are discussed and reviewed in this section.
3.1 Isolation Processes for Humin

Humin, in many isolation methods, is considered to be the organic compo-
nent of the residue following extractions with aqueous base (e.g., by the
IHSS method; Swift, 1996). However, in many cases (as referred to in
Section 2.2), such residues are composed of the humin materials in associ-
ation with soil inorganic colloids. Faced with a similar problem, geologists
have used a variety of chemical and physical procedures to isolate kerogen
(insoluble OM) from sedimentary rocks, and similar extraction methods
could be utilized to isolate the most insoluble fractions of humin. These
chemical methods are based on removing inorganic mineral matter using
reagents such as hydrochloric acid (HCl), hydrofluoric acid (HF), nitric
acid, lithium aluminum hydride, sodium borohydride, and zinc/HCl
a
It is important to appreciate that the word “humin” has two distinct uses and meanings in science and
these are not to be confused.
In Soil Science the word humin refers to that component of the soil organic matter which is insol-
uble in aqueous base as defined in this chapter.
In Chemistry the same word is used in organic chemistry to identify the dark, insoluble residue
formed when simple or complex saccharides are condensed or dehydrated under acidic conditions
and subjected to heating sometimes under pressure. Unfortunately, the same word has taken on another
meaning in the area of biofuels production often involving the use of carbohydrate-rich plant starting
materials. The word humin again refers to the insoluble residue remaining in the reaction vessel fol-
lowing a reaction process conducted at elevated temperature and pressure and can include plant com-
ponents and residues other than carbohydrates.
The compositions of these latter two materials differ from that of humin found in soil and the use of
the term must be carefully differentiated to avoid misunderstandings.
(Saxby, 1970). However, such chemicals can bring about alterations to the
functional groups of the OM (Saxby, 1970).
High concentrations of HF or HF/HCl solutions are effective for the
removal of the inorganic materials (metal oxides and aluminosilicates)
(e.g., Hatcher et al., 1980, 1985; Huc and Durand, 1977; Ishiwatari,
1985; Preston et al., 1989; Preston and Newman, 1995). Using modifica-
tions of this procedure humin can be recovered from soil by first extracting
the HSs and treating the residue with HF/HCl to remove mineral compo-
nents. However, it should be noted that high concentrations of HF can lead
to structural alteration of the organic components as well as the loss of poly-
saccharides (Gelinas et al., 2001).
The ability to study the composition and properties of amorphous mate-
rial usually requires that the substance be in solution in order to make a wide
variety of measurements and observations. The lack of research on humin
materials can be attributed to their insolubility (Rice, 2001) in solvent sys-
tems from which the solute is recoverable without compositional or struc-
tural alterations. Much attention has focussed in soil studies on the uses of
organic solvents for the isolation of humin. Any component of SOM that
is isolated in an organic solvent following exhaustive extraction with aque-
ous base is regarded as part of the humin fraction.
Several methods, involving aqueous and organic solvents, have been
investigated in order to isolate humin (Hayes, 1985, 2006; Rice, 2001;
Rice and MacCarthy, 1989; Song et al., 2008, 2011). Insignificant amounts
of OM are dissolved in low boiling organic solvents and the boiling points of
potentially useful dipolar aprotic solvents, such as dimethylformamide
(DMF) and dimethylsulfoxide (DMSO), are too high to make feasible the
recovery of the humin solutes by evaporation of the solvent. Thus, signif-
icant progress in awareness of the composition of humin was delayed until
solvent systems were found to isolate the humin from the soil mineral
colloids.
Rice and MacCarthy (1988, 1989) initiated a new era in humin research
when they introduced methylisobutyl ketone (MIBK), a mild solvent, to
separate the humin from the solid material remaining after alkaline extrac-
tion of HSs. They considered the humin fraction to consist of HA-like mate-
rials (HALMs), an organic fraction complexed to clay minerals (bound HAs)
with high C contents, of large molecules that are not soluble in the basic
media, and of insoluble bitumen and bound lipids (Rice and MacCarthy,
1989, 1990, 1992). To isolate the HALM, Rice and MacCarthy (1989) used
two immiscible liquids, aqueous base, and MIBK as the organic solvent
phase. The MIBK procedure can be regarded as a separation method, based

on the accumulation/partitioning at a liquid/liquid interface, and not as an
extraction process.
Almendros and his colleagues (Almendros and Sanz, 1989, 1992;
Almendros et al., 1981, 1991, 1996) applied the MIBK method, based on
that by Rice and MacCarthy, for the isolation of humin from forest soils.
They (Almendros et al., 1996) compared the compositions of the humin
products isolated by two different procedures from three forest soils. They
called the product from one isolation procedure “Inherited Humin” and
that from the other, using MIBK, the “Partitioning Procedure,” and they
carried out further studies on these fractions.
In developing this approach further, Hayes (1985, 2006) considered
organic solvents to be worthy of a systematic study for the isolation of
humin. He compared the structural and physical properties of a range of sol-
vents and then systematically compared the solvent properties of these with
0.1 M NaOH for the extraction of HAs. The best organic solvents had an
electrostatic factor value >140; a pKHB value >2; and dispersion (δp),
hydrogen bonding (δh), and proton acceptor (δb) values of the order of
or greater than 6, 5, and 5, respectively. DMSO, the best of the organic sol-
vents tested, satisfied all of these criteria. Hayes (1985, 2006) has suggested
mechanisms for the solubilization of HSs in DMSO. However, DMSO
alone was not effective in isolating the humin after the SOM had previously
been exhaustively extracted in aqueous base.
Tsutsuki and Kuwatsuka (1992) had used a DMSO + HCl (1.2%–3.2%,
v/v) mixture and isolated 9%–22% of humin from soils, but 43%–79% of
the isolates could be dissolved in base, indicating that large proportions
of these isolates could not be regarded as humin. The humin material
isolated in DMSO + 6% (v/v) HCl from a Mollisol soil by Clapp and
Hayes (1996, 1999) displayed solubility characteristics that resembled those
of FAs.
Relatively recently, Hayes and coworkers have developed an effective
solvent system based on DMSO for the isolation of humin components.
The HSs are first removed by exhaustively extracting soils in aqueous media
at pH 7, 10.6, and 12.6 (0.1 M NaOH). This is done to isolate HAs and FAs
sequentially on the basis of charge density and solubility differences. Then
the residual soil materials are exhaustively extracted with 0.1 M NaOH
+ 6 M urea, and up to 25% additional OM is extracted in that medium.
(All extractions at pH 10.6 and above were carried out under an atmosphere
of dinitrogen gas.) Interestingly, it was found that the extra amounts
extracted could be related to the soil management with only 1% additional

material isolated from soils in long-term cultivation, but up to 20% more was
isolated from adjacent soils in long-term grassland (Byrne, 2011).
High concentrations of urea are commonly used in biochemistry to dis-
rupt noncovalent bonds in proteins (Oh-Ishi and Maeda, 2002), and urea has
been used to break hydrogen bonds that aggregate humic fractions when
modern separation techniques are used (see, for example, Peuravuori
et al., 2004; Piccolo, 2002; Zhang and Lu, 1987).
In the classical operational definitions, the enhanced yields from the iso-
lates in the base/urea medium would be regarded as components of humin.
However, HA and FA materials isolated when the pH of the extract was
lowered to 1.0 were compositionally similar to those isolated at pH 12.6
(Song et al., 2008, 2011). It is probable that urea disrupts the forces that hold
humic components in aggregates and attached to clay/silt-sized particle sur-
faces. Although urea in aqueous solution is a mild solvent, it can react with
humic structures (Hayes et al., 1975).
The base/urea is removed from the residual soil (or from the soil clay
fraction) by dialysis and/or by washing in distilled water, and the retentates
are dried prior to further extraction with a DMSO system. The dry residual
soil, or the soil clay fraction, is exhaustively extracted with DMSO + 6% (v/v)
concentrated H2SO4 (DMSO/acid system) (Hayes, 2006; Song et al., 2008,
2011). Since DMSO is a poor solvent for anions (Martin and Hauthal, 1975),
it is important to H+-exchange the SOM when using this solvent. Hence a
DMSO + 6% concentrated H2SO4 system emerged as the solvent of choice
for the humin isolation process outlined.
DMSO and H2SO4 are both good solvents, and the DMSO + H2SO4
system combines solvent properties that can solvate a wide range of
organic substances with differing polarities. Due to the process of auto-
protolysis (Greenwood and Earnshaw, 1997), protons in H2SO4 are
highly mobile, allowing protonation of the conjugated bases (carboxylate
and phenolate functionalities in HSs), thereby increasing the solubilities of
recalcitrant organic components in DMSO. Exothermic reactions will not
take place when the DMSO/H2SO4 solvent system is applied to dry sam-
ples. The extracted humin is readily precipitated by adding distilled water.
A small amount of the extract (ca. 4%) stays in solution in the aqueous
medium.
The amounts of the recalcitrant SOM components associated intimately
with silt/clays (and not isolated in the base and base + urea systems), but iso-
lated in the DMSO + H2SO4 system, ranged from 32% in the case of a Terra
Preta ferrisol from the Amazon Region, to 65% for an Irish brown earth
(Song et al., 2008), to up to 90% for the Mollisol Soil Standard of the IHSS
(Song et al., 2008).
The DMSO + H2SO4 system for the isolation of humin arose from a
comprehensive study of the HSs and of humin in soils. However, should
the objective be to isolate the humin fraction an appropriate procedure
would be to H+-exchange the soil, exhaustively extract with 0.1 M
NaOH + 6 M urea, wash out the base/urea (or dialyze), dry, and then
exhaustively extract with DMSO + H2SO4, and proceed as described earlier.
3.2 Applications of NMR Spectroscopy for Studies

of the Compositions of Humins
Simpson et al. (2011) have described NMR spectroscopy as the most pow-
erful analytical instrumentation for the determination of complex organic
structures and interactions. Nowadays, NMR spectroscopy is an invaluable
tool for studies of the compositions and structures of HSs. Analyses can be
carried out on samples in the solid, liquid, or gaseous phases to provide infor-
mation about the immediate environment of each type of atom. Structural
information can be obtained from the energy splitting of nuclei which is
affected by the number and nature of chemical bonds as well as by the prox-
imity of adjacent atoms in space (Clapp et al., 2005). Electromagnetic (EM)
radiation with an energy corresponding to the gap between the spin states
can cause the nuclear magnets to flip from a low- to a high-energy state.
The nuclei of the atoms in different positions in the molecule give out
energy at slightly different frequencies as they drop back from a high- to
a low-energy state (called relaxation), indicative of the structure of the mol-
ecule. The difference in frequency between the induced signal and the exci-
tation frequency produces a free-induced decay (FID) plot (Dixon and
Larive, 1999). The FID is a time-domain signal that contains all the ampli-
tude and frequency information of each resonance in the NMR spectrum.
The Fourier transform (FT) algorithm is used to convert the FID into a fre-
quency domain (Keeler, 2013). It separates out signals from atoms in differ-
ent molecular environments and presents these as a spectrum that can be
interpreted. In order to obtain a good spectrum, it is usually necessary to
repeat the NMR experiment many times, especially for nuclei with low nat-
ural abundances (Keeler, 2013). We describe here only features of NMR
that are most relevant to the experiments that will be discussed.
Knicker (2011) has described the technical basis of NMR spectroscopy
that is highly relevant to the material in this communication. There follows a
brief outline of the types of information that can be obtained from applica-
tions of NMR to studies of the compositions of soil humin.
3.2.1 Diffusion Editing Relevant to 1H NMR and to Studies of Humin

The diffusion-editing (DE) capability of 1H NMR has relevance to our con-
siderations of humin. In DE, molecules are “spatially encoded” at the start of
the experiment and these are then “refocused” at the end of the experiment.
Species that diffuse or exhibit a high degree of motion during the experi-
ment are not refocused and are essentially “gated” from the final spectrum.
Thus the DE spectrum produced will contain only signals from species that
undergo little or no self-diffusion, and hence structures identified will be
macromolecular or in rigid domains, or both. The majority of the humin
signals remain after DE.
The pulsed gradient spin-echo (PGSE) method was developed by
Stejskal and Tanner (1965) for DE experiments, and it is described in detail
by Dixon and Larive (1999). The nuclei are magnetized with a 90-degree
pulse followed by a short delay (T1), and a gradient pulse is applied that
can encode the nuclear spins along the length of the sample.
Inverse diffusion editing (IDE) is a subspectrum of the total 1H spectrum.
The IDE experiment utilizes PGSE-based sequences along with the post-
processing difference spectroscopy to produce a spectrum that shows only
the small mobile molecules in the sample, i.e., those that exhibit rapid dif-
fusion (Courtier-Murias et al., 2012). The spectrum is obtained by sub-
tracting the DE spectrum (“large molecules”) from the unattenuated
spectrum containing both small and large molecules (conventional 1H spec-
trum). The result is a spectrum that contains only the most mobile/soluble/
small components (Courtier-Murias et al., 2012). This spectrum allows
identification of small mobile components in the sample that could be
masked in the other 1H spectra.
3.2.2 13C Nuclear Magnetic Resonance

The diverse range of 1H NMR experiments are useful for compositional
studies, but analysis using 13C nuclei gives a greater chemical shift dispersion
(0–240 ppm) that decreases a spectral overlap, and minor differences
between samples are easier to identify (Cardoza et al., 2004). In general,
one-dimensional (1D) 13C NMR experiments are carried out on samples
in a dry, solid state. Cross-polarization magic angle spinning (CPMAS)
13
C NMR is the most powerful experimental approach used to provide
direct information on the structural and conformational characteristics of
humic components (Conte et al., 2004; Piccolo and Conte, 1998; Preston,
1996; Wilson, 1987). Solid-state CPMAS 13C NMR spectroscopy has been
used to describe the composition of fractionated HSs and to help understand
their genesis, transformation, and degradation (Almendros et al., 2000;
Baldock et al., 1997; Conte et al., 2004; Lorenz and Preston, 2002;
Lorenz et al., 2000; Lu et al., 2000; Spaccini et al., 2000). A major advantage
of solid-state NMR is that no solvent is added. Therefore the sample does
not need to be soluble, samples are free from chemical interference, the spec-
trum does not have any resonances from the solvent, and the sample is easily
recovered. The disadvantages include problems that relate to the broadness
of the peaks (due to chemical shift anisotropy (CSA) and dipolar interac-
tions); low natural abundance of 13C (ca. 1.1%); difficulties in magnetizing
the carbon nuclei (due to a low gyromagnetic ratio); and interference from
paramagnetic species that may be present. The difficulties associated with
solid-state NMR have been successfully overcome using a combination
of high-frequency magic angle spinning (MAS) to remove CSA, broadband
proton-decoupling to remove dipolar and scalar coupling, and cross-
polarization (CP) for sensitivity enhancement by ca. 4 (Simpson et al.,
2011; von Philipsborn and M€ uller, 1986).
Conte et al. (2004) have reviewed CPMAS 13C NMR spectroscopy and
its applications to natural organic matter (NOM). The main points are
briefly summarized here to explain the basis of 13C CP NMR spectroscopy
that applies to the evidence and interpretations presented in this chapter.
3.2.2.1 CP and Spin Diffusion

1
H nuclei have a much larger gyromagnetic ratio than 13C, and this is
exploited in CP (Duer, 2002). Spin diffusion (dipolar interactions among
the 1H nuclei) is an alternative mechanism to CP (Scheler, 2002). 1H can
induce spin reorientation on any other 1H in close proximity and can sub-
sequently interact in a similar manner with either carbons or other protons.
3.2.2.2 Ramped amplitude CPMAS

This technique is now routinely used for solid-state 13C NMR. Ramped
amplitude (RAMP) CPMAS conditions are achieved when the amplitude
of one of the EM fields (1H or 13C) is varied during polarization transfer
(Samoson et al., 2001).
When running single-contact-time CPMAS 13C NMR experiments,
the most important parameters are acquisition time (AT) and recycle delay
(RD). The AT must be sufficiently long to avoid both signal loss and noise
increase, and the RD must be at least five times larger than the T1 (the time
constant for the relaxation process) (Wilson, 1987).
3.2.2.3 Chemical Shift Anisotropy

CSA affects the relaxation of molecules in solid-state NMR experiments and
can result in the spectra having very broad line shapes. CSA is not an issue in
solution-state NMR experiments where molecules continuously move. In
the solid-state, the chemical shift depends on the orientation of the bond
relative to the applied static magnetic field (Claridge, 2009; Popov and
Hallenga, 1991). CSA relaxation is related to the square of the applied
field; thus, it has greater significance at higher field strengths at which
broader line-widths are created, and thus the potential benefits of greater
dispersion and sensitivity provided by higher field strengths may be lost
(Claridge, 2009).
3.2.2.4 Spinning Side Bands

Spinning side bands (SSB) are “false” resonances positioned at chemical
shifts equal to the spinning frequency of the sample. These appear at both
the right and left sides of the center band and are not influenced by the
strength of the applied magnetic field (Stejskal and Memory, 1994). The
bands are easy to recognize by either measuring the distance between the
resonance (SSB) and the center band or observing their intensities (which
will decrease and their positions will move if the rotor spinning rate is chan-
ged, but the position and intensity of the center band will remain at the same
position (Conte et al., 2004)).
These bands become an issue in solid-state NMR if the rate of rotor spin-
ning frequency is lower than the frequency of the CSA (Conte et al., 2004;
Duer, 2002). SSBs can be eliminated by spinning the sample at a high rotor
spin velocity (ca. 15–20 kHz) and using RAMP pulse sequence to maximize
CP efficiency (Chefetz et al., 2002; Cook and Langford, 1998; Dai et al.,
2001, 2002; Dria et al., 2002; Kiem et al., 2000; Liu and Huang, 2002).
If the spectrometer has low spinning speeds, a pulse sequence called TOSS
(total suppression of side bands) (Duer, 2002; Lorenz and Preston, 2002;
Lorenz et al., 2000; Mao et al., 2000; Preston, 2001) can be utilized, but
these spectra may not allow for the C compositions to be quantified accu-
rately (Preston, 2001). A more simplistic approach is to mathematically sub-
tract the SSB, but that may not be accurate because of underestimation of
hidden/unidentified SSBs
3.2.2.5 Magic Angle Spinning

MAS was developed to counteract CSA and dipolar interactions. When the
sample is rotated about an axis inclined at 54.7 degrees relative to the direc-
tion of the applied magnetic field, all dipolar interactions are effectively
removed. That allows solid samples to behave in a similar way to solutions
in NMR spectroscopy (Knicker, 2011). Standard spinning frequencies range
from 12 to 35 kHz, and higher frequencies can remove intermolecular inter-
actions. Coupling MAS with CP enables high-resolution NMR spectra to
be obtained for 13C nuclei (Andrew, 1981). To eliminate the broadening
caused by the CSA/dipole interactions, the spinning speed frequency has
to be at least as high as the line broadening caused by these interactions
(Knicker, 2011).
3.2.2.6 Direct Polarization

Direct polarization (DP) (also called Bloch decay (BD) or single-pulse
experiment (SPE)) NMR involves direct excitation of the 13C nuclei,
and DPMAS quantifies the compositions of samples (Simpson et al.,
2011). DPMAS experiments are based on the suppression of 1H–13C dipolar
interactions and decreasing anisotropy using MAS. This technique is neces-
sary for samples with low/no protonation, e.g., aromatic cores of coal or
biochar (Franz et al., 1992; Freitas et al., 1999; Jurkiewicz and Maciel,
1995; Kidena et al., 1996; Maroto-Valer et al., 1996, 1998), but lengthy
experimental times are needed because of the low abundance of 13C. It
has also been used for humic samples (Conte et al., 2004; Mao et al.,
2000; Smernik and Oades, 1999, 2000a,b). DP of the 13C nuclei with long
relaxation delays is the only reliable method to prevent underestimation of
the C content in the sample (Berns and Conte, 2011; Johnson et al., 2005).
Unlike CPMAS, DPMAS can detect unprotonated carbons and can use high
spinning frequencies to eliminate the presence of SSBs. Poor sensitivity and
lengthy running times (because many scans are required to achieve good
signal to noise) are disadvantages in DP experiments (Xing et al., 1999).
3.2.2.7 Dipolar Dephasing

In dipolar dephasing (DD) experiments the signals that originate from any
nucleus directly bonded to hydrogen are removed. Thus, it gives indications
of the extents of replacement of hydrogen substituents in the aromatic
nucleus, and of the extents of fusions of aromatic structures, as in char/
biochar.
3.2.2.8 Dipolar and Quadrupolar Interactions

Dipolar interactions are the energy relationship between two magnetic
moments, and these interactions are most relevant to many nuclei with spin
1/2 (Claridge, 2009). Quadrupolar interactions are purely electrostatic
effects and are several orders of magnitude larger than dipolar interactions
(Popov and Hallenga, 1991). Dipolar and quadrupolar interactions can gen-
erate very broad lines in NMR spectra (Popov and Hallenga, 1991). When
two dipoles are in close proximity, they can attract or repel each other
depending on their relative orientations, and this results in faster relaxation
times (Claridge, 2009). The strength of the coupling is dependent on the
gyromagnetic ratio, the distance between the nuclei, as well as their orien-
tations (Laws et al., 2002). The spin–spin coupling interaction can be
suppressed by decoupling the unobserved nuclei, which is mostly 1H
(Knicker, 2011). The interactions only occur in nuclei with spin >1/2.
These nuclei have an electric quadrupole moment, which is a property sim-
ilar to the gyromagnetic ratio, and this can interact with the electric field
gradients (Schurko, 2009). Acquisition of NMR spectra of quadrupolar
nuclei is challenging because of the rapid relaxation of the nuclei, and gen-
erally very broad peaks are observed (Schurko, 2009).
3.2.2.9 Paramagnetic Species

Considerable quantities of OC are in strong associations with inorganic
components that can have high ash contents and high concentrations of
paramagnetic species (Gelinas et al., 2001). These paramagnetic species
broaden NMR signals and produce spectra with strongly overlapping reso-
nances. Additionally, these species can cause selective quenching of signal
intensity by decreasing relaxation times of specific C functional groups,
and OC in close to paramagnetic centers may be undetectable (Schmidt
et al., 1997). To improve the sensitivity and resolution of the NMR spec-
trum, it is strongly recommended to remove paramagnetic species (Conte
et al., 2001, 2004; Schmidt et al., 1997; Skjemstad et al., 1994).
HF and HF/HCl solutions dissolve silica (HF) and remove carbonates
(HCl; Gelinas et al., 2001). However, as already pointed out, concentrated
solutions of these acids can cause chemical alterations and considerable losses
of some labile OC components (Gelinas et al., 2001; Schmidt et al., 1997).
3.2.3 Comprehensive Multiphase NMR

High-resolution magic angle spinning (HRMAS) 1H NMR spectroscopy
was developed to study molecules in solution/swollen states. Components
in the solid phase are not observable (Shirzadi et al., 2008a,b) because the
probe has relatively low power handling, thus preventing the applications
of solid-state techniques for protons (Courtier-Murias et al., 2012). Follow-
ing on from the new insights provided using HRMAS NMR spectroscopy,
Simpson’s Research Group developed a complete or comprehensive
multiphase (CMP) probe (Courtier-Murias et al., 2012). The probe was
designed to study heterogeneous systems, such as soils and sediments that
can be composed of numerous phases that determine their environmental
properties (Courtier-Murias et al., 2012). Traditionally, each phase has been
studied separately using NMR spectroscopy. However, changing the natural
phases of the sample can lose important structural information; therefore,
CMP-NMR was developed to study all bonds in all phases in whole
unaltered natural samples (Courtier-Murias et al., 2012).
“The CMP-NMR probe is built with high power circuitry, MAS, fitted
with a lock channel, pulse field gradients, and is fully susceptibility matched”
(Courtier-Murias et al., 2012). Therefore, without compromising power
handling, the probe can allow the full range of solution-, gel-, and solid-state
experiments to be carried out using an HRMAS probe (Courtier-Murias
et al., 2012). This technology has provided new opportunities to study both
structures and interactions independently in each phase as well as interac-
tions between phases, within a heterogeneous sample (Courtier-Murias
et al., 2012), and will provide new insights into the composition, structure,
and behavior of NOM.
The CMP-NMR probe was not developed with the intention to replace
solution, solids, or HRMAS probes (Maas et al., 1996) but to complement
existing NMR experiments and to provide new possibilities to study in situ
interactions at interfaces and changes in conformations of structures
(Courtier-Murias et al., 2012). Currently, this probe is not available com-
mercially, but if successful, it holds the key to more in-depth studies on
whole samples. Future studies will allow the assessment of organic samples
with minimal preprocessing to provide insights into their composition and
their reactivity in the environment.
3.3 Two Dimensional NMR Spectroscopy

In order to gain an overview of the bulk chemical composition of a sample,
the spectrum can be divided into chemical shift regions assigned to the most
likely chemical groups that give rise to these resonances (Knicker, 2011).
Because of the complex nature of OM samples, it is difficult to identify
clear boundaries that separate the different C groups from each other,
and some overlapping can occur (Knicker, 2011). This problem can be
partially resolved by using multidimensional NMR spectroscopy, and two-
dimensional (2D) NMR provides valuable information about heterogeneous
samples because of the increased spectral dispersion, as well as the additional
connectivity information that allows detailed assignments of the chemical
functionalities and of the structural components present (Byrne et al.,
2010; Lam et al., 2007). Conventional 1D NMR spectra are plots of intensity
vs frequency, whereas in 2D the intensity is plotted as a function of two
frequencies, usually called F1 and F2. The position of each peak is specified
by two frequency coordinates corresponding to F1 and F2. A 2D NMR
experiment is acquired in a similar way to a 1D experiment; the main differ-
ence being that following the 90-degree pulse, there is a variable evolution
time prior to the next pulse (Balci, 2005).
The 2D NMR experiments are based on bond interactions (homonu-
clear and heteronuclear), through-space interactions, and diffusion.
Buddrus et al. (1989) first reported the application of 2D solution-state
NMR spectroscopy, and since then, there have been extensive develop-
ments in the area, many of which can be adapted for NOM studies. Several
techniques are available, which are suitable for a variety of nuclei, in a num-
ber of dimensions. Simpson and Simpson (2009) have identified the “Top-
Ten” NMR approaches for the study of NOM in solution. We refer here to
applications of 1H–1H and 1H–13C spectra that apply to the topics being
considered.
TOCSY homonuclear experiments. TOCSY (total correlation spectros-
copy, also called HOHAHA (homonuclear Hartmann–Hahn spectroscopy))
can help interpret the spectra of complicated molecules (Simpson, 2012),
especially those with large interconnected networks of spin couplings.
TOCSY provides information about protons that are in the same spin system
(a continuous chain of spin–spin coupled protons; Simpson, 2001). Magne-
tization from the first proton is transferred to the next proton and so forth.
The longer the mixing time, the greater is the likelihood that a cross-peak
from a spin that is many bonds away will be observed (Simpson, 2012). Tak-
ing a slice from the TOCSY experiment can show 1H 1D resonances in
greater detail, thus aiding the assignment of peaks.
Nuclear Overhauser effect spectroscopy (NOESY) relates to the relaxation of
one nuclear spin induced by a neighboring spin. It is observed as a change in
the intensity of one resonance when the intensity of a neighboring resonance
is perturbed (Gemmecker, 1999) and the effect is strongly influenced by the

internuclear distance (Williamson, 2006). NOESY provides information
about protons (or any other nuclei with spin > 0) that are in close proximity
and have magnetic dipole interactions, but are not connected by chemical
bonds (Balci, 2005). In order to determine the through-space relationship
between the protons, all the protons in a molecule must be irradiated one
by one that is a time-consuming process, and there can be issues arising from
protons whose chemical shifts are very close to each other (Balci, 2005). 1H
nuclei that may interact with one another through a dipolar relaxation pro-
cess will appear as a cross-peak in an NOESY spectrum. Because the cross-
peaks in NOESY spectra arise from spatial interactions, this type of spectros-
copy is used to study the configurations and conformations of molecules
(Pavia et al., 2009). Very large molecules have restricted mobility in solution
that allows additional time for the NOE interactions to develop (Pavia et al.,
2009). NOESY is useful for identifying groups in NOM fractions, such as
Ar–O–CH3 (methoxyl in lignin), that do not exhibit scalar through-bond
couplings in TOCSY (Simpson et al., 2011), and for confirmation of protein
resonances identified in 1D NMR methods.
Heteronuclear experiments. Heteronuclear correlation experiments are
especially valuable for correlations with those involving 1H–13C nuclei
(Simpson et al., 2011). These experiments decrease a spectral overlap due
to the additional dispersion provided by the 13C dimension, and that allows
assignments to be made that are not possible in the 1D NMR (Simpson,
2001). Heteronuclear spectra, unlike those for 2D homonuclear experi-
ments, do not exhibit a spectrum diagonal. A 1H–13C bond will produce
a resonance at the point of intersection of the 1H and 13C chemical shifts
(Simpson et al., 2002). Early 2D heteronuclear experiments used the heter-
onuclear multiple quantum coherence (HMQC) technique (Bax et al.,
1983), and this has been used in studies of HSs (Kingery et al., 2000;
Schmitt-Kopplin et al., 1998; Simpson et al., 2001, 2002). More recently,
heteronuclear single-quantum coherence (HSQC) has been developed
and shown to outperform HMQC for environmental samples (Simpson
et al., 2002, 2011). The HSQC experiment has an important advantage over
HMQC in that the 1H–1H multiples broaden the intersecting resonance in
either dimension, providing increased sensitivity and resolution (Simpson
et al., 2002). The HSQC experiment requires multiple refocusing periods
and the experiment requires very precise pulse calibration to minimize arti-
facts and optimize sensitivity (Simpson et al., 2002).
3.3.1 Applications of 1D and 2D NMR to Humin Studies

A number of 1D and 2D NMR experiments have provided new insights
into the compositions of NOM and of humin. DMSO–H2SO4 humin
extracts were studied by Simpson et al. (2007a) using conventional 1D
1
H and DE 1H NMR, and 2D TOCSY and 1H–13C HMQC NMR spec-
troscopy. The spectra showed that the major components in the humin iso-
lates were peptides, aliphatic species, carbohydrates, lignin, and some
components from bacterial inputs (e.g., peptidoglycan and lipoprotein).
Kelleher and Simpson (2006) had compared the multidimensional NMR
of HSs with those of biopolymers (e.g., bovine serum albumin, Kraft lignin,
cutin) and found that nearly all the resonances in the HS fractions also
occurred in the intact or degrading polymers. These and other NMR studies
are discussed in Section 4.
4. INFORMATION ABOUT THE ISOLATION AND

COMPOSITIONS OF HUMIN FROM APPLICATIONS
OF DIFFERENT NMR PROCEDURES
The key techniques that have facilitated advances in the understanding
of the composition of humin are improved isolation procedures and the
application of modern procedures used in NMR spectroscopy (see
Section 3). The capacity of the range of NMR techniques to provide sub-
stantial detailed knowledge about the chemical forms of carbon and hydro-
gen and related functionalities in the complex mixture of organic molecules
found in SOM has been outlined in the previous section. In this section, we
will review and summarize the growing body of evidence relating to humin
that differentiates it from humic substances.
4.1 Effects of Different Extraction and Fractionation

Procedures
It is important to know to what extent, if any, the compositions of the
humin materials are altered by the isolation and fractionation procedures
used to obtain the samples, and there have been few systematic studies of
the effects that the reagents and conditions used can have on the humin sub-
strates. Paramagnetic compounds in samples are known to greatly diminish
the quality of the solid-state 13C NMR spectra. Their presence leads to spec-
tra with strongly overlapping resonance lines and to selective quenching of
signal intensity by shortening the relaxation times of specific carbon func-
tional groups. As a result, interpretation of the spectra becomes complicated
(Schmidt et al., 1997). Hence, spectra are greatly improved by the removal
of iron.
Members of the Hayes group (Song et al., 2008, 2011) have investigated
the possible effects of the base/urea and DMSO/H2SO4 (acidified DMSO)
isolation processes on the compositions of the materials under study.
Organosolv lignin from Aldrich and microcrystalline cellulose (Avicel, from
Sigma-Aldrich) were subjected to sequential extractions with 0.1 M NaOH
+ 6 M urea followed by the acidified DMSO system as outlined for the soil
extraction process (Section 4.1). The VACP 13C NMR spectra in Fig. 1
show no substantial alterations to the compositions of the lignin sample
when compared with that subjected to the base/urea and the acidified
DMSO treatments. The FTIR spectra for all three samples were also the
same. In addition, the treatments did not give rise to compositional changes
to the cellulose, as determined by the NMR and FTIR analyses. This evi-
dence indicates that the chemical composition of the humin is unaffected by
the extraction procedure.
Fig. 1 Comparison of VACP 13C NMR spectra of (a) lignin and of the same lignin after
sequential treatment with: (b) 0.1 M NaOH + 6 M urea or, (C) with DMSO + 6% H2SO4. The
scale shows the 13C chemical shifts (ppm).
Hatcher et al. (1985) considered humin to be the residue after successive

extractions with benzene/methanol to remove lipids, dilute acid (1 M HCl),
and 0.5 M NaOH. The spectrum for humin isolated by Hatcher et al. (1985)
from an aerobic grassland soil using this procedure indicated that the treat-
ment had, to a large extent, removed the saccharide components that were
evident in the untreated samples. The major effect of the treatment was the
enhancement of the aliphatic signal in the humin, but the aromatic signal
was significant and there was evidence for lignin residues (140–150 ppm),
with possible evidence for methoxyl (56 ppm).
The CPMAS 13C NMR spectra for the “inherited humin” and “MIBK
humin” (see Section 3.1) of Almendros et al. (1996) are shown in Fig. 2.
From a visual inspection of the spectra, it is clear that all samples are highly
aliphatic, and that the O-alkyl functionality (60–90 ppm resonance) is a
major contributor to the compositions for all samples, regardless of the iso-
lation procedure used, or the soil source. This contrasts with spectra of
Fig. 2 CPMAS 13C NMR spectra of humin preparations from Oak (QUE), Beech (FAG), and
Pine (PIN) forest soils. The scales show the 13C chemical shifts (ppm). From Almendros, G.,
Guadalix, M.E., González-Vila, F.J., Martin, F., 1996. Preservation of aliphatic macromole-
cules in soil humins. Org. Geochem. 24, 651–659.
humin isolated by more modern procedures that will be discussed later in

this section. The anomeric C resonance at 105 ppm indicates that the
O-alkyl resonances were predominantly from carbohydrate. The dominant
peak at 73 ppm and the C4 resonance at 84 ppm are characteristics of cel-
lulose. The resonance around 146 ppm is the characteristic of guaiacyl lignin
(see Section 5.3), and the sharp resonance peak at 56 ppm is the characteristic
of the methoxyl of lignin.
The products from pyrolysis GC–MS and the hydrolyzable sugars con-
tents failed quantitatively to match the carbohydrate content suggested by
NMR. This caused the authors to consider that (a) “crystalline” or highly
structured cellulose, which has a degree of resistance to hydrolytic and pyro-
lytic degradation; or (b) polysaccharide forms fixed to the mineral surface; or
(c) para-carbohydrate structures generated by dehydration of carbohydrates
(Almendros et al., 1989) could be contained in the humin matrix. Analysis of
the individual sugars in the structures could give indications of their origins
(Hayes et al., 2008, 2012).
A visual inspection of the spectra in Fig. 2 reveals the significantly greater
abundances of the alkyl functionalities compared to that of the aromatic res-
onances (130–140 ppm), and that is especially evident in the cases of the iso-
lates from the beech (FAG) and pine forest (PIN) soils. The 33 ppm
resonance is indicative of ordered long-chain methylenic structures
(Hu et al., 1999). Pyrolysis gave a large yield of n-polymethylene structures
which Almendros et al. considered were comparable to those from resistant
aliphatic biopolymers and oil-generating kerogens.
Fig. 3 shows the NMR spectra of the HA extracted from the Mollisol
Soil Standard of the IHSS (Fig. 3A), and of the humin isolated following
demineralization of the clay isolate with HF (Fig. 3B). These spectra empha-
size clear differences between the HA and the humin fractions. The major
resonances in the HA arise from the aromatic and the carbonyl of carboxyl/
amide/ester functionalities, and the symmetrical shape of the aromatic res-
onance is suggestive of oxidized char, possibly from past vegetation fires.
Lesser contributions to the spectrum (Fig. 3A) are evident from carbohy-
drate (60–90 ppm), the methoxyl of lignin and/or peptide materials
(50–60 ppm), and from methylene (25–30 ppm) functionalities. The HA
from a Brazilian Terra Preta soil that had been amended with charred mate-
rials gave a very similar spectrum (not shown).
The spectrum for the demineralized clay (Fig. 3B) is dominated by the
aliphatic hydrocarbon resonances and with lesser though clear contributions
from carbohydrate, from methoxyl/peptide, from aromatic, and from
Fig. 3 VACP 13C NMR spectra for (A), the humic acid isolated from the IHSS Mollisol soil
standard, and (B), the VACP/MAS spectrum for the humin isolated from the silt–clay frac-
tion of the same soil following treatment with HCl/HF. The scales show the 13C chemical
shifts (ppm).
carboxyl/ester/amide entities. The shape of the resonance at 50–60 ppm

could suggest peptide structures, and the resonance for the methyl group
(15–20 ppm) supports that thesis, as will be considered further in connection
with observations in Figs. 6 and 7, for example. There is strong evidence for
carbohydrate (70–90 ppm and anomeric C around 105 ppm). The reso-
nance centered on 150 ppm could arise from O-aromatic structures of lignin
(though that is not confirmed by a sharp resonance at 56 ppm), but it could
also be indicative of tannin structures (Section 5.4). The evidence for car-
boxyl/ester/amide functionality (ca 175 ppm) in the case of the HA is likely
to be dominated by carboxyl, much of which will be associated with aro-
matic structures, but in the case of the humin, carboxyl is likely to arise from
fatty acid components, and much of this resonance is likely to arise from ester
functionalities, as is implicit in the interpretations of the 2D NMR exper-
iments (Section 4.4). Resonances for carbohydrate functionalities are also
evident for the humin fractions in Fig. 4, and in all of the humin samples
that we have studied, though the relative abundance of the resonance at
60–80 ppm and around 105 ppm (anomeric C) in the Mollisol humin is
unusual. The spectra demonstrate the differences between the compositions
of HAs with strong aromatic carboxylic and carbohydrate contents and of
humin dominated by aliphatic hydrocarbon structures.
The spectra for humin extracts in Fig. 4 are similar. The brown earth
(A) was isolated in acidified DMSO following prior exhaustive extraction
with base + 6 M urea; (B) was isolated from the HCl/HF digest of the clay
fraction that had been exhaustively extracted with acidified DMSO; and
(C) is the acidified DMSO extract of the Terra Preta soil. Notice the exten-
sive resonances at 33 ppm (indicative of ordered or “crystalline” methylene
according to Hu et al., 2000), and all spectra also have definite resonances
indicative of; carbohydrate (74 and 105 ppm), aromaticity (129 ppm, at
which char materials also resonate), and carbonyl of carboxyl/ester/amide
functionalities (at 173 ppm). The similarity between spectra A and
B indicates that the demineralization procedure had little effect on the com-
position of the humin component, and the material left associated with the
clay fraction following exhaustive extraction with DMSO–acid is essentially
the same as that extracted in the DMSO–acid.
The differences between the spectra are minor. Evidence for lignin res-
onances (at ca. 150 ppm) and that for methoxyl (at 56 ppm in the DD spec-
trum (Fig. 4A)) are somewhat stronger for the extract in DMSO–acid
(Fig. 4A) than that for the other samples. The DD spectra suggest little dif-
ferences in replacements of H in the aromatic rings (either through aromatic
substitution or through fused aromatic structures).
It is evident, though, surprising that, on the basis of the spectra in Figs. 3B
and 4A (B) and C, the compositions of the humin materials are so similar.
These humins were isolated using the same procedures from different soil
classes and formed in three different continents from different parent mate-
rials and under different climatic conditions.
Fig. 4 VACP 13C NMR and dipolar dephasing (DD) spectra of humins: (A) isolated using
acidified DMSO from an Irish brown earth (Oak Park, OP) soil; (B) isolated following treat-
ment with HCl/HF of the residual silt/clay of the brown earth following extraction with
acidified DMSO; and (C) extracted using acidified DMSO from the silt/clay fraction of a
Terra Preta soil after removal of the HSs using base/urea extractions. Scales show the 13C
chemical shifts (ppm).
4.2 Information From DE 1H NMR Spectra

Considerable additional information is provided by 1D 1H DE spectra. The
1D DE 1H spectra in Fig. 5 emphasize the difference between the base/urea
extract (which would be regarded as humin in the classical definitions) from
a Mollisol soil (Fig. 5a) and the acidified DMSO humin fraction from the
same soil (Fig. 5b). The 1D spectra had large water/urea signals in most cases,
and for that reason only DE spectra are shown in Fig. 5. The 1D 1H NMR
and the 1D DE spectra were similar, but some signals of small molecules
were greatly attenuated in the DE spectra.
Simpson et al. (2003, 2007a) have assigned spectral regions in 1H NMR.
General regions are indicated by brackets in Fig. 5 and are applicable to all
1
H NMR and DE 1H NMR spectra. These can be defined broadly as
(i) mainly aromatic and amide; (ii) signals from numerous moieties including
carbohydrate, peptides, and lignins; and (iii) aliphatic signals from different
chemical environments including various substituted α,β-protons to a func-
tionality in hydrocarbons, lipoprotein (LP), peptidoglycan (PG); long-chain
methylene in lipids, waxes, cuticles, etc.; and terminal CH3 groups. Some
more specific assignments are presented in Fig. 5a.
The spectrum of the precipitate from the base/urea extract is dominated
by contributions from peptides (see Fig. 5a); the double hump at 4–4.4 ppm
is indicative of peptides, especially when there is a large methyl resonance at
0.8 ppm, and H–N resonances are also present at ca. 6.5–8.1 ppm. Reso-
nances from cutins, waxes, and/or lipids; and lignin/carbohydrate is in
the 3–4.4 ppm band. These have previously been identified in classic HA
and FA fractions (Kelleher and Simpson, 2006; Simpson et al., 2002).
The spectrum of the acidified DMSO humin isolate (Fig. 5b) is dominated
by the large contribution from long-chain (CH2)n. The abundance of the
methylene functionality is such that the resonance is off scale in the spectrum
(Fig. 5B). The ratio between the main chain (CH2)n and methylene units in
an aliphatic chain, β to an acid or an ester, is closer to that in aliphatic bio-
polymers, such as cutins, and similar to that observed for the other humic
fractions (Simpson et al., 2003). This is consistent with the presence of ali-
phatic chains observed by solid-state VACP/MAS NMR spectroscopy,
which would be explained by the highly aliphatic nature of humin.
The data from the DE spectra reinforce the substantial contribution of
the methyl and methylene functionalities to humin and provide more infor-
mation about their linkages to other materials that would include waxes,
Fig. 5 1H diffusion-edited NMR of: (a) the extract precipitated at pH 1 from the Mollisol
soil extract in 0.1 M NaOH + 6 M urea following exhaustive extractions at pH 7, 10.6, and
12.6; and (b) DMSO humin extracted in DMSO + 6% H2SO4 (after prior exhaustive extrac-
tions in base and in base + urea). Assignments in (a) refer to: 1, amide; 2, phenylalanine;
3, aromatics in lignin; 4, anomeric protons in carbohydrates; 5, α-protons in proteins and
peptides; 6, methoxyl in lignin; 7, other carbohydrate protons; 8*, P–OCO–R methylene
units adjacent to the carbonyl in lipoproteins; 9, N-acetyl group in peptidoglycan; 10,
methylene units in an aliphatic chain β to an acid or ester; 11, methylene (CH2)n in ali-
phatic chains; 12, CH3 (note when this peak is large relative to 11, it often indicates
strong contributions from proteins, as in these examples). Assignments for spectrum
(b) indicate strong contributions from: aromatic and amide functionalities, carbohy-
drate, peptide and lignin-derived structures, lipoprotein (LP*) and peptidoglycan
(PG) structures, and a large contribution from aliphatic materials that would include
waxes, cutins, and lipids. Note: because of the addition of D2SO4 to solubilize DMSO
humin, deuterium exchanged the N–H to N–D, and so the amide resonance in the
DMSO humin is highly attenuated. *May contain signals from lipoprotein and/or
signals from fatty acids/cuticles. From Song, G., Novotny, E.H., Simpson, A.J., Clapp, C.E.,
Hayes, M.H.B. (2008). Sequential exhaustive extraction of a Mollisol soil, and characteriza-
tions of humic components, including humin, by solid and solution state NMR. Eur. J. Soil
Sci. 59, 505–516; see Simpson, A.J., Song, G., Smith, E., Lam, B., Novotny, E.H., Hayes, M.H.B.,
2007b. Unraveling the structural components of soil humin by use of solution-state nuclear
magnetic resonance spectroscopy. Environ. Sci. Technol. 41, 876–883 for more details.
cutins, and lipids (see Section 5). It also provides evidence for minor com-
ponents including lipoproteins and peptidoglycans.
In other studies employing similar experimental conditions, nearly all
signals are destroyed and only those from macromolecules survive
(Kelleher et al., 2006). Similarly, in classic humic and fulvic materials, the
vast majority of signals would be greatly attenuated under these conditions.
This disparity indicates that the components in the humin fraction are of
much greater size than those previously seen in FA and HA extracts and
are likely to be composed of macromolecules and/or very stable rigid
aggregates.
Humin isolated from the top 1 m of an estuarine sediment, using acid-
ified DMSO after exhaustive extraction with 0.1 M NaOH + 6 M urea
(Mylotte et al., 2015, 2016), was studied using DE and IDE 1H NMR.
The IDE indicated substantial methylene contributions to the composition
of this fraction. The DE spectrum (Fig. 6) illustrates the large contributions
made by both methyl and methylene groups to the structures of the solid- or
gel-like components. This study suggests that the aliphatic molecules in the
extracted humin consist of both small, “mobile” molecules, and large, rigid
molecules. In addition to the aliphatic signals, there are resonances charac-
teristic of carbohydrates and peptide/protein.
A Aliphatic
Carbohydrates, hydrocarbons
methoxyl, esters,
hydroxyl
Amide/aromatic
DMSO
8 6 4 2 0 (ppm)
1
Fig. 6 Diffusion-edited H NMR spectrum of the DMSO–H2SO4 humin extract from the
0 to 1 m depth of an estuarine core: (A) conventional 1H NMR spectrum showing all 1H
resonances; (B) inverse diffusion-edited (IDE) spectrum showing 1H resonances from
mobile/fast diffusing molecules; and (C) the diffusion-edited (DE) spectrum showing
1
H resonances from molecules with restricted diffusion (swollen materials and gels).
When the DE spectrum is expanded, evidence is provided for protein/

peptide and a variety of other functionalities (Fig. 7). The DE 1H spectrum
of the soluble humin (that dissolved in DMSO + 6% H2SO4) of the estuarine
core is stacked with the 1H spectrum for albumin in Fig. 7. It can be seen that
there is an overlap of key resonances in the two spectra. In particular, the
resonances in the aromatic amino acid region are similar for the humin
and the albumin, indicating the presence of the aromatic amino acids, phe-
nylalanine and tyrosine. The large CH3 peak in the albumin confirms that
proteins could contribute to the large CH3 peak identified in the DE spec-
trum, in Fig. 6, of the humin. A large contribution of protein has also been
observed in SOM and has been shown to result from living/dormant/and
dead microbial biomass (Simpson et al., 2007b). A more in-depth investiga-
tion of the DMSO/acid-soluble humin can be achieved by looking at the
Fig. 7 1H NMR spectrum of albumin, and the diffusion-edited (DE) 1H NMR spectrum of
soluble (in DMSO + 6% H2SO4) humin from the 0 to 1 m depth of an estuarine core.
Assignments are: 1, amide in peptides; 2, aromatic amino acids (• denotes phenylala-
nine, ▲ denotes tyrosine); 3, α-proton (peptides); 4, O-aromatics (methoxyl signal in
humin); 5, DMSO (solvent); 6, methylene adjacent to a carbonyl (R2–OCO–CH2–R1, some
appears to be in the form of lipoprotein, thus R2 would be a protein); 7, aliphatic meth-
ylene units γ to an acid or ester; 8, amino acid side chains; 9, aliphatic methylene (CH2);
10, methyl (CH3); and 11, silicate (Simpson et al., 2011). From Mylotte, R., Sutrisno, A.,
Farooq, H., Masoom, H., Soong, R., Hayes, M.H.B., Simpson, A.J., 2016. Insights into the com-
position of recalcitrant organic matter from estuarine sediments using NMR spectroscopy.
Org. Geochem. 98, 155–165.
1
H–1H and 1H–13C bonding/connectivities made possible using 2D NMR
experiments (Section 4.4).
4.3 Information From CMP-NMR for Studies of Humin

The CMP probe devised by Courtier-Murias et al. (2012), referred to in
Section 3.2.3, provides additional insights into the compositions of humin.
The probe allows the full range of solution-, gel-, and solid-state experi-
ments to be carried out using an HRMAS probe (Courtier-Murias et al.,
2012). An example (from Mylotte et al., 2016) of the usefulness of the
CMP 13C CPMAS technique for studies of humin is provided by the spectra
in Fig. 8A and B. Experiments were carried out on the dried samples and
after swelling with DMSO-d6. Spectra are better resolved after swelling
mainly as a result of decreased overlap in the CPMAS.
In CP, magnetization is passed from protons to carbons in proximity via a
dipole network, and strong dipole networks arise only in true solids and rigid
gels. In this way, CMP experiments selectively observe the solid/rigid gel
components while suppressing mobile components (Courtier-Murias
et al., 2012). Thus the CPMAS spectrum of the dry sample will give an
Fig. 8 13C cross-polarization magic angle spinning (CPMAS) solid-state NMR spectra of
samples from the top 1 m of an estuarine core. From the top (for A and B): (A) DMSO/
acid-soluble humin (SHU), insoluble humin (IHU), demineralized clay fraction (DCF), and
the whole clay sample (clay) (number of scans; SHU and IHU ¼ 4K, DCF ¼ 16k,
clay ¼ 80K). (B) Comprehensive multiphase (CMP) 13C NMR spectroscopy of the SHU,
IHU, and DCF humin samples (all samples ¼ 32K scans).
overview of the entire C in the sample, whereas CPMAS in the swollen state
will emphasize the domains that cannot be accessed by the solvent.
The clay sample (non-HF treated) with its associated humin material was
run only in the dried solid state (Fig. 8A). Despite the poor signal-to-noise
ratio, the spectrum of this fraction was sufficient to show that the profile of
the C distribution in the clay broadly matched that of the humin isolated in
the acidified DMSO and recovered following HF demineralization. The
clay was not run on the CMP probe because it was predicted that a useful
signal-to-noise ratio could not be obtained in a meaningful time (Masoom
et al., 2013). The less defined resonances in the untreated clay fraction are
attributable to the low concentration of organics compared with the SHU
(the soluble humin isolated in the acidified DMSO system), the IHU (the
insoluble humin isolated after the residual clay fraction was treated by
HCl/HF following extraction of the clay fraction in acidified DMSO),
and the DCF (the humin isolated after the clay fraction had been
demineralized).
Valuable information is obtained when the spectra for the dried samples
(Fig. 8A) are compared with the CMP spectra of those swollen in the
DMSO system (Fig. 8B). For example, the peaks at 30 and 33 ppm are
assigned to amorphous and crystalline (or ordered) (CH2)n, the main com-
ponents of waxes, lipids, including lipoprotein, or cutins, suberans (see
Section 5 for a more detailed description of these components), and fatty
acids/esters (Song et al., 2008). The crystalline (CH2)n resonance for the
SHU and the IHU in the CMP spectrum (Fig. 8B) is much better defined
relative to that for the similar solid-state spectra (Fig. 8A). That would indi-
cate that the crystallinity impedes access to the DMSO solvent system and
the solid state is preserved.
The dry, solid state and the CMP spectra have significant peaks in the
O-alkyl region (50–60 and 105 ppm resonances), which includes carbohy-
drates and because overlapping of the signals can also include peptides and
methoxyl. Two peaks are evident in the 50–65 ppm resonance in the
DCF solid state and CMP spectra (in Fig. 8A and B), and to a lesser extent
for the IHU. These could represent peptide and crystalline cellulose (65 ppm)
structures. The carbohydrate peak for the DCF and clay fractions is greater
than those for the SHU and IHU. The carbohydrate signal that remains in the
DCF CMP-NMR suggests the possible presence of crystalline cellulose or
large cellulose domains that cannot be completely penetrated by the solvent.
Carboxylates and aromatic components are attenuated in the CMP spec-
trum as these can be swollen (Fig. 8B). The carboxyl/ester functionalities
may arise, at least in part, from long-chain lipids/fatty acids/esters and could
be associated with the more rigid aliphatic structures.
4.4 Applications of 2D NMR Spectroscopy to Studies of Humin

Two-dimensional NMR spectroscopy offers significant advantages for the
analyses of complex humic structures. Increased signal dispersion into two
frequency dimensions greatly reduces resonance overlap. Cross-peaks in
2D NMR spectra indicate a range of connectivities defined by the kind
of NMR experiment carried out, allowing a probe of bonding interactions,
spatial relationships, and chemical exchanges (Simpson, 2001). A number of
2D experiments have provided new insights into the compositions
of NOM.
4.4.1 Total Correlation Spectroscopy

TOCSY is used to identify coupling between protons that are connected via
a bonded network and has applications in the study of the compositions of
humin. TOCSY peaks arise from the interactions of protons over numerous
bonds. Fig. 9 shows the TOCSY spectra of DMSO humin from the Mollisol
soil standard of the IHSS, and the proton couplings from the major func-
tional categories present have been highlighted in the figure. The major
connectivities can be summarized as coupling from amides in peptides; cou-
pling from double bonds from aromatic structures; coupling between
α-protons and amino acids side chains in peptides/proteins; coupling
between aliphatic compounds; and coupling between aliphatic alcohols
and ethers. However, a full interpretation of all of the cross-peaks can be
made only by combining information from a range of NMR techniques.
Some more detailed assignments can be found in the literature (Fan et al.,
2000; Kelleher and Simpson, 2006; Simpson, 2001).
A more clear-cut representation of the types of functionalities and com-
ponents in humin is evident in the TOCSY spectrum of Fig. 10 for the sol-
uble humin isolated in the DMSO acid system from 0 to 1 m depth of the
estuarine core for which data have been provided in Figs. 6–8. The aromatic
resonances are relatively weak (see region 1, Fig. 10). Region 2 has over-
lapping correlations from protons associated with sugars and amino acids.
Resonances in region 3 are mainly from correlations with amino acid side
chains. Regions 4 and 5 contain considerable overlap from a range of ali-
phatic residues including hydrocarbons and amino acid side chains. It is dif-
ficult to confidently assign structures because of the close proximity of the
resonances.
ppm
4 1
5
1 2
2
3
3
5
8 7 6 5 4 3 2 1 ppm
Fig. 9 Total correlation spectroscopy (TOCSY) of DMSO humin isolated from the Mollisol
soil. General assignments are: 1, aliphatic couplings; 2, couplings from aliphatic alcohols
and ethers (some amino acid side chains overlap in this region); 3, couplings between
α-protons and amino acid side chains in peptides/proteins (couplings from ester will
also overlap in this region); 4, couplings from double bonds; and 5, couplings from
amide in peptides (these couplings are weak as most of the amides have been
exchanged by the addition of D2SO4).
By taking a slice through the main chain (CH2)n resonance aliphatic

region, a more in-depth awareness is obtained from Fig. 10 of the hydrocar-
bon compositions. This slice shows all the functionalities primarily associ-
ated with long-chain hydrocarbons in the sample. Peaks shown in the
slice are consistent with resonances identified in cutin/cutan by
Deshmukh et al. (2005) and/or biological lipids (Simpson et al., 2007b).
The peak between 4.1 and 4.4 ppm is representative of methylenes attached
to the oxygen side of an ester, that at 3.3 ppm represents methylenes in
alcohols/ethers, that at 2.2 ppm represents methylenes in free fatty acids,
and peaks at 1.9 and 2.3 ppm are methylenes attached to the carbonyl
C]O side of an ester. Terminal methyls and main chain methylenes are
Fig. 10 Total correlation spectroscopy (TOCSY) of acidified DMSO-soluble humin (SHU)

from an estuarine core. Assignments for (A): 1, aromatics; 2, sugars, methine units bridg-
ing lignin aromatics, amino acids (α–β couplings); 3, methylene units adjacent to ethers,
esters, and hydroxyls in aliphatic chains and amino acids (α–β–γ couplings); 4, methy-
lene in aliphatic chains; and 5, CH3 units in amino acids and aliphatic chains (Simpson
et al., 2002). Assignments for (B), the 1H slice through the aliphatic region of the TOCSY
spectrum, are: 1, H attached to the O side of an ester; 2, methylene α to H in primary
alcohols; and 3, methylene bonded to (a) free fatty acids and (b) to the C]O side of
an ester (Deshmukh et al., 2005).
represented by the large, broad peaks between 0 and 1.8 ppm. Identification
of such aliphatic structures in that resonance region suggests inputs from
terrestrial higher plants to the organic materials in the recalcitrant SHU asso-
ciated with the sediments. However, it is difficult to differentiate between
microbial lipids and cuticular species, and this cannot be stated with
certainty.
4.4.2 HMQC Spectroscopy

HMQC experiments are applied widely to study single H–C bond correla-
tions in HSs. Heteronuclear 2D spectra do not exhibit a spectrum diagonal.
Cross-peaks arise from the coupling of a 13C with a 1H rather than coupling
between adjacent protons. A 1H bonded directly to a 13C atom will produce
a cross-peak at the point of interaction of the 1H and 13C chemical shifts
(Simpson, 2002).
An expansion of the aliphatic and the central regions of the spectrum of
HMQC in the 0–4.5 ppm region of the Mollisol DMSO humin is shown in
Fig. 11. Many components of the DMSO humin fractions that are masked or
Fig. 11 Heteronuclear multiple quantum coherence (HMQC) spectrum of the Mollisol

DMSO–H2SO4 humin fraction. Assignments are as follows: 1, aliphatic (for detailed
assignment of this region, see Simpson et al., 2007b); 2, methylene unit adjacent to
the carbonyl in lipids (including lipoproteins and cutins); 3, carbohydrate CH2; 4, carbo-
hydrate CH; 5, methoxyl; and 6, α H–C in peptides/proteins. These assignments are
based on spectra of known substances.
overlapped by 1H NMR and TOCSY are separated in the HMQC spectra.

Region 1 in Fig. 11 is crowded with aliphatic linkages, with numerous sig-
nals from lipids and side-chain protons in proteins/peptides. The resonances
seen at around 10–25 ppm in F2 (13C) and 0.5–0.9 ppm in F1 (1H) result
partly from terminal methyl groups from lipids. However, the majority of
signals could be from proteins/peptides. The methylene (CH2)n in aliphatic
chains and methylene units in aliphatic chains β and γ to acids or esters show
strong contributions in the region of 0.9–2 ppm (F1) and 25–45 ppm (F2).
Note that the small hump at region 1 is assigned to the N-acetyl group in PG,
the main component in bacterial cell walls. Label 2 in Fig. 11 may represent
components mainly from lipoprotein and signals from fatty acid/cuticular
materials. Labels 3 and 4 are resonances from CH2 and CH groups of car-
bohydrates. Label 5 indicates that a small contribution arises from methoxyl
in lignin-derived structures, and label 6 represents the α-protons in peptides/
proteins. Peptidoglycan can make contributions to the resonances from
methyl, carbohydrate, and peptide functionalities.
4.4.3 HSQC Spectroscopy

HSQC 2D NMR spectroscopy shows 1H–13C bonding in two dimensions.
This 2D dispersion allows overlapping signals to be better resolved and pro-
vides a more confident assignment of signals. Previous detailed assignments
from 2D NMR have been provided in the literature, and readers are referred
for further details to Kelleher and Simpson (2006) and Simpson et al. (2007c,
2011). As was seen for the 1D data, aliphatic hydrocarbon resonances also
dominate the HSQC (Fig. 12, region 1). The presence of protein, observ-
able in the 1D 1H NMR Fig. 5 and clearly evident in Fig. 7, is confirmed in
the HSQC by the identifications of amino acids and α-protons from peptides
(Fig. 12B, regions 4 and 7).
Carbohydrates are identified by the presence of signals from CH, CH2,
and anomeric protons (regions 2, 3, and 5 in the 2D spectrum, Fig. 12B).
The HSQC confirms that the “hump” in the 1D 1H NMR between 3
and 4 ppm arises predominately from carbohydrate signals (see Fig. 7) with
Fig. 12 Heteronuclear single-quantum coherence (HSQC) 2D NMR spectra of the

DMSO/H2SO4-soluble humin extract (SHU) from the 0 to 1 m depth of an estuarine core.
Assignments for (A) are: 1, aliphatic CH2 and CH3, including signals from various hydro-
carbons and from protein side chains; 2, CH2 from carbohydrate; 3, CH in carbohydrate;
4, α-protons in peptides; 5, anomeric protons in carbohydrates; 6, HC]CH; and 7, aro-
matic amino acids (Phe and Tyr); and assignments for (B), the expansion of the aliphatic
region of the HSQC, are: 1, CH3 from amino acid side chains and terminal hydrocarbon
groups; 2, aliphatic methylene to an acid or ester; 3, peptidoglycan (resonance is present
but not clear due to overlap at the vertical scaling shown); 4, methylene unit β to an acid
or ester; 5, (CH2)n; 6, methylene unit adjacent to carbonyl in lipids; and 7, DMSO. From
Mylotte, R., Sutrisno, A., Farooq, H., Masoom, H., Soong, R., Hayes, M.H.B., Simpson, A.J.,
2016. Insights into the composition of recalcitrant organic matter from estuarine sediments
using NMR spectroscopy. Org. Geochem. 98, 155–165.
the α-protons from protein/peptide superimposed on the left-hand side

with an apex from 4 to 4.7 ppm. There are resonances in the HSQC from
double bonds that indicate the presence of unsaturated aliphatic compounds.
Such compounds are present in both microbial lipids and components
derived from plant cuticles (Simpson et al., 2003, 2007b). Additional reso-
nances are evident when the aliphatic region of the spectrum is expanded
(Fig. 12B), as seen also for Fig. 10B. Contour levels are intense for the ali-
phatic CH2 due to the significant contribution of methylene to the humin
composition. Contour levels are also well defined for the methyl-containing
amino acids and confirm that a significant portion of the SHU has microbial
origins. It is postulated that there are peptidoglycan signals in the HSQC
(region 3 in Fig. 12B); and while a distinct resonance is more discernible
at lower plotting thresholds, a clear identification is difficult due to the over-
lapping of signals. It is difficult to clearly see lignin resonances in the HSQC,
although a relatively weak O–CH3 resonance is apparent, but the overlap
makes it difficult to confirm. α-Protons in amino acids and Ar–OCH3 res-
onate in very close proximity; therefore, it is useful to examine NOESY
spectra for more definitive identification of amino acids and lignin
structures.
4.4.4 Nuclear Overhauser Effect Spectroscopy

The NOESY spectrum for the same sample (Fig. 13) shows through-space
1
H–1H connectivities. As in the case of TOCSY (Fig. 10), both dimensions
show the 1H resonances of the sample. The NOESY experiment provides
additional information about the contributions of peptide and lignin to the
SHU. Resonances attributable to amino acids are clearly visible in NOESY
with strong resonances between 1–5 ppm and 7–8 ppm. This region arises
from correlations between amide (8–8.2 ppm), the α-protons in peptide/
protein (4–4.7 ppm), and various amino acid side chains (1.5–2 ppm).
There are also many other protein signals in regions 4 and 5 of the spectrum,
but these are more challenging to differentiate due to overlap. There are also
small but key lignin resonances in region 2 of the spectrum that result from
the spatial correlation between the methoxyl group on lignin and the aro-
matic protons (Simpson, 2001). The overlap of signals in regions 4 and 5
makes additional structural interpretation from the NOESY difficult.
Therefore, TOCSY NMR experiments that highlight coupling through
bonds tend to be less crowded and provide additional useful information
from these spectral regions.
Fig. 13 Nuclear Overhauser effect spectroscopy (NOESY) of the acidified DMSO-soluble

humin extract (SHU) from the 0 to 1 m depth of an estuarine core. Assignments are:
1, amino acids; 2, lignin aromatics (methoxyl–aromatic interactions); 3, lignin aromatic
structures (interactions between aromatic protons); 4, methylene and methyl units
(bonded to amino acids and hydrocarbon structures); and 5, mixture of long-chain
aliphatics and protein signals. Assignments are based on Simpson, A.J., Kingery, W.L.,
Spraul, M., Humpfer, E., Dvortsak, P., Kressebaum, R., 2001. Separation of structural com-
ponents in soil organic matter by diffusion ordered spectroscopy. Environ. Sci. Technol.
35, 4421–4425; Simpson, A.J., McNally, D.J., Simpson, M.J., 2011. NMR spectroscopy in envi-
ronmental research: from molecular interactions to global processes. Prog. Nucl. Magn.
Reson. Spectrosc. 58, 97–175.
The modern approaches reviewed in Section 3 highlight the importance

of isolation and separation of humin from the other components of NOM in
order to identify unambiguously its composition and defining characteris-
tics. This has helped to confirm the dominant contribution of aliphatic
hydrocarbons and has provided indications of compositional detail within
these structures. Furthermore, it has facilitated the identification of other
minor components, which, under normal circumstances, would be labile
if they were not protected within the humin/mineral matrix. We review
in Section 5 the possible origins of the sources of the humin materials.
5. POSSIBLE SOURCES OF HUMIN COMPONENTS

DERIVED FROM PLANT AND MICROBIAL SOURCES
As can be seen from Section 2.2, work carried out through most of the
20th century continued with the notion that humin was simply a less soluble
form of HA. However, toward the latter part of the 20th century, some evi-
dence was presented by various workers (as referenced in Sections 3 and 4) to
show that humins contained components whose chemical compositions dif-
fered significantly from that of HSs, particularly with respect to the presence
of increased aliphatic and decreased aromatic and carboxyl contents. In this
section, we consider the possible origins of such components.
In order to have an understanding of the origins of components of
humin, it is appropriate to consider the composition and structures of the
materials and the processes from which these could be derived. In some
instances the materials under consideration could give rise to HSs as well
as to humin. For completeness, a wide range of possible precursors will
be presented, but emphasis will be placed on the biomolecular structures that
are most likely to contribute to humin.
5.1 Cellulose
Cellulose, a tough, fibrous, water-insoluble biomolecule, located in cell
walls (O’Sullivan, 1997) of plants, algae, and fungi (De Leeuw and
Largeau, 1993; Peberdy, 1990), is abundant in vascular plants and can be pre-
sent at lower concentrations in algae and fungi.
Cellulose is composed of polyglucose units linked by β-(1 ! 4) glyco-
sidic bonds that form linear polymeric chains of over 10,000 glucose residues
(Teeri, 1997). Fig. 14 shows the repeating cellobiose unit that gives rise to
the cellulose chain. The chains attach to each other, held by H-bonding and
van der Waals forces (Teeri, 1997). These chains aggregate to form highly
ordered “crystalline” entities (Ibrahim et al., 2010). A single cellulose
“crystal” contains tens of glucan chains in parallel orientation although, as
CH2OH OH
O
O HO
HO O
O O
HO HOCH2
Fig. 14 The repeating unit of a cellulose chain (the cellobiose unit).
a natural polymer, it contains amorphous domains (Teeri, 1997). Cellulose

crystals in the fibers of higher plants associate with other cell-wall compo-
nents, such as lignin and hemicelluloses giving rise to even more complex
structures. Noncrystalline cellulose is more readily degraded by enzymes
(Beguin and Aubert, 1994; Perez et al., 2002), but the associations between
cellulose, hemicelluloses, and lignin can retard biodegradation (Perez et al.,
2002). Algal cellulose often contains sugars other than glucose, usually xylose
(Baldan et al., 2001; Mackie and Preston, 1974). Xylans and mannans are less
rigid biopolymers that can replace cellulose in the algal cell wall (Baldan
et al., 2001; McCandless, 1981).
Cellulose can be chemically or enzymatically hydrolyzed to simple sol-
uble sugars. Eubacteria and fungi are the main microorganisms involved in
the degradation of cellulose, although some anaerobic protozoa and slime
moulds are also capable of its degradation (Perez et al., 2002). Cellulose-
degrading microorganisms, along with noncellulose degrading organisms,
produce a suite of enzymes with different specificities that work together
to completely degrade cellulose, releasing CO2 and H2O under aerobic con-
ditions, and CO2, CH4, and H2O under anaerobic conditions (Beguin and
Aubert, 1994; Leschine, 1995; Perez et al., 2002). Cellulases hydrolyze the
β-(1 ! 4) linkages of cellulose to release cellobiose, which is subsequently
hydrolyzed by β-glucosidase releasing two glucose molecules (Perez et al.,
2002). Products of cellulose hydrolysis are available as carbon and energy
sources for cellulose-degrading microorganisms, or for other organisms liv-
ing in the environment where cellulose is being degraded (Perez et al.,
2002). Some of these organisms may produce substances that contribute
to humin.
5.2 Hemicelluloses
Hemicelluloses are heterogeneous biopolymers with xylan as the most abun-
dant component; they are the second most abundant type of polysaccharide
in nature (Saha, 2003). Relative to cellulose, hemicelluloses have lower
molecular weights and can be dissolved in aqueous alkali (Timell and
Syracuse, 1967). They have a role in filling the voids around cellulose fibrils
and providing couplings to the lignins and may also influence the aggrega-
tion of cellulose during the formation of the cell wall (Atalla et al., 1993).
Hemicelluloses are composed of a variety of sugar units that have a range
of structural linkages, i.e., α- or β-(1 ! 2, 1 ! 3, 1 ! 4, 1 ! 6) (Pettersen,
1984). Unlike cellulose, hemicelluloses are composed of pentoses (xylose
and arabinose), hexoses (mannose, glucose, and galactose), and sugar acids
(Saha, 2003). Hemicellulose compositions differ depending on their origins;
hardwood hemicelluloses contain mostly xylans, whereas softwood hemi-
celluloses contain mostly glucomannans (McMillan, 1993; Saha, 2003).
Xylans are heteropolysaccharides that have a homopolymeric backbone
composed of (1 ! 4)-linked β-D-xylopyranose units (Saha, 2003). In addi-
tion to xylose, xylans may also be composed of arabinose, glucuronic acid,
and acetic, ferulic, and p-coumaric acids (Saha, 2003). Unlike the linear cel-
lulose polymers, hemicelluloses have branches with short side chains com-
posed of different sugars that do not form aggregates, even when
cocrystallized with cellulose chains (Perez et al., 2002).
Because of the biodegradability of cellulose and of hemicelluloses in the
soil environment, we do not consider that they will be significant contrib-
utors to the humin fraction. However, it is possible that resilient fragments of
cellulose may be found in humin. It is also possible that more labile forms
will provide substrates for microorganisms, whose components contribute
to the compositions of humin (see, for example, Section 4.2).
5.3 Lignin
Lignin is a macromolecule of similar abundance to hemicellulose in woody
tissues and accounts for up to 30% of some secondary cell walls (Scheller
and Ulvskov, 2010). The secondary cell wall is defined as a thick layer
rich in lignin that strengthens and waterproofs the wall. It is formed inside
the primary cell wall, a thin, flexible, and extended layer of the cell wall
composed of cellulose, pectin, and hemicellulose. Lignin is an amorphous
heteropolymer composed of phenylpropane units (Fig. 15) joined together
by different types of linkages (Fig. 16; Perez et al., 2002) and is insoluble in
water. It has many functions, including structural support, and the provision
OH OH OH
OCH3 H3CO OCH3
OH OH OH
p-Courmaryl alcohol Coniferyl alcohol Sinapyl alcohol

Fig. 15 Structures of the phenylpropenoic alcohols involved in lignin biogenesis.
HOH2C CH2OH
HO O
OH
OCH3 OCH3
OCH3
HO O
H3CO O
OH H CO
3 O
O
O
OH
CH2OH
O
OH
H3CO H3CO
OH
O O
OH HO
HO H3CO OCH3
O OH
HO
OH
OCH3 O
HO
O OCH3 OH
H3CO OH OCH3
O O
OH
Fig. 16 Lignin from gymnosperms showing the different linkages between the phenyl-
propane units (Perez et al., 2002, p. 55).
of resistance against microbial attack and oxidative stress. The hydrophobic

nature of lignin shields the polysaccharides from degradation in the cell wall
by restricting the access of enzymes (Mansfield, 2009). The polymer is
thought to be synthesized by the generation of free radicals that are released
in the peroxide-mediated dehydrogenation of the three phenylpropanoic
alcohols: coumaryl alcohol (p-hydroxyphenyl propenol), coniferyl alcohol
(guaiacyl propenol), and sinapyl alcohol (syringyl propenol) shown in
Fig. 16 (Sánchez, 2009). Coniferyl alcohol is the major lignol component
of softwood lignins, whereas guaiacyl and syringyl alcohols are the major
components of hardwood lignins (Perez et al., 2002). There are a number
of other monolignols that can be present in low concentrations in some plant
species. The units in lignin polymers are linked by strong C–C and aryl–
ether linkages (Fig. 15) (Perez et al., 2002; Sánchez, 2009) that are difficult to
cleave.
The degradation of lignin, outlined in detail by Perez et al. (2002), is very
challenging because of its structural complexity, its high molecular weight,
and its insolubility. Enzymes (extracellular, oxidative, and unspecific) can
liberate highly unstable products (radicals) that undergo many additional
oxidative reactions to catalyze the initial steps of lignin depolymerization
(Perez et al., 2002). White rot fungi are the most efficient degraders of lignin
from wood (Perez et al., 2002). However, bacterially mediated lignin deg-
radation and the presence of lignin-degrading enzymes have been reported
for actinobacteria from the Streptomyces genus (Berrocal et al., 1997; Perez
et al., 2002). Peroxidases (lignin peroxidases and manganese-dependent per-
oxidases) and laccases (blue copper phenoloxidases) are involved in lignin
degradation mediated by white-rot fungi (Perez et al., 2002). Reductive
enzymes and aryl alcohol dehydrogenases also have major roles in the deg-
radation of lignin (Cullen, 1997; Perez et al., 2002). Thus, it can be seen that
there are a wide variety of mechanisms, involving microbial processes by
which the lignin components of plants can be transformed and degraded
in the soil environment.
Based on its chemical stability and resistance to decomposition, it might
be assumed that lignin and its decomposition products could make a signif-
icant contribution to humin. However, the low aromatic content of humin
would indicate that this is not so. Traces of altered lignin structures can occur
in humin fractions, but these are likely to arise from molecules trapped in the
hydrophobic matrix that is characteristic of humins, as will be referred to in
Section 6.
5.4 Tannins
Tannins are polyphenolic, secondary metabolites exclusive to higher plants.
They are found in the leaves, needles, and bark of many vascular plants. Tan-
nins are fourth in the order of abundance in terrestrial plants (after cellulose,
hemicellulose, and lignins) and have molecular weight values ranging from
500 to 3000 Da (Hagerman, 2011), and their overall input into the compo-
sition of humic substances may have been underestimated.
There are three major classes of tannins, and the base or monomer units
are gallic acid (I), flavone (II), and phloroglucinol (III) shown in Fig. 17. The
base units, in particular in the flavone-derived tannins, must be (additionally)
O OH O
OH HO O
O
HO OH
HO OH O OH
HO OH
OH O O
I II III IV
Fig. 17 Monomer units of tannins: (I) gallic acid; (II) flavone; (III) phloroglucinol;
(IV) ellagic acid.
Fig. 18 Structures of the most commonly occurring types of tannins are shown.
Gallotannins and ellagitannins are largely hydrolysable due to the presence of ester
groups in their structures, whereas complex and condensed tannins are either non-
or only weakly hydrolysable (Khanbabaee and van Ree, 2001).
heavily hydroxylated and polymerized in order to give the high molecular

weight polyphenol structures that characterize tannins.
Hydrolyzable tannins containing a sugar (mostly D-glucose) and phenolic
acids can be differentiated into gallotannins (Fig. 18), composed of a central
sugar unit esterified with gallic acid (structure I, Fig. 17), and ellagitannins
(Fig. 18), composed of the sugar and ellagic acid (structure IV, Fig. 17).
Condensed tannins (Fig. 18) are mainly composed of polyhydroxy-flavan-
3-ol with C–C bonds between C-4 and C-8, and between C-4 and C-6.
Condensed tannins are more heterogeneous because of the variety of func-
tional groups possible (K€ogel-Knabner, 2002). Almendros et al. (2000) using
13
C NMR spectroscopy have suggested that complex tannins are selectively
preserved during the composting of forest biomass.
As their name suggests, hydrolyzable tannins can be broken down hydro-
lytically (using hot water or tannases) into their components, whereas the
structures of condensed and of complex tannins make these more difficult
to separate (Khanbabaee and van Ree, 2001). It is difficult to quantify tan-
nins due to the complexity and the diversity of their chemical compositions.
In nature, it is reasonable to assume that hydrolyzable tannins are

decomposed more rapidly than condensed tannins.
Lignin and tannin compounds are the most likely sources of phenolic
compounds in HSs and humin, and these can be differentiated using
NMR spectroscopy (see Section 3.2). However, data presented in
Section 4 and referred to in Section 6 indicate that the contribution of tan-
nins to the composition of humin is likely to be minor.
However, the data in Section 4 show that there is strong evidence for
carbohydrate components in humin, but the evidence we have suggests that
the contribution of lignin and of lignin-derived materials is minor. Most
peaks for condensed tannins have a characteristic split peak at 144 and
154 ppm in the phenolic region, while that for guaiacyl lignin has a peak
or shoulder at 147 ppm (Lorenz et al., 2000). DD can provide confirmatory
evidence because of the overlap of the tannin resonances with the anomeric
C resonance (105 ppm) for carbohydrates. Confirmatory evidence can be
provided by DD NMR spectra; in this, the anomeric C resonance is lost.
Tannins also show a sharp peak at 130 ppm in CPMAS spectrum and this
is retained in the DD spectrum. The uses of these spectra provide evidence
for the presence of tannin-type materials in HSs.
5.5 Lipids
Lipids are a heterogeneous group of biomolecules that occur both in lower
and higher plants and in microorganisms. Eukaryotic lipids play only a minor
role in the compositions of recalcitrant SOM. Lipids (fats, oils, resins, waxes)
are water-insoluble molecules that have a diverse range of functions. They
act as an energy store and fuel cells, are components of membranes, and
can serve as hormones and intracellular second messengers (Hames and
Hooper, 2011). Fatty acids are the simplest lipids ((CH3–(CH2)n–COOH),
where n represents the number of repeating CH2 units), and these are
components of many more complex lipids, e.g., waxes, triacylglycerols,
glycerophospholipids, and sphingolipids (Horton et al., 2006). The relative
abundance of particular fatty acids varies with the type of organism; for
example, branched fatty acids are common components of bacterial mem-
branes (Horton et al., 2006).
Waxes are polyesters of long-chain fatty acids and long-chain mono-
hydroxylic alcohols in plant cell walls (Horton et al., 2006). The long-chain
aliphatic hydrocarbon components render waxes nonpolar. The main func-
tion of waxes is to provide a protective waterproof coating for plants (some
leaves and fruits) and animals (skin, fur, and feathers) (Horton et al., 2006).
Waxes appear to be minor components of bacteria (Dinel et al., 1990).
Lipoproteins are globular particles consisting of a hydrophobic core of
triacylglycerols and cholesterol esters surrounded by a coat of protein,
phospholipid, and cholesterol (in eukaryotes) (Hames and Hooper,
2011). Bacterial lipoproteins are responsible for various important cellular
functions, such as biogenesis and the maintenance of cell surface structures,
and the transport of substrates (Okuda and Tokuda, 2011). Other lipids
include steroids (mainly in eukaryotes, and rarely in prokaryotes), vita-
mins, and terpenes, all of which are classified as isoprenoids (Horton
et al., 2006).
Lipases are the enzymes involved in the hydrolysis of triacylglycerols to
fatty acids (Hames and Hooper, 2011). The availability of oxygen, microbial
populations, and the pH of the local environment will affect the decompo-
sition of lipids. For example, in acid soils, filamentous fungi and
acetomycetes are favored, whereas in alkaline soils, decomposition of lipids
is promoted by soil microorganisms that produce lipases, which hydrolyze
complex lipid molecules to forms more readily utilized by microorganisms
(Dinel et al., 1990).
5.5.1 Sources of Hydrocarbons and Lipids Associated With SOM

In the context of this discussion, the term “lipid” is used to describe both the
parent substances and the fatty compounds derived from these. The main
roles of lipids are in the protection and organizations of cells. Plants have
the largest range of lipids in comparison to bacteria and algae (Dinel
et al., 1990), and algae are a source of hydrocarbons, wax esters, and primary
alcohols (Dinel et al., 1990). The distribution of hydrocarbons in algae is a
homologous series of medium-chain alkanes ranging from C15 to C18, and
the distribution of alkanes is affected by abiotic factors, the algal species, and
the age of the organism (Dinel et al., 1990). Bacteria produce three main
categories of compounds: wax esters, aliphatic hydrocarbons, and extremely
complex, high molecular weight, polyfunctional compounds (Dinel et al.,
1990). The hydrocarbon composition of bacterial lipids may vary in relation
to C number distribution and the degree of saturation (Dinel et al., 1990).
Components in the lipids of plants include hydrocarbons, ketones, primary
alcohols, secondary alcohols, and wax esters (Dinel et al., 1990). Leaf waxes
are composed of long-chain fatty acids (C22 to C36) (Huang et al., 1998).
Grasset et al. (2009) found (using thermochemolysis) that lipids in clay-
associated OM were mainly short-chain fatty acids and hydrocarbons of
microbial origins. Insects are also a source of lipids in the soil environment as
their bodies are equipped with cuticular layers of hydrophobic materials to
prevent excessive desiccation; hydrophobic materials, for example, consti-
tute over 90% of the cuticular lipids of cockroaches (Dinel et al., 1990).
HSs and humin have large contents of aliphatic hydrocarbons that play an
important role in their long-term stabilization in the environment. The
presence of lipids does not conform to the classification of HSs, and their
removal is often regarded as a prerequisite for studies of the composition
of humic substances. Some of the lipid components are removable in non-
polar solvents; however, it is challenging to remove all of the lipid materials
because these can form strong associations with other components in the
humic fractions, especially the HAs, and in humin (Clapp et al., 2005).
Lipids show resistance to decomposition and are preserved for long periods
(Oro et al., 1965), as demonstrated by Eglinton et al. (1968) when they iso-
lated hydroxy fatty acids (10,16-dihydroxyhexadecanoic acid and
ω-hydroxy acids in the C16 to C24 range) from a 5000-year-old lacustrine
sediment. Accumulations of aliphatic compounds in SOM are also enhanced
when there is a high input of long-chain aliphatics in plant biomass, micro-
bial activity, and low soil pH (Bull et al., 2000; de Assis et al., 2011). The
evidence for contributions to humin structures of resistant long-chain ali-
phatic lipids/waxes, as provided by various NMR procedures, is discussed
in Section 4.
Lipids can be analyzed using mass spectrometry methods (LC/MS, GC/
MS, pyGC/MS, ESI-FTMS) and 1H and 13C NMR spectroscopy. Some
methods may require lipid extraction in organic solvents prior to character-
ization. Applications of pyGC/MS have significantly advanced our aware-
ness of long-chain hydrocarbons, esters, acids, and alcohols from the soil
environment. Alkanes, alkenes, fatty acid n-alkyl esters, and alkyl aromatics
were among the products identified by Schnitzer and Schulten (1995) from
soils using pyGC/MS (Clapp et al., 2005).
Given their resistance to decomposition, and their resulting persistence
in the environment, it is not surprising that components with origins in lipids
contribute significantly to the composition of humin.
5.6 Cutin and Cutan

There is ample evidence to indicate the presence of long-chain poly-
methylenic structures in the organic matter of soils and sediments
(Deshmukh et al., 2005). Some of these components are considered to
derive from plant cuticles containing the biopolymers cutin and cutan, and
the suberized parts of plants containing suberin (Nierop, 1998). Plant cuti-
cles are synthesized and secreted by the epidermis during plant development
(Kunst et al., 2005), and cutin and cutan are the most notable polymeric lipid
structures found in these cuticles (Deshmukh et al., 2003). Their function is
to provide the plant with a protective barrier from the external environment,
as well as separating different organs of the plant (Deshmukh et al., 2003;
Kolattukudy, 1980).
Cutin comprises the macromolecular frame of the plant cuticle in which
the low molecular weight waxes and fats are embedded, forming the cuticle.
It is described as the component that can be solubilized upon saponification
and can account for 40%–80% of the cuticular weight in plant organs
(Holloway, 1982; Kunst et al., 2005). It is composed of fatty acids that have
a chain length of 16- or 18-carbon ω-hydroxyacids, with hydroxyl or epoxy
groups in the mid-chain positions (Kunst et al., 2005). Cuticular waxes are
embedded in the cutin matrix and are “very long chain fatty acid
derivatives” that are readily removed using nonpolar solvents (Kunst
et al., 2005). These waxes are arranged into an intracuticular layer in close
association with the cutin matrix, and there is an epicuticular film exterior to
this (Kunst et al., 2005). Cutin forms a three-dimensional network, formed
by extensive ester cross-linking of the monomeric species (Walton and
Kolattukudy, 1972; Kolattukudy, 1984), that is associated with polysaccha-
rides and intracellular wax, as well as small amounts of phenolic compounds,
such as p-coumarate and ferulate (Holloway, 1982; Kunst et al., 2005). In
addition to ester functionalities, Deshmukh et al. (2005) observed epoxy
groups, free primary alcohols and carboxylic acid groups, as well as evidence
for α-branched fatty acids/esters in cutin. The α-branched carboxylic acids
offer opportunities for cross-linking and explain the presence of amorphous
chains in the cutin/cutan mixture (Deshmukh et al., 2005).
Early models depicted cutin monomers linked head to tail in a linear
manner via their primary functional groups and partially cross-linked through
their secondary hydroxyl groups (Kunst et al., 2005). More recently, glycerol
has been reported to be a major constituent of cutin accounting for up to 14%
of the total monomers (Graça et al., 2002; Kunst et al., 2005; Moire et al.,
1999). Solid-state NMR studies, using the leaves of lime trees and tomato
fruit cuticles, have shown that aliphatic, alkene, aromatic, keto, and ester
functionalities contribute to the compositions of cutin and demonstrated
the presence of cross-linking (Deshmukh et al., 2003; Fang et al., 2001;
Pacchiano et al., 1993; Zlotnik-Mazori and Stark, 1988).
Cutan is the resistant residue remaining after depolymerization of cutin

but is less well understood. Cutan was first detected as a nonsaponifiable
component, resistant to deesterification by alkaline hydrolysis. It is a com-
mon aliphatic biopolymer in cuticles of drought-adapted plants (Boom et al.,
2005) and increases in content in some plants as they reach maturity. Appar-
ently, during maturation of these plants, cutan replaces the cutin secreted in
the early stages of cuticle development (Schmidt and Sch€ onherr, 1982).
Deshmukh et al. (2005) have used both 1H and 13C NMR spectroscopy
to provide insights into its structure and composition. They describe cutan
(from Agave americana leaf ) as being composed mainly of free primary
hydroxyls and long-chain carboxylic acids that form ester linkages with tri-
hydroxylated benzene units (Fig. 19). They also identified benzenecarboxylic
acids that form ester links with fatty alcohols in the structure. The long-chain
Fig. 19 Structure proposed by Deshmukh et al. (2005) for Agave americana cutan, show-
ing various types of functional units. The values for n vary from 25 to 32 based on the
work by McKinney et al. (1996) and Schouten et al. (1998). Values for m are at least 6 to
allow sufficient remoteness from the carbonyl group to display a chemical shift assign-
able to L structures. Values of m + n are less than 31 (Deshmukh et al., 2005, p. 1083). For
assignments of structures attributable to A to L, see the Table associated with the
Figure in Deshmukh et al. (2005, p. 1075).
polymethylenic groups in cutan are shown to express a high degree of

“crystallinity” or rigidity (Collinson et al., 1994; Deshmukh et al.,
2005; Tegelaar et al., 1991, 1995; van Bergen et al., 1994). PyGC/MS
studies of A. americana cutan identified a homologous series of n-alkanes,
n-alk-1-enes, and α,ω-alkadienes (Nip et al., 1986a,b; Tegelaar et al.,
1989b) that they attributed to the thermal degradation of the poly-
methylenic chains (Deshmukh et al., 2005). Thermochemolysis (using
TMAH) of A. americana leaf cutan yields fatty acid methyl esters (C15–
C31, with a high concentration of C27–C31), as well as a number of
1,3,5-trimethoxylated benzene derivatives and benzene carboxylic acid
derivatives (Deshmukh et al., 2005; McKinney et al., 1996). The 13C
CPMAS NMR data and the pyGC/MS have been combined to suggest
a structure for cutan in which a backbone of 1,3,5-trihydroxylated aro-
matic rings (Deshmukh et al., 2005) forms ester linkages with fatty acids
(Deshmukh et al., 2005; McKinney et al., 1996). Fig. 19 provides an out-
line of the structure proposed by Deshmukh et al. (2005). The proposed
structure incorporates units that include α-branched carboxylic acids,
esterified secondary alcohols, free primary alcohols, olefinic linkages,
and aromatic structures with carboxylic acid and aryl-OH inclusions.
5.7 Suberin and Suberan

In addition to cutins, plants synthesize suberin (and suberan), a second type
of insoluble lipid polyester derived from fatty acids (Molina et al., 2006). The
function of suberins is to control the movement of water and solutes and to
provide strength to the cell wall (Molina et al., 2006; Nawrath, 2002).
Suberin is a major component of the outer bark cell walls and is organized
in a characteristic lamellar structure, as suggested in Fig. 20 (Gandini et al.,
2006). It does not have a distinct chemical structure as its compositions can
vary (Gandini et al., 2006). The monomer unit composition of suberin has
been determined for many plant species, but a detailed macromolecular
structure, and how it associates with other cell wall biopolymers, has not
been established (Gandini et al., 2006). Suberin is composed of both ali-
phatic and aromatic species (Bernards, 2002; Bernards et al., 1995;
Holloway, 1984; Kolattukudy, 2001); the aliphatic components are mostly
C16 to C28 ω-hydroxy fatty acids and C16 to C26 α,ω-dioic acids (these are
diagnostic for suberin) with little mid-chain oxygen functionality, and the
aromatic component is a “hydroxycinnamate-derived polymer, primarily
Fig. 20 The suberin model proposed by Bernards (2002); S indicates linkages to suberin
(see also Gandini et al., 2006, p. 881).
comprised of ferulic acid, N-feruloytyramine, cinnamic acid, p-coumaric

acid, or caffeic acid” (Bernards et al., 1995; Molina et al., 2006). Glycerol
has been shown to be an important component of suberin (Bernards,
1998; Gandini et al., 2006; Graça and Pereira, 1997, 1999, 2000a,b,
2002) because it can account for ca. 20% (by weight) of suberin in oak, cot-
ton, and potato (Graça and Pereira, 2000b,c; Moire et al., 1999; Molina
et al., 2006). The most recent model for the suberin macromolecule (based
on suberized potato cell walls) was proposed by Bernards (2002) (Fig. 20).
Solid-state NMR studies have shown that there are two distinct aromatic
domains in suberized cell walls; the first aromatic domain is located inside the
aliphatic domains and is composed mainly of hydroxycinnamates esterified
with glycerol or ω-hydroxyfatty acids. The second aromatic domain is a
lignin-like polymer that is composed of cross-linked hydroxycinnamic
acid-type molecules that are covalently bonded to the aliphatic suberin
(Gandini et al., 2006; Lopes et al., 2000; Pascoal Neto et al., 1996; Sitte,
1962). It is proposed that ether or ester bonds may form between suberin
and polysaccharides (Gandini et al., 2006; Lopes et al., 2000; Sitte, 1962;
Yan and Stark, 2000). Studies of suberin by Rocha et al. (2001) identified
both rigid and mobile aliphatic methylene species in suberin. Crystalline ali-
phatic suberin has been identified in Quercus suber cork (Bernards, 2002;
Gandini et al., 2006; Lopes et al., 2000).
Difficulties arise in estimating the suberin content because of its complex

macromolecular structure, and because of the similarity of the aromatic
domains in suberin and lignin (Bernards, 1998; Bernards, 2002; Gandini
et al., 2006; Kolattukudy, 1980; Kolattukudy, 2001; Kolattukudy and
Espelie, 1989). A solvent extraction step is necessary in studies of suberin
in order to isolate the low molecular weight components, followed by
the chemical scission of the ester bonds in the macromolecule (Gandini
et al., 2006; Kolattukudy and Espelie, 1989). Enzymes can be used to cleave
bonds between polysaccharides and aliphatic suberin (Rocha et al., 2001).
The isolated suberin polymers/fragments can be qualitatively and quantita-
tively characterized using GC/MS, FTIR, and NMR spectroscopy.
Suberan, a nonhydrolyzable biopolymer, is ubiquitous in the periderm
tissue of higher plants (Ogawa and Tanoue, 2003). Suberan, unlike suberin,
is not saponifiable, and therefore, it is defined as the residue remaining after
the saponification of suberin (Turner et al., 2013).
Suberans (and cutans) are suggested to be responsible for the enrichment
of aliphatic compounds in soils (Augris et al., 1998; Tegelaar et al., 1989c).
Therefore an awareness of the chemical nature of suberan is important for an
understanding of diagenetic and humification processes because the below-
ground plant biomass and the bark layer of trees make significant contribu-
tions to SOM (Turner et al., 2013).
Tegelaar et al. (1995) using pyGC/MS showed suberan to be composed
of a homologous series of n-alkanes, n-alk-1-enes, and α,ω-alkadienes that
ranged C6–C37, but mostly with chain lengths from C20 to C22. Suberan
retains ester functionality following saponification processes, and this is
attributed to the hydrophobic nature of the material (due to the poly-
methylene compositions) that provides some protection from extensive
alkaline hydrolysis (Deshmukh et al., 2005; McKinney et al., 1996;
Schouten et al., 1998; Turner et al., 2013). The data obtained by Turner
et al. (2013) for suberan indicate that the chain length, the dominant ester
functionality, and the functional groups present all have similar features to
those observed for suberin; however, suberan is structurally distinct. It is
possible that suberan is an artifact created during the isolation of suberin;
alternatively, it may be a distinct polymer that coexists with suberin
(Turner et al., 2013).
In the light of the information presented above about cutins, cutans,
suberins, and suberans, it is highly likely that these components will make
significant contributions to the humin fraction in soils. This will be referred
to in Section 6.
5.8 Latex Materials

About 10% of plant species exude latex products, and in many cases, com-
ponents in these have defensive roles against herbivorous predators (Konno,
2011). Latex is variously described as a “milky” suspension or emulsion of
particles in an aqueous fluid “although sometimes it may be a thin, clear,
yellow or orange, aqueous suspension.” Latex is produced in vessels or spe-
cial cells called laticifers, single cells, or strings of cells that form tubes, canals,
or networks in various plant organs. Well-known plants that produce latex
products are milkweeds (Asclepias spp.) which exude latex when the stems
are broken, and dandelion (Taraxacum officinale) that exudes latex through
its roots in defense of attack by the cockchafer larva (Melolontha melolontha).
Latex is a complex emulsion containing secondary metabolites including
proteins, lectins (chitin-binding proteins), chitinases, oxidases (polyphenol
oxidase and peroxidase), alkaloids, sugars, oils, tannins, resins, and gums.
They are the secondary metabolites that provide the active ingredients that
provide the mechanisms that defend plants against predators. Recently, it has
been shown that the sesquiterpene lactone taraxinic acid β-D-
glucopyranosyl ester (TA-G) is the active ingredient in the dandelion root
latex that is negatively associated with M. melolontha larval growth (Huber
et al., 2016).
It is plausible to consider that latex emulsions can incorporate the hydro-
phobic constituents of humin and transport these in the soil environment.
5.9 Algaenan
The algaenans are protective tissues in algae, but their exact physiological
function has not been determined (Blokker et al., 2006). Algaenan has been
described as an aliphatic, insoluble, and chemically resistant biopolymer
(Tegelaar et al., 1989a) that is a major component of the outer cell wall
in Botryococcus braunii (Berkaloff et al., 1983; Blokker et al., 1998a, 2000;
Derenne et al., 1989; Gatellier et al., 1993; Gelin et al., 1994; Kadouri
et al., 1988; Simpson et al., 2003), and a simplified proposed structure is
shown in Fig. 21. When algae reproduce, the parental cell wall is released
as a waste product that is predominantly composed of the resistant algaenan
biopolymer (Blokker et al., 1998b). These resistant biomolecules are ubiq-
uitous in freshwater green algae (de Leeuw and Largeau, 1993; Ogawa and
Tanoue, 2003) and can also be synthesized by cyanobacteria and marine
algae (Gelin et al., 1996, 1999; Largeau, 1995; Ogawa and Tanoue,
2003). Selective preservation of algaenan was also demonstrated by
O O O O
O (CH2)x CH CH (CH2)y C O (CH2)x CH (CH2)z C
O O O
O (CH2)x CH CH (CH2)y C O (CH2)x CH CH (CH2)y C
Fig. 21 Proposed simplified structure of algaenans (based on analyses of T. minimum,

S. communis, and P. boryanum) consisting of linear polyester chains cross-linked via
ether bonds (Blokker et al., 1998b, p. 1465).
Nguyen et al. (2003) who showed that insoluble, nonhydrolyzable algaenan

(from B. braunii) has a slower rate of degradation compared to diatoms
(predominantly silica-based cell walls; Kr€ oger et al., 1994), dinoflagellates
(principally composed of cellulose; Brock and Madigan, 1984), and cyano-
bacterium (predominantly composed of peptidoglycan and lipopolysaccha-
rides; Brock and Madigan, 1984). Despite comprising only a few percent of
the total biomass of algae, algaenan is selectively preserved during algal deg-
radation and becomes a major constituent in kerogen (Derenne et al., 1991;
Gillaizeau et al., 1996; Goth et al., 1988; Largeau et al., 1984, 1986). In a 2D
13
C–15N NMR spectroscopy study of degraded B. braunii, Zang et al. (2001)
found evidence to suggest that algaenan may be involved in the protection of
peptide bonds from enzymatic attack (Nguyen et al., 2003). Algaenan is
therefore of geochemical importance (Gelin et al., 1999; Largeau et al.,
1984, 1986) because of its persistence in the environment, and it represents
an important biomolecule in the sequestration of C.
Studies by Simpson et al. (2003) found that the hexane-insoluble botryals
(α-branched aldehydes originating from aldol condensation) isolated from
B. braunii are composed of a mixture of low molecular weight unsaturated
aliphatic aldehydes and unsaturated hydrocarbons with an average chain
length of about 40 carbons.
Spectroscopic methods, such as FTIR and solid-state 13C NMR, pro-
vide some information about the nature and functional groups of the
algaenan biopolymer, but the stability of the ether linkages has made it dif-
ficult to obtain information on the finer details of the structures of the build-
ing blocks (Blokker et al., 1998b). PyGC/MS has been the most successful
method to provide such information; however, thermal rearrangement
reactions can result in the loss of information about functional groups
(Blokker et al., 1998b). Ruthenium tetroxide (RuO4) oxidation, which
can cleave ether linkages (Ambles et al., 1996), coupled with GC/MS has
shown that α,ω-dicarboxylic acids, with either ester or ether bonds that link
these hydroxyacids to each other, are building blocks of algaenans (Schouten
et al., 1998). A proposed simplified structure by Blokker et al. (1998b) sug-
gests that double bonds may be biochemically oxidized and ether bonds
formed which in turn will cross-link the polymer (Fig. 21).
Traditionally the attention on algae has been largely directed toward
aquatic forms, but the existence of definite algal flora in soils is now well
recognized. Some of the soil species also occur in water (Shields and
Durrell, 1964), and the contributions of algae toward soil fertility are signif-
icant. Hence, algaenan could be a significant component of soil humin.
5.10 Bacteran
The term bacteran applies to a high level of insoluble material formed when
species of mycobacteria are subjected to drastic saponification and acid
hydrolysis. Allard et al. (1997) showed, by means of FTIR, 13C NMR, ther-
mal analysis (DTG), and KMnO4 degradation, that the products had similar
properties to melanoidin-like materials formed when monosaccharides and
amino acids from the cell walls of mycobacteria were subjected to the same
saponification and hydrolysis conditions. Thus the so-called bacteran mate-
rial may well be an artifact of the isolation process.
5.11 Glomalin
Glomalin is an abundant and persistent extracellular protein produced by
arbuscular mycorrhizal fungi discovered by Wright and Upadhyaya
(1996) and named as glomalin, after the source organism of phylum
“Glomeromycota.” Glomalin is reportedly a nonwater-soluble, highly per-
sistent glycoproteinaceous substance (Wright and Upadhyaya, 1998) pro-
duced in the mycorrhizal fungal cell walls, and it remains in soil after
hyphal death (Driver et al., 2005). Glomalin has been linked to aggregate
stability, to long-term C and N storage (Wright and Upadhyaya, 1998),
and it responds to land-use changes (Rillig et al., 2003). The role of glomalin
in the ecosystem is still unclear. Its hypothetical role has evolved from that of
an active secretion to enhance soil aggregation (Wright and Upadhyaya,
1996), or a hydrophobin that modifies water acquisition (Rillig, 2005), to
one that suggests that glomalin is specifically related to fungal metabolism,
and that its role as a persistent soil protein is fortuitous (Purin and
Rillig, 2007).
The process of isolating the material is harsh and involves autoclaving in a

sodium citrate buffer. It is now operationally defined as glomalin-related soil
protein (GRSP) and the term glomalin is reserved for the purified protein
gene product (Rillig, 2004). Based on its solubility characteristics, glomalin
would be a component of the humin fraction, but if it exists as a glycopro-
tein, it would need to be sorbed onto soil mineral colloids or refractory
organic substances.
5.12 Melanins
Haider and Martin (1967) (see also the extensive discussion in Hayes and
Swift, 1978 and in Clapp et al., 2005 to the work involving these authors
between 1967 and 1977) have shown in laboratory studies that fungal cul-
tures, such as E nigrum, Stachybotrys atra, S. chartarum, and Hendersonula
toruloidea, could form brown humic-like or melanin-type polymers from
phenols, quinones, and nonaromatic precursors. More recent work has
shown that fungi and some bacteria synthesize melanins that occur in cell
walls (Butler and Day, 1998). The melanin pigments contain protein, car-
bohydrate, lipids, and a polymeric core having various types of phenol, qui-
none, and indole monomers. Butler and Day (1998) have shown that white
rot fungi completely degrade melanins.
Although the lesser charged components of melanins may contribute to
soil humin, it is likely that the major components of melanins would be iso-
lated with the humic fractions
5.13 Black Carbon, Char/Biochar

Research on the organic geochemistry of black carbon (BC) was advanced
by the publication of Black Carbon in the Environment, by Goldberg (1985).
Interest in BC was accelerated when the work of Wim Sombroek (Woods
et al., 2009) made it evident that the fertility of the Terra Preta soils of the
Amazon Region could be attributed to what is now referred to as biochar.
Biochar is the name given to the char product formed from the pyrolysis of
biomass and used as a soil amendment. It exists as a continuum from slightly
charred, degradable biomass to charcoal, highly condensed, refractory soot
particles, graphitic black carbon, etc. All components of this continuum
have high carbon contents, are chemically heterogeneous, and are domi-
nated by condensed aromatic structures (Hedges et al., 2000; Masiello,
2004).
Char can be an important contributor to the total soil C in some

instances, and it can also be considered to be an important sink for
C when present in soils and sediments. Significant amounts of char have
been reported for Mollisol, Black Earth, and Chernozemic soils (Glaser
et al., 1998; Schmidt et al., 1999; Skjemstad et al., 2002). It has been reported
that the charred OC contributes up to 45% of the OC and up to 8 g kg1 in a
range of gray to black soils (Cambisol, Luvisol, Phaeozem, Chernozem, and
Greyzem) (Schmidt et al., 1999). The char content in the Elliott silt loam
Mollisol soil, the source of soil standard HA and FA fractions of the IHSS,
is ca. 6.6 g C kg1, or 23% of the total soil OC (Skjemstad et al., 2002). Song
et al. (2002) combined acid demineralization, base extraction, and dichro-
mate oxidation to isolate BC and kerogen from a humin material. Relative
contents of BC varied from 18.3% to 41.0% of the TOC in soils and sedi-
ments from an industrialized region of China.
The complexity of chars from different environments and the lack of
common methods for the study and analysis of BC/char make it difficult
to measure the proportion of this material in SOM (Novotny et al.,
2006, 2007). Thus, some of the soil BC or char contents may be over-
estimated (Simpson and Hatcher, 2004). Gonzalez-Perez et al. (2004)
reviewed the effect of fire on SOM, and they also discussed the BC stabili-
zation in soil and its bearing on the global carbon cycle. BC was found to be
accumulated in the <53 μm fraction and this pattern was sustained even after
tillage (Brodowski et al., 2006).
Using electrospray ionization Fourier transform ion cyclotron resonance
mass spectrometry (FT-ICR MS), Kramer et al. (2004) provided good evi-
dence for the existence of condensed fused aromatic ring systems in char-
enriched HAs extracted from a volcanic soil in Japan. Three different types
of highly carboxylated polycyclic aromatic compounds were identified in
the sample: linearly fused aromatic structures; aromatic structures linked
by carbon–carbon single bonds; and highly condensed aromatic structures.
The feature of these heavily carboxylated, hydrogen-deficient, condensed
aromatic structures is believed to be characteristic of black carbon-like
material.
Knicker et al. (1996) investigated the structural change of lignocellulosic
biomass from 15N-enriched ryegrass under controlled heating conditions
using solid-state 13C and 15N NMR spectroscopy. The solid-state 15N
NMR revealed amide-N in forms which are resistant to the thermal treat-
ment. Also, their stability toward further microbial degradation is increased
by the heating. The NMR spectra indicated that the carbohydrate fraction is
converted into condensed dehydrated material producing intense signals in
the aromatic region (Knicker et al., 2005a,b).
5.14 Summary of Section 5

In summary of this section, it is the view of the authors that because of their
wide distribution, chemical composition, and their degree of resistance to
microbial degradation, cutins, cutans, suberins, suberans, lipids, algaenans,
and bacterans, and latex exudates should make significant contributions to
insoluble and nonhydrolyzable components in HSs, especially to the humin
fractions. Some or all of these are very likely to be the major precursors by
selective preservation of aliphatic moieties in humin.
Cellulose, hemicellulose, glomalin, peptides, and latex materials can be
expected to degrade readily in soils and would need to be protected by sorp-
tion or associations with other persistent soil components in order to be
found to any significant extent in humin.
Altered lignins, tannins, and melanins make substantial contributions to
the extractable HA and FA, but make little contribution to the humin
fraction.
Char/biochar material, if present, will form part of the humin fraction
but because it was formed as the result of fire and has not been formed as
part of a biological process, it is best considered as a separate entity.
6. SUMMATION, SIGNIFICANCE, AND FUTURE

DIRECTIONS
In this section the main threads of the arguments and supporting infor-
mation presented will be drawn together and summarized. Then the signif-
icance and implications of our growing knowledge of the composition of
humin on the properties and behavior of SOM will be addressed. Following
this, a number of suggestions will be made about some of the issues that need
to be investigated further in order to improve our knowledge of humin and
to gain a better understanding the roles that it plays in soils.
6.1 Summation
As discussed in Sections 1 and 2, humin in soil has been operationally defined
as the fraction of the SOM that is insoluble in aqueous base. Traditionally, it
has been assumed that humin was similar in structure and composition to
humic substances but was insoluble in base, mainly because of its lower levels
of carboxyl groups leading to fewer sites that can be exchanged with highly
soluble, readily dissociating cations, resulting in lower charge density. Sim-
ilarly, it was believed that humin contained lower amounts of other oxygen-
containing functional groups and also had a higher molecular weight com-
pared to the humic substances, which were additional causes for its lack of
solubility in basic aqueous media.
Another reason advanced for the lack of solubility of humin was its inti-
mate association with clay minerals. Although this may well be the case in
some soils, it cannot be the cause in all soils because humin is found in sandy
soils and peats where clays or other finely divided minerals are absent.
The important consequence that arose from the lack of solubility of
humin was that it was not possible to utilize routine chemical and many
instrumental techniques to carry out compositional studies on this substantial
fraction of SOM.
A number of attempts were made to overcome the humin solubility
problem in the latter part of the 20th century. A significant input was made
by Rice and MacCarthy (1989) who introduced a “partitioning” approach
using an immiscible combination of MIBK and aqueous base. The humin-
rich fraction effectively accumulated at the interface of these immiscible
liquids and could then be collected. Another approach was to carry out
extractions using aqueous base in order to remove soluble organic com-
ponents and then to remove the mineral component of the soil leaving
the humin fraction behind. This was achieved using HCl/HF mixtures
to remove silicates and metal oxides, and this technique has been used
on whole soils from which the base-soluble organic materials had been
removed, and on soil components, especially the clay–silt fractions isolated
from such soils.
At around the same period of time the instrumental technique of solid-
state 13C NMR was being developed and refined. This proved to be an ideal
tool for the analysis of solid-state SOM samples. When applied to the sam-
ples of humin obtained from the procedures referred to above, the quality of
the NMR spectra was greatly improved and showed considerable enhance-
ments and in some cases the dominance of aliphatic hydrocarbon species.
However, from the spectral signals obtained, it was also evident that the
humin isolation techniques outlined above were still relatively crude and
that most of the samples studied still contained significant amounts of signals
from retained humic substances. Consequently, it was evident that
improvements in isolation methods and instrumental analyses would be

required to make further progress.
Fortunately, the types of advances required in both of these areas were
made in the first decade or so of this century. In terms of isolation, it became
clear that exhaustive extraction with aqueous base alone was not sufficient to
remove all of the potentially soluble humic substances probably because of
strong adsorption onto mineral surfaces, and/or intimate associations with
the humin components. It has been shown that aqueous base plus added urea
(made up to 6 M with respect to urea) was a very effective extractant for
removing most, if not all, of the remaining humic substances and other sol-
uble organic components. The result is that the residual organic material
could be regarded as being more representative of the true composition
of the humin fraction.
The next key development was the discovery that DMSO containing 6%
(by volume) of concentrated H2SO4 (referred to as “acidified DMSO” and
in this anhydrous system, it should be noted that the sulfuric acid is behaving
as a solvent) was found to be an effective solvent for substantial amounts of
the humin fraction. The extent of extraction of the humin from the clay
fraction varied from soil to soil. However, following HF/HCl treatment,
it was shown that the humin retained in the clay fraction gave the same
NMR spectrum as the humin extracted using acidified DMSO.
During the same period of time that the improved isolation techniques
for humin were being utilized, a range of highly sophisticated instrumental
analytical techniques were being developed for use in solid- and liquid-state
NMR. The application of these modern NMR techniques, which include
DE 1H NMR (Section 4.2), CMP (Section 4.3), and applications of 2D
NMR (Section 4.4), to the humin isolates has greatly enhanced our knowl-
edge of their compositions and their molecular structures, with a high level
of confidence.
For example, we now have a much greater knowledge of the major ali-
phatic hydrocarbon components of humin. It is a mixture of a range of ali-
phatic structures and long-chain fatty acids and esters, and there is also good
evidence for structures derived from a range of plant and microbial cuticular
and cell wall components. The evidence accumulated so far indicates that
the composition of the components identified showed little, if any, change
from the composition of the source material.
In most instances, much lower amounts of carbohydrate, peptide, and
lipoprotein materials (which could include glomalin and cellulose) were
associated with the humin fraction. Such materials would normally be read-
ily decomposed or transformed by microorganisms. Their survival, even in
small amounts, indicates that these minor components may be protected by
close association with the hydrophobic, largely aliphatic humin and/or with
the soil mineral colloids.
It is well understood that SOM is a highly complex mixture in which all
of the classes of compounds found in plants and other living organisms can be
found in varying amounts. All of these compounds are subject to decompo-
sition at different rates, following different pathways. During the transforma-
tion processes, some plant components persist largely unaltered for some
considerable time, whereas others undergo substantial transformations
and, as a result, are chemically different and bear no morphological resem-
blance to the starting materials.
A good example of these different types of component and pathways is
illustrated in Fig. 3 showing the 13C NMR spectra of an HA extract and
humin fraction isolated from the same soil. The HA is greatly modified
and is chemically distinct from its plant precursors. The spectra show strong
carboxyl and aromatic resonances and lesser amounts of carbohydrate and
aliphatic components. In contrast, the humin fraction is closely related to
the cuticular components of plants and its spectrum is dominated by the ali-
phatic carbon signals with much lower contributions from carboxyl, aro-
matic, and carbohydrate components. The observed low levels of
aromatic contributions to the composition of humin are puzzling because
aromatic moieties make significant contributions to the structures of com-
ponents of cuticular materials, such as cutin/cutan (Section 5.6) and
suberin/suberan (Section 5.7). It may well be that in the diagenesis pro-
cesses the more reactive aromatic functionalities are cleaved, or otherwise
degraded to leave the more hydrophobic long chain-associated hydrocar-
bon structures to persist, possibly in associations with the soil mineral
colloids.
Although it is clear that different components of plant residues and of
SOM decompose at different rates, it is also apparent that all of the compo-
nents must decompose eventually. If this were not so then, over time, the
nondecomposable component would totally dominate the composition of
the SOM (Jenkinson, 1981) and the total amount of SOM in the soil would
increase with time. This does not happen.
Nevertheless, the differential rates of decomposition of the components
of SOM are reflected in its overall composition. For example, from our
experience with mineral soils, around 20%–30% of the SOM is readily
extractable (of which the humic substances comprise 10%–20% of the

SOM). The insoluble remainder would be referred to as humin in the clas-
sical definitions. Much less detailed data are available about the composition
of this material compared to the humic substances, but based on currently
available verified data, it is apparent that 50%–70% of this fraction could
be derived from highly aliphatic cuticular components or other structures
of similar composition. For the most part, cuticular components make up
only a few percent of the composition of plants, yet they appear to make
up a large proportion of this residual organic matter in humin emphasizing
a slower rate of decomposition than the other plant components.
6.2 Significance of Humin Composition

If the composition of the humin fraction proposed here and elsewhere (see
Section 4 for relevant references) is found to be the norm, then there are
many consequences for our understanding of SOM and for the interpreta-
tion of its many contributions to the properties and behavior of soil. For
example, the retention of nutrient cations by negatively charged carboxylic
acid sites and the cycling of organic forms of nitrogen, sulfur, and phospho-
rus during the decomposition of SOM are commonly and correctly cited as
key functions of SOM. However, given the composition proposed for the
humin fraction, it is unlikely to play much of a role, if any, in these processes
which help to underpin soil fertility. This means that the humic substances
and associated organic components are almost entirely responsible for
undertaking these key functions.
Similarly, water retention by SOM is considered to be another important
contribution to the maintenance of soil fertility. The basis of this water
retention is the presence of the hydrophilic polar functional groups in humic
substances, carbohydrates, and other macromolecular organic acids plus the
associated hydrated cations. The hydrophobic nature of the humin fraction
indicates that it would play a little role in the retention of water and may well
be at least partly responsible water repellence or the nonwetting behavior in
certain soils. Conversely, the humin fraction is well suited to adsorb and
retain a range of hydrophobic lipid compounds (such as natural fats, oils,
and waxes). In addition, it is well suited to adsorb and retain nonpolar
biocides used for the control of weed pests and plant diseases as well as
other hydrophobic substances that might enter the soil and thereby help
to prevent their movement through the environment after use or accidental
contamination.
Another area where SOM is important is in formation and maintenance

of soil structure. Although some of the humin fraction is found to be closely
associated with the soil clay minerals, it is not possible to say at this stage
whether it is making a contribution to the formation or maintenance of soil
structure. Intuitively, it would be expected that the components with a
higher functional group content and greater flexibility would be more likely
to be involved in these processes.
A disturbing outcome of the spread of intensive modern agriculture has
been the general decline in SOM levels over a relatively short period of time.
This situation is of substantial concern if all of the components of the SOM
are declining at a similar rate. However, if the components that are largely
responsible for the important positive fertility contributions are
decomposing at a faster rate than components which make little contribu-
tion to overall soil fertility, then the situation is even worse.
If we briefly consider the wider environmental context of soil and SOM
beyond the agricultural context, the issues of global warming and the need
and the potential for carbon sequestration are most pressing. There is a gen-
eral awareness that SOM constitutes the largest terrestrial reservoir of carbon
within the Earth’s C-cycle. As might be expected given the difficulty of the
task, current estimates of total soil carbon vary quite widely, but it is not
unreasonable, for the sake of argument, to use 2000 Gt as a rough approx-
imation. On this basis, it is salutary to consider that if SOM levels could be
increased in surface soils by around 10% of their current values, then the
concentration of carbon dioxide in the atmosphere would be reduced to
preindustrial revolution levels (1850 AD). Although this would be a mam-
moth and probably unachievable task, it does indicate the significant impact
that could be made by implementing effective soil sequestration projects.
Certainly, soils where the SOM levels have been depleted to levels well
below their natural steady-state value as a result of agricultural practices
could well be targeted for attention. Given that humin makes up 50% or
more of the total SOM, it would be an important component of any carbon
sequestration strategy.
However, considerable amounts of plant residue would need to be added
together with changes in agricultural practices to achieve the required out-
come. But compared with other more complex and expensive technologies,
it represents a low-cost solution where the potential for action exists with
regard to both the land use and its day-to-day management already being
in place. In the context of this chapter, the resistant plant materials that
are the precursors of humin would probably result in longer turnover times
and more effective sequestration outcomes. Thus the use of plants with
higher levels of cuticular material, achieved by plant selection and breeding
without loss of product yield, would greatly enhance the value of this
approach.
The observations presented in this section underscore the importance of
SOM to soil fertility, nutrient and carbon cycling, and, in turn, agricultural
production. It is clear that it is important not only to maintain SOM at suf-
ficiently high levels but also to ensure that the necessary balance of the com-
ponents of SOM is achieved in order for it to perform its range of tasks
effectively.
6.3 Future Directions

It is important to understand that the information and concepts presented in
this chapter do not change the actual composition and functions of SOM per
se, but they do change our conceptions and understanding of that compo-
sition and, thereby, our ability to interpret and predict its behavior in a wide
range of climatic, vegetation, and management systems. In addition, this
improved understanding of the composition of SOM will provide a poten-
tial basis for greater manipulation of SOM composition for specific purposes,
such as improving soil fertility in one situation or increasing carbon seques-
tration in another by modifying vegetation and its composition.
In order to reach this situation, there is much more work to be done.
Initially, a much wider range of soils need to be examined to determine
the size of the humin fraction and to determine whether its composition
is generally the same as that indicated in this chapter, i.e., a dominance of
alkyl hydrocarbon and the extents to which these components reflect the
long-term vegetation and/or the biological activity of the soil. It will also
be instructive to carry out chemical analyses of the minor components of
the humin, particularly the identification of the component sugars and
amino acids in the carbohydrate and protein fractions, respectively. This
information can be used to determine whether these components originate
in plants or microorganisms or both (Hayes et al., 2008, 2012) and the pro-
portions of each.
Another worthwhile set of studies would be to explore differences arising
from different management systems, different vegetation, different climates,
etc. For example, measurements should be made of the total SOM levels and
the related amounts and compositions of the humin fraction in soils
maintained under continuous pasture or native vegetation and compared
to the same soils subjected to long-term cultivation. Similarly, comparisons

should be made between soils in arid/semiarid regions and those from more
humid areas to assess the effects of climate and vegetation. It will be impor-
tant also to measure the amounts and changes in composition in humin frac-
tions at different depths in the soil profile.
Allied to this work could be a range of studies of the nature and compo-
sition of the dominant vegetation in order to assess the influence of plants
types that naturally contain higher or lower levels of cuticular materials
on the composition of the resultant SOM and the proportion and compo-
sition of the humin fraction.
In addition to the studies outlined above, it will also be important to
ascertain the stability or resistance to decomposition of the humin fraction.
This work could take two main directions, one of which is to study the rate
of decomposition of the humin fraction using (potentially very long-term)
incubation experiments. For such incubation studies, it would be useful to
include the humin isolated from a soil by extraction compared with the same
humin retained in the soil following the removal of the base-soluble humic
fractions. This would help to assess the impact of clay–organic interactions
on the rate of decomposition. The processes involved in the decomposition
of humin may well involve the same types of organisms that are capable of
decomposing heavy oil deposits to remediate contaminated soils.
Another approach to this type of study is to use radiocarbon studies to
determine in a wide range of soils the age and turnover rates for the humin
fraction relative to the age of other fractions and the SOM as a whole. Such
work would involve carbon dating using natural radiocarbon levels and
decomposition studies could be carried out using labeled samples. In the case
of natural radiocarbon studies, it would be highly desirable to be able to
access historical soil samples which were taken before 1944 and have been
sealed and carefully stored since then.
A puzzling aspect of the work carried out on humin so far is the close
association that is observed with the clay fraction in some soils. Given the
refractory nature, resistance to decomposition, and insolubility of these
materials, it is not easy to understand how this association with the clay frac-
tion might occur. One possibility is that the cuticular materials are commi-
nuted by soil organisms to very small particle sizes and simply become an
inseparable part of the clay-sized soil fraction. Another possibility is that
the hydrophobic, highly aliphatic compounds are formed and released as
part of the biological decomposition process and adsorbed onto adjacent clay
minerals surfaces in a similar way that clays are able to remediate oil spillages.
However, prima facie, surface adsorption of such compounds onto clays and
other minerals would be difficult to achieve because adsorption onto a sur-
face requires that the sorbed species needs to be in solution before it can
move to the surface and be adsorbed. That would be an unlikely occurrence
for hydrophobic substances in the aqueous soil environment.
An alternative mechanism that may allow the adsorption of hydrophobic
species to occur onto mineral surfaces in soils is through the formation of an
emulsion in which small particles of hydrophobic material are suspended
within an aqueous medium. Such emulsion suspensions occur naturally as
latex within many plants (see Section 5.8), and it is possible that similar sus-
pensions could occur within the cells or bodies of the soil microorganisms
and mesofauna that are capable of decomposing the humin fraction and are
also in contact soil mineral particles and colloids. When such emulsion sus-
pensions are released from the cells or bodies of dead or dying organisms the
opportunity exists for the suspended hydrophobic materials to move to and
be adsorbed onto clay or mineral surfaces. Detailed studies to explore this
possible mechanism would greatly enhance our understanding of many
clay–organic interactions and also offer possible explanations not only as a
sorption mechanism per se but also in the formation of nonwetting and
water-repellent soils.
These are just a few of the many studies that need to be carried out to
increase our knowledge about humin its origins and its contributions in
the soil and the wider environment.
A final consideration arising from this review is the current description of
humin as a component of the soil humic substances. This description states
that: “To be regarded as a humic substance in the classical definitions, humin
should bear no morphological resemblance to the materials of origin, should
not include biological molecules that are not covalently linked to the humic
matrix, and be insoluble in water at all pH values (Aiken et al., 1985; Rice
and MacCarthy, 1988).” Prior to this, in regularizing the descriptions of
SOM fractions, Hayes and Swift (1978) have stated that in addition to having
no morphological resemblance to their precursors, humic substances should
also have undergone transformations resulting in major changes in chemical
composition and structure. Since there is a clear possibility that cuticular and
related plant components may well persist in soil in largely chemically
unaltered forms, it will be necessary to reconsider this status of humin as
a component of humic substances.
REFERENCES
Aiken, G.R., McKnight, D.M., Wershaw, R.L., MacCarthy, P., 1985. An introduction to
humic substances in soil, sediment, and water. In: Aiken, G.R., McKnight, D.M.,
Wershaw, R.L., MacCarthy, P. (Eds.), Humic Substances in Soil, Sediment, and Water:
Geochemistry, Isolation and Characterization. John Wiley & Sons, New York, NY, pp. 1–9.
Allard, B., Templier, J., Largeau, C., 1997. Artificial origin of mycobacterial bacterin. For-
mation of melanoidin-like artefact macromolecular material during the usual isolation
process. Org. Geochem. 27, 697–703.
Almendros, G., González-Vila, F.J., 1987. Degradative studies of a soil humin fraction.
Sequential degradation of inherited humin. Soil Biol. Biochem. 19, 513–520.
Almendros, G., Sanz, J., 1989. Compounds released from humic acid upon BF3-MeOH
transesterification. Sci. Total Environ. 81/82, 51–60.
Almendros, G., Sanz, J., 1992. A structural study of alkyl polymers in soil after perborate deg-
radation of humin. Geoderma 53, 79–95.
Almendros, G., Polo, A., Dorado, E., 1981. Isolation and characterization of the non-
extractable humin fraction from different soils. Agrochimica 25, 479–491.
Almendros, G., Sanz, J., Sobrados, I., 1989. Characterisation of synthetic carbohydrate-
derived humic-like polymers. Sci. Total Environ. 81 (82), 91–98.
Almendros, G., Sanz, J., González-Vila, F.J., Martin, F., 1991. Evidence for a polyalkyl
nature of soil humin. Naturwissenschaften 78, 359–362.
Almendros, G., Guadalix, M.E., González-Vila, F.J., Martin, F., 1996. Preservation of ali-
phatic macromolecules in soil humins. Org. Geochem. 24, 651–659.
Almendros, G., Dorado, J., González-Vila, F.J., Blanco, M.J., Lankes, U., 2000. 13C NMR
assessment of decomposition patterns during composting of forest and shrub biomass.
Soil Biol. Biochem. 32, 793–804.
Ambles, A., Grasset, L., Dupas, G., Jacquesy, J.C., 1996. Ester and ether bond cleavage in
immature kerogens. Org. Geochem. 24, 681–690.
Anderson, D.W., Paul, E.A., Arnard, R.J., 1974. Extraction and characterization of humus
with reference to clay-associated humus. Can. J. Soil Sci. 54, 317–323.
Andrew, E.R., 1981. Magic angle spinning. Int. Rev. Phys. Chem. 1 (2), 195–224.
Ash, H.B., 1941. Translation of De Re Rustica Published by LJM Columella Prior to 70 CE
Loeb Classical Library. Harvard University Press, Cambridge, MA.
Atalla, R.H., Hackney, J.M., Uhlin, I., Thompson, N.S., 1993. Hemicelluloses as structure
regulators in the aggregation of native cellulose. Int. J. Biol. Macromol. 15, 109–112.
Augris, N., Balesdent, J., Mariotti, A., Derenne, S., Largeau, C., 1998. Structure and origin
of insoluble and non-hydrolyzable, aliphatic organic matter in a forest soil. Org. Geo-
chem. 28, 119–124.
Balci, M., 2005. Basic 1H-13C-NMR Spectroscopy. Elsevier, Amsterdam.
Baldan, B., Andolfo, P., Navazio, L., Tolomio, C., Mariani, P., 2001. Cellulose in algal cell
wall: an “in situ” localization. Eur. J. Histochem. 45, 51–56.
Baldock, J.B., Oades, J.M., Nelson, P.N., Skene, T.M., Golchin, A., Clarke, P., 1997.
Assessing the extent of decomposition of natural organic materials using solid-state
13
C NMR spectroscopy. Aust. J. Soil Res. 35, 1061–1083.
Bax, A., Griffey, R.H., Hawkins, B.L., 1983. Correlation of proton and nitrogen-15 chem-
ical shifts by multiple quantum NMR. J. Magn. Reson. 55, 301–315.
Beguin, P., Aubert, J.P., 1994. The biological degradation of cellulose. FEMS Microbiol.
Rev. 13, 25–58.
Berkaloff, C., Casadevall, E., Largeau, C., Metzger, P., Peracca, S., Virlet, J., 1983. Hydro-
carbon formation in the green alga Botryococcus braunii. Part 3. The resistant polymer of
the walls of the hydrocarbon-rich alga B. braunii. Phytochemistry 22, 389–397.
Bernards, M.A., 1998. The macromolecular aromatic domain in suberized tissue: a changing
paradigm. Phytochemistry 47, 915–933.
Bernards, M.A., 2002. Demystifying suberin. Can. J. Bot. 80, 227–240.

Bernards, M.A., Lopez, M.L., Zajicek, J., Lewis, N.G., 1995. Hydroxycinnamic acid-
derived polymers constitute the polyaromatic domain of suberin. J. Biol. Chem.
270, 7382–7386.
Berns, A.E., Conte, P., 2011. Effect of ramp size and sample spinning speed on CPMAS 13C
NMR spectra of soil organic matter. Org. Geochem. 42, 926–935.
Berrocal, M., Rodrıguez, J., Ball, A.S., Perez-Leblic, M.I., Arias, M.E., 1997. Solubilization
and mineralisation of 14C lignocelluloses from wheat straw by Streptomyces cyaneus CECT
3335 during growth in solid-state fermentation. Appl. Microbiol. Biotechnol.
48, 379–384.
Berzelius, J., 1839. Lehrbuch der Chemie, third ed. (W€ ohler, Trans.). Arnoldische
Buchhandlung; Leipzig (available from the Harvard University Library, Cambridge,
MA).
Blokker, P., Sinninghe Damste, J.S., De Leeuw, J.W., Van den Ende, H., 1998a. Structural
comparison of algal microfossils with extant algaenans using ruthenium tetroxide degra-
dation. Mineral. Mag. 62A, 170–171.
Blokker, P., Schouten, S., van den Ende, H., de Leeuw, J.W., Hatcher, P.G., Sinninghe
Damste, J.S., 1998b. Chemical structure of algaenans from the fresh water algae
Tetraedron minimum, Scenedesmus communis and Pediastrum boryanum. Org. Geochem.
29, 1453–1468.
Blokker, P., Schouten, S., de Leeuw, J.W., Sinninghe Damste, J.S., van den Ende, H., 2000.
A comparative study of fossil and extant algaenans using ruthenium tetroxide degrada-
tion. Geochim. Cosmochim. Acta 64, 2055–2065.
Blokker, P., Van den Ende, H., De Leeuw, J.W., Versteegh, G.J.M., Sinninghe Damste, J.S.,
2006. Chemical fingerprinting of algaenans using RuO4 degradation. Org. Geochem.
37, 871–881.
Boom, A., Sinninge Damste, J., Deleeuw, J., 2005. Cutan, a common aliphatic biopolymer
in cuticles of drought-adapted plants. Org. Geochem. 36, 595.
Brock, T.D., Madigan, M.T., 1984. Biology of Microorganisms. Prentice Hall, Englewood
Cliffs.
Brodowski, S.B., John, H.F., Amelung, W., 2006. Aggregate occluded black carbon in soil.
Eur. J. Soil Sci. 57, 539–546.
Buddrus, J., Burba, P., Herzog, H., Lambert, J., 1989. Quantitation of partial structures of
aquatic humic substances by one- and two-dimensional solution 13C nuclear magnetic
resonance spectroscopy. Anal. Chem. 61, 628–631.
Bull, I.D., Van Bergen, P.F., Nott, C.J., Poulton, P.R., Evershed, R.P., 2000. Organic geo-
chemical studies of soils from the Rothamsted classical experiments—V. The fate of lipids
in different long-term experiments. Org. Geochem. 31, 389–408.
Burdon, J., 2001. Are the traditional concepts of the structures of humic substances realistic?
Soil Sci. 166, 752–769.
Butler, M.J., Day, A.W., 1998. Fungal melanins: a review. Can. J. Microbiol. 44, 1115–1136.
Byrne, C.M.P., 2011. Studies of the organic matter components in Irish grassland soils and
their drainage waters. University of Limerick, Ireland. PhD Thesis.
Byrne, C.M.P., Hayes, M.H.B., Humar, R., Novotny, E.H., Lanigan, G., Richards, K.G.,
Fay, D., Simpson, A.J., 2010. Compositional changes in the hydrophobic acids frac-
tion of drainage water from different land management practices. Water Res.
44, 4379–4390.
Cardoza, L.A., Korir, A.K., Otto, W.H., Wurrey, C.J., Larive, C.K., 2004. Applications of
NMR spectroscopy in environmental science. Prog. Nucl. Magn. Reson. Spectrosc.
45, 209–238.
Chaney, K., Swift, R.S., 1984. The influence of organic matter on aggregate stability of some
British soils. J. Soil Sci. 35, 223–230.
Chefetz, B., Salloum, M.J., Deshmukh, A.P., Hatcher, P.G., 2002. Structural components of
humic acids as determined by chemical modifications and carbon-13 NMR, pyrolysis-
and thermochemolysis-gas chromatography/mass spectrometry. Soil Sci. Soc. Am. J.
66, 1159–1171.
Cheshire, M.V., Cranwell, P.A., Falshaw, C.P., Floyd, A.J., Haworth, R.D., 1967. Humic
acid—II: Structure of humic acids. Tetrahedron 23, 1669–1682.
Clapp, C.E., Hayes, M.H.B., 1996. Isolation of humic substance from an agriculture soil
using a sequential and exhaustive extraction process. In: Clapp, C.E., Hayes, M.H.B.,
Senesi, N., Griffith, S.M. (Eds.), Humic Substances and Organic Matter in Soil
and Water Environments: Characterization, Transformations and Interactions.
International Humic Substances Society, Inc., Minnesota, pp. 3–11.
Clapp, C.E., Hayes, M.H.B., 1999. Characterization of humic substances isolated from clay-
and silt-sized fractions of a corn residue amended agricultural soil. Soil Sci. 164, 899–913.
Clapp, C.E., Hayes, M.H.B., Simpson, A.J., Kingery, W.L., 2005. Chemistry of soil organic
matter. In: Tabatabai, M.A., Sparks, D.L. (Eds.), Chemical Processes in Soil. Soil Science
Society of America, Madison, WI, pp. 1–150.
Claridge, T.D.W., 2009. High-Resolution NMR Techniques in Organic Chemistry.
Elsevier, Hungary.
Cloos, P., Badot, C., Herbillon, A., 1981. Interlayer formation of humin in smectites. Nature
289, 391–393.
Collinson, M.E., van Bergen, P.F., Scott, A., de Leeuw, J.W., 1994. The oil-generating
potential of plants from coal and coal-bearing strata through time: a review with new
evidence from carboniferous plants. In: Scott, A.C., Fleet, A.J. (Eds.), Coal and Coal-
Bearing Strata as Oil-Prone Source Rocks. Geological Society, London, pp. 31–67. Spe-
cial Publication No. 77.
Conte, P., Piccolo, A., van Lagen, B., Buurman, P., Hemminga, M.A., 2001. Effect of resid-
ual ashes on CPMAS-13C NMR spectra of humic substances from volcanic soils.
Fresenius Environ. Bull. 10, 368.
Conte, P., Spaccini, R., Piccolo, A., 2004. State of the art of CPMAS 13C-NMR spectros-
copy applied to natural organic matter. Prog. Nucl. Magn. Reson. Spectrosc.
44, 215–223.
Cook, R.L., Langford, C.H., 1998. Structural characterization of a fulvic acid and a humic
acid using solid-state ramp-CP-MAS 13C nuclear magnetic resonance. Environ. Sci.
Technol. 32, 719–725.
Courtier-Murias, D., Farooq, F., Masoom, H., Botana, A., Soong, R., Longstaffe, J.G.,
Simpson, M.J., Maas, W.E., Fey, M., Andrew, B., Struppe, J., Hutchins, H.,
Krishnamurthy, S., Kumar, R., Monette, M., Stronks, H.J., Hume, A., Simpson, A.J.,
2012. Comprehensive multiphase NMR spectroscopy: basic experimental approaches to
differentiate phases in heterogeneous samples. J. Magn. Reson. 217, 61–76.
Cullen, D., 1997. Recent advances on the molecular genetics of ligninolytic fungi.
J. Biotechnol. 53, 273–289.
Dai, X.L., Ping, C.L., Candler, R., Haumaier, L., Zech, W., 2001. Characterization of soil
organic matter fractions of Tundra soils in Arctic Alaska by carbon-13 nuclear magnetic
resonance spectroscopy. Soil Sci. Soc. Am. J. 65, 87–93.
Dai, X.Y., Ping, C.L., Michaelson, G.J., 2002. Characterizing soil organic matter in Arctic
tundra soils by different analytical approaches. Org. Geochem. 33, 407–419.
Dakyn, H.G., 1897. The Works of Xenophon. MacMillan, London.
Darwin, C., 1881. The Formation of Vegetable Mould, Through the Action of Worms,
With Observations on Their Habits. John Murray, London.
de Assis, C.P., González-Vila, F.J., Jucksch, I., González-Perez, J.A., Neves, J.C.L.,
Lani, J.L., de Sá Mendonça, E., 2011. Lipid abundance and composition of a humic
oxisol as a function of land use. Sci. Agric. (Piracicaba, Braz.) 68 (2), 230–236.
de Leeuw, J.W., Largeau, C., 1993. A review of macromolecular organic compounds that
comprise living organisms and their role in kerogen, coal and petroleum formation. In: -
Engel, M.H., Macko, S.A. (Eds.), Organic Geochemistry. Plenum Press, New York,
pp. 23–72.
Deherain, P., 1902. Traite de Chimieagricole, second ed. Masson, Paris.
Derenne, S., Largeau, C., Casadevall, E., Berkaloff, C., 1989. Occurrence of a resis-
tant biopolymer in the L race of Botryococcus braunii. Phys. Chem. Chem. Phys.
28, 1137–1142.
Derenne, S., Largeau, C., Casadevall, E., Berkaloff, C., Rousseau, B., 1991. Chemical evi-
dence of kerogen formation in source rocks and oil shales via selective preservation of
thin resistant outer walls of microalgae: origin of ultralaminae. Geochim. Cosmochim.
Acta 55, 1041–1050.
Deshmukh, A.P., Simpson, A.J., Hatcher, P.G., 2003. Evidence for cross-linking in tomato
cutin using HR-MAS NMR spectroscopy. Phytochemistry 64, 1163–1170.
Deshmukh, A.P., Simpson, A.J., Hadad, C.M., Hatcher, P.G., 2005. Insights into the struc-
ture of cutin and cutan from Agave americana leaf cuticle using HRMAS NMR spectros-
copy. Org. Geochem. 36, 1072–1085.
Dinel, H., Schnitzer, M., Mehuys, G.R., 1990. Soil lipids: origin, nature, content, decom-
position and effect on soil physical properties. In: Bollag, J.M., Stotzky, G. (Eds.), Soil
Biochemistry, vol. 6. Marcel Dekker, New York, pp. 397–429.
Dixon, A.M., Larive, C.K., 1999. NMR spectroscopy with spectral editing for the analysis of
complex mixtures. Appl. Spectrosc. 53 (11), 426A–440A.
Dokuchaev, V.V., 1883. Russian chernozem (Russkiy chernozem). also (1949) soch. 3, Izd.
Akad. Nauk SSSR.
Dria, K.J., Sachleben, J.R., Hatcher, P.G., 2002. Solid-state carbon-13 nuclear magnetic res-
onance of humic acids at high magnetic field strengths. J. Environ. Qual. 31, 393–401.
Driver, J.D., Holben, W.E., Rillig, M.C., 2005. Characterization of glomalin as a hyphal wall
component of arbuscular mycorrhizal fungi. Soil Biol. Biochem. 37, 101–106.
Duer, M.J., 2002. Essential techniques for spin-1/2 nuclei. In: Duer, M.J. (Ed.), Solid-State
NMR Spectroscopy: Principles and Applications. Blackwell Science, Oxford,
pp. 73–110.
Eglinton, G., Hunneman, D.H., McCormick, A., 1968. Gas chromatographic-mass spectro-
metric studies of long chain hydroxy acids. III. The mass spectra of the trimethylsilyl
ethers of aliphatic hydroxy acid methyl esters. A facile method of double bond location.
Org. Mass Spectrom. 1, 593–611.
Ellis, G.P., 1959. The Maillard reaction. Adv. Carbohydr. Chem. 14, 63–124.
Enders, C., 1942. pH-abhangigkeit und gleichgewichtsreaktion der aufspaltung der zucker
zu methylglyoxal. Biochem. Z. 312, 349–353.
Enders, C., 1943a. pH-abhangigkeit und gleichgewichtsreaktion der aufspaltung der zucker
zu methylglyoxal II. Mitteilung. Biochem. Z. 314, 265–269.
Enders, C., 1943b. Uber den chemismus der huminsauerbildung unter physiologischen
bedingungen. IV. Mitteilung. Die rolle der mikroorganismen bei den
humifizierungsvorgangen. Biochem. Z. 315, 259–292.
Enders, C., Fries, G., 1936. Zur analogie von melanoidinen und huminsauren. Kolloid Z.
76, 289–291.
Enders, C., Marquardt, R., 1941. Methylglyoxal als zwischenprodukt der melanoidin und
karmelbildung. Naturwissenschaften 29, 46–47.
Enders, C., Sigurdsson, S., 1943. Uber den chemismus der huminsauerbildung unter phys-
iologischen bedingungen V. Mitteilung. Die einleitende phase der huminsaurebildung.
Eine aldolkondensation van methylglyoxal. Ber. Deutsch. Chem. Ges. 76, 560–563.
Enders, C., Sigurdsson, S., 1947. Uber den chemismus den huminsaurenbildung unter phys-
iologischen bedingungen VII. Mitteilung. Biochem. Z. 318, 44–46.
Enders, C., Tschapek, M., Glane, R., 1948. Vergleichende untersuchungen einiger kolloider
eigenschaften von naturlichen huminsauren und synthetischen melanoidinen. Kolloid Z.
110, 240–244.
Fan, T.W.-M., Higashi, R.M., Lane, A.N., 2000. Chemical characterization of a chelator-
treated soil humate by solution-state multinuclear two-dimensional NMR with FTIR
and pyrolysis-GC-MS. Environ. Sci. Technol. 34, 636–646.
Fang, X., Qiu, F., Wang, H., Mort, A.J., Stark, R.E., 2001. NMR studies of molecular struc-
ture in fruit cuticle polyesters. Phytochemistry 57, 1035–1042.
Flaig, W., Beutelspacher, H., Reitz, E., 1975. Composition and physical properties of humic
substances. In: Gieseking, J.E. (Ed.), In: Soil Components, vol. 1. Springer-Verlag,
Berlin, New York, pp. 1–219.
Franz, J.A., Garcia, R., Linehan, J.C., Love, G.D., Snape, C.E., 1992. Single-pulse excitation
carbon-13 NMR measurements on the Argonne premium coal samples. Energy Fuels
6, 598–602.
Freitas, J.C.C., Bonagamba, T.J., Emmerich, F.G., 1999. 13C high-resolution solid-state
NMR study of peat carbonization. Energy Fuels 13, 53–59.
Fuchs, W., 1931. Die Chemie der Kohle. Springer, Berlin.
Gandini, A., Pascoal Neto, C., Silvestre, A.J., 2006. Suberin: a promising renewable resource
for novel macromolecular materials. Prog. Polym. Sci. 31 (10), 878–892.
Gatellier, J.P.L.A., De Leeuw, J., Damste, J.S.S., Derenne, S., Largeau, C., Metzger, P.,
1993. A comparative study of macromolecular substances of a coorongite and
cell walls of the extant alga Botryococcus braunii. Geochim. Cosmochim. Acta 57,
2053–2068.
Gelin, F., De Leeuw, J.W., Damste, J.S.S., Derenne, S., Largeau, C., Metzger, P., 1994. The
similarity of chemical structures of soluble aliphatic polyaldehyde and insoluble algaenan
in the green microalga Botryococcus braunii race A as revealed by analytical pyrolysis. Org.
Geochem. 21, 423–435.
Gelin, F., Boogers, I., Noordeloos, A.A.M., Sinninghe Damste, J.S., Hatcher, P.G., de
Leeuw, J.W., 1996. Novel, resistant microalgal polyethers: an important sink of organic
carbon in the marine environments? Geochim. Cosmochim. Acta 60, 1275–1280.
Gelin, F., Volkman, J.K., Largeau, C., Derenne, S., Damste, J.S.S., De Leeuw, J.W., 1999.
Distribution of aliphatic, nonhydrolyzable biopolymers in marine microalgae. Org. Geo-
chem. 30, 147–159.
Gelinas, Y., Baldock, J.A., Hedges, J.I., 2001. Demineralization of marine and freshwater
sediments for CP/MAS 13C NMR analysis. Org. Geochem. 32, 677–693.
Gemmecker, G., 1999. The NOE effect. Online, available at: http://www.spectroscopynow.
com/userfiles/sepspec/file/specNOW/Tutorials/chem843-6.pdf (accessed 23/03/2014).
Gillaizeau, B., Derenne, S., Largeau, C., Berkaloff, C., Rosseau, B., 1996. Source organisms
and formation pathway of the kerogen of the G€ oyn€ uk oil shale (Oligocene, Turkey) as
revealed by electron microscopy, spectroscopy and pyrolysis. Org. Geochem.
24, 671–679.
Glaser, B., Haumaier, L., Guggenberger, G., Zech, W., 1998. Black carbon in soils: the use of
benzenecarboxylic acids as specific markers. Org. Geochem. 29, 811–819.
Goldberg, E.D., 1985. Black Carbon in the Environment. Wiley, New York.
Gonzalez-Perez, J.A., Gonzales-Vila, F.S., Gonzalo, A., Knicker, H., 2004. The effect of fire
on soil organic matter—a review. Environ. Int. 30, 855–870.
Goth, K., de Leeuw, J.W., P€ uttmann, W., Tegelaar, E.W., 1988. Origin of Messel oil shale
kerogen. Nature 336, 759–761.
Graça, J., Pereira, H., 1997. Cork suberin: a glyceryl based polyester. Holzforschung
51, 225–234.
Graça, J., Pereira, H., 1999. Glyceryl-acyl and aryl-acyl dimers in Pseudotsuga menziesii bark
suberin. Holzforschung 53 (4), 397–402.
Graça, J., Pereira, H., 2000a. Methanolysis of bark suberins: analysis of glycerol and acid
monomers. Phytochem. Anal. 11, 45–51.
Graça, J., Pereira, H., 2000b. Diglycerolalkenedioates in suberin: building units of a
poly(acylglycerol) polyester. Biomacromolecules 1 (4), 519–522.
Graça, J., Pereira, H., 2000c. Suberin structure in potato periderm: glycerol, long-chain
monomers, and glyceryl and feruloyl dimers. J. Agric. Food Chem. 48, 5476–5483.
Graça, J., Pereira, H., 2002. Suberin structure in potato periderm: glycerol, long-chain
monomers and glyceryl and feruloyl dimers. J. Agric. Food Chem. 48 (11),
5476–5483.
Graça, M.A.S., Pozo, J., Canhoto, C., Elosegi, A., 2002. Effect of eucalyptus plantations and
detritus, decomposers, and detritivores in streams. Sci. World J. 2, 1173–1185.
Grasset, L., Martinod, J., Plante, A.F., Amblès, A., Chenu, C., Righi, D., 2009. Nature and
origin of lipids in clay size fraction of a cultivated soil as revealed using preparative the-
rmochemolysis. Org. Geochem. 40, 70–78.
Greenwood, N.N., Earnshaw, A., 1997. Chemistry of the Elements, second ed.
Butterworth-Heinemann, Oxford.
Guo, L.S., Gifford, R.M., 2002. Soil carbon stocks and land use change: a meta analysis.
Global Change Biol. 8, 345–360.
Hagerman, A.E., 2011. The Tannin Handbook. The Hagerman Laboratory, Miami
University, Oxford.
Haider, K., Martin, J.P., 1967. Synthesis and transformation of phenolic compounds by Epi-
coccum nigrum. Soil Sci. Soc. Am. Proc. 31, 766–772.
Hames, D., Hooper, N., 2011. Biochemistry, fourth ed. Garland Science, New York,
pp. 27–47.
Hatcher, P.G., VanderHart, D.L., Earl, W.L., 1980. Use of solid-state 13C NMR in structural
studies of humic acids and humin from Holocene sediments. Org. Geochem. 2, 87–92.
Hatcher, P., Breger, I., Maciel, G., Szeverenyi, N., 1985. Geochemistry of humin. In:
Aiken, G.R., McKnight, D.M., Wershaw, R.L., MacCarthy, P. (Eds.), Geochemistry,
Isolation and Characterization. John Wiley and Sons, New York, pp. 275–302.
Haworth, R.D., 1971. The chemical nature of humic acids. Soil Sci. 111, 71–79.
Hayes, M.H.B., 1960. Subsidence and humification in peats. The Ohio State University.
PhD Dissertation.
Hayes, M.H.B., 1985. Extraction of humic substances from soils. In: Aiken, G.R.,
McKnight, D.M., Wershaw, R.L., MacCarthy, P. (Eds.), Humic Substances in Soil,
Sediment, and Water; Geochemistry, Isolation and Characterization. John Wiley and
Sons, New York, pp. 329–362.
Hayes, M.H.B., 2006. Solvent systems for the isolation of organic components from soils.
Soil Sci. Soc. Am. 70, 986–994.
Hayes, M.H.B., 2009. Evolution of concepts of environmental natural nonliving organic
matter. In: Senesi, N., Xing, B., Huang, P.M. (Eds.), Biophysico-Chemical Process
Involving Natural Nonliving Organic Matter in Environmental Systems. In: IUPAC
Series on Biophysico-Chemical Process in Environmental Systems, vol. 2. Wiley,
Chichester, pp. 1–39.
Hayes, M.H.B., Swift, R.S., 1978. The chemistry of soil organic colloids. In: Greenland, D.J.,
Hayes, M.H.B. (Eds.), The Chemistry of Soil Constituents. Wiley, Chichester,
pp. 179–320.
Hayes, M.H.B., Swift, R.S., 1990. Genesis, isolation, composition and structures of soil
humic substances. In: DeBoodt, M.F., Hayes, M.H.B., Herbillon, A. (Eds.), Soil Col-
loids and Their Associations in Aggregates. Plenum, New York, pp. 245–305.
Hayes, M.H.B., Swift, R.S., Wardle, R.E., Brown, J.K., 1975. Humic materials from an
organic soil: a comparison of extractants and of properties of extracts. Geoderma
13, 231–245.
Hayes, M.H.B., Dawson, J.E., Mortensen, J.L., Clapp, C.E., Hausler, M.J., 1984. Compar-
ison of synthetic ‘humic-like’ substances with soil humic acids. Volunteered Papers 2nd
International Conference IHSS (Birmingham). pp. 157–165.
Hayes, M.H.B., Swift, R.S., Byrne, C.M., Simpson, A.J., 2010. The isolation and charac-
terization of humic substances and humin from grey brown podzolic and gley grassland
soils. In: Gilkes, R.J., Prakongkep, N. (Eds.), Proceedings of the 19th World Congress of
Soil Science: Soil Solutions for a Changing World, Brisbane, Australia, 1–6 August,
pp. 198–201.
Hayes, T.M., Hayes, M.H.B., Skjemstad, J.O., Swift, R.S., 2008. Studies of compositional
relationships between organic matter in a grassland soil and its drainage waters. Eur. J. Soil
Sci. 59, 603–616.
Hayes, T.M., Hayes, M.H.B., Swift, R.S., 2012. Organic matter components in extracts and
in drainage waters from a soil under long-term cultivation. Org. Geochem. 52, 13–21.
Hedges, J.I., Keil, R.G., 1995. Sedimentary organic matter preservation: an assessment and
speculative synthesis. Mar. Chem. 49, 81–115.
Hedges, J.I., Eglinton, G., Hatcher, P.G., Kirchman, D.L., Arnosti, C., Derenne, S.,
Evershed, R.P., K€ ogel-Knabner, I., de Leeuw, J.W., Littke, R., Michaelis, W.,
Rullk€ otter, J., 2000. The molecularly-uncharacterized component of nonliving organic
matter in natural environments. Org. Geochem. 31 (10), 945–958.
Hodge, J.E., 1953. Chemistry of browning reactions in model systems. Agric. Food Chem.
1, 928–943.
Holloway, P.J., 1982. The chemical constitution of plant cutins. In: Cutler, D., Alvin, K.,
Price, C. (Eds.), The Plant Cuticle. Linnean Society Symposium Series, Academic
Press, London, pp. 45–85.
Holloway, P.J., 1984. Cutins and suberins, the polymeric plant lipids. In: Mangold, H.K.,
Zweig, G., Sherma, J. (Eds.), In: CRC Handbook of Chromatography: Lipids,
vol. 1. CRC Press, Florida, pp. 321–345.
Hoppe-Seyler, F., 1889. Uber huminsubstanzen, ihre emstenung and ihre eigenschaften. 2.
Physiol. Chem. 13, 66–121.
Horton, H.R., Moran, L.A., Scrimgeour, K.G., Perry, M.D., Rawn, J.D., 2006. Principles
of Biochemistry, fourth ed. Pearson Prentice Hall, New Jersey, pp. 52–253.
Hu, W.G., Mao, J., Schmidt-Rohr, K., Xing, B., 1999. NMR evidence for crystalline
domains in humin substances. In: Ghabbour, E.A., Davies, G. (Eds.), Understanding
Humic Substances, Advanced Methods, Properties and Applications. The Royal
Society of Chemistry, Cambridge, pp. 63–68.
Hu, W.G., Mao, J., Xing, B., Schmidt-Rohr, K., 2000. Poly(methylene) crystallites in humic
substances detected by nuclear magnetic resonance. Environ. Sci. Technol. 34, 530–534.
Huang, Y., Eglinton, G., Van der Hage, E.R.E., Boon, J.J., Bol, R., Ineson, P., 1998. Dis-
solved organic matter and its parent organic matter in grass upland soil horizons studied
by analytical pyrolysis techniques. Eur. J. Soil Sci. 49, 1–15.
Huber, M., Epping, J., Gronover, C.S., Fricke, J., Aziz, Z., Brillatz, T., Swyers, M.,
Kollner, T.G., Vogel, H., Hammerbacher, A., Triebwasser-Freese, D., Robert, C.A.,
Verhoeven, K., Preite, V., Gershenzon, J., Erb, M., 2016. A latex metabolite benefits
plant fitness under root herbivore attack. PLoS Biol. 14 (1) e1002332.
Huc, A.Y., Durand, B.M., 1977. Occurrence and significance of humic acids in ancient
sediments. Fuel 56, 73–80.
Ibrahim, M.M., Agblevor, F.A., El-Zawawy, W.K., 2010. Isolation and characterization of
cellulose and lignin from steam-exploded lignocellulosic biomass. BioResources 5 (1),
397–418.
Ishiwatari, R., 1985. Geochemistry of humic substances in lake waters. In: Aiken, G.R.,
McKnight, D.M., Wershaw, R.L., MacCarthy, P. (Eds.), Geochemistry, Isolation and
Characterization. John Wiley & Sons, New York, pp. 147–180.
Jenkinson, D.S., 1981. The fate of plant and animal residues in soil. In: Greenland, D.J.,
Hayes, M.H.B. (Eds.), The Chemistry of Soil Processes. Wiley, Chichester, UK,
pp. 505–561.
Jobbagy, E.G., Jackson, R.B., 2000. The vertical distribution of soil organic carbon and its
relation to climate and vegetation. Ecol. Appl. 10, 423–436.
Johnson, M.G., 1995. The role of soil management in sequestration of soil carbon. In:
Lal, R. et al., (Ed.), Soil Management and the Greenhouse Effect. Lewis Publishers,
Boca Raton, FL, pp. 351–364.
Johnson, C.E., Smernik, R., Siccama, T.G., Kiemle, D.K., Xu, Z.H., Vogt, D.J., 2005.
Using C-13 nuclear magnetic resonance spectroscopy for the study of northern hard-
wood tissues. Can. J. Forest Res. 35, 1821–1831.
Jurkiewicz, A., Maciel, G.E., 1995. 13C NMR spin-lattice relaxation properties and quan-
titative analytical methodology of 13C NMR spectroscopy for coals. Anal. Chem.
67, 2188–2194.
Kadouri, A., Derenne, S., Largeau, C., Casadevall, E., Berkaloff, C., 1988. Resistant
biopolymer in the outer walls of Botryococcus braunii, B race. Phytochemistry
27, 551–557.
Keeler, J., 2013. Understanding NMR Spectroscopy, second ed. John Wiley & Sons,
Singapore, p. 13.
Kelleher, B.P., Simpson, A.J., 2006. Humic substances in soils: are they really chemically dis-
tinct? Environ. Sci. Technol. 40, 4605–4611.
Kelleher, B.P., Simpson, M.J., Simpson, A.J., 2006. Assessing the fate of transformed plant
residues in the terrestrial environment using HR-MAS NMR spectroscopy. Geochim.
Cosmochim. Acta 70, 4080–4094.
Khan, D.V., 1945. A method of isolating the insoluble fraction (humin) from podzolic soils.
Dokl. vsesoyuz. Akad. s.-kh. Nauk Lenina, 7–8.
Khanbabaee, K., van Ree, T., 2001. Tannins: classification and definition. Nat. Prod. Rep.
18, 641–649.
Kidena, K., Murata, S., Nomura, M., 1996. Studies on the chemical structural change during
carbonization process. Energy Fuels 10 (3), 672–678.
Kiem, R., Knicker, H., K€ orschen, M., K€ ogel-Knabner, I., 2000. Refractory organic carbon
in C-depleted arable soil, as studied by 13C NMR spectroscopy and carbohydrate anal-
ysis. Org. Geochem. 31, 655–668.
Kingery, W.L., Simpson, A.J., Hayes, M.H.B., Locke, M.A., Hicks, R.P., 2000. Structural
analysis of humic substances by homonuclear and heteronuclear two-dimensional NMR.
Soil Sci. 165, 483–494.
Kleber, M., Johnson, M.G., 2010. Advances in understanding the molecular structure of soil
organic matter: implications for interactions in the environment. Adv. Agron.
106, 77–142.
Knicker, H., 2011. Solid state CPMAS 13C and 15N NMR spectroscopy in organic geo-
chemistry and how spin dynamics can either aggravate or improve spectra interpretation.
Org. Geochem. 42, 867–890.
Knicker, H., Almendros, G., Gonzales-Vila, F.J., Martin, F., Ludemann, H.D., 1996. 13C
and 15N NMR spectroscopic examination of the transformation of organic nitrogen
in plant biomass during thermal treatment. Soil Biol. Biochem. 28, 1053–1061.
Knicker, H., Gonzales-Vila, F.J., Polvillo, O., Gonzales, A., Almendros, G., 2005a. Fire-
induced transformation of C- and N-forms in different organic soil fractions from a Dys-
tric Cambisol under a Mediterranean pine forest (Pinus pinaster). Soil Biol. Biochem.
37, 701–718.
Knicker, H., Totsche, K.U., Almendros, G., Gonzalez-Vila, F.J., 2005b. Condensation
degree of burnt peat and plant residues and the reliability of solid-state VACP MAS
13
C NMR spectra obtained from pyrogenic humic material. Org. Geochem.
36, 1359–1370.
K€ogel-Knabner, I., 2002. The macromolecular organic composition of plant and microbial
residues as inputs to soil organic matter. Soil Biol. Biochem. 34, 139–162.
Kolattukudy, P.E., 1980. Biopolyester membranes of plants: cutin and suberin. Science
208, 990–1000.
Kolattukudy, P.E., 1984. Biochemistry and function of cutin and suberin. Can. J. Bot.
62, 2918–2933.
Kolattukudy, P.E., 2001. Polyesters in higher plants. In: Babel, W., Steinb€ uchel, A. (Eds.),
Advances in Biochemical Engineering/Biotechnology. In: Biopolyesters, vol. 71.
Springer, Berlin, pp. 1–49.
Kolattukudy, P.E., Espelie, K.E., 1989. Chemistry, biochemistry, and function of suberin
and associated waxes. In: Rowe, J. (Ed.), Natural Products of Woody Plants, Chemical
Extraneous to the Lignocellulosic Cell Wall. Springer, Berlin, pp. 304–367.
Konno, K., 2011. Plant latex and other exudates as plant defense systems: roles of various
defense chemicals and proteins contained therein. Phytochemistry 72, 1510–1530.
Kononova, M.M., 1961. Soil Organic Matter: Its Nature, Its Role in Soil Formation and in
Soil Fertility. Pergamon Press, Oxford.
Kononova, M.M., 1966. Soil Organic Matter: Its Nature, Its Role in Soil Formation and in
Soil Fertility, second English ed. Pergamon Press, Oxford.
Kononova, M.M., 1975. Humus of virgin and cultivated soils. In: Gieseking, J.E. (Ed.),
In: Soil Components, vol. 1. Springer-Verlag, Berlin, New York, pp. 475–526.
Kramer, R.W., Kujawinski, E.B., Hatcher, P.G., 2004. Identification of black carbon
derived structures in a volcanic ash soil humic acid by Fourier transform ion cyclotron
resonance mass spectrometry. Environ. Sci. Technol. 38, 3387–3395.
Kr€oger, N., Bergsdorf, C., Sumper, M., 1994. A new calcium binding glycoprotein family
constitutes a major diatom cell wall component. EMBO J. 13, 4676–4683.
Kunst, L., Samuels, A.L., Jetter, R., 2005. The plant cuticle: formation and structure of epi-
dermal surfaces. In: Murphy, D.J. (Ed.), Plant Lipids: Biology, Utilisation and Manipu-
lation. Blackwell Publishing, Cornwall, pp. 270–302.
Lam, B., Baer, A., Alaee, M., Lefebvre, B., Moser, A., Williams, A.J., Simpson, A.J., 2007.
Major structural components in freshwater dissolved organic matter. Environ. Sci.
Technol. 41, 8240–8247.
Largeau, C., 1995. Formation of refractory organic matter from biological precursors. In: -
Zepp, R.G., Sonntag, C.H. (Eds.), Role of Nonliving Organic Matter in the Earth’s
Carbon Cycle. John Wiley & Sons, Chichester, pp. 275–292.
Largeau, C., Casadevall, E., Kadouri, A., Metzger, P., 1984. Formation of a Botryococcus-
derived kerogens. Comparative study of immature Torbanite and the extent alga
Botryococcus braunii. Org. Geochem. 6, 327–332.
Largeau, C., Derenne, S., Casadevall, E., Kadouri, A., Sellier, N., 1986. Pyrolysis of immature
Torbanite and of the resistant biopolymer (PRB A) isolated from extant Botryococcus braunii.
Mechanism of formation and structure of torbanite. Org. Geochem. 10, 1023–1032.
Laws, D.D., Bitter, H.M.L., Jerschow, A., 2002. Methoden der festk€ orper NMR
spektroskopie in der chemie. Angew. Chem. 114 (17), 3224–3259.
Leschine, S.B., 1995. Cellulose degradation in anaerobic environments. Annu. Rev.
Microbiol. 49, 399–426.
Liu, C., Huang, P.M., 2002. Role of hydroxyl-aluminosilicate ions (proto-imogolite sol) in
the formation of humic substances. Org. Geochem. 33, 295–305.
Lopes, M.H., Gil, A.M., Silvestre, A.J., Pascoal Neto, C., 2000. Composition of suberin
extracted upon gradual alkaline methanolysis of Quercus suber cork. J. Agric. Food Chem.
48, 383–391.
Lorenz, K., Preston, C.M., 2002. Characterization of high-tannin fractions from humus by
carbon-13 cross-polarization and magic-angle spinning nuclear magnetic resonance.
J. Environ. Q. 31, 431–436.
Lorenz, K., Preston, C.M., Raspe, S., Morrison, I.K., Feger, K.H., 2000. Litter decompo-
sition and humus characteristics in Canadian and German spruce ecosystems: information
from tannin analysis and 13C CPMAS NMR. Soil Biol. Biochem. 32, 779–792.
Lu, X.Q., Hanna, J.V., Johnson, W.D., 2000. Source indicators of humic substances: an ele-
mental composition, solid state 13C CP/MAS NMR and Py-GC/MS study. Appl. Geo-
chem. 15, 1019–1033.
Maas, W.E., Laukien, F.H., Cory, D.G., 1996. Gradient, high resolution, magic angle sample
spinning NMR. J. Am. Chem. Soc. 118, 13085–13086.
Mackie, W., Preston, R.D., 1974. Cell wall and intercellular region polysaccharides. In:
Stewart, W.D.P. (Ed.), Algal Physiology and Biochemistry. Blackwell Scientific
Publishing, Oxford, pp. 40–85.
Maillard, L.C., 1912. Action des acides amines sur les sucre; formation des melanoidins par
voie methodique. C. R. Acad. Sci. Paris 15, 66–68.
Maillard, L.C., 1916. Synthese des matieres humiques par action des acides aminessur les
sucres reductours. Ann. Chim. 5, 258–317.
Maillard, L.C., 1917. Identite des materies humiques de syntheses avec les materies humiques
naturelles. Ann. Chim. 7, 113–152.
Mansfield, S.D., 2009. Solutions for dissolution-engineering cell walls for deconstruction.
Curr. Opin. Biotechnol. 20 (3), 286–294.
Mao, J.D., Hu, W.G., Schmidt-Rohr, K., Davies, G., Ghabbour, E.A., Xing, B., 2000.
Quantitative characterization of humic substances by solid-state carbon-13 nuclear mag-
netic resonance. Soil Sci. Soc. Am. J. Soil 64, 873–884.
Maroto-Valer, M.M., Andresen, J.M., Rocha, J.D., Snape, C.E., 1996. Quantitative solid-
state 13C NMR measurements on cokes, chars and coal tar pitch fractions. Fuel
75, 1721–1726.
Maroto-Valer, M.M., Atkinson, C.J., Willmers, R.R., Snape, C.E., 1998. Characterization
of partially carbonized coals by solid-state 13C NMR and optical microscopy. Energy
Fuels 12 (5), 833–842.
Martin, D., Hauthal, H.G., 1975. Dimethyl Sulphoxide. Van-Nostrand-Reinhold, New York
(E. S. Halberstadt, Trans.).
Martin, J.P., Richards, S.J., Haider, K., 1967. Properties and decomposition and binding
action in soil “humic acid” synthesized by Epicoccum nigrum. Soil Sci. Soc. Am. Proc.
31, 657–662.
Masiello, C.A., 2004. New directions in black carbon organic geochemistry. Mar. Chem.
92, 201–213.
Masoom, H., Courtier-Murias, D., Farooq, H., Soong, R., Simpson, M.J., Maas, W.,
Kumar, R., Monette, M., Stronks, H., Simpson, A.J., 2013. Rapid estimation of nuclear
magnetic resonance experiment time in low-concentration environmental samples.
Environ. Toxicol. Chem. 32 (1), 129–136.
McCandless, E.L., 1981. Polysaccharides of the seaweeds. In: Lobban, C.S., Wynne, M.J.
(Eds.), The Biology of the Seaweeds, Botanical Monographs 17. Blackwell Scientific
Publishing, Oxford, pp. 559–588.
McKinney, D.E., Bortiatynski, J.M., Carson, D.M., Clifford, D.J., de Leeuw, J.W.,
Hatcher, P.G., 1996. Tetramethylammonium hydroxide (TMAH) thermochemolysis
of the aliphatic biopolymer cutan: insights into the chemical structure. Org. Geochem.
24, 641–650.
McMillan, J.D., 1993. Pretreatment of lignocellulosic biomass. In: Himmel, M.E.,
Baker, J.O., Overend, R.P. (Eds.), Enzymatic Conversion of Biomass for Fuel Produc-
tion. American Chemical Society, Washington, DC, pp. 292–323.
Moire, L., Schmutz, A., Buchala, A., Yan, B., Stark, R.E., Ryser, U., 1999. Glycerol is a
suberin monomer. New experimental evidence for an old hypothesis. Plant Physiol.
119, 1137–1146.
Molina, I., Bonaventure, G., Ohlrogge, J., Pollard, M., 2006. The lipid polyester composi-
tion of Arabidopsis thaliana and Brassica napus seeds. Phys. Chem. Chem. Phys.
67, 2597–2610.
M€uller, P., 1887. Studien űber die natűrlichen humusformen und deren einwirkung auf veg-
etation und boden. Springer, Berlin.
Mylotte, R., Verheyen, T.V., Reynolds, A., Dalton, C., Patti, A.F., Chang, R.R.,
Burdon, J., Hayes, M.H.B., 2015. Isolation and characterisation of recalcitrant organic
components from an estuarine sediment core. J. Soils Sediments 15, 211–224.
Mylotte, R., Sutrisno, A., Farooq, H., Masoom, H., Soong, R., Hayes, M.H.B.,
Simpson, A.J., 2016. Insights into the composition of recalcitrant organic matter from
estuarine sediments using NMR spectroscopy. Org. Geochem. 98, 155–165.
Nawrath, C., 2002. The biopolymers cutin and suberin. In: Somerville, C.R.,
Meyerowitz, E.M. (Eds.), The Arabidopsis Book. American Society of Plant
Biologists, Rockville, MD, pp. 1–14.
Nguyen, R.T., Rodger Harvey, H., Zang, X., van Heemst, J.D.H., Hetenyi, M.,
Hatcher, P.G., 2003. Preservation of algaenan and proteinaceous material during the
oxic decay of Botryococcus braunii as revealed by pyrolysis-gas chromatography/mass spec-
trometry and 13C NMR spectroscopy. Org. Geochem. 34, 483–497.
Nierop, K.G.J., 1998. Origin of aliphatic compounds in a forest soil. Org. Geochem.
29, 1009–1014.
Nip, M., Tegelaar, E.W., de Leeuw, J.W., Schenk, P.A., Holloway, P.J., 1986a. A new non-
saponifiable highly aliphatic and resistant biopolymer in plant cuticles. Evidence from
pyrolysis and 13C NMR analysis of present-day and fossil plants. Naturwissenschaften
73, 579–585.
Nip, M., Tegelaar, E.W., Brinkhuis, H., de Leeuw, J.W., Schenk, P.A., Holloway, P.J.,
1986b. Analysis of modern and fossil plant cuticles by Curie point Py-GC and Curie
point Py-GC-MS: recognition of a new, highly aliphatic and resistant biopolymer.
Org. Geochem. 10, 769–778.
Novotny, E.H., Hayes, M.H.B., deAzevedo, E.R., Bonagamba, T.J., 2006. Characterisation
of black carbon-rich samples by 13C solid state nuclear magnetic resonance.
Naturwissenschaften 93, 447–450.
Novotny, E.H., deAzevedo, E.R., Bonagamba, T.J., Cunha, T.J.F., Madari, B.E.,
Benites, V.M., Hayes, M.H.B., 2007. Studies of the compositions of humic acids from
Amazonian dark earth soils. Environ. Sci. Technol. 41, 400–405.
O’Sullivan, A.C., 1997. Cellulose: the structure slowly unravels. Cellulose 4 (3), 173–207.
Ogawa, H., Tanoue, E., 2003. Dissolved organic matter in oceanic waters. J. Oceanogr.
59, 129–147.
Oh-Ishi, M., Maeda, T., 2002. Separation techniques for high-molecular-mass proteins.
J. Chromatogr. B 771, 49–66.
Okuda, S., Tokuda, H., 2011. Lipoprotein sorting in bacteria. Ann. Rev. Microbiol.
65, 239–259.
Oro, J., Nooner, D.W., Zlatkis, Z., Wikstrom, S.A., Barghoorn, E.S., 1965. Hydrocarbons
of biological origin in sediment about 2 billion years old. Science 148, 77–79.
Pacchiano Jr., R.A., Sohn, W., Chlanda, V.L., Garbow, J.R., Stark, R.E., 1993. Isolation
and spectral characterization of plant-cuticle polyesters. J. Agric. Food Chem.
41, 78–83.
Pascoal Neto, C., Cordeiro, N., Seca, A., Domingues, F., Gandini, A., Robert, D., 1996.
Isolation and characterization of a lignin polymer of the cork of Quercus suber L.
Holzforschung 50 (6), 563–568.
Pavia, D.L., Lampman, G.M., Kriz, G.S., Vyvyan, J.R., 2009. Introduction to Spectroscopy,
fourth ed. Brooks/Cole, USA.
Peberdy, J.F., 1990. Fungal cell walls—a review. In: Kuhn, P.J., Trinci, A.P.J., Jung, M.J.,
Goosey, M.W., Copping, L.G. (Eds.), Biochemistry of Cell Walls and Membranes in
Fungi. Springer, Berlin, pp. 5–30.
Perez, J., Munoz-Dorado, J., de la Rubin, T., Martinez, J., 2002. Biodegradation and biological
treatments of cellulose, hemicelluloses and lignin: an overview. Int. Microbiol. 5, 53–63.
Pettersen, R.C., 1984. The chemical composition of wood. In: Wood, R.M. (Ed.), The
Chemistry of Solid Wood. American Chemical Society, Washington, pp. 57–126.
Peuravuori, J., Lepane, V., Lehtonen, T., Pihlaja, K., 2004. Comparative study for separation
of aquatic humic substances by capillary zone electrophoresis using uncoated, polymer
coated and gel-filled capillaries. J. Chromatogr. A 1023, 129–142.
Piccolo, A., 2002. The supramolecular structure of humic substances: a novel understanding
of humus chemistry and implications in soil science. Adv. Agron. 75, 57–134.
Piccolo, A., Conte, P., 1998. Advances in nuclear magnetic resonance and infrared spectros-
copies of soil organic particles. In: Huang, P.M., Senesi, N., Buffle, J. (Eds.), Structure
and Surface Reactions of Soil Particles. John Wiley & Sons, New York, pp. 183–250.
Popov, A.I., Hallenga, K., 1991. Modern NMR Techniques and Their Application in
Chemistry. Marcel Dekker, New York.
Preston, C.M., 1996. Application of NMR to soil organic matter analysis: history and pros-
pects. Soil Sci. 161, 144–166.
Preston, C.M., 2001. Carbon-13 solid-state NMR of soil organic matter—using the tech-
nique effectively. Can. J. Soil Sci. 81, 255–270.
Preston, C.M., Newman, R.H., 1992. Demonstration of spatial heterogeneity in the organic
matter of de-ashed humin samples by solid-state 13C CPMAS NMR. Can. J. Soil Sci.
72, 13–19.
Preston, C.M., Newman, R.H., 1995. NMR of de-ashed soil humin in a second-growth
Douglas-fir stand of coastal British Columbia. Geoderma 68, 229–241.
Preston, C.M., Ripmeester, J.A., 1982. Application of solution and solid-state 13CNMR to
four organic soils, their humic acids, fulvic acids, humins, and hydrolysis residues. Can. J.
Spectr. 27, 99–105.
Preston, C.M., Schnitzer, M., Ripmeester, J.A., 1989. A spectroscopic and chemical inves-
tigation on the de-ashing of a humin. Soil Sci. Soc. Am. J. 53, 1442–1447.
Purin, S., Rillig, M.C., 2007. The arbuscular mycorrhizal fungal protein glomalin: limita-
tions, progress, and a new hypothesis for its function. Pedobiologia 51, 123–130.
Ramann, E., 1888. Die Arbeiten von Post űber Schlamm. Moor, Torf, und Humus. Landw.
Jb. 17, 405.
Rice, J.A., 2001. Humin. Soil Sci. 166, 848–857.
Rice, J.A., MacCarthy, P., 1988. Comments on the literature of the humin fraction of
humus. Geoderma 43, 65–73.
Rice, J.A., MacCarthy, P., 1989. Isolation of humin by liquid-liquid partitioning. Sci. Total
Environ. 81/82, 61–69.
Rice, J.A., MacCarthy, P., 1990. A model of humin. Environ. Sci. Technol. 24, 1875–1877.
Rice, J.A., MacCarthy, P., 1992. Disaggregation and characterization of humin. Sci. Total
Environ. 117 (118), 83–88.
Rillig, M.C., 2004. Arbuscular mycorrhizae, glomalin, and soil aggregation. Can. J. Soil Sci.
84, 355–363.
Rillig, M.C., 2005. A connection between fungal hydrophobins and soil water repellency?
Pedobiologia 49, 395–399.
Rillig, M.C., Ramsey, P.W., Morris, S., Paul, E.A., 2003. Glomalin, an arbuscular-mycor-
rhizal fungal soil protein, responds to land-use change. Plant Soil 253, 293–299.
Rocha, S.M., Goodfellow, B.J., Delgadillo, I., Neto, C.P., Gil, A.M., 2001. Enzymatic iso-
lation and structural characterisation of polymeric suberin of cork from Quercus suber L.
Int. J. Biol. Macromol. 28, 107–119.
Russell, E.W., 1950. Soil Conditions and Plant Growth, eighth ed. Longmans, London
(Chapter 1).
Russell, E.W., 1973. Soil Conditions and Plant Growth, tenth ed. Longmans, London
(Chapter 1).
Russell, J.D., Vaughan, D., James, D., Fraser, A.R., 1983. An IR spectroscopic study of soil
humin and its relationship to other soil humic substances and fungal pigments. Geoderma
29, 1–12.
Saiz-Jimenez, C., De Leeuw, J.W., 1986. Chemical characterization of soil organic matter
fractions by analytical pyrolysis-gas chromatography-mass spectrometry. J. Anal. Appl.
Pyrol. 9, 99–119.
Saha, B.C., 2003. Hemicellulose bioconversion. J. Ind. Microbiol. Biotechnol. 30, 279–291.
Samoson, A., Tuherm, T., Past, J., 2001. Ramped-speed cross polarization MAS NMR.
J. Magn. Reson. 149, 264–267.
Sánchez, C., 2009. Lignocellulosic residues: biodegradation and bioconversion by fungi.
Biotechnol. Adv. 27, 185–194.
Saxby, J.D., 1970. Isolation of kerogen in sediments by chemical methods. Chem. Geol.
6, 173–184.
Scheler, U., 2002. Solid polymers. In: Duer, M.J. (Ed.), Solid-State NMR Spectroscopy:
Principles and Applications. Blackwell Science, Oxford, pp. 483–511.
Scheller, H.V., Ulvskov, P., 2010. Hemicelluloses. Plant Biol. 61, 263.
Schmidt, H.W., Sch€ onherr, J., 1982. Development of plant cuticles—occurrence and role of
non-ester bonds in cutin of Clivia miniata Reg. leaves. Planta 156, 380–384.
Schmidt, M.W.I., Knicker, H., Hatcher, P.G., K€ ogel-Knabner, I., 1997. Improvement of
13
C and 15N CPMAS NMR spectra of bulk soils, particle size fractions and organic mate-
rial by treatment with 10% hydrofluoric acid. Eur. J. Soil Sci. 48, 319–328.
Schmidt, M.W.I., Skjemstad, J.O., Gehrt, E., K€ ogel -Knabner, I., 1999. Charred organic
carbon in German chernozemic soils. Eur. J. Soil Sci. 50, 351–365.
Schmitt-Kopplin, P., Hertkorn, N., Schulten, H.-R., Kettrup, A., 1998. Structural changes
in a dissolved soil humic acid during photochemical degradation processes under O2 and
N2 atmosphere. Environ. Sci. Technol. 32, 2531–2541.
Schnitzer, M., 1978. Humic substances: chemistry and reactions. In: Schnitzer, M.,
Khan, S.U. (Eds.), Soil Organic Matter, vol. 8. Elsevier, The Netherlands, pp. 1–64.
Schnitzer, M., 2000. A lifetime perspective in the chemistry of soil organic matter. Adv.
Agron. 68, 1–58.
Schnitzer, M., Schulten, H.R., 1995. Analysis of organic matter in soil extracts and whole
soils by pyrolysis-mass spectrometry. Adv. Agron. 55, 168–217.
Schouten, S., Moerkerken, P., Gelin, F., Baas, M., de Leeuw, J.W., Sinninghe Damste, J.S.,
1998. Structural characterization of aliphatic, non-hydrolyzable biopolymers in freshwa-
ter algae and leaf cuticle by ruthenium tetroxide degradation. Phytochemistry 49 (4),
987–993.
Schreiner, O., Shorey, E.C., 1910. Chemical nature of soil organic matter. USDA Bureau
Soils Bull. 74, 2–48.
Schuffelen, A.C., Bolt, G.H., 1950. Some notes on the synthesis of humic compounds.
Overfruk Uit Het. Landbouwkundig Titzschrift 62 ste Jaargang No. 4/5.
Schurko, R., 2009. Introduction to Solid State NMR. Online, available at: http://mutuslab.
cs.uwindsor.ca/schurko/ssnmr/ (accessed 30/01/2014).
Shields, L.M., Durrell, L.W., 1964. Algae in relation to soil fertility. Bot. Rev. 30,
92–128.
Shirzadi, A., Simpson, M.J., Kumar, R., Baer, A.J., Xu, Y.P., Simpson, A.J., 2008a. Molec-
ular interactions of pesticides at the soil–water interface. Environ. Sci. Technol.
42, 5514–5520.
Shirzadi, A., Simpson, M.J., Xu, Y.P., Simpson, A.J., 2008b. Application of saturation trans-
fer double difference NMR to elucidate the mechanistic interactions of pesticides with
humic acid. Environ. Sci. Technol. 42, 1084–1090.
Shmuk, A.A., 1924. The chemical nature of soil organic matter. Byull. Pochvoveda, 5–7(as
cited by Kononova 1961).
Shmuk, A.A., 1930. Zur Frage vom Chemismus der Organischen Stoffe des Bodens.
Pedology 25, 5–48.
Simpson, A., 2001. Multidimensional solution state NMR of humic substances: a practical
guide and review. Soil Sci. 166, 795–809.
Simpson, A.J., 2002. Determining the molecular weight, aggregation, structures and inter-
actions of natural organic matter using diffusion ordered spectroscopy. Magn. Reson.
Chem. 40, S72–S82.
Simpson, A.J., Simpson, M.J., 2009. Nuclear magnetic resonance analysis of natural organic
matter. In: Senesi, N., Xing, B., Huang, P.M. (Eds.), Biophysio-Chemical Processes
Involving Natural Non-Living Organic Matter in Environmental Systems. John
Wiley & Sons, New Jersey, pp. 589–650.
Simpson, A.J., Kingery, W.L., Spraul, M., Humpfer, E., Dvortsak, P., Kressebaum, R., 2001.
Separation of structural components in soil organic matter by diffusion ordered spectros-
copy. Environ. Sci. Technol. 35, 4421–4425.
Simpson, A.J., Salloum, M.J., Kingery, W.I., Hatcher, P.G., 2002. Improvements in the
two-dimensional nuclear magnetic resonance spectroscopy of humic substances.
J. Environ. Qual. 31, 388–392.
Simpson, A.J., Kingery, W.L., Hatcher, P.G., 2003. The identification of plant derived struc-
tures in humic materials using three-dimensional NMR spectroscopy. Environ. Sci.
Technol. 37, 337–342.
Simpson, A.J., Song, G., Smith, E., Lam, B., Novotny, E.H., Hayes, M.H.B., 2007a.
Unraveling the structural components of soil humin by use of solution-state nuclear mag-
netic resonance spectroscopy. Environ. Sci. Technol. 41, 876–883.
Simpson, A.J., Simpson, M.J., Smith, E., Kelleher, B.P., 2007b. Microbially derived inputs to
soil organic matter: are current estimates too low? Environ. Sci. Technol.
41, 8070–8076.
Simpson, A.J., Song, G., Smith, E., Lam, B., Novotny, E.H., Hayes, M.H.B., 2007c.
Unravelling the structural components of soil humin by use of solution-state nuclear
magnetic resonance spectroscopy. Environ. Sci. Technol. 41, 876–883.
Simpson, A.J., McNally, D.J., Simpson, M.J., 2011. NMR spectroscopy in environmental
research: from molecular interactions to global processes. Prog. Nucl. Magn. Reson.
Spectrosc. 58, 97–175.
Simpson, J.H., 2012. Organic Structure Determination Using 2-D NMR Spectroscopy:
A Problem Based Approach, second ed. Academic Press, USA, pp. 151–153.
Simpson, M.J., Hatcher, P.G., 2004. Overestimations of black carbon in soils and sediments.
Naturwissen 91, 436–440.
Simpson, M.J., Johnson, P.C.E., 2006. Identification of mobile aliphatic sorptive domains in
soil humin by solid-state 13C nuclear magnetic resonance. Environ. Toxicol. Chem.
25, 52–57.
Sitte, P., 1962. Zum feinbau der suberinschichten im flaschenkork. Protoplasma
54, 555–559.
Skjemstad, J.O., Clarke, P., Taylor, J.A., Oades, J.M., Newman, R.H., 1994. The removal of
magnetic materials from surface soils. A solid state 13C CP/MAS NMR study. Aust. J.
Soil. Res. 32, 1215–1229.
Skjemstad, J.O., Reicosky, D.C., Wilts, A.R., McGowan, J.A., 2002. Charcoal carbon in
U.S. agricultural soils. Soil Sci. Soc. Am. J. 66, 1249–1255.
Smernik, R.J., Oades, J.M., 1999. Effects of added paramagnetic ions on the 13C CP/MAS
NMR spectrum of a de-ashed soil. Geoderma 89, 219–248.
Smernik, R.J., Oades, J.M., 2000a. The use of spin counting for determining quantitation in
solid state 13C NMR spectra of natural organic matter: 1. Model systems and the effects of
paramagnetic impurities. Geoderma 96, 101–129.
Smernik, R.J., Oades, J.M., 2000b. The use of spin counting for determining quantitation in
solid state 13C NMR spectra of natural organic matter: 2. HF-treated soil fractions.
Geoderma 96, 159–171.
Song, J.Z., Peng, P.A., Huang, W.L., 2002. Black carbon and kerogen in soils and sediments.
1. Quantification and characterization. Environ. Sci. Technol. 36, 3960–3967.
Song, G., Novotny, E.H., Simpson, A.J., Clapp, C.E., Hayes, M.H.B., 2008. Sequen-
tial exhaustive extraction of a Mollisol soil, and characterizations of humic com-
ponents, including humin, by solid and solution state NMR. Eur. J. Soil Sci.
59, 505–516.
Song, G., Hayes, M.H.B., Novotny, E., Simpson, A.J., 2011. Isolation and fractionation of
soil humin using alkaline urea and dimethylsulphoxide plus sulfuric acid.
Naturwissenschaften 98, 7–13.
Spaccini, R., Piccolo, A., Haberauer, G., Gerzabek, M.H., 2000. Transformation of
organic matter from maize residues into labile and humic fractions of three European
soils as revealed by 13C distribution and CPMAS-NMR spectra. Eur. J. Soil Sci.
51, 583–594.
€
Sprengel, C., 1826. Uber pflanenzhumus, humuss€aure and humuss€aure slaze. Kastne’s Arch.
Ges. Naturlehre 8, 145–229.
Stejskal, E.O., Memory, J.D., 1994. High Resolution in the Solid State. Fundamentals of
CP/MAS. Oxford University Press, New York.
Stejskal, E.O., Tanner, J.E., 1965. Spin diffusion measurements: spin echoes in the presence
of a time-dependent field gradient. J. Chem. Phys. 42 (1), 288.
Stevenson, F.J., 1982. Humus Chemistry: Genesis, Composition, Reactions. Wiley,
New York.
Stevenson, F.J., 1994. Humus Chemistry: Genesis, Composition, Reactions, second ed.
Wiley, New York.
Stockmann, U., Adams, M., Crawford, J., Field, D., et al., 2013. The knowns, known
unknowns and unknowns of sequestration of soil organic carbon. Agric. Ecosyst. Envi-
ron. 162, 80–99.
Swift, R.S., 1996. Organic matter characterization. In: Sparks, D.L. (Ed.), Methods of Soil
Analysis. Part 3. Chemical Methods. In: Soil Science Society of America Book Series,
vol. 5. Soil Science Society of America, Madison, WI, pp. 1011–1070.
Teeri, T.T., 1997. Crystalline cellulose degradation: new insight into the function of
cellobiohydrolases. Trends Biotechnol. 15 (5), 160–167.
Tegelaar, E.W., De Leeuw, J.W., Derenne, S., Largeau, C., 1989a. A reappraisal of kerogen
formation. Geochim. Cosmochim. Acta 53, 3103–3106.
Tegelaar, E.W., de Leeuw, J.W., Largeau, C., Derenne, S., Schulten, H.-R., Muller, R.,
Boon, J.J., Nip, M., Sprenkels, J.C.M., 1989b. Scope and limitations of several pyrolysis
methods in the structural elucidation of a macromolecular plant constituent in the leaf
cuticle of Agave americana L. J. Anal. Appl. Pyrol. 15, 29–54.
Tegelaar, E.W., de Leeuw, J.W., Saiz-Jimenez, C., 1989c. Possible origin of aliphatic moi-
eties in humic substances. Sci. Total Environ. 81/82, 1–17.
Tegelaar, E.W., Kerp, H., Visscher, H., Schenck, P.A., de Leeuw, J.W., 1991. Bias of the
paleobotanical record as a consequence of variations in the chemical composition of
higher vascular plant cuticles. Paleobiology 17, 133–144.
Tegelaar, E.W., Hollman, G., van der Vegt, P., de Leeuw, J.W., Holloway, P.J., 1995.
Chemical characterization of periderm tissue of some angiosperm species: recognition
of an insoluble, non-hydrolyzable, aliphatic biomacromolecule (Suberan). Org. Geo-
chem. 23, 239–250.
Theng, B.K.G., 1979. Formation and Properties of Clay-Polymer Complexes. Elsevier,
Amsterdam.
Timell, T.E., Syracuse, N.Y., 1967. Recent progress in the chemistry of wood hemicellu-
loses. Wood Sci. Technol. 1, 45–70.
Trusov, A.G., 1914. The humification of compounds which are constituents of plants. Sel.
Khoz. Lesovod. Oct., Nov. As cited by Kononova (1966).
Trusov, A.G., 1915. The humification of compounds which are constituents of plants. Sel.
Khoz. Lesovod. April, July, Nov. As cited by Kononova (1966).
Trusov, A.G., 1916a. The humification of compounds which are constituents of plants. Sel.
Khoz. Lesovod. March, Sept., Oct., Nov. As cited by Kononova (1966).
Trusov, A.G., 1916b. Some chemical-biological processes occurring during the humification
of plant residues. Zh. Opyt. Agron. 17. As cited by Kononova (1966).
Tsutsuki, K., Kuwatsuka, S., 1992. Characterization of humin-metal complexes in a buried
volcanic ash soil profile and a peat soil. Soil Sci Plant Nutr. 38, 297–306.
Turner, J.W., Hartman, B.E., Hatcher, P.G., 2013. Structural characterisation of suberan iso-
lated from river birch (Betula nigra) bark. Org. Geochem. 57, 41–53.
Tyurin, I.V., Gutkina, E.L., 1940. Data of studies on the nature of “humins” of chernozem.
Trudy pochv. Inst. Docuchaeva 23, 41.
van Bergen, P.F., Scott, A.C., Barrie, P.J., de Leeuw, J.W., Collinson, M.E., 1994. The
chemical composition of upper carboniferous pteridosperm cuticles. Org. Geochem.
21, 107–112.
von Philipsborn, W., M€ uller, R., 1986. 15N NMR spectroscopy—new methods and appli-
cations. Angew. Chem. Int. Ed. 25, 383–413.
von Post, H., 1862. Studien uber die koprogenen erdbildungen unser tage. Landw. Jb.
17, 405. Cited by Ramann (1888).
Waksman, S.A., 1936. Humus, Origin, Chemical Composition, and Importance in Nature.
Tindall and Cox, London.
Waksman, S.A., Iyer, K.R.N., 1932. Contribution to our knowledge of the chemical nature
and origin of humus: I. On the synthesis of the “humus nucleus”. Soil Sci. 34, 43–69.
Waksman, S.A., Iyer, K.R.N., 1933. Contribution to our knowledge of the chemical nature
and origin of humus: IV. Fixation of proteins by lignins and formation of complexes
resistant to microbial decomposition. Soil Sci. 36, 69–82.
Wallerius, J.G., 1761. Agriculturae fundamenta chemical spz. De homo Diss, Uppsala.
Walton, T.J., Kolattukudy, P.E., 1972. Demonstration of the structures of cutin monomers
by a novel depolymerisation procedure and combined gas-chromatography and mass
spectrometry. Biochemistry 11, 1885–1897.
Williams, V.R., 1939. Soil Science and the Basis of Agriculture. Sel’Khozgiz (Pochvovedenie
s osnovami zemledeliya). As cited by Kononova (1966).
Williamson, M.P., 2006. The nuclear Overhauser effect. In: Webb, G.A. (Ed.), Modern
Magnetic Resonance. Springer, Netherlands, pp. 409–412.
Wilson, M.A., 1987. NMR Techniques and Applications in Geochemistry and Soil Chem-
istry. Pergamon Press, Oxford.
Woods, W.I., Teixeira, W.G., Lehmann, J., Steiner, C., WinklerPrins, A., Rebellato, L.,
2009. Amazonian Dark Earths: Wim Sombroek’s Vision. Springer, Berlin.
Wright, S.F., Upadhyaya, A., 1996. Extraction of an abundant and unusual protein from soil
and comparison with Hyphal protein of Arbuscular Mycorrhizal fungi. Soil Sci.
161, 575–586.
Wright, S.F., Upadhyaya, A., 1998. A survey of soils for aggregate stability and glomalin, a
glycoprotein produced by hyphae of arbuscular mycorrhizal fungi. Plant Soil
198, 97–107.
Xing, B., Mao, J., Hu, W.-G., Schmidt-Rohr, K., Davies, G., Ghabbour, E.A., 1999. Eval-
uation of different solid-state 13C NMR techniques for characterising humic acids. In:
Davies, G., Ghabbour, E.A. (Eds.), Understanding Humic Substances: Advanced
Methods, Properties and Applications. The Royal Society of Chemistry, Cambridge,
pp. 49–61.
Yan, B., Stark, R.E., 2000. Biosynthesis, molecular structure, and domain architecture of
potato suberin: a C-13 NMR study using isotopically labelled precursors. J. Agric. Food
Chem. 48 (8), 3298–3304.
Zang, X., Nguyen, R.T., Harvey, H.R., Knicker, H., Hatcher, P.G., 2001. Preservation
of proteinaceous material during the degradation of the green alga Botryococcus braunii:
a solid state 2D 15N 13C NMR spectroscopy study. Geochim. Cosmochim. Acta
65, 3299–3305.
Zhang, D., Lu, S., 1987. An assessment of the separation and analysis of humic substances by
isoelectric focusing (IEF) method. Sci. Total Environ. 62, 89–96.
Zlotnik-Mazori, T., Stark, R., 1988. Nuclear magnetic resonances studies of cutin, an insol-
uble plant polyester. Macromolecules 21, 2412–2417.
CHAPTER THREE
Effects of Drought Stress on

Morphophysiological Traits,
Biochemical Characteristics, Yield,
and Yield Components in Different
Ploidy Wheat: A Meta-Analysis
Jian-Yong Wang*,†, You-Cai Xiong*,1, Feng-Min Li*,
Kadambot H.M. Siddique†, Neil C. Turner†,1
*State Key Laboratory of Grassland Agroecosystems, School of Life Sciences, Institute of Arid Agroecology,
Lanzhou University, Lanzhou, China
†
1
Corresponding authors: e-mail address: xiongyc@lzu.edu.cn; neil.turner@uwa.edu.au
Contents
1. Introduction 140
1.1 Wheat Domestication, Breeding, and Production 140
1.2 Drought and Climate Change 142
1.3 Meta-Analysis 144
2. Methodology 145
2.1 Database Construction 145
2.2 Composition of the Database 147
2.3 Sources of Variation 149
2.4 Statistical Analyses 150
3. Responses to Drought Stress in Different Ploidy Wheat 151
3.1 Yield and Yield Components 151
3.2 Plant Architecture, Biomass Allocation, and Physiological Traits 156
3.3 Root Environment (Pot or Field) and Wheat Type (Spring
or Winter) 160
3.4 Drought Stress at Different Phenological Stages 166
4. Implications and Future Directions 167
Acknowledgments 169
References 169
Abstract
Increases in the yield of wheat, a major cereal crop grown in semiarid and temperate
regions of the world, are often limited by drought. In this chapter, we quantitatively
evaluated the effects of drought stress on the morphophysiological and biochemical

140 Jian-Yong Wang et al.
characteristics, growth and biomass partitioning, and yield formation of diploid (2n), tet-
raploid (4n), and hexaploid (6n) wheat using a meta-analysis of published data. The study
synthesized results from 303 papers published before May 2015, taking into account
wheat ploidy level, rooting environment (pots or field), spring or winter type, and drought
stress at different phenological stages. Drought stress reduced yields more in 2n wheat
than 4n and 6n wheat due to a greater reduction in grain number and seed size; reduced
aboveground biomass more in 2n and 4n than 6n wheat, but reduced root biomass more
in 6n than 2n and 4n wheat; and gas exchange more in 4n wheat than 6n wheat; while
major biochemical parameters increased more in 2n and 4n wheat than in 6n wheat.
Across all ploidy levels, drought stress in the reproductive phase affected grain yield more
than drought stress in the vegetative stage. Wheat grown in pots with limited rooting
capacity had a greater reduction in yield, yield components and biomass, and greater
increases in stress-induced biochemical parameters, than plants grown in the field.
Drought stress reduced yields of spring wheat more than winter wheat. Domestication
and selection of higher ploidy wheat have reduced the adverse effects of drought stress
on yield and yield components, optimized biomass allocation toward higher seed yields,
and reduced stress-related physiological and biochemical responses.
1. INTRODUCTION
1.1 Wheat Domestication, Breeding, and Production
Although cereals have been collected for food for more than 20,000 years
before the present (BP, noncalibrated 14C years), wheat is generally considered
to have been domesticated from wild forms between 11,000 and 9500 years
BP in the West Asia region of southeastern Turkey and northern Syria, the
so-called cradle of agriculture (Lev-Yadun et al., 2000) where wild einkorn
and emmer wheat still grow today. The wild diploid (2n, 2x ¼ 2n ¼ 14, where
2x is the number of chromosomes in each somatic cell and n is the basic chro-
mosome number) einkorn wheat (Triticum boeoticum Boiss. and Triticum urartu
Tumanian ex Gandilyan) is the A-genome donor of hexaploid wheat, and
wild tetraploid (4n, 2x ¼ 4n¼ 28) emmer wheat [Triticum turgidum ssp.
dicoccoides (K€
orn) Aschers and Graebner Thell.] is considered the result of a
hybridization event several thousand years ago between T. urartu and wild
goat grass that is closely related with Aegilops speltoides Taush. (2n,
2x ¼ 2n ¼ 14) (IWGSC, 2014; Petersen et al., 2006). The domestication of
wild emmer wheat gave rise to domesticated T. turgidum L. ssp. dicoccum
(Schrank ex Sch€ ubl.) Thell. and the free-threshing T. turgidum L. spp. durum
(Desf.) Husn. (or T. durum Desf.) grown mainly in Mediterranean climatic
regions for pasta and semolina products, and still subject to further breeding
and selection. A second hybridization event (under domestication) between
Drought Stress in Wheat: A Meta-Analysis 141
T. turgidum and Aegilops tauschii Coss. gave rise to the hexaploid (6n,
2x¼ 6n ¼ 42) bread wheat T. aestivum L. that has subsequently been subject
to considerable breeding and selection. The polyploid genome of T. aestivum
comprises three subgenomes, AA from T. urartu, BB from A. speltoides, and
DD from A. tauschii (Fig. 1). The draft gene sequence of T. aestivum cv. Chi-
nese Spring has recently been published (IWGSC, 2014). Domestication
involved the selection for (i) nonshattering, to reduce losses from shattering
of the spike by wind and during harvesting, and (ii) naked grains, that is
the glumes surrounding the grain are not tightly held against the grain (hulled
grain), but are easily removed by threshing (Dubcovsky and Dvorak, 2007).
In the 1940s, studies revealed that A. tauschii carries the D-genome of
hexaploid wheat (Kihara and Lilienfeld, 1949; McFadden and Sears,
1946); these authors synthesized many 6n-amphidiploid wheat between
T. dicoccoides and 2n-species of Aegilops, using colchicine (Kihara and
Lilienfeld, 1949). The so-called synthetic 6n wheat has since been used to
genetically improve hexaploid wheat worldwide. Through interspecific
hybridization of the original donors of the wheat genome, the production
Fig. 1 Schematic diagram of wheat evolution and domestication. Developed with infor-
mation from Dong, Y.C., Zheng, D.S. (Eds.), 1999. China wheat genetic resources. China
Agriculture Press, Beijing, pp. 58–112 (in Chinese) and IWGSC (International Wheat Genome
Sequencing Consortium), 2014. A chromosome-based draft sequence of the hexaploid
bread wheat (Triticum aestivum) genome. Science 345, 1251788.
of synthetic bread wheats is one of the strategies employed at the Interna-

tional Maize and Wheat Improvement Center (CIMMYT) to solve the lack
the genetic diversity (Dreccer et al., 2007). Since the early 1990s, CIMMYT
has produced more than 1100 synthetic hexaploid wheat accessions
(CIMMYT, 2016), thereby increasing the source of genetic diversity,
especially of the D-genome, for wheat breeding (Tang et al., 2015). Syn-
thetic wheats have been systematically introduced into Australia from
Mexico in the last two decades (Dreccer et al., 2007) and evaluated for their
potential to improve the adaptation of local hexaploid wheat cultivars, par-
ticularly for drought and heat stress tolerance (Ogbonnaya et al., 2007;
Pradhan et al., 2012).
Wheat is among the most widely cultivated crops in the world, currently
grown on more than 215 million hectares annually, producing more than
725 million metric tons of grain in 2014 (FAOSTAT, 2015) and serving
as a staple food for 30% of the world’s population. Hexaploid (6n) wheat
accounts for about 95% of global wheat production, while tetraploid (4n)
wheat accounts for the other 5% (IWGSC, 2014). Because of the widespread
improvements in 4n and 6n wheat, the cultivation of 2n wheat is almost
nonexistent. The increase in ploidy of 4n and 6n wheat and subsequent
breeding and selection has resulted in increased yields, increased quality,
and better adaptation to a wide range of environments. For the increasing
global population to enjoy food security, yields need to increase further
in response to the growing demand. According to CIMMYT (2016), con-
sumers will require 60% more wheat by 2050 than today. Sustaining a stable
supply of wheat in the future is going to be challenging as arable land and
water resources for agriculture decrease, while temperatures rise and
drought events become more frequent and intense (McKersie, 2015;
Turner and Meyer, 2011).
1.2 Drought and Climate Change

Drought is one of the most important factors reducing crop yields, and affect-
ing growth and development (Araus et al., 2008; Farooq et al., 2014; Turner
et al., 2014). When water resources are limited, the life-cycle shortens, which
reduces the size of organs such as leaves, tillers, spikes, the number of spikelets,
and the ratio of spike dry weight to total dry weight in cereals, and alters phys-
iological and biochemical processes resulting in low yields (Lascano et al.,
2001). Wheat is grown from temperate to tropical environments and, in many
regions, experiences water shortage during some part of the growing season,
particularly in semiarid and subtropical areas (Moustafa et al., 1996). Drought

may affect wheat growth at all phenological stages, but not all growth stages
are equally sensitive to water stress (Farooq et al., 2014). Based on the scale of
growth stages in cereals designed by Feekes, the growth of wheat can be
divided into three stages: preanthesis (vegetative period, from emergence to
Feekes 10.5), anthesis (from Feekes 10.5 to Feekes 10.53), and postanthesis
(reproductive period, from Feekes 10.53 to Feekes 11.4) (Zadoks et al.,
1974). Drought during early growth reduces germination and emergence,
and decreases tiller number, plant height, and leaf area, while at anthesis
and during grain filling, drought reduces the percentage of fertile spikelets
and number of grains (Farooq et al., 2014), possibly as a result of reduced pol-
lination (Nawaz et al., 2012). During grain filling, water stress also affects the
accumulation of starch and protein in the kernels, thereby reducing grain size
and affecting grain yield and quality (Ahmadi and Baker, 2001; Wardlaw and
Willenbrink, 2000).
It is clear that plant survival and growth performance in water-limited
environments depend on a range of adaptive mechanisms (Begg and
Turner, 1976; Turner, 1979, 1986, 1997), such as phenological plasticity, sto-
matal control of water loss, water uptake from the soil, and osmotic adjust-
ment by the active accumulation of sugars, ions, free amino acids, and
proline (Loss and Siddique, 1994; Sultan, 1995; Turner and Jones, 1980).
The pattern of biomass allocation is an important physiological indicator of
an adaptive strategy in plants, especially in resource-limited environments.
It is closely linked with the size and morphological construction of crop organs
(Siddique et al., 1989). Over the long period of selection and breeding of
wheat by farmers and plant breeders, complex morphophysiological and
biochemical mechanisms and biomass allocation strategies have been incor-
porated to cope with intermittent and terminal water shortage (Turner,
1986, 1997).
In semiarid and subtropical regions, climate change is widely predicted to
increase the frequency and intensity of drought, accompanied by higher
temperatures and higher CO2 concentration (Stocker et al., 2013). The
increased concentration of CO2 in the atmosphere will increase growth
in C3 species such as wheat, but there is general consensus that crop yields
will decrease as a result of more rapid crop development and reduced crop
fecundity at higher temperatures, and an increased incidence of water stress
with reduced rainfall (Asseng et al., 2011; Turner and Meyer, 2011; Yadav
et al., 2011). While acclimation to high temperatures and adaptation to
water shortage may have less impact than the 50% reduction in yields
predicted in many semiarid and subtropical areas, an analysis of the impact of

drought on wheat yields and the factors affecting on yield is clearly
warranted if food security is to be guaranteed. In this chapter, we report
the results of a meta-analysis on the impact of drought on wheat yields,
and the factors that influence yields not only in 6n wheat but also in 2n
and 4n wheat to determine whether wheat with lower levels of ploidy have
traits that will benefit the yield of 6n wheat when exposed to drought.
1.3 Meta-Analysis
A meta-analysis is a quantitative approach that combines and compares the
results from numerous studies to summarize the range of projected outcomes
in the literature and assess consensus (Gurevitch and Hedges, 1999). One
advantage of meta-analysis over traditional literature surveys is the ability
to report the effect of sample size and provide confidence intervals, discrim-
inate statistically significant subsets of data, and compare variation within and
among categories of studies (Ainsworth et al., 2002; Curtis and Wang,
1998). Meta-analysis was initially used to bring together large data sets gen-
erated by medical studies worldwide; since the 1990s, it has been developed
as a methodology in the fields of ecology, agriculture, and biology (Feng
et al., 2009). Meta-analysis has been used to examine the responses of dif-
ferent crop species to biotic and/or abiotic stresses, for example, studies
on the effects of elevated CO2 or O3 in soybean (Ainsworth et al., 2002)
and wheat (Feng et al., 2008; Wang et al., 2013), heat stress on ecosystem
nitrogen pools (Bai et al., 2013), nitrogen fertilizer application on plant root
traits (Li et al., 2015), arbuscular mycorrhizal fungi on wheat and other plants
(Pellegrino et al., 2015), drought stress on food legumes (Daryanto et al.,
2015), and humic substances, limited water, and phosphorus supply on
plant growth (Rose et al., 2014; Suriyagoda et al., 2014). Meta-analysis
has also been used to assess changes in soil organic carbon with the cultiva-
tion of arable land (Zhao et al., 2015). The flood of experimental results in
the literature and the increasing concern of global-scale issues have made
meta-analysis a necessary tool for modern scientific research (Osenberg
et al., 1999). The recent development of the response ratio (defined as
the treatment mean effect divided by the control mean effect) of
meta-analysis has provided a meaningful metric for reviewing data on plant
responses to stress (Gurevitch and Hedges, 1999), as the response to a stress
treatment relative to a control treatment allows comparisons at a range of
sites that have different responses among the control treatments.
As one of the three major cereal crops (following maize and rice), wheat
has been widely studied, and the changes in physiological parameters and
yield as a result of drought stress have been extensively reported in 6n wheat
genotypes, and to a lesser extent in 4n genotypes. However, the large data-
base of studies in 6n wheat on yield formation, biomass partitioning, and
physiological and agronomic performance under drought stress has not been
analyzed and compared, and certainly not compared with 2n and 4n wheat.
To summarize and synthesize the results of the numerous studies on phys-
iological and agronomic traits, and yield formation in wheat, we conducted a
comprehensive meta-analysis to: (1) quantitatively evaluate the effect of
drought stress on grain yield, yield components, and morphophysiological
and biochemical traits; (2) determine how dry matter partitioning changes
under drought stress; (3) evaluate the effects of drought stress in different
rooting environments (pots or field), wheat types (spring or winter), and
at various growth stages (vegetative, anthesis, and grain filling); and (4) eval-
uate the mechanisms of adaptation to drought in different ploidy wheat.
2. METHODOLOGY
2.1 Database Construction
A database was built of drought stress on yield and its components, mor-
phophysiological, biochemical, and agronomic traits among different ploidy
wheat by surveying peer-reviewed literature within the Web of Science
(ISI, USA, http://apps.webofknowledge.com/) and Google Scholar
(http://scholar.google.com.au). The keywords used for the search were:
drought stress OR water deficit OR water stress AND wheat, to identify
articles that reported the targeted research content (Fig. 2). The year of pub-
lication was not restricted, but publications after May, 2015 were not
included. The articles were screened on the basis of the following criteria:
(1) studies had to be published in English, (2) studies had to include pair-wise
control and experimental treatments, (3) data had to have at least one mor-
phophysiological, biochemical, gas exchange, yield or yield component
parameter, and (4) data had to include means (M), sample size (N), and a
measure of variance [standard deviation (SD), standard error (SE), or coef-
ficient of variation (CV)] for both control and treatment groups. Studies that
were excluded were those in which: (1) the data had median (not mean)
results or statistical differences between control and treatment groups as F
values, P values, or least significant difference values instead of SD or SE
values, (2) the sample size for treatment and/or control groups was not clear,
Using the Web of Science, using ‘drought * water deficit

* water stress’ AND ‘wheat’ as keywords, N > 7000 articles
were identified
Step 1: Screened these articles based on

title/abstract/keywords
Total N = 963 articles were downloaded
Step 2: Fast read full-text articles, N = 627

were excluded because they did not fit the
criteria of this study
N = 336 articles were eligible
Step 3: Carefully read full-text articles, N = 33

were excluded because they had the same
authors/same experiments while published in
different Journals
Articles used in meta-analysis (N = 303)
Data extraction, data analysis, and figures drawn
Prepare for publication
Fig. 2 The flowchart of the process of building the database and meta-analysis.
(3) the period of drought stress was less than 48 h, and (4) the data were pre-
viously or more completely reported in another article (to avoid data dupli-
cation). Based on these criteria, the meta-analysis database was constructed
from 303 publications (Fig. 2). In addition to genetic ploidy level, root envi-
ronment (pot or field), wheat type (spring or winter), and study location, the
means of yield and yield components, morphophysiological, biochemical

and agronomic traits for drought stress, and control groups were extracted
from each study. The mean values, SD or SE, and number of observations
were gathered from the text, tables, and/or figures from each publication. In
cases where the data were only available from the figures, these were digi-
tized, and means and variance were obtained using the software Getdata
Graph Digitizer (http://getdata-graph-digitizer.com/). When more than
one degree of drought stress was applied in a study, the treatment represent-
ing a severe drought stress was selected.
2.2 Composition of the Database

The database comprised 303 publications from studies at experimental sites
across 45 countries (Fig. 3): 163 publications from Asia, 73 from Europe,
33 from North America, 15 from Africa, 13 from Australia, and 6 from
South America. The database was divided into five classes based on plant
growth parameters (Table 1): (1) yield, yield components, and biomass
Fig. 3 Distribution of the experiments reported in the articles analyzed in this

meta-analysis.
Table 1 Classification of Yield, Yield Components, Morphophysiological, Biochemical,

and Agronomic Characteristics, Water Relations, and the Period Exposed to Drought
Stress as Reported in this Meta-Analysis Study
Parameter
General Category Abbreviation Description
Yield, yield components, and Y Yield
biomass components
TKW Thousand kernel weight
HI Harvest index
GN Number of grains
AB DW Aboveground dry weight
Root DW Root dry weight
Stem DW Stem dry weight
Ear DW Ear dry weight
Leaf DW Leaf dry weight
Total DW Total dry weight
Plant architecture LA Leaf area
PH Plant height
EL Ear length
TN Tiller number
Gas exchange parameters and Pn Leaf net photosynthetic rate
chlorophyll concentration
gs Stomatal conductance
T/E Transpiration/
evapotranspiration
Ci Intercellular CO2 concentration
Fv/Fm Quantum efficiency of PSII
photochemistry in dark-adapted
state
Chl a Chlorophyll a concentration in
leaves
Chl b Chlorophyll b concentration in
leaves
Chl a + b Total chlorophyll concentration
in leaves
Table 1 Classification of Yield, Yield Components, Morphophysiological, Biochemical,

and Agronomic Characteristics, Water Relations, and the Period Exposed to Drought
Stress as Reported in this Meta-Analysis Study—cont’d
Parameter
General Category Abbreviation Description
Water relations, water use RWC Leaf relative water content
efficiency, and biochemical
LWP Leaf water potential
components
Sugar Soluble sugar concentration in
leaves
Proline Proline concentration in leaves
MDA Malondialdehyde concentration
in leaves
WUEG Water use efficiency for grain
Period exposed to drought stress W drought Drought stress across whole
growing season
V drought Drought stress at the vegetative
stage
A drought Drought stress over the anthesis
period
R drought Drought stress at the
reproductive stage
components; (2) plant architecture (leaf area, plant height, ear length, tiller
number); (3) gas exchange components and chlorophyll content; (4) water
relations, water use efficiency, and biochemical parameters; and (5) period
exposed to drought stress.
2.3 Sources of Variation

In the meta-analysis, we did not distinguish between different species and
subspecies of wheat. The reported species in the 2n ploidy wheat that were
combined were T. boeoticum, T. monococcum, and A. tauschii (Fig. 1). In the 4n
wheat, we did not distinguish between the wild T. turgidum L. ssp. dicoccoides
and the domesticated T. turgidum L. ssp. dicoccum or T. durum. In 6n wheat, all
the genotypes were T. aestivum. To explain the variation in the response of
wheat to drought, the following categories were identified: (1) wheat with
different levels of ploidy (2n, 4n, and 6n); (2) wheat type (spring or winter);
(3) rooting environment (field or pots); (4) timing and duration of the
imposed drought stress (vegetative growth, around anthesis, reproductive

growth, and whole growth period). Where the wheat type was not reported,
the type was identified from the website of the Research Program on Wheat
(http://wheatatlas.org/) using the name of the cultivar and planting location.
If the wheat type was still not clear, the data were only used for those parts of
the meta-analysis in which the type of wheat did not need to be specified.
2.4 Statistical Analyses

To reduce variation in the meta-analysis, the natural logarithm (ln) of the
response ratio (R), termed effect size, was calculated to measure the response
of wheat to drought stress. For a given variable, R was calculated as the ratio
of its value in the drought-stress treatment (XE) to that in the well-watered
control treatment (XC) and the effect size calculated as Eq. (1):
E
X
lnR ¼ ln C
¼ ln X E ln X C (1)
X
The variation (v) of ln R was approximated using Eq. (2):
2 2
ðS E Þ ðSC Þ
ν lnR ¼ + (2)
N E ðX E Þ2 N C ðX C Þ2
where SE is the treatment standard deviation, SC is the control standard devi-
ation, NE is the treatment sample size, and NC is the control sample size
(Rosenberg et al., 2000). If values were reported as SE or coefficient of var-
iation (CV), these were converted to SD using Eqs. (3) and (4):
pffiffiffiffiffi
SD ¼ SE N (3)
SD ¼ CVð%Þ X (4)
where X is the treatment mean value. Several studies did not report values of
SD or SE. In these cases, we calculated the average CV within each data set
and then back calculated SE from the average CV.
The meta-analysis procedure followed the methods described by Curtis
and Wang (1998). A mixed-effects model was employed to determine
whether drought stress significantly affected each variable using the statistical
software MetaWin 2.1 with a resampling of 9999 iterations (Rosenberg
et al., 2000). Confidence intervals on the estimated effect size were gener-
ated using the bootstrapping test. When there are fewer than 20 observations,
confidence limits based on bootstrapping tests are wider than the standard
confidence limits (Gurevitch and Hedges, 1999; Li et al., 2015). If the 95%
bootstrapping confidence interval values of the effect size for a variable did
not overlap zero, the effect of the drought treatment on the variable
was considered significant; otherwise, it was not considered significant.
Means of the different categorical variables were considered significantly dif-
ferent from one another if their 95% bootstrapping confidence intervals did
not overlap (Ainsworth et al., 2002; Wang et al., 2013). Analyses of the
parameters in Table 1 were performed to test differences in the response
of different ploidy wheat to drought under different environmental and
experimental conditions. To ease interpretation, the results of the ln R
(effect size) were back transformed and reported as a percentage change
((R 1) 100) under drought stress. Negative values indicate a reduction
in a plant variable to drought stress, whereas positive values indicated an
increase. If the value was equal to zero, drought stress did not affect that plant
variable. Meta-analysis assumes that studies are independent (Gurevitch and
Hedges, 1999) and free from publication bias (Rosenberg et al., 2000), and
we considered this to be the case in the meta-analysis reported here.
We use the homogeneity statistic Q, an estimate of the among-study var-
iance, to test whether the variances were significantly different or not; if
P < 0.05 (tested against a chi-square distribution) then the data were consid-
ered to be heterogeneous and further analyzed by single factor categorical
analysis. When conducting categorical analyses, total heterogeneity of effect
sizes among studies (QT) was generated and partitioned into heterogeneity
within categorical variables (QW), and heterogeneity between categorical
variables (QB). Comparison between categorical variables was examined
by QB (Chandrasekaran et al., 2014; Curtis and Wang, 1998).
3. RESPONSES TO DROUGHT STRESS IN DIFFERENT

PLOIDY WHEAT
3.1 Yield and Yield Components
Across all studies and all ploidy levels, drought stress significantly decreased
grain yield, yield components, harvest index (HI), plant height, leaf area, and
dry weight (DW) compared with the well-watered treatment (Fig. 4). Yield
decreased by 53%, in diploid (2n) wheat, which was significantly greater
than the 42% and 39% reductions in tetraploid (4n) and hexaploid (6n)
wheat, respectively (Fig. 4; Table 2). Thousand kernel weight (TKW)
decreased by 21%, 12%, and 16% in 2n, 4n, and 6n wheat, respectively, indi-
cating that water stress reduced the seed size significantly more in 2n wheat
Fig. 4 Effect of drought stress on the yield, yield components, biomass components,
plant height, leaf area, tiller number, and water use efficiency for grain in different ploidy
wheat. Symbols (2n, ▲; 4n, ; 6n, ■) represent the mean percentage change with
drought stress relative to well-watered conditions, and bars show the 95% confidence
intervals. The number of studies and the number of measurements are shown in paren-
theses. Abbreviations are given in Table 1.
than 4n and 6n wheat (QB ¼ 10.05, P < 0.05; Table 2). Grain number
decreased by 54%, 29%, and 16% in 2n, 4n, and 6n wheat, respectively, with
a significant difference between ploidy levels (QB ¼ 55.96, P < 0.001;
Table 2). The water use efficiency for grain (WUEG) remained unaffected
by drought stress in 6n wheat (95% CIs were 6% to 3%), but significantly
increased in 2n wheat (16%) and significantly decreased in 4n wheat (46%).
Drought stress reduced all the yield components, but the yield compo-
nent most affected by drought varied with ploidy level. Grain number and
size (TKW), particularly grain number, decreased most in 2n wheat under
drought stress. Drought reduced grain number to a greater degree than seed
size in 2n and 4n wheat, but not in 6n wheat. As a result of natural selection,
grain number is far more plastic than seed size, and high plasticity of grain
number and narrow variability of seed size is generally consistent with evo-
lutionary and genetic considerations among plants (Pedró et al., 2012;
Sadras, 2007). However, the meta-analysis suggests that evolution under
Table 2 Between-Group Heterogeneity (QB) of Wheat Variables as a Result of Drought

Stress and Ploidy Level, Wheat Type (Spring or Winter), Root Environment (Pot or Field),
and Period Exposed to Drought
Ploidy Drought Stress
Variables Level Ploidy Rooting Wheat Types Period
Y 2n 34.46*** — — 2.79
4n 20.62*** 0.9 95.43***
6n 276.53*** 570.55*** 693.84***
TKW 2n 10.05* — — 4.72*
4n 33.16*** 3.07 4.46
6n 65.19*** 21.96*** 62.27***
GN 2n 55.96*** — — 0.93
4n 2.36 20.77** 6.17
6n 26.34*** 32.03*** 49.69***
HI 2n 2.49 — — 9.46***
4n 3.43 34.5** 53.03***
6n 15.08*** 1.03 8.93*
WUEG 2n 547.37*** — — —
4n 5.19 — 3.52
6n 65.53*** 19.11*** 91.89***
AB DW 2n 74.92*** — — —
4n 3.44 — —
6n 45.69*** 38.28*** —
Root 2n 8.25* — — —
DW
4n — — —
6n 6.31* 20.69***
Stem 2n 29.85*** — — —
DW
4n 4.36 — —
6n 0.29 10.09*** —
Ear DW 2n 10.22*** — — —
4n 25.96*** — —
6n 7.25* 3.32 —
Continued

and Period Exposed to Drought—cont’d
Total 2n 25.34*** — — —
DW
4n 0.03 — —
6n 8.16** 7.55** —
LA 2n 57.46*** — — —
4n 6.71** — —
6n 0.97 0.07 —
PH 2n 1.21 — — —
4n 0.38 — —
6n 0.17 35.39*** —
EL 2n 18.88*** — — —
4n 0.26 — —
6n 1.47 0.45 —
TN 2n 11.43*** — — —
4n 45.46*** 12.77* 20.54**
6n 0.49 62.24*** 52.09***
Pn 2n 31.26*** — — —
4n — — —
6n 0.01 1.98 —
Ci 2n 20.01*** — — —
4n 9.92*** — —
6n 1.38 0.73 —
gs 2n 11.08*** — — —
4n 17.69*** — —
6n 2.02 1.01 —
T/E 2n 7.38* — — —
4n — — —
6n 0.33 18.91*** —

and Period Exposed to Drought—cont’d
Fv/Fm 4n 29.01*** 4.95* 8.4** —
6n 0.48 2.75 —
Chl a + b 2n 0.15 — — —
4n 0.82 1.55 —
6n 1.41 13.95*** —
Chl a 4n 0.17 — — —
6n 3.66* 11.78*** —
Chl b 4n 2.47* — — —
6n 0.37 16.75*** —
RWC 2n 17.63*** — — —
4n 3.25 3.56 —
6n 2.2 23.38*** —
LWP 4n 85.74*** 0.14 — —
6n 5.09* 1.55 —
Sugar 2n 28.91*** — — —
4n — — —
6n 12.71*** 16.04*** —
Proline 2n 7.57** — — —
4n 12.54*** — —
6n 2.53 8.65*** —
MDA 2n 1.08 — — —
4n — — —
6n 0.96 10.97*** —
Statistical significance is reported at P < 0.05 (*), P < 0.01 (**), and P < 0.001 (***).
Variable abbreviations are described in Table 1.
domestication from 2n to 6n appears to have weakened natural selection

leading to relatively narrow grain numbers, while it reinforced natural selec-
tion leading to relatively narrow grain size in wheat, with the magnitude of
the reduction in grain number and seed size decreasing with increasing
ploidy level. The smaller reduction (narrow variability, low plasticity) in
seed size in 4n and 6n wheat than in 2n wheat under drought stress may have
resulted from the greater transport efficiency from the preanthesis reserve
pools to the grain in 4n and 6n wheat (Pheloung and Siddique, 1991).
Drought stress can cause reproductive failure, but cereals crops have devel-
oped an adaptive response by accumulating reserves in different organs such
as stems and roots in the vegetative phase and redistributing the assimilates to
the ear in the reproductive phase. Thus, when the current photosynthesis is
inhibited by drought, grain filling becomes more dependent on mobilized
preanthesis stem reserves (Chaves et al., 2002) that buffer the grain yield
against environmental stresses during grain filling, particularly in wheat
(Bidinger et al., 1977; Blum, 1998).
3.2 Plant Architecture, Biomass Allocation, and

Physiological Traits
For plant architecture, leaf area decreased by 73%, in 2n wheat, significantly
greater than the 50% and 30% reductions in 4n and 6n wheat, respectively, as
a result of drought stress (Fig. 4; Table 2). Drought stress reduced plant
height by a similar amount (19%–24%) in all ploidy levels (Fig. 4;
Table 2). Tiller number decreased by 15%, 18%, and 26% in 2n, 4n, and
6n wheat, respectively (Fig. 4), indicating that 6n wheat had a significantly
greater reduction than 2n and 4n wheat under drought stress (QB ¼ 11.43,
P < 0.001; Table 2). Ear length decreased by 16%, 5%, and 8% in 2n, 4n, and
6n wheat, respectively; 2n wheat had a significantly greater reduction than
the other ploidy levels (QB ¼ 18.88, P < 0.001), while there was no signif-
icant difference between 4n and 6n wheats (Fig. 4). Drought stress signifi-
cantly reduced the DW in all ploidy levels (Fig. 4). In general, the DW of all
organs except the roots decreased the least in 6n wheat (Fig. 4). For example,
aboveground DW decreased by 42%, 42%, and 27%, ear DW decreased by
54%, 31% and 23%, and root DW decreased by 14%, 16%, and 29% in 2n,
4n, and 6n wheat, respectively (Fig. 4; Table 2). These results indicate that
under drought stress, lower ploidy level wheat had significantly more bio-
mass allocated to the root and less biomass allocated to the ear, indicating that
drought stress induced markedly different biomass distribution patterns in
different ploidy wheat with root allocation taking preference over allocation
to the ear in 2n wheat.
Across all studies, drought stress significantly decreased the leaf water
relations [relative water content (RWC) and leaf water potential (LWP)],
gas exchange parameters [photosynthetic rate (Pn), stomatal conductance
(gs), intercellular CO2 concentration (Ci), transpiration/evapotranspiration
rate (T/E), the quantum efficiency of PSII (Fv/Fm)] and chlorophyll con-
centrations (chl a, chl b, and chl a + b) (Fig. 5). The reduction in RWC as a
result of drought stress was least in 2n wheat (13%), significantly smaller than
the 24% and 20% in 4n and 6n wheat, respectively. While there were no
measurements of LWP in 2n wheat, the decrease in LWP was significantly
greater (QB ¼ 85.74, P < 0.001; Table 2) in 6n wheat (48%) than in 4n wheat
(19%) (Fig. 5). Drought stress reduced Pn by 33% and 40% in 2n and 6n
wheat, respectively, and 65% in 4n wheat, which was significantly greater
than the reduction in 2n or 6n wheat. The reduction in gs was similar at
63% and 74% in 2n and 4n wheat, respectively, while the reduction was
Fig. 5 Effect of drought stress on physiological, biochemical, gas exchange characteris-

tics, and chlorophyll concentration in different ploidy wheat. Symbols (2n, ▲; 4n, ; 6n, ■)
represent the mean percentage change with drought stress relative to well-watered
conditions, and bars show the 95% confidence intervals. The number of studies and
the number of measurements are shown in parentheses. Abbreviations are given in
Table 1.
56% in 6n wheat, significantly less than 4n wheat, but not less than 2n wheat
(Fig. 5; Table 2). Drought reduced Ci the most in 2n wheat (38%), while the
reduction was smaller in 4n wheat (14%) and 6n wheat (9%). T (or E)
decreased by 42%, 61%, and 43% in 2n, 4n, and 6n wheat, respectively;
in this case the largest reduction was in 4n wheat. As with Pn, Fv/Fm
decreased more in 4n wheat than in 6n wheat, even though the reduction
was small (7%) compared with the other parameters (Fig. 5; Table 2). For all
the gas exchange parameters except Ci, 4n wheat had significantly greater
reductions than 6n wheat, indicating drought affects the leaf gas exchange
in 4n wheat more than 6n wheat. The reduction in gas exchange parameters
varied widely in 2n wheat (Fig. 5). There was no significant difference in the
reduction of chl a and chl a + b between ploidy levels which decreased by
26% and 25%, respectively. However, there was a significantly greater
reduction of chl b in 6n wheat than 4n wheat (Fig. 5; Table 2). In contrast
to the reductions in water relations and gas exchange, drought stress signif-
icantly increased the biochemical parameters. In leaves, the soluble sugar
concentration increased by 90% and 120% in the 2n and 4n wheat, signif-
icantly more than the 35% in 6n wheat; proline concentration increased by
493% in 4n wheat, significantly more than the 274% and 213% in 2n and 6n
wheat; while the increased malondialdehyde (MDA) concentrations did not
differ significantly (mean 100%) between ploidy level (Fig. 5). Thus, overall,
biochemical parameters increased more in the 2n and 4n wheat than in 6n
wheat in response to drought stress.
Yield losses from drought stress have often been attributed to a reduction
in photosynthetic activity and a lower supply of assimilates to support repro-
ductive development and seed growth (Feng et al., 2008). However, this
meta-analysis showed that as ploidy level increased from 2n to 4n to 6n,
yields did not decline by the same amount as Pn, rather with leaf area, indi-
cating that the reduction in leaf area was a key driver of the reduction in
grain yield with different ploidy level. Nevertheless, the yields of different
ploidy wheats were associated with the whole-plant assimilation rate
(Pn leaf area per plant) which was driven largely by leaf area (Wang
et al., 2017). Indeed, crop evolution since domestication has been driven
by the selection of desired traits at the phenotypic level, e.g., flowering time,
plant height, leaf area, tiller number, grain number, and grain size (Cattivelli
et al., 2008). In addition to leaf area, it is accepted that ear photosynthesis can
make a major contribution to final grain yield, especially under drought
stress (Kriedemann, 1966; Tambussi et al., 2005). The 4n and 6n wheats
had smaller reductions in ear length than the 2n wheat, which can be a
significant photosynthetic contributor to grain filling in water-limited envi-

ronments (Fig. 4).
Unlike animals, plants cannot move to wetter areas as drought intensity
and frequency increase, but must adapt to the drier environment. Knowl-
edge of phenotypic traits that contribute to improved yields under stress
is fundamental to understanding the complex physiological and genetic
mechanisms of wheat adaptability (Reynolds et al., 2005). Mechanisms of
adaptation include a flexible response to the immediate environment; plants
can vary both the number and size of parts and can differentially allocate and
reallocate resources to various tissues (Kumar et al., 2016; Sultan, 1995).
Under drought stress, increased plant tolerance and grain yield may result
from changes in plant resource allocation patterns. This meta-analysis
showed that biomass allocation patterns change considerably with increasing
in ploidy level. In 2n and 4n wheat, aboveground biomass decreased signif-
icantly more than belowground biomass under drought stress, and the
reduction in total biomass was mainly caused by the loss of aboveground
biomass (Fig. 4). However, in 6n wheat, the total biomass reduction was
caused by loss of both the aboveground and belowground biomass. Further-
more, aboveground biomass decreased more in 2n and 4n wheat than 6n
wheat, while the decrease in belowground biomass was less than that of
6n wheat, indicating that different ploidy wheats have different biomass allo-
cation strategies under drought stress. A previous allometric analysis showed
that as the ploidy level increased from 2n to 6n with evolution under domes-
tication, less biomass was allocated to roots and more biomass was allocated
to shoots (Wang et al., 2017). Qin et al. (2012) investigated the aboveground
vs belowground biomass relationship in different ploidy wheat and also
found that evolution under domestication had resulted in an increased bio-
mass allocation to shoots compared to roots.
The meta-analysis showed that 6n wheat had the greatest reduction
(26%) in tiller number, but the smallest reduction (37%) in stem biomass
and grain number (16%), presumably because the reduction in tiller number
arose from a reduction in infertile (secondary) tillers under drought stress.
From this perspective, the growth redundancy was less in 6n wheat than
2n and 4n wheat. According to Ma et al. (2008), a reduction in infertile tillers
decreases water use, which could explain why WUEG only decreased by 1%,
despite a 39% reduction in grain yield. In contrast, the 2n and 4n wheat had a
greater reduction in stem and aboveground biomass, and had a greater
reduction in grain number, but not tiller number. The analysis showed that,
under drought stress, 2n and 4n wheat mainly reduced the number of fertile
tillers (main shoot and primary tillers), but still retained a high number of
smaller tillers. The retention of more small tillers would increase the poten-
tial for survival but also increase the competition for limited water resources
(Ma et al., 2008). It is likely that the 2n and 4n wheat retain some of the
characteristics of undomesticated wheat growing under natural conditions
(Qin et al., 2012).
During the vegetative phase, wheat plants produce many primary and
high-order tillers, some of which are sterile (Kebrom and Richards,
2013). During the transition to the reproductive phase, tiller formation
ceases and tiller senescence begins (Hay, 1986). In this study, as the ploidy
level increased, the maintenance of tiller number decreased (Fig. 4). At
anthesis and in the reproductive phase, tiller number in the 4n wheat chan-
ged little even under drought stress (data not shown), while in 6n wheat tiller
number decreased significantly presumably because, in addition to the effect
of drought stress, tiller mortality was inherently higher in 6n wheat than in
2n and 4n wheat. In drought-prone environments, some consider that the
small (infertile) tillers are detrimental in cereals because they do not contrib-
ute directly to grain yield and often compete with the main shoot for assim-
ilates and water, thereby reducing its yield. Many results support the concept
that there is a yield advantage associated with reduced tillering, particularly
in water-limited environments (Donald, 1968; Islam and Sedgley, 1981; Ma
et al., 2008), but tiller senescence during the reproductive stage could arise
from diversion or reallocation of resources to the ear and then to the grain, so
that infertile tillers act as a source of assimilates for grain filling, particularly
with terminal drought. This may explain why 6n wheat had the highest
reduction in tiller number and the smallest reduction in HI and yield as a
result of the drought treatments.
3.3 Root Environment (Pot or Field) and Wheat Type (Spring

or Winter)
In this study, the root environment, that is whether the wheat was grown in
pots or in the field, significantly affected the reduction in yield (Fig. 6). In the
available literature, the number of measurements with 2n wheat grown in
the field was limited, so that most of the data in Fig. 6 is limited to 4n
and 6n wheat. In general, in 4n wheat, yield, tiller number, leaf area, and
ear DW decreased significantly more in pots than in the field, but not
WUEG, HI, grain number, ear length, plant height, stem DW, aboveground
DW, or total DW. In 6n wheat, yield, WUEG, HI, seed size (TKW), grain
number, ear DW, root DW, aboveground DW, and total DW decreased
Fig. 6 Effect of drought stress on yield, yield formation, plant height, leaf area, tiller
number, water use efficiency for grain, and biomass allocation in different ploidy wheat
grown in pots or in the field. Symbols (pot, ■; field, □) represent the mean percentage
change at drought stress relative to well-watered conditions and bars show the 95%
confidence intervals. The number of studies and the number of measurements are
shown in parentheses. Abbreviations are given in Table 1.
significantly more in pots than in the field, but not tiller number, ear length,
plant height, leaf area, or stem DW (Fig. 6). For example, for the yield and
yield components, drought stress reduced yield more in pots than in the field
in both 4n wheat (52% vs 42%, QB ¼ 20.62, P < 0.001) and 6n wheat (46%
vs 34%, QB ¼ 276.53, P < 0.001) (Fig. 6; Table 2), despite no significant var-
iation in LWP or Pn (Fig. 7). Grain number decreased by 29% on average in
both pots and the field in 4n wheat but decreased significantly more in pots
(21%) than in the field (15%) in 6n wheat (QB ¼ 26.34, P < 0.001). In both
4n and 6n wheat, the reduction in grain number was not associated with a
reduction in ear length which always decreased by less than 10% and did not
differ whether the wheat was grown in pots or the field (Fig. 6). In 4n wheat,
the reduction in tiller number was significantly higher (QB ¼ 45.46,
P < 0.001) in pots (59%) than in the field (16%), but this was not in the case
for 6n wheat (QB ¼ 0.49, P ¼ 0.61). In 4n wheat, the average reduction in
HI was 19% in pots and field (QB ¼ 3.43, P ¼ 0.15), while in 6n wheat, the
reduction in pots (19%) was significantly greater (QB ¼ 15.08, P < 0.001)
Fig. 7 Effect of drought stress on physiological, biochemical, gas exchange character-

istics, and chlorophyll concentration in different ploidy wheat grown in pots or in field.
Symbols (pot, ■; field, □) represent the mean percentage change in drought stress rel-
ative to well-watered conditions and the bars show the 95% confidence intervals. The
number of studies and the number of measurements are shown in parentheses. Abbre-
viations are given in Table 1.
than the field (10%). Drought stress significantly reduced WUEG by more
than 40% in 4n wheat in both pots and the field, while in 6n wheat, WUEG
was higher in the drought treatment than the well-watered treatment in the
field, and decreased by less than 10% when grown in pots. For plant archi-
tecture, in both 4n and 6n wheat, drought stress reduced plant height, leaf
area, and ear length by similar amounts when grown in pots or the field
(Fig. 6), except for leaf area in 4n wheat which decreased significantly more
in pots than the field (Fig. 6; Table 2). For DW, the ear DW decreased by
60% and 41% in 4n wheat, and 36% and 20% in 6n wheat in pots and the
field, respectively. In 6n wheat, drought stress reduced root DW by 29% and
16%, aboveground DW by 35% and 23%, and total DW by 40% and 32% in
pots and the field, respectively (Fig. 6; Table 2). In 2n wheat, drought
imposed in pots reduced ear DW by 54%, which was significantly higher
than that for 6n wheat (Fig. 6).
Among the physiological and biochemical parameters (except LWP

and leaf soluble sugar concentration), there was no significant difference
between 6n wheat grown in pots or in the field, whereas several gas
exchange parameters (gs, Ci, Fv/Fm) decreased more and leaf proline con-
centration increased more in 4n wheat grown in pots than the field (Fig. 7;
Table 2). Overall, the physiological, biochemical, and gas exchange param-
eters were more sensitive to the root environment than morphological and
biomass parameters (Figs. 6 and 7).
In the field, water deficits do not always act alone but can be associated
with high temperature, wind, high irradiance, other soil-related constraints,
and biotic stresses. Under experimental conditions other stresses are mini-
mized as far as possible, but the effects of water stress are difficult to separate
from some of these other stresses in the field. Nevertheless, in this analysis,
over all ploidy levels, the performance of wheat in the field under drought
stress was better than that in pots. This may be because plants in the field are
usually subjected to a more gradual stress than in pots as water deficits slowly
develop and the plants are able to access water deep in the soil profile.
A gradual imposition of stress is likely to lead to osmotic adjustment and less
cellular damage, thus enabling plants to tolerate even more intense stress
(Begg and Turner, 1976; Chen et al., 2004; Turner and Jones, 1980). While
it is possible to induce a slow drying regime by the use of pots with soil vol-
umes per plant similar to those in the field and by addition of small amounts
of water to ensure a slow daily drying rate (see Pang et al., 2017), drought
experiments in pots usually deplete the available water rapidly due to the
limited soil volume and the finite amount of water available. Further, many
pot studies analyzed in this study added a limited volume of water to the soil
at regular (often daily) intervals to maintain the soil water content in the pot
at a particular level, but this effectively restricts the roots to a small volume of
wet soil in the pot, a condition rarely experienced in the field, and imposes a
drought stress that is significantly greater than the field.
In both 4n and 6n wheat, spring wheat was more affected by
drought stress than winter wheat (Figs. 8 and 9). The grain yield in 6n wheat
decreased by 45% and 28% in spring and winter wheat types (QB ¼ 570.55,
P < 0.001), respectively, but in 4n wheat grain yield did not differ signifi-
cantly (about 43% in the two types) (Fig. 8). In 4n wheat, the wheat type
did not affect the reduction in seed size as a result of drought stress
(QB ¼ 3.07, P ¼ 0.18), but in 6n wheat seed size decreased more in spring
wheat than winter wheat (QB ¼ 21.96, P < 0.001). In 4n wheat, the reduc-
tion in HI in spring wheat was significantly greater than in the winter wheat
Fig. 8 Effect of drought stress on yield, yield components, biomass components, plant
height, leaf area, tiller number, and water use efficiency for grain in spring and winter
wheat types at different ploidy levels. Symbols (spring, ; winter, ) represent the mean
percentage change under drought stress relative to well-watered conditions, and bars
show the 95% confidence intervals. The number of studies and the number of
measurements are shown in parentheses. Abbreviations are given in Table 1.
(QB ¼ 34.5, P < 0.01), while in 6n wheat, the reduction in HI was not sig-
nificantly different between spring and winter types. Grain number also
decreased more under drought in spring than winter wheat, with reductions
of 30% and 7% in 4n wheat and 20% and 11% in 6n wheat in spring and
winter types, respectively. In 4n winter wheat, drought stress did not affect
tiller number, while tiller number in spring wheat decreased by 16%; in con-
trast in 6n wheat, tiller number decreased more in winter wheat (57%) than
spring wheat (18%). Due to the limited data, the remaining parameters were
only analyzed in 6n wheat. Total DW, aboveground DW, root DW, stem
DW, and plant height all decreased more with drought stress in spring than
winter wheat, but the reduction in leaf area, ear length, and ear DW did not
significantly differ between wheat types (Fig. 8; Table 2).
The greater yield reductions in 6n spring wheat than 6n winter wheat
under drought stress were not associated with differences in Pn, gs, Ci,
Fig. 9 Effect of drought stress on physiological, biochemical, gas exchange character-

istics, and chlorophyll concentration in spring and winter wheat types at different ploidy
levels. Symbols (spring, ; winter, ) represent the mean percentage change under
drought stress relative to well-water conditions and bars show the 95% confidence
intervals. The number of studies and the number of measurements are shown in paren-
theses. Abbreviations are given in Table 1.
Fv/Fm, RWC, or LWP between types, but were associated with lower leaf
chl a, b and total chlorophyll concentrations, lower leaf proline concentra-
tion, and a greater increase in soluble sugar and MDA concentration in
spring wheat than winter wheat (Fig. 9; Table 2).
Classification into spring or winter wheat depends on the requirements
for vernalization; winter wheat normally requires 30–60 days of low temper-
atures (0–5°C) to initiate the reproductive phase, while spring wheat has no
such requirement. Spring wheat, as the name implies, is usually planted in
April or May (spring in the northern hemisphere), grows over summer,
and is harvested in August (late summer in the northern hemisphere). In
countries that experience mild winters, such as Australia, South Asia, South
Africa, the Middle East, and the lower latitudes, spring wheat is often sown
in autumn, grows slowly over the cooler winter months, and is harvested in
late spring or early summer (Curtis et al., 2002). Winter wheat needs to be
sown in autumn, is vernalized during winter, and harvested the following
spring or early summer. In this meta-analysis, drought stress affected spring

wheat more than winter wheat; for example, biomass decreased by 35%–
45% in 6n spring wheat, but only 0%–20% in 6n winter wheat. Notably,
root biomass was less affected by drought stress in winter wheat (Fig. 8).
Moreover, drought stress reduced tiller number more, but grain number
and yield less, in winter wheat than in spring wheat, suggesting that the
reduction in tiller number in winter wheat arose largely from a reduction
in infertile tillers. A reduction in infertile tillers as a consequence of drought
stress will result in a reduction in water consumption and growth redun-
dancy which may explain the increased WUEG in winter wheat (Fig. 8).
Previous studies demonstrated that anthesis and grain filling in wheat are
the most sensitive stages to environmental stresses such as drought, heat,
and ozone (Farooq et al., 2014; Feng et al., 2009; Wang et al., 2013). Spring
wheat is vulnerable during summer when drought stress and high temper-
atures are more likely, leading to a shorter grain filling period and a reduced
grain number and size, while winter wheat usually reaches anthesis and grain
filling in spring before the severe heat stress and greater drought frequency of
summer.
3.4 Drought Stress at Different Phenological Stages

Drought stress can occur in one or any combination of three recognized
growth stages [vegetative growth (from ear emergence to Feekes 10.5),
around anthesis (from Feekes 10.5 to 10.53), and reproductive growth (from
Feekes 10.53 to 11.4) (Zadoks et al., 1974)]. Although data are not shown, in
this study the effect of drought at one or a combination of stages on yield was
analyzed in the different ploidy wheat. When drought stress was imposed
over the whole growth period from sowing to maturity, yield decreased
by 49% in all three ploidy levels. Drought stress imposed at anthesis or in
the reproductive phase reduced yields by 32%–40%, while drought stress
imposed in the vegetative phase reduced yields by 31% and 22% in 4n
and 6n wheat, respectively, which was significantly lower than that during
the whole growth period. In 6n wheat, which had the most studies, drought
stress at anthesis and during the reproductive phase reduced tiller number by
about half that in the vegetative and whole growth periods (16% vs 32%),
while grain size decreased significantly more with drought stress at anthesis
and during the reproductive phase (21%) than during the vegetative phase
(6%). Surprisingly, the imposition of drought at different times had little
effect on grain number and ear length in 4n or 6n wheat, partly because
of the large variation among studies in yield components measured. Never-

theless, the meta-analysis suggested that drought stress at anthesis affected
grain number the most, while seed size was largely determined at both
anthesis and during grain filling.
The extent of the yield reduction with drought stress depends not only
on the magnitude of the stress but also on the plant growth stage when the
stress develops. In this study, across all ploidy levels, tiller number decreased
the most when drought stress occurred during vegetative growth when til-
lers develop. Drought stress during the vegetative stage, but not at later
stages, not only had the greatest reduction in tiller number per plant but also
in plant height and leaf area, suggesting that tillering and leaf area develop-
ment cannot recover even if the stress was relieved from anthesis, thereby
affecting growth and development in the reproductive phase. Additionally,
water shortage during vegetative development reduces the level of carbohy-
drate reserves in the stem, which is considered a significant factor affecting
flower and grain development under terminal stress (Chen et al., 2004; Palta
et al., 1994). Although early drought stress reduced tiller number the most,
drought stress during the reproductive phase had a greater effect on yield.
Many studies have reported that moisture stress at flowering (anthesis) results
in floret sterility as a consequence of pollen infertility (Briggs et al., 1999).
Sink strength can be reduced by drought stress during anthesis by a reduction
in endosperm cell number and metabolic activity (Nezhadahmadi et al.,
2013), and ultimately reduced grain number. Drought stress during the
reproductive phase directly influences kernel development and shortening
the reproductive phase reduces the assimilate supply and translocation to
grain. While there is compensation for developing grains of primary tillers
by assimilates from secondary tillers, even if these secondary tillers have an
ear (Palta et al., 1994), the grains in the later-formed tillers can be smaller
(Turner et al., 1994), possibly due to the reduced number of endosperm cells
and/or the number of amyloplasts initiated, resulting in shriveled grains
(Nayyar and Walia, 2004). This may explain why seed size decreased at
all ploidy levels under drought stress (Farooq et al., 2014; Pedró et al., 2012).
4. IMPLICATIONS AND FUTURE DIRECTIONS

Plants have developed appropriate developmental strategies to ensure
their survival and reproduction under drought-stress conditions. Evolution
under domestication from 2n to 4n to 6n wheat and subsequent breeding has
emphasized improved yields rather than survival. The meta-analysis showed
that drought stress has consistent negative effects on yield, yield components,
biomass, HI, plant height, leaf area, but not always WUEG. Evolution and
breeding in dryland environments have reduced the effects of drought stress
on yield and yield components of wheat, but not all traits selected are nec-
essarily beneficial in all drought-prone environments. For example, 2n and
4n wheat, but not 6n wheat, had greater biomass allocation to roots under
drought stress, but whether this is a beneficial or negative trait is not clear and
may depend on the amount and distribution of precipitation during the
growing season. Reducing root biomass by root pruning increased the yield
of winter wheat in the field (Fang et al., 2010), and modern high-yielding
spring wheat genotypes had less root biomass and a lower root-to-shoot ratio
than older lower-yielding genotypes (Siddique et al., 1990; Song et al.,
2009). This suggests that more assimilates allocated to roots result in fewer
resources allocated to grain (Passioura, 1983; Siddique et al. 1990; Song
et al., 2009), but simulation analysis indicates that faster root growth and
deeper roots benefit yield in environments and soils where water is left deep
in the profile (Asseng et al., 2011). Due to the constrained root environment
of pots, wheat was more affected by drought stress in pots than in the field,
and this needs to be considered when evaluating the effect of drought in pot
studies, particularly when using pot studies for phenotyping plants for their
performance in the field. Moreover, the maintenance of soil water contents
at less than field capacity in pots by daily watering is not a meaningful
drought stress treatment compared with the field as a portion of the soil is
effectively maintained at a soil water content near 100% field capacity while
the rest of the soil dries and roots die, so that root DW decreases to a similar
proportion as the proportion of moist soil. Using large volumes of soil per
plant and slowing daily water use to more realistic rates of soil drying will
provide more meaningful data on the responses to drought stress (Pang
et al., 2017). In this analysis, data from short-term drought stress treatments,
less than 48 h, were not included, but many of the pot studies utilized daily
soil watering to a proportion of 100% field capacity which may explain the
greater effect of drought stress in pots than in field.
Domestication can limit and act as a bottleneck for diversity in crops
(Abbo et al., 2003). In wheat, selection for loss of spike shattering and
free-threshing of dehulled grain (Dubcovsky and Dvorak, 2007) likely lim-
ited the gene pool, although the hybridization events that led to polyploid
4n and 6n wheat helped to reestablish some diversity that is important in its
adaptation to a wide range of environments. However, subsequent breeding
and selection have been largely restricted within this limited gene pool.
Recent developments in genomics that allows the identification of useful

alleles from landraces and early domesticated 2n and 4n wheat has revived
interest this “wild” gene pool as a source of traits, including drought resis-
tance traits, for incorporation into new genotypes of cultivated 4n and 6n
wheat. For example, a breeding program at CIMMYT has taken 4n lines
of wheat and crossed them with goat grass (A. tauschii) to produce a series
of “synthetic” wheat lines that are being evaluated for a range of biotic
and abiotic stress tolerances and other beneficial traits (CIMMYT, 2016;
Ogbonnaya et al., 2007; Pradhan et al., 2012). While this meta-analysis
did not include any of these evaluations, hence the difficulty in identifying
those traits that have been incorporated from the wild progenitors of modern
4n and 6n wheat, it does indicate that there are traits in 2n wheat that may
increase the drought resistance of durum (T. durum) and bread (T. aestivum)
wheat. It is important to evaluate not only the synthetic wheat genotypes but
also their parents, so that the traits and genes that confer abiotic stress resis-
tance, such as drought and heat resistance, can be identified and incorporated
into otherwise well-adapted and agronomically superior cultivars with desir-
able grain quality characteristics.
ACKNOWLEDGMENTS
The authors thank Yingxia Liu and Saiyong Zhu for assistance with data extraction and
analysis, Jian Zhang for assistance with the ArcGIS used for Fig. 3, Fei Mo and Pufang Li
for the constructive comments on an earlier version of this manuscript. The research was
funded by the International Cooperation Program of Ministry of Science and Technology
of China (2015DFG31840), Natural Science Foundation of China (31570415), State
Technology Support Program (2015BAD22B04), Fundamental Research Funds for the
Central Universities of China (lzujbky-2015-br02), the Overseas Masters Program of
Ministry of Education (Ms2011LZDX059), Chinese Scholarship Council (CSC), and the
Institute of Agriculture at The University of Western Australia.
REFERENCES
Abbo, S., Berger, J., Turner, N.C., 2003. Evolution of cultivated chickpea: four bottlenecks
limit diversity and constrain adaptation. Funct. Plant Biol. 30, 1081–1087.
Ahmadi, A., Baker, D.A., 2001. The effect of water stress on the activities of key regulatory
enzymes of the sucrose to starch pathway in wheat. Plant Growth Regul. 35, 81–91.
Ainsworth, E.A., Davey, P.A., Bernacchi, C.J., Dermody, O.C., Heaton, E.A., Moore, D.J.,
Morgan, P.B., Naidu, S.L., Ra, H.Y., Zhu, X.G., Curtis, P.S., Long, S.P., 2002.
A meta-analysis of elevated CO2 effects on soybean (Glycine max) physiology, growth
and yield. Global Change Biol. 8, 695–709.
Araus, J.L., Slafer, G.A., Royo, C., Serret, M.D., 2008. Breeding for yield potential and stress
adaptation in cereals. Crit. Rev. Plant Sci. 27, 377–412.
Asseng, S., Foster, I., Turner, N.C., 2011. The impact of temperature variability on wheat
yields. Global Change Biol. 21, 997–1012.
Bai, E., Li, S., Xu, W., Li, W., Dai, W., Jiang, P., 2013. A meta-analysis of experimental
warming effects on terrestrial nitrogen pools and dynamics. New Phytol. 199, 441–451.
Begg, J.E., Turner, N.C., 1976. Crop water deficits. In: Sparks, D.L. (Ed.), Advances in
Agronomy, vol. 28. Academic Press, San Diego, pp. 161–217.
Bidinger, F., Musgrave, R.B., Fischer, R.A., 1977. Contribution of stored pre-anthesis
assimilate to grain yield in wheat and barley. Nature 270, 431–433.
Blum, A., 1998. Improving wheat grain filling under stress by stem reserve mobilization.
Euphytica 100, 77–83.
Briggs, K.G., Kiplagat, O.K., Johnson-Flanagan, A.M., 1999. Effects of pre-anthesis mois-
ture stress on floret sterility in some semi-dwarf and conventional height spring wheat
cultivars. Can. J. Plant Sci. 79, 515–520.
Cattivelli, L., Rizza, F., Badeck, F.W., Mazzucotlli, E., Mastrangelo, A.M., Francia, E.,
Marè, C., Tondelli, A., Stanca, A.M., 2008. Drought tolerance improvement in crop
plants: an integrated view from breeding to genomics. Field Crops Res. 105, 1–14.
Chandrasekaran, M., Boughattas, S., Hu, S.J., Oh, S.H., Sa, T., 2014. A meta-analysis
of arbuscular mycorrhizal effects on plants grown under salt stress. Mycorrhiza
24, 611–625.
Chaves, M.M., Pereira, J.S., Maroco, J., Rodrigues, M.L., Ricardo, C.P.P., Osório, M.L.,
Carvalho, I., Faria, T., Pinheiro, C., 2002. How plants cope with water stress in the field?
Photosynthesis and growth. Ann. Bot. 89, 907–916, London.
Chen, K.M., Gong, H.J., Chen, G.C., Wang, S.M., Zhang, C.L., 2004. Gradual drought
under field conditions influences the glutathione metabolism, redox balance and energy
supply in spring wheat. J. Plant Growth Regul. 23, 20–28.
CIMMYT (International Maize and Wheat Improvement Center), 2016. Wheat in the
World. http://wheat.org/wheat-in-the-world/(accessed 26.05.16.).
Curtis, P.S., Wang, X., 1998. A meta-analysis of elevated CO2 effects on woody plant mass,
form, and physiology. Oecologia 113, 299–313.
Curtis, B.C., Rajaram, S., Macpherson, H.G. (Eds.), 2002. Bread Wheat: Improvement and
Production. Food and Agriculture Organization of the United Nations, Rome.
Daryanto, S., Wang, L., Jacinthe, P.A., 2015. Global synthesis of drought effects on food
legume production. PloS One 10, e0127401.
Donald, C.M., 1968. The breeding of crop ideotypes. Euphytica 17, 385–403.
Dreccer, M.F., Borgognone, M.G., Ogbonnaya, F.C., Trethowan, R.M., Winter, B., 2007.
CIMMYT-selected derived synthetic bread wheats for rainfed environments: yield eval-
uation in Mexico and Australia. Field Crops Res. 100, 218–228.
Dubcovsky, J., Dvorak, J., 2007. Genome plasticity a key factor in the success of polyploidy
wheat under domestication. Science 316, 1862–1866.
Fang, Y., Xu, B.C., Turner, N.C., Li, F.M., 2010. Grain yield, dry matter accumulation and
remobilization, and root respiration in winter wheat as affected by seeding rate and root
pruning. Europ. J. Agron. 33, 257–266.
FAOSTAT (Food and Agriculture Organization of the United Nations Statistics Division),
2015. http://faostat3.fao.org/browse/Q/QC/E (accessed 26.05.16.).
Farooq, M., Hussain, M., Siddique, K.H.M., 2014. Drought stress in wheat during flowering
and grain-filling periods. Crit. Rev. Plant Sci. 33, 331–349.
Feng, Z., Kobayashi, K., Ainsworth, E.A., 2008. Impact of elevated ozone concentration on
growth, physiology, and yield of wheat (Triticum aestivum L.): a meta-analysis. Global
Change Biol. 14, 2696–2708.
Feng, Z., Kobayashi, K., Wang, X., Feng, Z., 2009. A meta-analysis of responses of wheat
yield formation to elevated ozone concentration. Chinese Sci. Bull. 54, 249–255.
Gurevitch, J., Hedges, L.V., 1999. Statistical issues in ecological meta-analyses. Ecology
80, 1142–1149.
Hay, R.K.M., 1986. Sowing date and the relationships between plant and apex development
in winter cereals. Field Crops Res. 14, 321–337.
Islam, T.M.T., Sedgley, R.H., 1981. Evidence for a ‘uniculm effect’ in spring wheat (Triticum
aestivum L.) in a Mediterranean environment. Euphytica 30, 277–282.
IWGSC (International Wheat Genome Sequencing Consortium), 2014. A chromosome-based
draft sequence of the hexaploid bread wheat (Triticum aestivum) genome. Science
345, 1251788.
Kebrom, T.H., Richards, R.A., 2013. Physiological perspectives of reduced tillering and stu-
nting in the tiller inhibition (tin) mutant of wheat. Funct. Plant Biol. 40, 977–985.
Kihara, H., Lilienfeld, F., 1949. A new synthesized 6x-wheat. Hereditas 35 (S1), 307–319.
Kriedemann, P., 1966. The photosynthetic activity of the wheat ear. Ann. Bot. 30, 349–363.
Kumar, U., Laza, M.R., Soulie, J.-C., Pasco, R., Mendez, K.V.S., Dingkuhn, M., 2016.
Analysis and simulation of phenotypic plasticity for traits contributing to yield potential
in twelve rice genotypes. Field Crops Res. 202, 94–107. http://dx.doi.org/10.1016/j.
fcr.2016.04.037.
Lascano, H.R., Antonicelli, G.E., Luna, C.M., Antonicelli, G.E., Luna, C.M.,
Melchiorre, M.N., Gómez, L.D., Racca, R.W., Trippi, V.S., Casano, L.M., 2001. Anti-
oxidant system response of different wheat cultivars under drought: field and in vitro
studies. Funct. Plant Biol. 28, 1095–1102.
Lev-Yadun, S., Gopher, A., Abbo, S., 2000. The cradle of agriculture. Science 288, 1602.
Li, W., Jin, C., Guan, D., Wang, Q., Wang, A., Yuan, F., Wu, J., 2015. The effects of sim-
ulated nitrogen deposition on plant root traits: a meta-analysis. Soil Biol. Biochem.
82, 112–118.
Loss, S.P., Siddique, K.H.M., 1994. Morphological and physiological traits associated with
wheat yield increases in Mediterranean environments. In: Sparks, D.L. (Ed.), Advances
in Agronomy, vol. 52. Academic Press, San Diego, pp. 229–276.
Ma, S.C., Xu, B.C., Li, F.M., Huang, Z.B., 2008. Ecological significance of redundancy in
tillers of winter wheat (Triticum aestivum) and effect of reducing redundancy on water use
efficiency. Acta Ecol. Sin. 28, 321–326 (in Chinese with English abstract).
McFadden, E.S., Sears, E.R., 1946. The origin of Triticum spelta and its free-threshing hexa-
ploid relatives. J. Hered. 37, 107–116.
McKersie, B., 2015. Planning for food security in a changing climate. J. Exp. Bot. 66 (12),
3435–3450.
Moustafa, M.A., Boersma, L., Kronstad, W.E., 1996. Response of four spring wheat cultivars
to drought stress. Crop Sci. 36, 982–986.
Nawaz, F., Ahmad, R., Waraich, E.A., Naeem, M.S., Shabbir, R.N., 2012. Nutrient uptake,
physiological responses, and yield attributes of wheat (Triticum aestivum L.) exposed to
early and late drought stress. J. Plant Nutr. 35, 961–974.
Nayyar, H., Walia, D.P., 2004. Genotypic variation in wheat in response to water stress and
abscisic acid-induced accumulation of osmolytes in developing grains. J. Agron. Crop
Sci. 190, 39–45.
Nezhadahmadi, A., Prodhan, Z.H., Faruq, G., 2013. Drought tolerance in wheat. Scientific
World J. 2013, 1–12.
Ogbonnaya, F.C., Ye, G.Y., Trethowan, R., Dreccer, F., Lush, D., Shepperd, J.,
Ginkel, M.V., 2007. Yield of synthetic backcross-derived lines in rainfed environments
of Australia. Euphytica 157, 312–336.
Osenberg, C.W., Sarnelle, O., Goldberg, D.E., 1999. Special feature: meta-analysis in ecol-
ogy: concepts, statistics, and applications. Ecology 80, 1103.
Palta, J.A., Kobata, T., Turner, N.C., Fillery, I.R., 1994. Remobilization of carbon and
nitrogen in wheat as influenced by post anthesis water deficits. Crop Sci. 34, 118–124.
Pang, J.Y., Turner, N.C., Khan, T., Du, Y.-L., Xiong, J.-L., Colmer, T.D., Devilla, R.,
Stefanova, K., Siddique, K.H.M., 2017. Response of chickpea (Cicer arietinum L.) to ter-
minal drought: leaf stomatal conductance, pod abscisic acid concentration, and seed set.
J. Exp. Bot. http://dx.doi.org/10.1093/jxb/erw153.
Passioura, J.B., 1983. Roots and drought resistance. Agric. Water Manage. 7, 265–280.
Pedró, A., Savin, R., Slafer, G.A., 2012. Crop productivity as related to single-plant traits at
key phenological stages in durum wheat. Field Crops Res. 138, 42–51.
€
Pellegrino, E., Opik, M., Bonari, E., Ercoli, L., 2015. Responses of wheat to arbuscular
mycorrhizal fungi: a meta-analysis of field studies from 1975 to 2013. Soil Biol. Biochem.
84, 210–217.
Petersen, G., Seberg, O., Yde, M., Berthelsen, K., 2006. Phylogenetic relationships of
Triticum and Aegilops and evidence for the origin of the A, B, and D genomes of common
wheat (Triticum aestivum). Mol. Phylogenet. Evol. 39, 70–82.
Pheloung, P.C., Siddique, K.H.M., 1991. Contribution of stem dry matter to grain yield in
wheat cultivars. Aust. J. Plant Physiol. 18, 53–64.
Pradhan, G.P., Prasad, P.V.V., Fritz, A.K., Kirkham, M.B., Gill, B.S., 2012. Effects of
drought and high temperature stress on synthetic hexaploid wheat. Funct. Plant Biol.
39, 190–198.
Qin, X.L., Niklas, K.J., Qi, L., Xiong, Y.C., Li, F.M., 2012. The effects of domestication on
the scaling of below- vs. aboveground biomass in four selected wheat (Triticum; Poaceae)
genotypes. Am. J. Bot. 99, 1112–1117.
Reynolds, M.P., Mujeeb-Kazi, A., Sawkins, M., 2005. Prospects for utilising plant-adaptive
mechanisms to improve wheat and other crops in drought- and salinity-prone environ-
ments. Ann. Appl. Biol. 146, 239–259.
Rose, M.T., Patti, A.F., Little, K.R., Brown, A.L., Jackson, W.R., Cavagnaro, T.R., 2014.
A meta-analysis and review of plant-growth response to humic substances: practical
implications for agriculture. In: Sparks, D.L. (Ed.), Advances in Agronomy, vol. 124.
Academic Press, San Diego, pp. 37–89.
Rosenberg, M.S., Adams, D.C., Gurevitch, J. (Eds.), 2000. MetaWin: Statistical Software for
Meta-Analysis. Sunderland, Massachusetts, USA.
Sadras, V.O., 2007. Evolutionary aspects of the trade-off between seed size and number in
crops. Field Crops Res. 100, 125–138.
Siddique, K.H.M., Kirby, E.J.M., Perry, M.W., 1989. Ear:stem ratio in old and modern
wheat varieties; relationship with improvement in number of grains per ear and yield.
Field Crops Res. 21, 59–78.
Siddique, K.H.M., Belford, R.K., Tennant, D., 1990. Root:shoot ratios of old and modern,
tall and semi-dwarf wheats in a Mediterranean environment. Plant Soil. 121, 89–98.
Song, L., Li, F.M., Fan, X.W., Xiong, Y.C., Wang, W.Q., Wu, X.B., Turner, N.C., 2009.
Soil water availability and plant competition affect the yield of spring wheat. Europ. J.
Agron. 31, 51–60.
Stocker, T.F., Qin, D.H., Plattner, G.K., Alexander, L.V., Allen, S.K., Bindoff, N.L.,
Breon, F.M., Church, J.A., Cubasch, U., Emori, S., Forster, P., Friedlingstein, P.,
Gillett, N., Gregory, J.M., Hartmann, D.L., Jansen, E., Kirtman, B., Knutti, R.,
Kumar, K.K., Lemke, P., Marotzke, J., Masson-Delmotte, V., Meehl, G.A.,
Mokhov, I.I., Piao, S., Ramaswamy, V., Randall, D., Rhein, M., Rojas, M.,
Sabine, C., Shindell, D., Talley, L.D., Vaughan, D.G., Xie, S.P., 2013. Technical sum-
mary. In: Stocker, T.F., Qin, D.H., Plattner, G.K., Tignor, M., Allen, S.K.,
Boschung, J., Nauels, A., Xia, Y., Bex, V., Midgley, P.M. (Eds.), Climate Change
2013: The Physical Science Basis. Contribution of Working Group I to the Fifth Assess-
ment Report of the Intergovernmental Panel on Climate Change. Cambridge
University Press, Cambridge, UK/New York, NY.
Sultan, S.E., 1995. Phenotypic plasticity and plant adaptation. Acta Bot. Neerlandica.
44, 363–383.
Suriyagoda, L.D., Ryan, M.H., Renton, M., Lambers, H., 2014. Plant responses to limited
moisture and phosphorus availability: a meta-analysis. In: Sparks, D.L. (Ed.), Advances in
Agronomy, vol. 124. Academic Press, San Diego, pp. 133–200.
Tambussi, E.A., Nogues, S., Araus, J.L., 2005. Ear of durum wheat under water stress: water
relations and photosynthetic metabolism. Planta 221, 446–458.
Tang, Y., Rosewarne, G.M., Li, C., Wu, X., Yang, W., Wu, C., 2015. Physiological factors
underpinning grain yield improvements of synthetic-derived wheat in southwestern
China. Crop Sci. 55, 98–112.
Turner, N.C., 1979. Drought resistance and adaptation to water deficits in crop plants.
In: Mussell, H., Staples, R.C. (Eds.), Stress Physiology in Crop Plants. John Wiley,
New York, pp. 343–372.
Turner, N.C., 1986. Crop water deficits: a decade of progress. In: Sparks, D.L. (Ed.),
Advances in Agronomy, vol. 39. Academic Press, San Diego, pp. 1–51.
Turner, N.C., 1997. Further progress in crop water relations. In: Sparks, D.L. (Ed.),
Advances in Agronomy, vol. 58. Academic Press, San Diego, pp. 293–338.
Turner, N.C., Jones, M.M., 1980. Turgor maintenance by osmotic adjustment: a review and
evaluation. In: Turner, N.C., Kramer, P.J. (Eds.), Adaptation of Plants to Water and
High Temperature Stress. John Wiley & Sons, New York, pp. 87–103.
Turner, N.C., Meyer, R., 2011. Synthesis of regional impacts and global agricultural adjust-
ments. In: Yadav, S.S., Redden, R.J., Hatfield, J.L., Lotze-Campden, H., Hall, A.E.
(Eds.), Crop Adaptation to Climate Change. John Wiley & Sons, New York,
pp. 156–165.
Turner, N.C., Prasertsak, P., Setter, T.L., 1994. Plant spacing, density, and yield of wheat
subjected to post anthesis water deficits. Crop Sci. 34, 741–748.
Turner, N.C., Blum, A., Cakir, M., Steduto, P., Tuberosa, R., Young, N., 2014. Strategies
to increase the yield and yield stability of crops under drought—are we making progress?
Funct. Plant Biol. 41, 1199–1206.
Wang, L., Feng, Z., Schjoerring, J.K., 2013. Effects of elevated atmospheric CO2 on phys-
iology and yield of wheat (Triticum aestivum L.): a meta-analytic test of current hypoth-
eses. Agr. Ecosyst. Environ. 178, 57–63.
Wang, J.Y., Turner, N.C., Liu, Y.X., Siddique, K.H.M., Xiong, Y.C., 2017. Effects of
drought stress on morphological, physiological and biochemical characteristics in differ-
ent ploidy wheat genotypes. Funct. Plant Biol. 44, 219–234.
Wardlaw, I.F., Willenbrink, J., 2000. Mobilization of fructan reserves and changes in enzyme
activities in wheat stems correlate with water stress during kernel filling. New Phytol.
148, 413–422.
Yadav, S.S., Redden, R., Hatfield, J.L., Lotze-Campen, H., Hall, A. (Eds.), 2011. Crop
Adaptation to Climate Change. John Wiley & Sons, New York, USA.
Zadoks, J.C., Chang, T.T., Konzak, C.F., 1974. A decimal code for the growth stages of
cereals. Weed Res. 14, 415–421.
Zhao, X., Zhang, R., Xue, J.F., Pu, C., Zhang, X.Q., Liu, S.L., Chen, F., Lal, R.,
Zhang, H.L., 2015. Management-induced changes to soil organic carbon in China: a
meta-analysis. In: Sparks, D.L. (Ed.), Advances in Agronomy, vol. 134. Academic
Press, San Diego, pp. 1–50.
CHAPTER FOUR
Delineation of Soil Management

Zones for Variable-Rate
Fertilization: A Review
Said Nawar*, Ronald Corstanje*, Graham Halcro*, David Mulla†,
Abdul M. Mouazen*,{,1
*Cranfield Soil and AgriFood Institute, School of Environment, Energy and AgriFood, Cranfield University,
Bedfordshire, United Kingdom
†
University of Minnesota, Saint Paul, MN, United States
{
Precision Soil and Crop Engineering Group, Faculty of Bioscience Engineering, Ghent University, Ghent,
Belgium
1
Corresponding author: e-mail address: abdul.mouazen@ugent.be
Contents
1. Introduction 176
2. Conceptual Framework and Main Steps of MZs 179
2.1 Mapping Properties Useful as MZs Proxies 181
2.2 Individuation of Homogeneous Areas 189
2.3 Finding the Optimal Number of Classes 190
2.4 Delineating Final MZs 191
2.5 Assessing the Effectiveness of the Classification 191
3. MZ Delineation Approaches 197
3.1 MZ Delineation Based on Farmer Knowledge 197
3.2 MZ Delineation Based on Geomorphology 200
3.3 MZ Delineation Based on Traditional Soil Chemical Analyses 201
3.4 MZ Delineation Based on Soil Class 203
3.5 MZ Delineation Based on Yield Maps 204
3.6 MZ Delineation Based on Crop Coverage 204
3.7 MZ Delineation Based on Proximal Soil Sensors and Data Fusion 208
4. Application Maps for VR Fertilization 220
5. Opportunities for Evaluation MZs by Cost/Benefit Analysis 221
6. A Case Study: Site-Specific N Application for Oil Seed Rape (OSR) Crop Based on
MZ Delineation With Multisensor and Data Fusion Approach 223
6.1 Experimental Site 223
6.2 Results and Discussion 230
7. Conclusions 233
Acknowledgments 236
References 236

176 Said Nawar et al.
Abstract
Different methods of management zone (MZ) delineation have been established over
the past 2 decades based on approaches, which have been largely constrained by the
available data collection methods that are often time consuming and expensive. This
situation is being changed by recent advances in sensor technology, making a huge
amount of data available. Advances in computing power make it possible to analyze
and utilize this large amount of data. These current advances in technology are grad-
ually turning MZ maps into commercially viable agricultural products for large-scale
adoption. The aim of this paper is to provide a critical overview of MZ delineation
approaches for precision agriculture applications, and to compare and contrast tradi-
tional with advanced sensing technologies for delineating MZs. This review illustrates
how recent development in sensing technologies, geostatistical analysis, data fusion,
and interpolation techniques have improved precision and reliability of MZ delineation,
making it a viable strategy in commercial agriculture.
Studies from the last decade showed that when MZ delineation techniques are
used for variable-rate nutrient application, farm efficiency increased when this is
compared to traditional uniform-rate application methods. This improved farm
production efficiency is accompanied by a reduction in environmental impacts.
Implementation of MZ therefore often provides financial and environmental benefits,
and we can foresee an increase in the diffusion and application of precision agriculture
techniques in the near future.
1. INTRODUCTION
Economic and environmental pressures on farming have rapidly
increased over the last decades. The increasing demand for food from a
growing population (Baudron and Giller, 2014), together with an increased
demand for energy derived from agricultural products, is putting an enor-
mous pressure on agricultural productivity (Foley et al., 2011). At the same
time, there is a growing trend in the environmental challenges the world is
facing, increasing the need for environmental conservation practices (Foley
et al., 2011). These two rapidly growing needs are extremely difficult to
satisfy simultaneously, and this is multiplying the challenges agriculture is
going to face in the current century. The scientific community is responding
to these challenges by developing sustainable ways for improving the
efficiency of agricultural inputs, in order to maximize production and at
the same time minimize environmental impacts.
A promising possibility is to vary the application of inputs according to
soil and crop requirements, an approach called precision agriculture or
precision farming. This approach is rapidly being adopted, motivated by a
Delineation of Soil Management Zones 177
need to increase in efficiency of agriculture under economic, sociological,

environmental, and entrepreneurial pressures (Pierpaoli et al., 2013). Soil
is an extremely complex and highly variable medium (Phillips, 2001) and
this poses significant challenges. However, there are other sources of vari-
ability attributable to, for instance topography, land use and fine-scale
weather conditions, which all affect crop growth and yield. Precision
farming embraces such variability.
Traditional farm management uses a whole-field approach, in which
each field is treated as a homogeneous area (Srinivasan, 2006) and the
variability in soil, topography, local weather conditions, and land use is
not considered. Inputs are applied uniformly across the field. This manage-
ment approach is attractive to growers because it is relatively quick and easy
to implement, but implies an inefficient application of inputs. This ineffi-
cient application carries some unwanted explicit economic costs, but also
several hidden environmental costs. Any unused nutrients not retained in
the soil are lost to the environment through leaching, runoff, and gaseous
emissions. The excessive use of fertilizers increases the contamination levels
in soil and ground water and represents a hidden cost for society (Hyyti€ainen
et al., 2011; Rodriguez et al., 2011). Furthermore, under application of
fertilizers has negative influences on crop growth, and subsequently yield.
Customized and spatially accurate nutrient applications can greatly reduce
these agronomic, economic, and environmental losses.
Farmers have been using the precision agriculture approach informally
for thousands of years. Familiarity allowed them to identify specific areas of
a field, which regularly provided high or low yields (Fleming et al., 2000b).
In the past, experience or lore may have guided the application of an
appropriate remediation. The agronomic experience was and is still con-
sidered by the farmers as the key success of farming today. However, a
more objective approach might be developed by decreasing soil variability
through classifying a field into areas of broad similarity. These classes can be
defined as management zones (MZs) and can be used as a baseline for most
farming decisions. MZs are defined as subregions of a field that have a rel-
atively homogeneous combination of yield-limiting factors, for which a
single rate of a specific crop input is appropriate to attain maximum
efficiency of farm inputs (Vrindts et al., 2005). Also, MZ can be defined
as a subregion of a field that is relatively homogeneous with respect to
soil-landscape attributes (Haghverdi et al., 2015). MZs are usually
generated from a single data layer or combinations of data layers, including
yield maps, topography, soil sampling data, aerial photographs, apparent
electrical conductivity (ECa), and canopy images assessing crop growth

variability. The resulting MZ should be simple, stable over years, accurate,
and inexpensive to identify, and can be classified into areas that need to
be managed differently (Khosla et al., 2010). The availability and adoption
of relatively cheap and accurate positioning systems led to modern
farm machinery that can now apply localized inputs at adjustable rates.
Thus, several distinct zones within existing fields can be managed despite
the presence of spatial variability in many yield-limiting factors, and a
proper identification of MZ is one of the keys to an efficient application
of inputs.
Research into soil fertility-based MZ has largely focused on characteriz-
ing within-field variability of yield-limiting factors, but this objective in soil
poses several challenges. The detection of such soil-limiting factors must be
economically viable, and it will therefore be likely based on some proxy
variables. The correlation of these proxy variables with yield responses is
therefore one of the key points where the focus should be to develop
effective precision farming approaches. Areas of similarity need to be derived
from proxy factors so that each zone is not only significantly different from
the others, but also reflects well the conditions affecting crop responses in
order to plan specific interventions with accuracy.
Early research into the benefits of using MZ to increase yield (Mulla
et al., 1992) and/or nitrogen-use efficiency (NUE) (Khosla and Alley,
1999) were generally positive and encouraging. Over years, delineation
techniques were developed, which used more complex assessments of soil
fertility variation and are orientating more and more toward a multivariate
approach (e.g., historical yield, soil and crop characteristics, topography,
weather, and within-season measures of vigor) (Guastaferro et al., 2010).
These developments have improved the performance of MZ, as compared
with traditional MZ delineation methods that are based only on soil or crop
properties. In addition, the adoption of new sensing technologies for
characterizing spatial variability in soil, e.g., gamma ray (i.e., Castrignanò
et al., 2012), electromagnetic induction (EMI) (i.e., King et al., 2005),
and visible and near-infrared (vis–NIR) spectroscopy (i.e., Mouazen and
Kuang, 2016) has gained vast attention of scientists in the last decade.
Furthermore, measurement of variability in crop growth with proximal
of satellite assets to delineate MZs based on spectral indices such as normal-
ized difference vegetation index (NDVI) (i.e., Inman et al., 2008) has meant
that a wide range of yield-limiting factors in the soil and crop properties can
be measured rapidly at finer resolutions.
Generally, there is very limited information in the literature focusing

on a cost–benefit analysis of the MZ–variable-rate (VR) fertilization, as
compared to uniform-rate (UR) fertilization. Nevertheless, the success of
MZ in agricultural research has led over the last decades to the establishment
of technically advanced commercial precision agriculture services (Kaivosoja
et al., 2014). Precision farming based on MZ seems therefore commercially
viable, and further research into improving delineation techniques could
provide further economic and efficiency benefits.
This review attempts to discuss the main approaches adopted for
soil fertility-based MZ delineation, focusing on variable-rate application
(VRA) of nutrients. A case study on the use of MZ for site-specific nitrogen
fertilization based on high sampling resolution data on soil will be discussed
and reported, as an example.
2. CONCEPTUAL FRAMEWORK AND MAIN STEPS OF MZs

With recent advancements in information technologies, remote and
proximal sensing and geospatial analyses supported by global positioning
systems, it is increasingly possible to identify and analyze the temporal
and spatial variability within fields to maximize the yield and protect the
environment (Duffera et al., 2007; Li et al., 2008). The efficiency of the
whole agricultural production can be achieved not only by increasing
resource use efficiency, but also should generate less amount of waste,
improve gross margin, and reduce environmental impacts (National
Research Council, 1997). The MZ concept is different from the traditional
farming system in that it mainly considers the management of within-field
variability aiming at increasing input use efficiency, increasing yield, and also
protecting the environment (Adhikari et al., 2009). In order to improve farm
input use efficiency and enhance yield in environmentally friendly condi-
tions, a new paradigm of farming is required. This has to focus on the
fine-tuning of agricultural production system inputs (e.g., seed, nutrients,
water, pesticide, etc.) based on smaller management units (Adhikari et al.,
2009), established by dividing a field into subunits according to actual
variability in the system. MZ is proven to result in economic efficiency
through reducing the cost of crop production, by improving resources
use efficiency (Koch et al., 2004). This can be achieved by using appropriate
applications of farm management information system (FMIS) (Adhikari
et al., 2009) (Fig. 1), to inform VRA of farm resources based on predictive
models of multiple data sources. Input data include a geospatial database
Economic Optimization Environmental

viability protection
Reduce inputs
Decision
VRA support
system
(DSS)
Predictive
models Improved
control
Site-specific Management
management information
management
system (MIS)
Proximal (ICM) zones (MZs)
sensing
Spatial
database and
GIS, networks
GPS, RS,
UAV
Increased efficiency
Improved gross Less impact on

Less waste
margin environmental
Fig. 1 Components of precision agriculture and the conceptual framework of

site-specific management practice. Adopted from Srivastava, S., 2009. Space Inputs
for Precision Agriculture: Scope for Proto-Type Experiments in the Diverse Indian
Agro-Ecosystems. Indian Space Research Organization, Bangalore, India. http://www.
geospatialworld.net/article/space-inputs-for-precision-agriculture-scope-for-proto-type-
experiments-in-the-diverse-indian-agro-ecosystems/ (accessed 01.06.2016).
acquired for example by proximal soil sensing (PSS) methods (e.g., vis–NIR
spectroscopy), remote sensing, and unmanned aerial vehicle (UAV) images
of canopy and soil, topography, and climatic data. The FMIS includes
decision support systems (DSS), and associated geodatabase of crop, soil,
and climate. The final output is improved farm gross margin, and reduced
farm waste and environmental impacts, which is in line with National
Research Council (1997) and Fig. 1.
The development of proximal soil sensors that could provide fine-scale
data together with increasingly powerful, affordable computers, and running
statistical and GIS software, allow nowadays the use of sophisticated
geostatistical analysis to delineate accurate MZ. In a paper outlining the
practical definition and interpretation of potential MZ, Whelan and
McBratney (2003) suggested five steps to delineate and verify zones
delineated using high-resolution soil and landscape data: (1) gather relevant
data layers, (2) interpolation, (3) clustering, (4) determine the confidence
interval (CI) for zone partitioning, and (5) directed soil sampling to
assess soil-related causes of between-zone variability. This functional
description can be rearranged as (1) mapping properties useful as MZ
proxies, (2) individuation of homogeneous areas (clustering), (3) finding
the optimal number of classes, (4) delineating final MZs, and (5) assessing
the effectiveness of the classification.
2.1 Mapping Properties Useful as MZs Proxies

The first step in MZ creation is mapping of properties involved in the
analysis. Numerous interpolation and approximation methods were
developed to predict values of an attribute at unsampled points (e.g.,
Burrough and McDonnell, 1998; Wackernagel, 2003; Webster and
Oliver, 2007) and provide continuous spatial data which is critical in preci-
sion agriculture (PA). This is also necessary when the available dataset does
not provide a sufficient coverage of the studied area (Burrough and
McDonnell, 1998). In such cases, spatial interpolation methods provide tools
to accomplish such task by estimating the values of an attribute at unsampled
sites using data from the observation points within the same region. How-
ever, the interpolation of data depends on the resolution of sampling points,
which varies depending on the sampling method. Traditional soil and crop
sampling is based on low sample resolution data collected at typically one
composite sample per 1–3 ha. With current advancements in sensing tech-
nologies, observations can now be collected with proximal sensors of differ-
ent technologies (Kuang et al., 2012) or by remote sensing. New proximal
sensors allow the collection of high spatial sampling resolution >1500–2000
readings per ha (Mouazen et al., 2007), enabling the exploration of spatial
variability at fine scale. These two methods of data collection will influence
the requirements of mapping and MZ delineation.
Given the continuous nature of soil variability, point data from soil
samples collected at a 1 sample per 1–3 ha require interpolation. Two
methods of interpolation have commonly been chosen in the research papers
reviewed here, namely, inverse distance weighting (IDW) and kriging. Both
methods primarily depend on the distributional nature of the data, which
can be examined by producing a table of descriptive statistics (Wang
et al., 2009). Generally, this includes the minimum, maximum, mean,
median, standard deviation (SD), and coefficient of variation (CV) for each
property. Often a histogram or skewness and kurtosis values are examined to
indicate if the data have a normal distribution.
If the number of data points is too low to determine spatial

dependency, i.e., less than 100 (Webster and Oliver, 2007) or the data
are not normally distributed, then IDW is generally chosen. This is a deter-
ministic method where unknown point values are weighted according to
their distance from the known values, as expressed by Eq. (1):
X
N
w ðdi Þzi
i¼1
f ðx, yÞ ¼ (1)
XN
w ðdi Þ
i¼1
where w(d) is the weighting function, Zi is the data value point at i, and di is
the distance from point i to (x,y). If the data are skewed positively then func-
tions such as square roots, or log(x) can be applied in order to transform the
data to normality.
With a sufficient number of points and where spatial dependence exists,
it is possible to use a geostatistical approach called kriging. Kriging assumes
that the spatial covariation in the phenomenon is stationary throughout the
surface (e.g., the same pattern of variation can be observed at all locations on
the surface, the mean and variance do not change spatially). The spatial var-
iation is quantified by the semivariogram that can be estimated by the sample
semivariogram calculated from the sample data, as expressed in Eq. (2):
1 X
N ðhÞ
1
γ ðhÞ ¼ fZ ðxi Þ Z ðxi + hÞg2 (2)
2 N ðhÞ i ¼ 1
where Z is a datum at a particular location, h is the distance between ordered

data, and N(h) is the number of paired data at a distance h. Through this
method, the observed spatial autocorrelative structure can be obtained, to
which an authorized semivariogram model is fitted.
Kriging is a more robust method of interpolation than IDW and is com-
monly used as data density increases (e.g., Castrignanò et al., 2012; Mouazen
and Kuang, 2016; Shaddad et al., 2015). A critical advantage of geostatistics
over IDW is the ability to assess the uncertainty in unsampled values, which
can be represented as a map of the probability of exceeding critical values for
soil quality (Castrignanò et al., 2002). The result of uncertainty assessment
can be combined with expert knowledge for decision making in precision
farming MZs (Kitanidis, 1997). There are various types of kriging, i.e.,
simple kriging (SK), ordinary kriging (OK), universal kriging (UK),
indicator kriging, block kriging, regression kriging, factorial kriging (FK),

and cokriging. Of these, OK is most commonly used to determine MZ.
All kriging estimators are variants of the basic Eq. (3), written as follows:
X
N
Z^ ðx0 Þ ¼ μ + λi ðZ ðxi Þ μðx0 ÞÞ (3)
i¼1
where μ is a known stationary mean, assumed to be constant over the whole

domain and calculated as the average of the data (Wackernagel, 2003). The
parameter λi is kriging weight, which is obtained from the variogram model
and depends on the distance between the interpolated point and the neigh-
boring measured point, N is the number of sampled points used to make the
estimation and depends on the size of the search window, and μ(x0) is the
mean of samples within the search window.
The kriging weights are estimated by minimizing the variance, as
follows:
h 2 i
var Z^ ðx0 Þ ¼ E Z^ ðx0 Þ Zðx0 Þ
X
N X N X
N
¼ λi λj C xi , xj + C ðx0 , x0 Þ 2 λi C ðxi , x0 Þ (4)
i¼1 j¼1 i¼1
where Z(x0) is the true value expected at point x0, N represents the number
of observations to be included in the estimation, and C(xi, xj) ¼ Cov[Z(xi),
Z(xj)] (Isaaks and Srivastava, 1989).
2.1.1 Simple Kriging (SK)

The estimation of SK is based on a slightly modified version of Eq. (3),
written as follows:
!
X
N X
N
Z^ ðx0 Þ ¼ λi Z ðxi Þ + 1 λi μ (5)
i¼1 i¼1
where μ is a known stationary mean. The parameter μ is assumed constant

over the whole domain and calculated as the average of the data
(Wackernagel, 2003). SK is used to estimate residuals from this reference
value μ given a priori and is sometimes referred to as “kriging with known
mean” (Wackernagel, 2003). The parameter μ(x0) in Eq. (3) is replaced by
the stationary mean μ in Eq. (5). The number of sampled points used to
make the estimation in Eq. (5) is determined by the range of influence of
the semivariogram (Burrough and McDonnell, 1998). Because SK does not

XN
have a nonbias condition, 1 λ is not necessarily 0, the greater the
i¼1 i
XN
value of 1 λ , the more the estimator will be drawn toward the
i¼1 i
XN
mean, and in general the value of 1 λ increases in relative poorly
i¼1 i
sampled regions (Boufassa and Armstrong, 1989). SK assumes second-order
stationary that is constant mean, variance, and covariance over the domain or
the region of interest (Wackernagel, 2003; Webster and Oliver, 2007).
Because such an assumption is often too restrictive, OK (no a priori mean)
is most often used (Burrough and McDonnell, 1998).
2.1.2 Ordinary Kriging (OK)

OK is one of the most common and basic kriging methods (Meul and Van
Meirvenne, 2003). At an unsampled location Χ 0, Z is estimated by:
X
n
Z*ðx0 Þ ¼ λi Z ðxi Þ (6)
i¼1
where Z*(x0) is the estimated value of the random variables (RV) Z at the
unsampled location x0 and λi are the n weights allocated to the observation
points Z(xi). The weights λi whole to one to guarantee unbiased conditions
and they are found by minimizing the estimation variance. The RV Z(x) can
be divided into a two components, namely, trend m(x) and a residual R(x):
Z ðxÞ ¼ mðxÞ + RðxÞ (7)
OK assumes stationarity of the mean and considers the trend component
m(x) to be a constant, but unknown, value. Nonstationary conditions are
considered by limiting the domain of stationarity to a local neighborhood
and moving it across the study area. The residual component R(x) is
modeled as a stationary RV with zero mean and under the assumption of
intrinsic stationarity, its spatial dependence is given by the semivariance
γ R(h):
1
γ R ðhÞ ¼ E fRðx + hÞ RðxÞg2 (8)
2
Assuming a constant mean m(x), Eq. (8) is equivalent to:
1
γ ðhÞ ¼ E fΖ ðx + hÞ Ζ ðxÞg2 (9)
2
The first major applications of OK in soil studies emerged in the early

1980s (Burgess and Webster, 1980). OK has played a major role in efficiently
predicting and mapping soil properties (e.g., Lopez-Granados et al., 2005;
Meul and Van Meirvenne, 2003; Sumfleth and Duttmann, 2008).
A major limitation of OK is that it requires a large amount of data to define
the spatial autocorrelation and exploit the relationship between environ-
mental variables and soil properties (Meul and Van Meirvenne, 2003).
2.1.3 Universal Kriging (UK)

UK known as the kriging with a trend (KT) (Matheron, 1969). It is an
extension of OK by incorporating the local trend within the neighborhood
search window as a smoothly varying function of the coordinates. UK esti-
mates the trend components within each search neighborhood window and
then performs SK on the corresponding residuals. UK assumes the model to
be written as follows:
Z ðxÞ ¼ mðxÞ + RðxÞ (10)
where m(x) is not constant, but varies smoothly within the local neighbor-
hood, representing a local trend. The trend m(x) is recalculated within each
local neighborhood. This trend component is modeled as a weighted sum of
known functions fi(x) and unknown coefficients ai, and i ¼ 0, …, L (Journel
and Rossi, 1989):
X
L
mðxÞ ¼ ai fi ðxÞ (11)
i¼0
UK has been the commonly used method to accommodate the trend

(sometimes known as changing drift) in a soil variable. UK is a combination
of the standard model of multiple-linear regression (MLR) and the
geostatistical method of OK (Webester, 1994). The intrinsic random func-
tion of order k (IRF-k) has been utilized to suit the varying nature of the
trend in a regionalized soil variable (McBratney et al., 1991). The variable
k represents the order of polynomial trends, where k ¼ 0 means constant
drift, and the IRF-k is equivalent to OK system. If k ¼ 1 it means that we
have linear drift, and where k ¼ 2 refers to having a quadratic drift. But,
where there is no trend but deterministic relationships with some known
or readily available and inexpensive covariates (CLORPT factors, CL as cli-
mate, O as organisms, R as relief, P as parent material, and T as time) or other
easy-to-measure soil variables, UK has been successfully used to predict the

target soil variable (Brus and Heuvelink, 2007; Hu et al., 2004).
2.1.4 Factorial Kriging (FK)

FK is designed to determine the origins of the value of a continuous attribute
(Goovaerts, 1997). It models the experimental semivariogram as a linear
combination of a few basic structure models to represent the different factors
operating at different scales (e.g., local and regional scales). FK can decom-
pose the kriging estimates into different components such as nugget,
short-range, long-range, and trend, and such components could be filtered
in mapping if considered as noise. For example, the nugget component at
sampled points could be filtered to remove discontinuities (peaks) at the
sampled points, while the long-range component could be filtered to
enhance the short-range variability of the attribute. FK assumes that noise
and the underlying signal are additive and that the noise (error) is homosce-
dastic, in which values are the same across all values of the independent
variables.
FK was first introduced to soil science by Wackernagel (1988) and
Goovaerts (1992). The assumption behind FK is that many of the soil vari-
ables have the same communality that enables principal component analysis
(PCA) of the variance–covariance matrices of the variables, which are them-
selves associated with spatial scales (Goovaerts, 1992). A noteworthy issue
with FK is the linearity assumption which is regularly not met by many soil
variables.
2.1.5 SK With Varying Local Means (SKlm)

SKlm is an extension of SK by replacing the stationary mean with known
varying means at each point that depends on the secondary information
(Goovaerts, 1997). If the secondary variable is categorical, the primary local
mean is the mean of the primary variable within a specific category of the
secondary variable. If it is continuous, the primary local mean is a function
of the secondary variable or can be acquired by discretizing it into classes. SK
is then used to produce the weights and estimates.
2.1.6 Kriging With an External Drift (KED)

KED is similar to UK but incorporates the local trend within the neighbor-
hood search window as a linear function of a smoothly varying secondary
variable instead of as a function of the spatial coordinates (Goovaerts,
1997). The trend of the primary variable must be linearly related to that
of the secondary variable. This secondary variable should vary smoothly
in space and is measured at all primary data points and at all points being
estimated.
2.1.7 Cokriging
Cokriging is the multivariate extension of kriging that permits the incorpo-
ration of all the more promptly accessible and inexpensive attributes in the
prediction process. Numerous instances occur in soil survey where the
CLORPT factors are readily available and/or cheap to obtain. For more
efficiency in prediction of the expensive target soil variable, soil forming fac-
tors or remote sensing data can be used in cokriging the target soil
variable-sampled at fewer locations, into dense grid nodes. This is termed
as heterotopic cokriging (HCOK) (Wackernagel, 1995) in comparison with
isotopic cokriging, in which both target variable and covariables should be
available at all sample locations. In the collocated cokriging (CCK), the
covariates are available at all interpolation points, even though the target var-
iable exists at only a few locations. This is regularly the case with using exter-
nal variables such as landform attributes derived from a 3D representation of
a terrain’s surface known as a digital elevation model (DEM) (Odeh et al.,
1995) or remote sensing data for predicting the target soil variable. Incom-
plete heterotopy includes cases where there is some coincidence between
locations of the target variables and the covariables. The latter is the regular
case when other soil covariates are used (McBratney and Webster, 1983).
2.1.8 Principal Component Kriging (PCK)

PCK applies PCA to a few (Nv) secondary variables to generate Nv orthog-
onal or uncorrelated PCA components (Goovaerts, 1997). OK is then
applied to each of the components to get principal component (PC) esti-
mates. This means that Nv semivariograms of PCs are estimated, that each
of the Nv PCs is estimated separately at each locations. The final estimate of
zi at s is then reconstituted as a linear combination of the PC estimates
weighted by their loadings and plus the local attribute mean. In contrast
to cokriging, in PCK only those data locations where all Nv variables of
zi at s are observed can be considered.
2.1.9 Collocated Cokriging (CCK)

CCK is a variant of CK (Goovaerts, 1997). It only uses the single secondary
datum of any given type closest to the point being estimated. Like CK, CCK
can also have several variants like simple collocated cokriging (SCCK), and
ordinary collocated cokriging (OCCK). CCK is proposed to overcome
problems, such as screening effects of samples of the secondary variables close
to or collocated with the point of interest. This situation arises when the
sample densities of the secondary variables are much higher than that of
the primary variable. OCCK is also the preferred method for categorical soft
information.
2.1.10 Probability Kriging (PK)

PK is the cokriging of the indicator data using the rank-order transform as a
secondary variable (Goovaerts, 1997). The indicator data are values of 0 or 1.
The rank-order transform is the standardized ranks that are the rank order of
each datum of the primary variable divided by sample size. Replacing the
values of the primary variable in CK by indicator data and using the
rank-order transform as the secondary variable in CK would result in a
PK estimator.
2.1.11 Disjunctive Kriging (DK)

DK is used for the primary variable that the conventional transformations
(e.g., logarithm or square rooting) cannot yield a near-normal distribution.
In DK, the primary variable is transformed into Hermite polynomials, which
are a series of normally distributed subvariables that are kriged separately.
The resultant estimates are summed to give the DK estimator (Gaus
et al., 2003). DK also gives an estimate of the conditional probability that
an arbitrary variable situated at a point, or its average value over a block
in two-dimensional space, exceeds certain thresholds. DK produces a
nonlinear unbiased, distribution-dependent estimator with the characteris-
tics minimum variance of errors (Burrough and McDonnell, 1998). The
theory of DK and examples of its practical application are described by
Oliver et al. (1996).
2.1.12 Model-Based Kriging (MBK)

MBK was developed by Diggle et al. (1998). This method embeds the linear
kriging methodology within a more general distributional framework that is
characteristically similar to the structure of a generalized linear model.
A Bayesian approach is adopted and implemented via the Markov chain
Monte Carlo (MCMC) methodology, to predict arbitrary functionals of
an unobserved latent process while making an appropriate allowance for
the uncertainty in the modeling process (Moyeed and Papritz, 2002). This
method was further illustrated in Diggle and Ribeiro (2007) and involves
heavy computational demands (Moyeed and Papritz, 2002).
2.2 Individuation of Homogeneous Areas

This step, given the continuous nature of soil variation, is the most challeng-
ing and the one that presents more choices to be made, and usually requires
subjecting the data to some sort of clustering procedure in order to achieve
objectivity. Several techniques have evolved in the last century for
unsupervised machine classification, letting the machine alone decide the
boundaries based on a specific algorithm, and achieving therefore repeatabil-
ity by avoiding human subjectivity.
Given the multivariate nature of data, a possibility is to use PCA, a tech-
nique that compresses the variability of a multivariate dataset into few com-
ponents and identifies correlated attributes (e.g., Dragovic and Onjia, 2006).
Generally the first few PCs explain most of the variance in a target property.
It is often possible to identify the property or group of correlated properties
in each PC, which assists in identifying the most influential factors. Cluster-
ing algorithms can be applied to the PC layers.
One clustering approach is hierarchical clustering (Ruß and Kruse,
2011), which builds classes based on distance connectivity between pairs
of observation (often Euclidean or Mahalanobis distance). Algorithms can
proceed both in an assimilative and a dissimilative way. A recent develop-
ment of this technique applies bootstrapping in order to assess the probability
associated with each class (Suzuki and Shimodaira, 2006).
The k-means clustering algorithm aims at partition n observations into k
clusters, in which each observation belongs to the cluster with the nearest
mean. The software used for this technique often allows for k to be set
by the user or it can be randomly set. Fuzzy sets theory (Zadeh, 1965)
can be applied to k-means clustering to extend the technique. Fuzzy clus-
tering allows each point to be a member of more than one class, defined
by a certain degree of overlapping membership. The degree of membership
is related inversely to the distance of the cluster center and how much weight
is given to the closest center. This method is less sensitive to outliers than
k-means and allows a greater degree of flexibility (Fu et al., 2010;
McBratney and Odeh, 1997; Tagarakis et al., 2013).
An alternative method of grouping property data is to formulate a soil
fertility index (SFI), e.g., a fertility index which summarizes the main vari-
ables which influence soil fertility. This index can be calculated based on the
integration of several physical and chemical soil indicators into a quantitative
value. Examples of methods used to estimate the SFI are PC regression anal-
ysis or CV. When the SFI is calculated, it can be mapped and classified to
delineate MZ based on a classification criterion (Ortega and Santibáñez,

2007). Moral et al. (2011) employed the Rasch model to agglomerate five
soil properties with different units into a uniform analytical framework to get
representative measures of soil fertility potential. Then, kriged estimates
were employed to map soil fertility potential and MZs were delineated using
an equal-size classification. A minimum dataset which sufficiently assesses
the fertility, function, and quality of soil was used to produce a fertility index
for sugarcane production in Brazil (Viscarra Rossel et al., 2010b). In this
study, boosting decision tree rules (Quinlan, 1993) were used to classify soil
into three fertility classes.
More recently, k-means clustering was compared with the
self-organizing map (SOM) as a popular neural network models based on
unsupervised learning (Kohonen, 1995) for delineating MZ maps for VR
N application. Furthermore, a hybrid SOM algorithm in combination with
k-means was compared with k-means in terms of cluster separation and MZ
formation based on data fusion of NDVI and soil parameters. The cluster
centers of the hybrid SOM and k-means algorithm show better separation
of clusters when compared with the standard k-means algorithm (Pantazi
et al., 2015). This recent findings illustrate that there are more opportunities
to improve the quality of MZs delineation based on combination of data
fusion modeling and clustering methods.
2.3 Finding the Optimal Number of Classes

If fuzzy clustering has been used for classification then the optimal number of
classes can be derived from both the fuzziness performance index (FPI), a
measure of the degree to which different classes share membership and nor-
malized classification entropy (NCE), an estimation of the amount of disor-
ganization from a specified number of classes. For k-means clustering, it is
possible to examine the variance reduction (commonly in yield or soil con-
centration) for a range of values of k. The optimum number of classes should
have the highest reduction in variance (Frogbrook and Oliver, 2007) and
may also be chosen to be suboptimal, and rather determined by the purpose
of the MZ delineation. For example, if the aim is VR nutrient application,
the researcher often desires to set a number of application rates (high,
medium, and low) according to the fertility/productivity zones delineated.
Specifying a smaller number of classes can also provide a smaller number of
potential MZs, which are more contiguous and within the spreading
capabilities of farm machinery. Cluster maps using a range of classes can also
be compared to classified maps of yield responses. The measure of agreement
between the two can be tested by a real agreement and kappa coefficient
(Guastaferro et al., 2010).
2.4 Delineating Final MZs

Early research used hand-drawn MZ delineated according to farmer knowl-
edge. This was a subjective but simple and quick method suitable for a small
number of data layers. Actually, the earliest MZ (starting in 1986) in the
United States were delineated using a customized GIS system that generated
electronic maps that were burned onto EPROMs in computers on board
variable fertilizer spreaders. In Europe, early practitioners of PA were
prevented from using this approach because of patent laws, so they resorted
to hand-drawn MZ.
If a clustering map is produced, the boundaries between clusters can form
the potential MZs. Although this may be sufficient to provide final MZ,
many small and irregular clusters can be produced when merging a large
number of high-resolution data layers and as a result of edge effects. These
may be undesirable for practical management purposes and can be removed
by merging classes or by resampling the data with a smoothing function
(Frogbrook and Oliver, 2007).
2.5 Assessing the Effectiveness of the Classification

Effectiveness of the MZ delineation technique can be assessed by several
criteria. The measure of variance reduction of the MZ when compared
to the within-field variance can be calculated (Ortega and Santibáñez,
2007). Alternatively, for VR fertilization nutrients can be applied based
on a recommendation map derived from data associated with the MZ. Then,
a cost–benefit analysis of input–output can be conducted, based on the
nutrients used (input), and yield obtained (output) (Halcro et al., 2013;
Sarangi et al., 2015). If costs for the practical implementation of the delin-
eation methods are known, these can be included in any cost–benefit analysis
for comparison. It is to be hoped that future research into MZ delineation
methods are tested in a practical farming environment in a side-by-side com-
parison with the farmer’s normal approach. The following sections explore
the most common MZ approaches that have been described in the literature
(Table 1).
Table 1 Management Zone (MZ) Approaches, Its Criteria, Techniques for Data Collection, Classification, and the Performance Indicator With
the Economic Assessment
Classification Performance Economic
MZ Target Measurement Method Properties Used Technique Indicator Perfo. A. References
Fertility based Chemical analyses N Ext., P EXt., pH, Threshold Grain quality, Missing Mulla et al.
OM, water content values yields (1992)
Fertility based Chemical analyses pH, OM, nutrient Fuzzy c-means Missing Missing Wang et al.
storage (2009)
Fertility based Chemical analyses OM, nutrient storage, Fuzzy c-means Missing Missing Yao et al.
salinity, alkalinity, (2014)
water, yields
Soil properties Soil survey maps Available soil maps Soil survey N efficiency Missing Khosla and
Alley (1999)
Soil properties LIDAR, NO3-N maps Available soil maps Soil survey NA Missing Franzen et al.
(2002)
Soil properties Soil survey profiles Available soil maps Model NA Missing Van Alphen
and
Stoorvogel
(1999)
Soil properties Soil survey profiles, yields Available soil surveys, Supervised FPI, NCE, Missing Brock et al.
+ fertility based yields fuzzy c-means agreement (2005)
between
classifications
Soil Landscape attributes DEM Fuzzy c-means Variance Missing MacMillan
geomorophology explained et al. (2000)
Soil Landscape attributes, DEM, bulk density, NA Variance Missing Nolan et al.
geomorophology chemical analyses OM, clay, water explained (2000)
content
Fertility based Yield sensor Yields Correlation Accuracy of Missing Lamb et al.
predictions (1997)
Fertility based Yield sensor Yields Various Accuracy of Present Milne et al.
k-means predictions (2012)
clustering
Fertility based Farmer knowledge, Aerial photography, Ward’s Accuracy of Missing Fleming et al.
remote sensing, chemical OM, nutrient storage clustering predictions (2000a)
analyses
Fertility based Farmer knowledge, Aerial photography, NA Accuracy of Missing Fleming et al.
remote sensing, chemical OM, nutrient storage predictions (2000b)
analyses
Fertility based Farmer knowledge, Aerial photography Farmer Economic Present Koch et al.
remote sensing knowledge performances (2004)
Fertility based Farmer knowledge, Aerial photography, Farmer Accuracy of Missing Mzuku et al.
Remote sensing soil physical properties knowledge predictions (2005)
Fertility based Remote sensing, Aerial photography Farmer Yields Missing Hornung
harvesting, chemical yields, OM, CEC, knowledge, et al. (2006)
analyses texture k-means
clustering
Continued
Table 1 Management Zone (MZ) Approaches, Its Criteria, Techniques for Data Collection, Classification, and the Performance Indicator With
the Economic Assessment—cont’d
Classification Performance Economic
MZ Target Measurement Method Properties Used Technique Indicator Perfo. A. References
Fertility based Farmer knowledge, Aerial photography, Farmer Yields Present Khosla et al.
harvesting, chemical yields, OM, CEC, knowledge, (2008)
analyses texture k-means
clustering
Soil properties Electrical measurements ECa NA NA Missing Sudduth
et al. (1995)
Soil properties Electrical measurements, ECa, OM, nutrient Unsupervised Accuracy of Missing Johnson et al.
chemical analyses storage, salinity clustering predictions (2001)
Soil properties Electrical measurements, ECa, aerial Farmer Accuracy of Missing Fleming et al.
+ fertility based farmer knowledge, photography, farmer knowledge, predictions (2004)
chemical analyses knowledge, nutrient k-means
storage clustering
Soil properties Electrical measurements, ECa, yields, DEM Unsupervised Accuracy of Missing Fraisse et al.
+ fertility based harvesting, remote sensing clustering predictions (2001)
Soil properties Electrical measurements, ECa, yields Unsupervised Accuracy of Missing Johnson et al.
+ fertility based yield sensor clustering predictions (2003)
Soil properties Electrical measurements, ECa, yields Unsupervised Accuracy of Missing Kitchen et al.
+ fertility based yield fuzzy c-means predictions (2005)
Soil properties Electrical measurements, ECa, yields, DEM, k-means Accuracy of Missing Frogbrook
harvesting, remote sensing, OM, nutrient storage clustering predictions and Oliver
chemical analyses (2007)
Soil properties Electrical measurements, ECa, yields Fuzzy c-means Accuracy of Missing Peralta et al.
yield sensor predictions (2015)
Soil properties Remote sensing, electrical ECa, nutrient storage, NA Economic Present Godwin
+ fertility based measurements, yield sensor NDVI effectiveness et al. (2003)
Soil properties Remote sensing, yield Aerial NDVI and k-means Accuracy of Missing Inman et al.
+ fertility based sensor photography, yields clustering predictions (2008)
Soil properties Remote sensing, electrical Aerial NDVI, ECa, soil Fuzzy c-means Accuracy of Missing Tagarakis
+ fertility based measurements, chemical properties predictions et al. (2013)
analyses
Soil properties Spectroscopy and remote Vis–NIR spectra, Boosting Accuracy of Missing Viscarra
+ fertility based sensing DEM decision trees predictions Rossel et al.
(2010b)
Soil properties Spectroscopy and NDVI Vis–NIR spectra, k-means Accuracy of Present Halcro et al.
+ fertility based sensor NDVI clustering predictions (2013)
Perfo. A., performance analysis; N Ext., nitrogen extract; P EXt., phosphorus extract; OM, organic matter; LIDAR, light detection and ranging; CEC, cation exchange
capacity; ECa, apparent electrical conductivity; NDVI, normalize difference vegetation index; DEM, digital elevation model.
The key conclusions of information in Table 1 are the following:

1) Techniques for delineating MZs are mostly based on a single soil or crop
property or a combination of several properties that are known to affect
crop productivity and yield (Khosla et al., 2010). These can be divided
into three main approaches. The first approach is based on soil and/or
relief information, derived from topographic maps, direct soil sampling,
noninvasive soil sampling by EMI or electrical resistivity equipment, and
soil organic matter or crop canopy characteristics measured by remote
sensing (Ortega and Santibáñez, 2007). The second approach is based
on yield maps, combining data from several seasons (e.g., historic data),
while the third approach is the integration of the two previous
approaches and considers soil and/or relief information plus the use of
yield maps.
2) Although farmer knowledge-based MZs appear to be effective in iden-
tifying different MZs, ground verification is needed to develop accurate
VRA maps from the zones. The need for confirmation of specific soil
characteristics in MZ delineation was reported also by King et al.
(2005), based on the analysis of resulting yield map. Authors concluded
that EMI measurement may well be a cost-effective method for identi-
fying and mapping within-field MZ.
3) Soil and crop sensing have been used extensively for delineation of MZs.
The most common soil attribute produced by proximal sensing is soil
ECa based on EMI sensors. NDVI is the most common crop parameter
used in MZs delineation, which can be derived from satellite, airplane,
or UAV imagery, or can be created using commercial optical crop sen-
sors such as Crop Circle (Holland Scientific & AgLeader), Yara
N-Sensor ALS (Active Light Source) (Yara & agricon), and Gre-
enSeeker (N-Tech & Trimble).
4) The most common procedure that has been used for clustering analysis is
k-means or fuzzy k-means method (Minasny and McBratney, 2002;
Ortega and Santibáñez, 2007). However, as stated earlier, the estimation
of optimal number of clusters can be defined by FPI, NCE (Brock et al.,
2005; Li et al., 2008), and/or the previous knowledge of the farmer
(Galambošová et al., 2014).
5) Cost–benefit analysis is missing from majority of these studies and few
studies conducted a cost–benefit analysis for evaluation of the perfor-
mance of MZs (e.g., Halcro et al., 2013; Khosla et al., 2008; Koch
et al., 2004). For example, in order to confirm precision application
of VR N fertilization, a cost–benefit analysis comparing UR and VR

was performed (Halcro et al., 2013). More information about this
study is provided in the case study presented in this review. Cost–benefit
analysis is essential to convince farmers and growers to adopt VR
technologies, otherwise it is doubtful that farmers will invest in technol-
ogies for implementation of precision agriculture.
3. MZ DELINEATION APPROACHES
3.1 MZ Delineation Based on Farmer Knowledge
Farmers and growers are in a position to know the past production history of
their fields and are familiar with areas, which consistently produce higher
and lower yields. There have been many approaches, which used farmer
knowledge as one layer of data input for MZ delineation. For example,
Fleming et al. (2000a) evaluated farmer-defined MZ maps for VRA of fer-
tilizer. The paper proposed using farmer’s knowledge, as an alternative to
clustering based on grid sampling, by encouraging farmers to draw vector
lines on gray-scale soil photographs to delineate zones of high, medium
and low productivity. These divisions were based primarily on soil color
and producer knowledge of the topography and management history.
The knowledge-based zones were compared to the spatial variation of
yield-limiting soil factors (texture, OM, ECa pH, P, zinc (Zn), K, and N)
and previous years yield. Farmer-defined MZs were mostly in agreement
with MZ delineated by clustering based on soil properties, but presented also
some inconsistencies. A zone classed as medium fertility by the farmer actu-
ally contained the highest levels of yield-limiting soil and crop factors. The
authors conclude that additional ground truthing is needed before using
these zones for VRA.
Subsequently, an evaluation of farmer-delineated MZ and grid soil sam-
pling was conducted for VR nitrogen application alongside UR treatments
by Fleming et al. (2000b). The farmer-delineated zones of high productivity
were those containing high levels of OM and clay, which are associated with
higher water holding capacity (WHC). The authors found that the MZ and
grid-sampling approach were not significantly different concerning the
overall yield. However, MZ creation based on farmer knowledge was a
much cheaper approach than grid sampling and it was recommended as via-
ble alternative.
A comparison of VR N application strategies was investigated by Khosla

et al. (2002). The aim was to calculate the agricultural efficiency (AE) of
applying more fertilizer to high productivity zones and less to low produc-
tivity zones. Comparison for N requirement was made between zones
defined according to the farmer knowledge of own land, nitrogen levels
from analysis of grid soil sampling, and a “yield goal” calculated by a semi-
empiric equation considering soil OM, nitrate, and expected (potential)
yields. The results showed that in one of the two sites studied, VR
N management through site-specific farmer-defined MZ led to increased
yield. This was achieved while reducing the total N fertilizer applied when
compared to grid-sampling and yield-goal (potential yield) approaches. The
authors reported no significant yield difference between the high- and
medium-fertility MZ in the second study site. They suggested that
N may not have been a prime yield-limiting factor here and plan to develop
other techniques of MZ delineation on the site.
In a continuation of the research into the effectiveness of farmer-based
MZ delineation a study on the economic feasibility of four N application
strategies was conducted over 3 years on the same fields (Koch et al.,
2004). The results showed that VR N application based on a separate yield
goal for each MZ was significantly more cost effective than the grid-based
strategy due to the unfeasible costs associated with grid sampling and high
N fertilizer use. It provided additional net returns from 18.21 to 29.57
USD$ per ha over the uniform N strategy. This result could be further
improved if cultivation and fertilizer applications were conducted in one
operation. Farm profitability can be improved by VR over UR
N application and the authors suggest that it could also be more environ-
mentally friendly and sustainable due to the increased N-use efficiency.
Generally, farmer knowledge-based MZ approach was found to have advan-
tages of cost and speed over the traditional soil sampling-based delineation
approaches. However, they do not provide growers with information about
the variability of the soil properties, which may be limiting the yield.
Mzuku et al. (2005) examined the spatial variability of soil physical prop-
erties (e.g., bulk density (BD), organic carbon (OC), sand, silt, porosity, and
moisture content (MC)) across site-specific farmer knowledge-based MZ
produced from the commercially available AgriTrak Professional software
(Fleming et al., 2000a), which works by combining aerial photography
and farmer knowledge about site topography and fertility. The authors
found that the MZ contained significant differences in several measured soil
physical properties. The high fertility MZs were consistently separable from
the low fertility MZs. Soil property trends generally followed the produc-
tivity potential of the MZ delineated using bare-soil color, farmer percep-
tion of topography, and the farmer’s past crop and soil management
experience. The authors concluded that low and high productivity areas
can be reliably separated using farmer-based determined MZ.
Potential improvements in the performance of farmer-based MZ, made
by including soil properties and additional data layers such as a satellite
imagery, topographic properties, and the previous season’s yield map, were
subsequently assessed (Hornung et al., 2006). Farmer-based MZs were cre-
ated by AgriTrak Professional software as previously described. This first
technique, soil color-based management zone (SCMZ) relied on
bare-soil imagery, topography, and farmer’s experience. The second tech-
nique, designated as yield-based MZ (YBMZ), involved delineating zones
in GIS using the same layers as SCMZ plus kriged data layers of OM, CEC,
clay, sand, and silt from grid soil sampling and the previous year’s yield
map. All property maps were resampled to the same resolution of a
10 m grid. Classification was achieved through k-means clustering, pro-
ducing three classes. The resulting clusters were smoothed to reflect the
capabilities of the fertilizer applicator. Nitrate fertilizer was applied in three
rates: one was derived from a yield-goal approach, the second was half this
recommendation and the final was a control of zero fertilizer. Results from
the 3-year experiment showed that the SCMZ performed relatively better
than the YBMZ technique in terms of correctly classifying grain-yield
potential for each MZ. These researchers suggested that the relatively poor
performance of the YBMZ technique may be attributed to the lack of
weights assigned to the different data layers. They proposed further inves-
tigations into weighting input factors according to their influence in the
MZ delineation process in order to improve accuracy. The smoothing
used to reflect the capabilities of the spreader also introduced inaccuracies.
An economic comparison of these two approaches can be found in Khosla
et al. (2008). Neither of these approaches performed well in terms of areal
agreement with the three zones produced from clustering the yield data.
This suggests that more innovative approaches to characterize fertility var-
iation should be investigated. More recent study, Heijting et al. (2011)
suggested the utilization of farmer knowledge for the determination of
MZ as a starting point for mapping field variation. An integration of such
knowledge with other techniques seems therefore a viable option.
3.2 MZ Delineation Based on Geomorphology

Topographical variations have been utilized extensively for inferring soil
classification from local geomorphology and have a recognized influence
on soil fertility. Elevation maps or DEMs enabled early researchers to
derive a wide range of landform properties. MacMillan et al. (2000)
employed DEMs to produce a fuzzy classification of 15 landform catego-
ries through a landform segmentation model, which attempted to explain
the significant differences in soil properties and management requirements
over a Canadian prairie landscape. Simplifying the 15 categories into three
generalized classes resulted in spatially coherent zones, which explained
meaningful differences in soil properties (e.g., OM, pH, and soil depth)
and wheat yield. They concluded that this automated method was well
suited for defining management units for precision agriculture over a wide
variety of landforms.
Four landscape-based MZ delineation methods were compared on
strongly rolling topography by Nolan et al. (2000). In the first MZ delin-
eation approach, the farmer, who had grown crops on the study site for
30 years, was asked to draw areas which should be managed differently.
The farmer knowledge-based classes consisted of “knolls,” “steep side
slopes,” “bottom areas,” and “other areas.” The second MZ delineation
approach was to use only elevation data to classify the field into “high,”
“medium,” “low,” and “very low” classes. Two further terrain-based
modeling approaches were used for this comparison. Model B was based
on four landscape classes, namely, “shoulder-slopes,” “back-slopes,”
“foot-slopes,” and “level.” Model A refined the previous terrain-based
model by including relative elevation data. The classes derived were
“upper,” “mid,” “lower,” and “depression.” The grid size was 10 m2
across a 81 ha field. Performances of the four techniques were measured
according to their ability to separate yield, BD, OM, clay, and WHC var-
iations within the two cropping years. There were mixed results from
examining how the MZ characterized the soil property variations. Each
of the four schemes was best at explaining different properties. The man-
agement units delineated by “Model A” explained 51% and 35% of the
yield variation in the two study years, while farmer-delineated zones were
the next best, explaining 37% and 22%. The authors conclude that utility of
these landscape-based models may vary at different sites and from year to
year. Again, landscape attributes alone seem unable to account for the
many factors affecting fertility (e.g., pH, OC, clay, and MC). Nevertheless,
the wide availability of DEM data and the diffusion of laser sensors make
geomorphometry, now seen as a scientific discipline in its own (Pike et al.,
2009), a potentially valuable tool for improving the precision in the
definition of MZ when utilized together with other data.
3.3 MZ Delineation Based on Traditional Soil Chemical

Analyses
The major limitation of MZ delineation based on traditional laboratory
analyses, is the prohibitive cost of relying on large number of samples
needed to explore within-field variability at proper spatial or temporal
scales. Early MZ delineation approaches were limited to using properties
from readily available data sources. Commonly these were
low-resolution soil surveys, topographical information, aerial photo-
graphs, and farmer knowledge. Mulla et al. (1992) employed the results
of laboratory chemical analysis of soil samples, collected at approximately
6 ha for the delineation of fertility-based MZ of winter wheat. They
determined soil fertility as a combination of available nitrogen (N),
extractable phosphorus (PEXT), and available water in soil and produced
three fertility-based MZs. Suitable application rates of nitrate and phos-
phate fertilizers were chosen for each MZ. Grain quality was found to be
significantly improved in the VR strips, as compared to UR strips. How-
ever, there were no significant differences in wheat yield between the
UR and VR strips even though two of the three VR zones received
51 kg N ha1 less than the adjacent UR strip. This early study indicated
that NUE can be increased by applying fertilizer according to the
requirements of the soil rather than “blanket” coverage. This study
pointed out therefore the effectiveness of MZ-based VR fertilization,
although not proving the economic viability of the method.
In more recent times, soil fertility-based MZ approach was
implemented in a Chinese tobacco field (Wang et al., 2009) based on
the analyses of 81 samples aligned on a 100 m grid for pH, OM, total nitro-
gen (TN), alkalytic nitrogen (AN), available phosphorus (PAV), available
potassium (KAV), and CEC. The spatial distribution of these soil properties
was mapped by kriging interpolation. Since many of these yield-limiting
factors (e.g., soil properties) were correlated, PCA was used to simplify the
variability. The first two PC scores were subjected to a fuzzy c-means clus-
tering algorithm, a soft clustering algorithm (McBratney and Odeh, 1997;
Panda et al., 2012). The optimal number of classes was obtained by
minimizing FPI and NCE. Three soil fertility classes were found to be suf-
ficient. The results showed that among the three MZs, there was a statis-
tically significant difference (P < 0.05) in levels of pH, OM, PAV, KAV, and
CEC. No cost–benefit analysis was performed and authors did not attempt
to examine how well the MZ reflected actual productivity levels by com-
paring them with historic yield variation. Authors recommended includ-
ing within-season growth variations during MZ delineation in order to
improve N fertilizer application.
A slightly different set of soil properties and clustering technique was
chosen by Fu et al. (2010), where 100 soil samples from a 25 ha paddy field
were analyzed for TN, available N (NAV), total phosphorus (PTOT), PAV,
OM, and KAV. OK and ArcGIS 9.0 software were used to interpolate the
spatial variation of unsampled areas and map each property. Fuzzy clustering
analysis optimized by PSO was conducted in order to find the best number
of classes. The rational partition number was found to be two classes. This
figure was determined by applying two performance indices, namely, sep-
aration coefficient and separation entropy on a range of possible divisions
(2–6). Effectiveness of MZ was tested by examining the single property var-
iances within and between the two delineated zones. All properties except
PAV were found to be very significantly different. This result implies that
spatial variability of soil nutrients can be characterized by this approach
and the fertility zones could be used for VR fertilizer application, which
was not considered.
More recently a similar approach was utilized to delineate site-specific
MZ (SSMZ) by linking yields with different soil properties, namely, OM,
nutrient storage, salinity and alkalinity, water retention, and permeation
(Yao et al., 2014). Soil properties were determined by grid sampling and sub-
sequent laboratory analyses of the selected properties, and soil MZs were
determined by fuzzy c-means clustering (defined in this paper as fuzzy
k-means) applied on the first four PCs. The optimal number of classes was
again determined by minimization of FPI and MPE and was found to be
two classes. The selected four PCs explained 90% of the recorded variance,
and it was possible to distinguish them as “organic nutrient supply component”
(PC1, 24.88% variance), “soil compaction component” (PC2, 21.38% variance),
“mineral nutrient supply component” (PC3, 17.42% variance), and “soil saliniza-
tion component” (PC4, 13.63% variance). The resulting MZs were found to
reflect the rotation systems already applied in the fields, confirming the suit-
ability of the approach. However, also in this case no performance indicator
was proposed, and in particular no economic evaluation was considered.
3.4 MZ Delineation Based on Soil Class

Other early sources of information on the spatial variability of soil are soil
classification maps. Khosla and Alley (1999) utilized a georeferenced soil
map of a 36 acre test field, based on a coarse (order II) USDA soil survey
for site-specific N management. The authors hypothesized that crop
response to N depends on soil textural class and WHC, hence soil maps
can be used for optimization of N application. Soil type boundaries were
used to delineate four MZ, and nitrate fertilizer was applied at four different
rates according to the potential yield for each soil type. Total amount of fer-
tilizer used was reduced to 13% by adopting this approach. Among the four
classes, only the grain yield on the less fertile soils was less than the yield goal.
In the entire remaining three classes, grain yield exceeded the yield goal.
Overall N-use efficiency was increased from 1.2:1 pounds of N per bushel
of grain produced to a ratio of 1:1.
Franzen et al. (2002) examined delineating site-specific N MZs by
directed soil sampling based on topographical data and compared these with
MZ from coarse (order II) and fine (order I) USDA soil survey maps. MZ
delineated from order I maps were comparable with MZ delineated by
direct soil analysis, whereas MZ from order II maps differed often substan-
tially. This study pointed out some shortcomings and possible risks associated
with MZ delineation from soil maps.
Soil profile classification in addition to soil samples analyzed for N, tex-
ture, OM, and lime, were used also in a model-based approach to define
management units (van Alphen and Stoorvogel, 1999). Their model consid-
ered soil, water, and plant interactions in two study fields, taking account of
standard N application, simulated yield, known N leaching, and residual N.
Three classes were delineated using Jenks’ natural breaks to minimize class
variance. Management units were delineated using three classes and three
levels for each indicator variable. This method would have been problem-
atical to use for actual VRA, since the zones produced were noncontiguous,
irregularly shaped and some were quite rather small to match the speed of
response from VR applicators.
In a later work comparing yield-based MZ to order II soil maps Brock
et al. (2005) concluded that soil maps alone were insufficient to reliably iden-
tify crop productivity MZ. A factor in this inference was that within-field
topographical variations can influence yields on soil of the same series. It
may be concluded that soil maps alone do not contain enough information
to characterize soil variation sufficiently well at field scale.
3.5 MZ Delineation Based on Yield Maps

In precision agriculture, it is generally recognized that the spatial variation in
yield can reflect variation in within-field soil fertility or more often MC. The
situation is complicated concerning temporal variation of yields, which
might be really high from year to year, due to, e.g., differences in weather
and not necessarily correlated to spatial variation of soil properties. An inves-
tigation by Lamb et al. (1997) showed that knowledge of previous years yield
could only account for 4%–42% of the grain yield variability for a following
year. They concluded that to account for the complex patterns in yield var-
iability at least 5 years of yield data should be used to delineate MZ.
The advent of yield sensors on combined harvesters provided a cheap and
readily available source of yield data to researchers, allowing for more data,
and improving the effectiveness of yields as a mean to characterize MZ. In
the previously described paper, Brock et al. (2005) derived fertility-based
MZ from high-resolution multiyear yield data in order to deal with
year-to-year variations from climate, pests, and disease to produce stable
zones. More recently, Milne et al. (2012) investigated several possible clus-
tering approaches based on past yields data, selecting soft (fuzzy) methods as
the most suitable. Nevertheless, they found little difference between the
nitrogen response function of the different zones, concluding that a classi-
fication of MZ based on yields alone is not enough for being economically
profitable, and suggested to utilize additional, soil-related parameters.
Given the availability of yield data, nowadays the majority of recent
research uses yield data as an additional layer of information to characterize
fertility variation. There is also a huge potential for development of remote
sensing techniques to provide high-resolution data on yields (e.g., by
predicting yield potential) and crop canopy characteristics, for example air-
borne sensors in combination with UAV (Link et al., 2013). These data will
be soon cheap (e.g., free imagery from recently launched Sentinel 2) to col-
lect and potentially extremely valuable for being integrated in MZ delinea-
tion. Yield data at various resolutions are also among the most common
agronomic data recorded. Although yield data alone are not directly suitable
for determination of MZ, their availability and low cost makes them a valu-
able possibility for improving the effectiveness of MZ delineation based on
other information.
3.6 MZ Delineation Based on Crop Coverage

Information about crop cover indicated as NDVI or leaf area index (LAI)
can be collected by means of ground-based vehicle mounted proximal
sensors (e.g., CropCircle, Yara, and others), UAV or by satellite remote

sensing imagery. It is calculated using the following equation:
ðλNIR λR Þ
NDVI ¼ (12)
ðλNIR + λR Þ
where λNIR is the reflectance from NIR wavelengths (760 nm) and λR is the
reflectance from red wavelengths (680 nm).
MZ delineation based on several landscape attributes including soil color,
elevation, and EC was reported by Schepers et al. (2004). However, the
authors observed significant temporal changes in yield spatial patterns even
in irrigated fields. They concluded that use of these MZ in, for example,
VRA of N would only have been appropriate in three out of five seasons.
In addition, the use of the static soil-based MZ concept that relies on aggre-
gation of the measured landscape attributes is unlikely to be adequate for
VRAs across years exhibiting temporal variability. They suggested that an
alternative strategy would be to combine MZ with crop-based, in-season
remote sensing systems. This would provide crop data, which reflect the
current climatic conditions, soil N supply, and make VRAs such as N, more
effective. VRA of N fertilizer based on calculations of shoot density, or
NDVI (Eq. 12), from aerial photographs have shown promising results
(Godwin et al., 2003). When most fertilizer was applied to areas of lower
shoot density (low NDVI), this resulted in measurable economic benefits.
Inman et al. (2008) used yield data to compare SCMZ with those derived
by adding NDVI data from remotely sensed aerial imagery. The results
showed that early season NDVI had the potential to be useful for manage-
ment of irrigated maize, since it had a stronger areal agreement than SCMZ.
NDVI was found to explain 25%–82% of the grain yield variability in the six
sites (three irrigated fields were planted with three different maize hybrids),
though this was inconsistent. In general, the authors found no compelling
evidence that including NDVI data would improve their usual SCMZ
approach. No other crops were tested in this research.
A one hectare commercial vineyard field in Greece was used to evaluate
MZ delineated for VR irrigation and fertilizer application (Tagarakis et al.,
2013). ECa data were gathered by using an EM38 instrument in vertical
mode using a 4 m transect separation. Elevation was provided by a
Real-Time Kinematic GPS (RTK-GPS) survey. Crop canopy levels
(NDVI) were supplied by a CropCircle ACS210 system (Holland Scientific,
USA) and were measured on five occasions during the season. Yield was
measured by weighing the grape harvest at points logged with a GPS. Grape
quality was measured by an electronic refractometer. Data for each property

was interpolated by kriging to a 5 5 m2 grid. Parameters that were not
“normally” distributed were subjected to logarithmic transformation
beforehand. Spherical and exponential semivarigram models were found
to best fit the spatial variation of the data. Fuzzy clustering was used to group
the data into classes. The number of classes was determined by the FPI and
NCE. The optimal number of classes was calculated to be 2 or 3. The results
showed that a combination of ECa and elevation data correlated well with
canopy properties and yield and these properties were used to delineate
MZ maps.
In recent years, the availability and price of acquiring satellite imagery
have made it an attractive source for soil and crop cover data. Li et al.
(2007) used NDVI data calculated from a SPOT5 satellite image of a
10.5 ha cotton field, as a layer for MZ delineation. NDVI and four soil prop-
erties including EC, OM, TN, and CEC were found to be the main con-
tributing variables in two PCs which explained the majority of the yield
variance. Fuzzy c-means clustering of the PCs, followed by FPI and
NCE analyses, led to three MZs, which could characterize the spatial var-
iation of soil chemical properties and crop productivity. More recently,
Chang et al. (2014) followed a similar approach using a GreenSeeker hand-
held ground-based sensor to determine MZ for a tobacco field in China.
They used a 20-m grid-sampling scheme with 101 points on the field and
collected soil samples from the top 20 cm at each point before tobacco plant-
ing. The NDVI data were measured at five growth stages of the tobacco
growth cycle at the location of each sample point. Yield mapping was carried
out at the harvest. Using stepwise multiple analysis, they identified the key
yield-limiting factors as either soil properties or NDVI measured at different
growth stages. Soil properties such as OM, AP, and Fe were used for
yield-based MZ delineation, in comparison with MZ delineated using can-
opy parameters as indicated by NDVI during knee-high and flowering
stages. Fuzzy c-means clustering was performed in order to determine the
optimal number of the clusters. The authors found that five MZs were opti-
mal for either of the two methods. A variance analysis was used to assess the
effectiveness of both methods to characterize the spatial variation in soil
properties and crop productivity. The results of the variance analysis indi-
cated the heterogeneity of soil properties and yield among the different
MZs. They concluded that it is feasible to use an active canopy sensor to
delineate MZs for tobacco-planting fields.
In comparison to satellite imagery, UAV acquires images at higher tem-

poral (e.g., daily acquisitions) and spatial resolutions (e.g., centimeters),
which allows innovative applications in PA (Primicerio et al., 2012;
Zhang and Kovacs, 2012). The restrictions known for applications of remote
sensing systems in farm management include: (a) the gathering and delivery
of images in an exceedingly timely manner, (b) the shortage of high spatial
resolution, image interpretation, and data extraction issues, and (c) the com-
bination of those data with agronomic knowledge into expert systems
(Du et al., 2008). Unfortunately, the remote sensing data most readily avail-
able are the medium resolution satellite imagery (e.g., LandsatTM, ASTER,
SPOT5), which are only useful for large-scale studies. While Sentinel-2 is
characterized by a high spatial resolution (10, 20, and 60 m), the revisit
period is limited to 10 days with 1 satellite and 5 days with 2 satellites. More-
over, even the newest higher resolution satellite imagery (e.g.,
WorldView-2 & 3 and GeoEye-1) cannot provide high frequency data
for emergent situation (e.g., nutrient stress monitoring, disease) with a lim-
ited 1–2-day revisit period. Furthermore, acquiring a useful satellite image
requires cloud free days throughout the growing season.
Finally, it is suggested (Griffin et al., 2004) that one main reason for the
low adoption rate of remote sensing images in PA is the unavailability of
reliable economic estimates of return from the application of remotely
sensed images. It is believed by some (Lamb et al., 2008; Stafford, 2000) that
the limitations of remotely sensed imagery from satellites such as cost, avail-
ability, and processing have made their applications prohibiting and thus
nonpractical for small areas (e.g., field scale). Consequently, UAVs could
be an inexpensive and more practical substitute for satellite and general avi-
ation aircraft for high-resolution remotely sensed data. Moreover, UAVs are
useful tools for remote sensing scientists and farmers (Gago et al., 2015). In
recent years, small commercial UAVs (<50 kg) (Laliberte and Rango, 2011;
Rokhmana, 2015) have been available for environmental and PA applica-
tions. Images captured using UAV are characterized by a very high spatial
resolution of centimeters and cost effectiveness allowing for higher monitor-
ing frequencies. Therefore, UAV acquired images could be a practical alter-
native to aircraft aerial photos and high-resolution satellite images (Gago
et al., 2015). Although UAVs have not been used broadly in the PA, its
applications in MZ delineation are becoming increasingly more apparent
in the literature (Magalhães et al., 2013). Such studies may provide insight
as to how useful these instruments may be for various MZ endeavors.
3.7 MZ Delineation Based on Proximal Soil Sensors

and Data Fusion
Traditionally, soil conditions of agricultural fields are characterized by a
limited number of samples being pooled into one “representative” sample,
on which a number of soil properties are analyzed in the laboratory using
traditional wet chemistry. A similar single-value approach is followed to
characterize other growing conditions and to determine crop characteristics
and yield. Increasing the number of samples and analyze them individually to
characterize the within-field variability is often prohibitively laborious and
expensive. To overcome the bottleneck of data acquisition by traditional
means, soil inventory has evolved toward PSS. This is according to the
paradigm “Measure more, less precise.” Or in other words, bring the laboratory
measurements into the field to increase the spatial coverage, but at the cost of
measurement precision.
The increased need for precise data and rapid methods for better soil
management promoted the development of sensors for soil measurement
to complement or replace more conventional laboratory methods
(Viscarra Rossel et al., 2010a,b). PSS has gained more attention in PA, some-
times for delineation of MZ. PSS provides rapid, inexpensive, and
high-resolution soil data that can be used to better understand and quantify
the spatial and temporal variability within a field (Kuang et al., 2012), which
is anticipated to lead to more precise delineation of MZ. Recently, PSS in
PA has been developed utilizing a wide range of sensor technologies (Kuang
et al., 2012), which include single, multisensor platforms, and wireless
sensors networks, in order to fully control farm resources site specifically
as illustrated in Fig. 2. In the next section, we further describe the soil
sampling aspects and the application of PSS data fusion techniques in
delineating MZs.
3.7.1 Sensor Sampling for MZ Delineation

Sampling for high-resolution data based on PSS can be divided into two
sampling methods, e.g., sensor sampling and calibration sampling
(de Gruijter et al., 2010). The former is required if there is limited informa-
tion, where PSS can produce high-resolution data. This type of sampling
aims to identify spatial variability with high sampling resolution over the
studied area. Calibration sampling is used to produce a predictive model
(e.g., kriging interpolation) over the studied area. Typically, sensor sampling
is taken at fixed intervals using a vehicle while driving along straight parallel
Fig. 2 Remote and proximal sensors platforms and location of sensors in PA. Source:
DFG, German Research Foundation, 2014. Long-Term Perspectives and Infrastructure in
Terrestrial Research in Germany – A Systemic Approach. Strategy Paper. German
Research Foundation, Bonn, Germany. http://www.dfg.de/download/pdf/dfg_im_profil/
gremien/senat/agraroekosystemforschung/strategiepapier_infrastruktur_en.pdf (accessed
10.03.2016).
lines, thus resulting in a regular grid of sample points. The structure of the
grid sampling (shape and size) should be defined as it will be used for
predicting the target variable, which is subjected to a cost constraint. In cases
where there is a multisensor mounted on a vehicle, the data collected are
subjected to postprocessing to transform data into numerical values.
Ultimately, PSS produces fine-resolution spatial data that can be
obtained using online (real-time) measurements (Adamchuk et al., 2011)
at relatively low cost (McBratney et al., 2005), revealing detailed spatial
patterns of measured parameters (e.g., electrical, optical, mechanical, elec-
trochemical, acoustic, and pneumatic) (Adamchuk et al., 2004; Kuang
et al., 2012). In this context, EMI sensors that measure variations in ECa
across the field indicate variations in soil MC and clay content (CC), and
diverse other properties (Abdu et al., 2008; Hossain et al., 2010; Peralta
and Costa, 2013; Saey et al., 2009; Sun et al., 2011). Therefore, ECa has
been defended as a proxy for soil CC (Chen et al., 2004) and as a primary
variable to characterize soils (Meirvenne et al., 2013). Furthermore, ECa has
been identified as a key variable describing field variation for the delineation
of MZ (Meirvenne et al., 2013). However, EMI and electrical resistivity
sensors are reported to be of limited use for measuring soil chemical and
fertility parameters, e.g., OM, total nitrogen, and CEC (Kuang et al.,
2012). Among PSS technologies, vis–NIR is one of the most promising
technologies to quantify the spatial variability in key soil chemical and
fertility parameters (Kuang et al., 2012; Stenberg et al., 2010) and can be
successfully used to provide data for delineation of MZs for VR fertilization
(Halcro et al., 2013). In addition, other PSS technologies, e.g., ground
penetration radar (GPR) and gamma ray have been reported recently in
the literature for successful delineation of MZs, although EMI and ER
sensors are the most common technologies used for this purpose. More
details about the use of the EMI, GPR, gamma ray, and vis–NIR spectros-
copy for the delineation of MZs are described in the following sections.
3.7.2 MZ Delineation Based on ECa Measurement

EMI is based on Faraday’s law used in physics and is a contactless noninvasive
method. de Jong et al. (1979) reported that the use of EMI for mapping
subsurface geology by injecting electrical current into the soil started in
the beginning of 20th century. In agriculture, the EMI technique was first
introduced in the late 1970s for salinity appraisal (Corwin and Rhoades,
1982; de Jong et al., 1979; Rhoades and Corwin, 1981; Williams and
Baker, 1982). Nowadays, this technique is mature and has become a
commonly used practice for quick characterization of in-field variability
of various soil properties (e.g., Kuang et al., 2012).
EMI has been used in agriculture for the last 2 decades. Sudduth et al.
(1995) demonstrated that topsoil depth could be quantified from measure-
ments of soil EC (measured in milliSiemens per meter, mS m1). Areas with
deep topsoil (>60 cm) generally provide low readings and are often found at
lower elevations. If a clay pan is close to the surface and topsoil is thin
(<20 cm), this corresponds to areas of high EC. Mobile sensors that measure
ECa through EMI became a relatively cheap and quick method of assessing
fine-scale soil textural variation over large areas. Soil conductivity is a
function of several factors, namely, MC, salinity, BD, temperature, and per-
centage clay. It was soon adopted as both a method to direct soil sampling to
areas of varying soil textures (Johnson et al., 2001) and as a source of

fine-scale data which proved more effective than using farmer-defined
MZ (Fleming et al., 2004).
Fraisse et al. (2001) suggested that previous MZ delineation methods
which involved traditional soil sampling were too costly for practical users,
at the high-resolutions necessary for effective mapping of within-field spatial
variation. Also, soil survey maps used inconsistent and arbitrary classification
schemes, which were not intended for MZ delineation. They therefore
developed an unbiased method of delineating MZ using bulk soil ECa
and topographic attributes by employing clustering algorithms in a GIS.
Selected topographic attributes were derived from elevation data at two sites
of 36 and 28 ha in area. ECa data were collected from a survey conducted
using an EM38 (Geonics Ltd., Canada) in a vertical mode with an effective
measurement depth of 1.5 m with positional data supplied by a DGPS. Yield
data were obtained from a commercial yield sensing system on board of a
combine harvester. PCA was used to determine the most important vari-
ables, which were found to be elevation, slope, and ECa. Unsupervised clus-
tering of these three layers was performed using the ArcInfo GIS software
“Isocluster” function for a range of specified classes (2–6). Normalized yield
was calculated for each cluster delineated zone for the five study years for the
specified number of classes. This enabled the identification of the optimal
number of MZ for each field, which could effectively minimize the
within-zone yield variance. Different years produced different optimal
MZ numbers and the authors suggested this to be dependent on the weather
conditions and crop planted. In general the appropriate numbers of MZ in
the two fields were 5 and 4. While this technique was successfully used to
delineate MZ from ECa and topographical data, it was mentioned that
future work should include a method of weighting yield maps from years
with “typical” weather patterns. It was also pointed out that this approach
could easily cope with additional data layers from remotely sensed images,
disease maps, and other soil chemical properties, since it can be automated in
GIS software.
Attempts to use soil ECa by itself or with selected landscape properties
and historical yield data to produce temporally stable MZ resulted in mixed
findings. EC shallow (ECSH) and deep (ECDP) measurements (0–30 and
0–90 cm, respectively) were used to derive four MZ in a 250 ha semiarid
cropping system (Johnson et al., 2003). The MZ performance was evaluated
based on 2 years of yield data. Within-zone wheat yield means were
negatively correlated with ECSH and positively correlated with ECDP.

Within-zone corn yield means showed no consistent relationship with
ECSH but positive correlation with ECDP. The authors concluded that
MZ based on unsupervised classification of ECSH could be used for calcu-
lating nutrient inputs and for deriving prescription maps for fertilizer, pes-
ticide, and seed. However, maps of ECa were not effective in explaining
yield variability since the soil properties determining ECa may or may
not be the yield-limiting factors in different crops. Also, seasonal weather
interactions can alter soil–yield relationships. A similar conclusion has been
reached by Moral et al. (2010), based on a similar technical approach. From
their study, it was concluded that ECa measurements can be used with suc-
cess as auxiliary variable for soil mapping and MZ determination, but the
correlations with soil fertility are really variable and sometimes poor.
A more recent attempt by Peralta et al. (2015) has utilized ECa measure-
ments coupled with terrain elevation and soil depth to delineate MZs.
The fuzzy k-means was used for clustering and the optimal number of
MZs were defined when each of NCE and FPI reach the minimum values.
Also, ECa and elevation data were successfully used to delineate productivity
zones on clay pan soil fields (Kitchen et al., 2005). Furthermore, MZ derived
from yield data were compared to those produced from combining yield,
elevation, and ECa data on several fields in southern England (Frogbrook
and Oliver, 2007). This paper highlighted the increasing use of geostatistical
techniques in data analysis, interpolation of properties, and delineation of
MZs. Several techniques were required to merge and interpolate data from
different sensors, years, and crops. Trend was detected and removed in both
the ECa and elevation data, since its existence violates the assumptions on
which geostatistics is based. The yield data came from different crops and
had to be standardized to a zero mean and unit variance. The yield, eleva-
tion, and ECa data were not collocated and had to be kriged to a grid size of
approximately 14 m. Spatially constrained nonhierarchical clustering was
used in preference to an agglomerative hierarchical approach in order to
improve the spatial contiguity of the clusters. This is helpful for locating soil
sampling sites used in MZ validation and nutrient application by machinery.
Results showed that MZ produced from spatially weighted data on ECa,
yield, and elevation could be effective in land management. Validation of
the zones by variance analysis of soil sample properties showed that the
MZs characterized the variance effectively. The authors also recommend
that land managers are consulted for their perception of potential MZs when
classification is undertaken.
To generate potential MZ, Moral et al. (2010) used a high-resolution

georeferenced soil ECa measurements, carried out by the a Veris 3100
electrical conductivity sensor at two depths shallow (ECSH, 0–30 cm)
and deep (ECDP, 0–90 cm). Both of ECa measurements were related to
other variables associated with soil fertility as CEC, pH, and TN, provid-
ing good correlations. Consequently, they considered ECa measurements
as an indicator of soil fertility parameters. Thus, soil texture variables (clay,
coarse sand, and fine sand) and both ECa measurements were considered
to delineate MZ maps. They used the regression kriging technique with
ECSH data as auxiliary variable to predict both clay and sand contents at all
unsampled locations within the field. In consequence, spatial distribution
of the main soil texture variables was accurately and rapidly performed by
using the OK algorithm. Using PCA, they found that the first two PCs
explained near completely the total variance and were chosen with the
aim of defining different potential MZs within the field. Then, a fuzzy
c-means classification procedure was used and, as a result of the clustering
analysis, two homogeneous zones were delineated with low or high CC,
which coincided with areas of low and high ECa, respectively. They
suggested further studies to compare the proposed subfield regions,
MZs, with yield maps, to make clearer the agronomic significance of this
classification.
Literature showed that EMI is of limited capability to quantify spatial var-
iation of key soil chemical and fertility parameters (e.g., TN, OC, P, K, and
micronutrients) (Kuang et al., 2012), since it is simultaneously affected by
soil salinity, temperature, BD, OM, and texture. Therefore, a PSS technol-
ogy capable to quantify accurately these soil properties is a key requirement
for improving the quality of MZ delineated for site-specific fertilizer
applications.
3.7.3 MZ Delineation Based on Other Proximal Soil Sensors

There are a plenty of sensor platforms that can provided a various dataset
in support for MZ delineation and precision applications of farm resources
(Fig. 2). The most common sensors for MZ delineation are vis–NIR
spectroscopy, passive gamma ray, and GPR (Kuang et al., 2012; Yao
et al., 2014). They have the potential to increase our understanding of
the complexity of soil–crop–water system. They also provide high-
resolution data on key soil properties to enable a successful delineation
of MZ based on fine-scale information of within-field variability. This
multilayer information is provided online at low cost and speedy way as

compared to the traditional sampling and laboratory analysis.
Gamma ray is a relatively new soil sensing technique that measures
gamma radiation emitted from the natural decay of radioactive isotopes that
are present in all soils (Castrignanò et al., 2012; Viscarra Rossel et al., 2007).
The main radioisotopes in soils are potassium (K), uranium (U), and thorium
(Th), which are associates with certain constituents in the soil. Information
about mineralogy, weathering, chemical properties of soil can be obtained.
Viscarra Rossel et al. (2007) reported that a significant amount of
preprocessing is often required to reveal relationships between the
gamma-ray spectra and the soil data. Several studies indicating that
gamma-ray sensing can be used to predict key soil properties, i.e., CC
and CEC (De Benedetto et al., 2012; Huang et al., 2014; Pracilio et al.,
2006; Rodrigues et al., 2015; Taylor et al., 2010).
Mapping of soil elemental isotopes can be performed by installing a γ-ray
sensor on a vehicle. Viscarra Rossel et al. (2007) in their research used a
GR320 portable γ-ray spectrometer mounted in a wooden cradle on the
front of a four-wheel-drive vehicle for online field measurements. In soil
surveying, the value of γ-ray spectrometry lies principally in the fact that dif-
ferent rock types contain varying amounts of radioisotopes of K, U, and Th,
as do the soil profiles to which they weather (Dickson and Scott, 1997).
Approximately 95% of the measurable γ-radiation is emitted from the upper
0.5 m of the profile (Gregory and Horwood, 1961). The attenuation of
γ-rays through the soil varies with BD and water content (Taylor
et al., 2002).
Rampant and Abuzar (2004) used gamma-ray spectrometer counts
coupled with EMI and DEM technologies to identify yield MZs. The deci-
sion tree classifier was used to predict soil types and yield data from the var-
ious combinations of geophysical and terrain attributes. Individually the
geophysical data were relatively poor predictors of the soil and yield infor-
mation, misclassifying 18%–53% of the trial area. Combined, the geophys-
ical data misclassified only 7%–16% of the area. Terrain attributes were
found to predict soil types very well, misclassifying only 2.3% of the area.
However, terrain attributes alone misclassified 6% of area when predicting
yield zones and over 11% when predicting the yield classes for an individual
year. Using all of the geophysical and terrain data, soil types could be
predicted very well, with less than 2% of the area misclassified, and they
could also predict yield zones quite well, misclassifying 5% of the area.
The predictions of yield for an individual year were always worse than for
soil types and yield zones.
Castrignanò et al. (2012) integrated multisensor data as gamma ray, EMI
measured with EM38 and EM31, and RTK-GPS data, coupled with
geostatistics approach for delineating areas of homogeneous soil. They
showed the potential of gamma-radiometric sensing by estimating a rela-
tionship for crop available soil potassium (K) from the gamma-ray signal.
The geophysical survey was carried out on a 80-ha cropping field in
Corrigin, Western Australia. Seventy-seven soil samples were collected at
the nodes of a 100 100 m2 grid and analyzed for different properties.
The EM38 and EM31 data were strongly correlated with each other
and so were gamma-ray counts from Th, U, and all element total count
(TC). The multisensor data were split into four subgroups, based on
their similarities: (1) EMI data; (2) gamma-radiometric counts K; (3)
gamma-radiometric counts from Th, U, and TC, and (4) RTK-GPS height.
Each group of data was separately analyzed using geostatistical techniques.
The multicollocated cokriging was used to joint and interpolate the soil sam-
ples and geophysical data. They reported that the EM31 and EM38 maps
were similar to gamma-U, Th, and TC maps, suggesting that they reflected
the same soil properties, but were somewhat different from the gamma-K
maps. They used the first two PCs of the geophysical data to partition the
field into homogeneous areas. To test the utility of geophysical survey for
K recommendations, they demonstrated the spatial association between the
maps of the estimates of available soil K content and gamma-K counts vari-
ables. This was done by using different agreement coefficients and a regression
model and correlated errors was estimated between the two variables.
Another promising soil sensing technique is GPR, which has been
increasingly used as noninvasive proximal sensing technology to generate
an image of subsurface soil and assist in identifying its physical properties
(Yao et al., 2014). The principle of GPR is based on the transmission and
reflection of high frequency (106–109 Hz) EM waves in the soil. Transmit-
ter and receiver antennas are moved across the soil surface to survey subsur-
face properties. Because GPR measurements depend on the large contrast
between the dielectric constants of water, air, and minerals, GPR can be
used to measure variations in soil water content (e.g., Lambot et al.,
2004). The resolution of GPR images can be varied through the use of dif-
ferent antennae frequencies. Typically, higher frequencies increase the res-
olution at the expense of depth of penetration. The penetration depth of
GPR measurements is affected by the EC of the soil. Good penetration

depth of up to about 15 m can be achieved in dry sandy soils. Groundwaves
do not require a reflective interface and so can be applied in many soil envi-
ronments. Since water content is often influenced by soil texture, ground-
wave measurements have also been used to map variations in soil texture at
the field scale (Grote et al., 2003). The groundwave sampling depth is fre-
quency dependent, so multifrequency groundwave data could be used to
map the shallow, three-dimensional water content distribution
(Galagedara et al., 2005; Grote et al., 2010). GPR type (air-launched
GPR) uses the magnitude of the reflection from the ground surface to mea-
sure the dielectric permittivity and has a sampling depth of less than 5 cm,
but the accuracy of the data is greatly diminished by vegetation, uneven soil
surfaces, and vertical variations in water content (Serbin and Or, 2003).
In agricultural areas, GPR has been used to identify soil vertical struc-
tures (Gish et al., 2002), water table depth (Hengari et al., 2013), measure
soil MC (Huisman et al., 2003; Lunt et al., 2005), assess soil salinity
(Al Hagrey and M€ uller, 2000), nitrogen loss (Walthall et al., 2001), delineate
soil compaction zones (Petersen et al., 2005), and evaluate soil pollution
(Van Meirvenne et al., 2014). Recently, GPR has evolved from tool for
measuring soil properties to a means of mapping spatial variability of the
measured soil properties for applications in PA. De Benedetto et al.
(2013a) in their research combined multisensor data including GPR,
EMI, hyperspectral spectroscopy (HS), and remote sensing for MZ delinea-
tion. The data were classified into four groups based on: (1) bulk ECa from
EMI data, (2) amplitude of GPR signal data, (3) the first PC relating to five
EM bands (green, yellow, red, red edge, and NIR) of hyperspectral reflec-
tance data, and (4) the vegetation indices (NDVI, normalized difference red
edge (NDRE), and NIR/Green) calculated from the WorldView-2 satellite
imagery. Ordinary cokriging was used to analyze and interpolate the data at
the nodes of a grid. To obtain spatially contiguous clusters, they used a com-
bined approach of clustering based on multivariate geostatistics and a non-
parametric density function algorithm, applied to the overall multisensor
dataset of the estimates. The full approach allowed to identify three homog-
enous areas (clusters). In particular, cluster 1 was delineated with the lowest
values of ECa and GPR amplitude, and the highest red PC values. The other
two clusters were delineated with the highest values of green, yellow, red
edge, and NIR PCs for cluster 2, and the highest values of ECa and vege-
tation indices for cluster 3. They reported that the delineation might be
related to the intrinsic spatial variability of soil and the health status of plants.
They concluded that the approach can be used to produce a prescription

map for site-specific management.
When a soil sample is illuminated with a light source, molecules vibrate,
leading to absorption of light, to various degrees, with a specific energy
quantum corresponding to the difference between two energy levels. As
the energy quantum is directly related to frequency, the resulting absorption
spectrum produces a characteristic shape that can be used for analytical pur-
poses (Stenberg et al., 2010). The fundamental vibrations in the mid-
infrared (MIR) region result in overtones and/or combinations in the
NIR region. In the visible (vis) range (400–780 nm), absorption bands
related to soil color are due to electron excitations, which assist in the mea-
surement of OM and MC in particular. However, in the NIR range, the
overtones of OH and overtones and/or combinations of C–H + C–H,
C–H + C–C, OH+ minerals and N–H are important for the detection of
OM, MC, clay minerals, and total nitrogen (Mouazen et al., 2010;
Stenberg et al., 2010).
A study by Mouazen et al. (2005) described development of one of three
(Christy, 2008; Mouazen, 2006; Shibusawa et al., 2001) online soil sensors
that can collect mega soil reflectance spectra (>500 spectra per ha) using a
vis–NIR spectrophotometer mounted on a tractor. These online vis–NIR
sensor platforms, which measure soil diffuse spectral reflectance have been
successfully used to map a variety of yield-limiting soil properties (e.g.,
Kodaira and Shibusawa, 2013; Kuang and Mouazen, 2013; Kuang et al.,
2015; Kweon et al., 2013; Mouazen and Kuang, 2016) and show potential
for inclusion in future MZ delineation techniques. In general, better calibra-
tion models can be derived for soil properties with direct spectral responses
in the NIR range, which include OC, MC, and clay (Kuang et al., 2012;
Stenberg et al., 2010). However, properties with indirect spectral
responses, i.e., Ca, CEC, pH, P, K, and Mg can also be measured, but with
lower accuracies (Kodaira and Shibusawa, 2013; Marı́n-González et al.,
2013). This can be attributed to the correlation of these properties with
other soil properties, having direct spectral responses in the NIR range
(Kuang et al., 2012; Stenberg et al., 2010). Disadvantages of the use of
vis–NIR sensors include the variable prediction accuracy of the models
derived, and the expense of the equipment and the expert knowledge
needed to process the survey data into property maps.
Only limited research has been reported so far on the use of online
vis–NIR spectroscopy sensors to delineate MZs for VRAs, majority of
which are listed earlier (Mouazen and Kuang, 2016; Mouazen et al., 2014).
For instance, Mouazen et al. (2014) implemented online multisensor and data
fusion approach for the delineation of MZs of soil WHC for site-specific irri-
gation. A multisensor platform was used, which consisted of a load cell to mea-
sure subsoiler draft, a wheel gage to measure depth, and vis–NIR
spectrophotometer for simultaneous measurement of soil properties,
namely OC, CC, BD, plasticity index, and MC. Partial least squares
(PLS), artificial neural network (ANN), and MLR analyses were carried
out on the six named soil properties to derive maps of WHC. They dis-
tinguished a spatial similarity between calculated WHC and measured
available water (AWC) maps. Accordingly, they recommended the
multisensors and data fusion as a new approach to optimize irrigation
scheduling. Also, Mouazen and Kuang (2016) utilized the online vis–
NIR spectroscopy coupled with PLSR and OK for VRA of P in a
21 ha field in Duck End Farm, Bedfordshire, UK. The experiment and
online scanning were carried out over 3 years (2011, 2012, and 2013) after
crop harvest. They reported that the online measurement accuracy of
P was acceptable and the VRA of P2O5 after crop harvest in year 2
improved the uniformity of the spatial distribution of P, measured in year
3 with the online soil sensor. The CV of P in the field was reduced from
26%, to 25%, to 16% in 2011, 2012, and 2013, respectively showing sig-
nificant improvement in the uniformity of P spatial distribution across the
field. They concluded that the online vis–NIR soil sensor is an effective
tool to manage and minimize within-field variation in P in arable crops.
A case study on MZ delineation for VR N application will be described
in detail later based on a report from Halcro et al. (2013).
3.7.4 Multisensor and Data Fusion

Data integration or data fusion is needed to enhance the effectiveness of
techniques for mechanically or semiautomatically converting data from
diverse bases and dissimilar scales into a representation that offers efficient
support for individual or computerized decision making. Data fusion, within
this framework is a multidisciplinary field based on numerous varied areas
such as information theory, signal processing, statistical evaluation and sup-
position, and synthetic intelligence (Khaleghi et al., 2013).
In soil science, although research on individual PSSs has demonstrated
potential, no solitary single sensor (or method) is capable of completely char-
acterizing soil complexity. Soils offer numerous scales of spatial and temporal
variation that can be monitored through different sensor systems. Every sen-
sor presents an exclusive perspective on the soil forming factors, giving
insight into temporally, and spatially varied soil systems. Therefore, selecting
or developing a sole common sensor to function underneath these circum-
stances is virtually infeasible (Grunwald et al., 2015). Sensor fusion can be a
sensible option to incorporate diverse variation across scales (both vertically
and horizontally) and dissimilar soil properties so as to deal with this chal-
lenge. There are three key kinds of sensor fusion: (1) proximal sensor fusion,
in which just proximal sensors are applied jointly; (2) remote and proximal
sensor fusion, in which proximal sensor(s) are employed simultaneously by
remote sensor(s) (RS); and (3) remote sensing fusion, whereby just RS are
combined. With all these three methods extra environmental covariates are
utilized along fusion of sensor information to model soils (Grunwald et al.,
2015). In recent years, there has been an increasing amount of published
research on the implementation of proximal sensor fusion in agronomy.
In contrast with individual sensors, proximal sensor fusion has enhanced
the evaluation of multiple properties of soil at multiple scales (Al-Asadi
and Mouazen, 2014; Mahmood et al., 2012), delineation of MZs
(Castrignanò et al., 2012), and so has enabled the characterization, approx-
imation, and mapping of numerous soil and crop features (e.g., Kweon et al.,
2013; Mahmood et al., 2012) across depths (De Benedetto et al., 2013b).
Multisensor and data fusion have lately been popular in PA due to the neces-
sity for characterizing and mapping field-scale soil property differences that
inform site-specific management. This can be attributed to the ease in
attaching numerous sensors on a tractor or vehicle and moving across a field.
Additional information on fusion of proximal sensor data can be found in
literature such as Viscarra Rossel et al. (2010a) and Grunwald et al. (2015).
The decision on which set of sensor needs to be combined within this
system depends on the application in question and practical considerations.
As some sensors are comparatively simple to combine (e.g., ECa sensors),
others might need modified hardware, software (e.g., vis–NIR, gamma
ray, GPR), regular calibration (e.g., vis–NIR–MIR sensors), and compound
data processing and understanding (e.g., GPR). De Benedetto et al. (2013b)
highlighted the intricacy of GPR data processing and recommended its
application simply in situations where specific spatial constructions are antic-
ipated within the subsoil, such as soil layering because of tillage (Jonard
et al., 2013).
Remote and proximal sensors fusion are more regularly used in digital
soil mapping at local scales, in which proximal sensors are employed to make
possible or harmonize field sampling (e.g., Gomez et al., 2008; Wetterlind
et al., 2008), and RS are used to offer a variety of environmental covariates
(e.g., reflectance of bare soils; vegetation indices). Example uses of

proximal–RS fusion were offered by De Benedetto et al. (2013a), who mer-
ged an ECa field sensor with vegetation indices obtained from Worldview-2
and GeoEye-1 satellite imagery to outline homogeneous soil–crop supervi-
sion areas in Italy.
Fusion of RS is used mainly to integrate (merge) different sensor reso-
lutions (spatial or spectral) to assess soil properties and classes and improve
the level of information obtained from these sensors (Grunwald et al.,
2015). Particularly, fusion of RS increases the spectral efficiency when dif-
ferent sensors addressing different regions of the spectrum that are effective
in spectral segmentation (e.g., visible, infrared, thermal, and microwave)
(Metternicht and Zinck, 2003). For instance, Metternicht (1996) merged
(fused) Landsat TM (vis–NIR bands) and JERS-1 SAR data (microwave)
in his study of soil salinity in Cochabamba, Bolivia. He achieved a better
discrimination of salinized areas with a 35%–40% increase in accuracy com-
pared with results from individual sensors. More examples on multisensor
and data fusion implemented in precision agriculture are described in
Section 3.7.3.
4. APPLICATION MAPS FOR VR FERTILIZATION

Application (or prescription) maps are used to manage the VRA of
inputs by stating how much to use and where they should be placed.
Fleming et al. (2004) stated that application maps are essential for effective
VRA management. The examples of MZ-derived VRA strategies for fertil-
izer found in the available literature are almost exclusively from research in
the United States (Fleming et al., 2004; Khosla et al., 2002; Koch et al.,
2004). The application rates used by researchers are often based on homog-
enous recommendations derived from an algorithm based on expected
yield (EY) of a crop. EY figures can be obtained from state universities
(e.g., Nebraska (Fleming et al., 2000b) or Washington State (Mulla et al.,
1992)). Khosla and Alley (1999) reported that farmers often use the EY from
the most productive areas of the field for N rate calculations. This is a sub-
optimal strategy, which leads to low NUE figures (Khosla et al., 2002).
Researchers have typically used rates above and below this recommendation
based on the fertility or yield potential of MZs.
Mulla et al. (1992) used MZ based on the potential yield calculated based
upon available water, the expected precipitation in the growth season, and
the water needed for crop to produce grain in order to calculate N and
P fertilizer requirements, whereas Khosla et al. (2002) used MZ based on
EY (which is the same as yield goal). Godwin et al. (2003) investigated
the appropriate rates of N application that are determined from average
rainfall, crop type, previous management practice, and average soil type
for the field. No account was taken of within-field variability. The rates used
by Godwin et al. (2003) varied between 25% and 30% above and below the
recommended rate. The researchers chose these levels to provide measurable
differences when comparing UR and VR N applications (Welsh et al.,
2003). Mulla et al. (1992) used 25% above and 70% below the UR
N application. Khosla et al. (2002) used 30% variation to contrast the per-
formance of different SSMZ approaches with soil grid-sampling and UR
approaches. Khosla et al. (2008) compared VR and UR of N using three
treatments of (1) no N added, (2) recommended N (based on yield poten-
tial), and (3) 50% less N of the recommended N. Fleming et al. (2000b) var-
ied N applications by 25%. Clearly, since there are no clear guidelines for
VRA, researchers have generally used experience and judgment to decide
on suitable rates.
An application map design must also take account of the capabilities of
the machinery and experimental goals (Welsh et al., 2003). Comparison strip
experiments are a common approach (Mulla et al., 1992) to compare
between VR and UR applications. Welsh et al. (2003) favored strip treat-
ments to ensure that UR and VR treatments covered comparable soil var-
iations. Also, the correct fertilizer application required the treatment width
to be the same, or half, the tramline width. This spacing also allowed the
combined harvester to collect a full header without including the bare soil
of the wheel tracks.
5. OPPORTUNITIES FOR EVALUATION MZs

BY COST/BENEFIT ANALYSIS
Contrary to “uniform” fertilizer application, VRA permits treatments
to be done with variation of the quantities used inside the equivalent parcel.
Fertilizers and pesticides are merely applied where and when they are essen-
tial and within the needed quantities for each zone. Having this in mind, it is
possible to acknowledge the fact that PA can bring obvious ecological ben-
efits and fruitful potential input of PA to better sustainability of farming pro-
duction procedures.
Many researchers have studied the economic impacts of PA (Ancev et al.,

2005; McBratney et al., 2005; Tekin, 2010). For example, Tekin (2010)
examined the potential economic benefits of VRA in a study conducted
in Turkish wheat production. In this study, an investment appraisal and par-
tial budgeting analysis were used to determine applicable conditions for
farmers. It was concluded that applying N fertilizer based on soil variability
could be economically justified with 1%–10% yield increases and 4%–37%
savings in fertilizer. Moreover, the key factor for PA implementation is the
degree of variability; with higher variability leading to easier implementation
(Tekin, 2010). The benefits that VRA can present in PA appear to be a lot
clearer, this is because PA enhances productive aspects for the farmers (fer-
tilizers, water, crop protection items), decreasing expenses, and minimizing
the ecological influence (Arnó et al., 2009). Despite study of the economical
impacts in PA, only few studies have involved a cost and benefit analysis of
MZs and VRA (e.g., Halcro et al., 2013; Khosla et al., 2008; Koch et al.,
2004). For example, Koch et al. (2004) evaluated the economics of four
N management strategies by assessing the net–return sensitivity of each
N management strategy as influenced by changes in grain yield and com-
modity prices. They analyzed the N management strategies under farmer
vs custom-applied scenarios. The N management strategies were uniform,
grid based, SSMZ–constant yield goal (SSMZ-CYG), and SSMZ–variable
yield goal (SSMZ-VYG). They found that the three site–years consistently
indicated that less total N fertilizer (6%–46%) was used with the
SSMZ-VYG N management strategy when compared with uniform
N management. Net returns from the SSMZ-VYG N management strategy
were $18.21–29.57 ha1 more than uniform N management. They con-
cluded that the VR N application utilizing SSMZs are more economically
feasible than conventional uniform N application. Khosla et al. (2008) pref-
ormed an economic analysis of MZs in a field that had been treated with four
N management strategies using enterprise budgets that were created by using
the Profit and Loss Enterprise Budget software (v.2.0). Gross revenue was
calculated based on prices of $138 (USD) per mg maize grain, and $0.16
per kg N. To calculate the net returns, the difference between total operat-
ing costs (including ownership costs) and gross revenue was calculated.
Then, weighted mean net returns were calculated based on the proportions
of MZs across the entire field as the size of each MZs was not equal. In two
out of three site–years, the variable yield-goal strategy resulted in the largest
net returns compared with the UR. Net returns ranged from $188 to $679
(USD) ha1. Also, they found that using the SSMZs as a basis for VR
N fertilizer application improved economic returns by as much as $679
(USD ha1). They concluded that SSMZs are a practical approach to precise
N management in the Western Great Plains region of the United States.
Overall, cost–benefit analysis has proved that VRA is more profitable
compared with UR application through different cases with different fertil-
izers. However, this analysis has so far been limited in the PA, due to the
detailed data required to perform such analysis. These data include inputs
and outputs of the agricultural process as well as the cost of the PA technol-
ogies involved in this process.
6. A CASE STUDY: SITE-SPECIFIC N APPLICATION FOR

OIL SEED RAPE (OSR) CROP BASED ON MZ
DELINEATION WITH MULTISENSOR AND DATA
FUSION APPROACH
The objective of this case study was to compare between three
approaches of delineating MZs for site-specific N fertilizer application.
The success of innovative approaches (VR2) was determined by the reduc-
tion in fertilizer application and improvement in yield, compared to the
farmer’s normal UR and traditional (VR1) application.
6.1 Experimental Site

The study site Horn End Field was a 22 ha located in Duck End Farm,
Wilstead, Bedfordshire, UK (Fig. 3). The field is normally under annual crop
rotation system of winter wheat, winter barley, and winter OSR. The soil is
characterized by fine texture and varies from clay to clay loam, sandy clay
loam, and loam. The topography of the area is rather flat with an elevation
that varies between 30 and 38 m. The study took place over two cropping
seasons (2011–12) with OSR.
Fig. 4 shows experimental plots with strips of 24 m wide and approxi-
mately 405 m long. The order of VR1, VR2, and UR treatments was ran-
domly chosen. Five sets of three treatments were spread over the width of
the field. Edge effects were reduced by placing the study away from the vehi-
cle turning areas at the end of each tramline (Fig. 4). The “VR1, VR2” and
UR applications were applied as follows.
6.1.1 UR-Based on the Farmers Usual Method

The field was treated as a single MZ with a UR N fertilizer calculated in
accordance with the UK Department of Environment, Food and Rural
Affairs (DEFRA) recommendations for specific crop, soil type, and
off-take strategy (i.e., a total of 220 kg ha1 nitrogen) (Table 2).
Fig. 3 Location of Duck End Farm and study Horn End Field.
VR 1 UR
Study
VR 2 area
Fig. 4 The 22 ha study area divided into parallel treatment plots of 24 m wide and
about 405 m long. Source: Halcro, G., Corstanje, R., Mouazen, A.M., 2013. Site-specific
land management of cereal crops based on management zone delineation by proximal
soil sensing. In: J. Stafford (Ed.), Precision Agriculture 2013, Proceedings of the 10th
European Conference on Precision Agriculture (pp. 475–481). Wageningen Academic
Publishers, Wageningen.
Table 2 Assignment of Fertilizer Application Rates

Total N Application
Method Class MZ Relative Fertility Variable Rate (kg ha21)
UR n/a n/a N 220
VR1 4 Highest N 24% 184
2 Medium N 220
3 Lowest N + 24% 255
VR2 2 Highest N 24% 184
5 High medium N 12% 202
4 Medium N 220
3 Low medium N + 120 238
1 Lowest N + 24% 255
6.1.2 Traditional Variable-Rate (VR1) MZ Delineation Based

on Standard Commercial Approach
An EM1 survey was conducted across the field in 2011, a few months after
seeding with winter wheat. The ECa data were gathered by a Dualem
25 (Dualem, Canada) proximal sensor towed behind a quad bike. Sensor
data were imported into ArcGIS (ESRl, USA) and mapped to show spatial
variation in the soil. A total of 30 random soil samples were taken from the
study field with a minimum separation distance of 15 m to provide wide
coverage. Laboratory chemical analyses were carried out to measure level
of pH, phosphorous (P), potassium (K), and magnesium (Mg). Maps for each
soil property were produced by IDW interpolation.
The ECa, pH, P, K, and Mg layers were combined using k-means clus-
tering (Statistica—StatSoft, USA) to group areas of similar fertility. The out-
put was exported into ArcGIS software for visualization and spatial analysis.
Small and irregular-shaped clusters were merged with neighbors of similar
fertility, which resulted in the delineation of three fertility MZ classes. This
MZ map provided the basis for the fertilizer application map. The relative
fertility of each zone was decided by analysis of the cluster means of the input
properties, together with historical yield and NDVI maps. The
medium-fertility zone was assigned an application rate identical to the
UR application (e.g., 220 kg ha1). The most fertile zone was assigned
an N rate of 24% below this and the lower zone was 24% higher (Table 2).
6.1.3 Innovative Variable-Rate (VR2) MZ Delineation Based on Proximal

Sensor Surveys
This innovative approach employs high-resolution data on soil properties
and crop growth collected with a suite of proximal sensors, as explained
in the next section.
6.1.3.1 Online Sensor Measurement and Soil Sampling

A spectral survey in diffuse reflectance mode using an online vis–NIR sensor
platform (Mouazen, 2006) was performed in the field after the 2011 harvest.
The sensor hardware consisted of a subsoiler, attached to a metal frame
which was set up on the three-point linkage of a tractor. An optical probe
is attached to the backside of the subsoiler to collect soil spectra in diffuse
reflectance mode using an “AgroSpec” mobile, fiber type, vis–NIR spectro-
photometer (Tec5 Technology for Spectroscopy, Germany) with a mea-
surement range of 305–2200 nm. Online measurements were carried out
at 2 km h1 down to 15 cm depth, and consisted of 21 parallel traverses spa-
ced approximately 20 m apart. More information about the online soil sen-
sor can be found in Quraishi and Mouazen (2013).
A total of 60 soil samples were collected from the bottom of the survey
trenches opened by the subsoiler. These samples were used to validate the
online vis–NIR measurement. Soil samples were sent to the laboratory
where they were stored frozen. Subsequently analyses of TN, OC, pH,
P, K, Mg, Ca, CEC, and MC were performed using standard methods.
6.1.3.2 Crop Growth and Yield Measurements

Crop growth data were logged as NDVI values collected with a boom
mounted CropCircle ACS-210 system (Holland Scientific, USA) on April
6, 2011. The reflectance data were logged together with the GPS coordinates
and NDVI was automatically calculated for each data point. Yield data from the
2011–12 season harvest were collected by the onboard yield sensor and GPS
system of the farmer’s combine harvester (New Holland, CX8070 model).
6.1.3.3 Laboratory Chemical and Optical Analyses

Each sample was divided into two parts: one part was dried for 24 h at 60°C
and the other part was left fresh (wet). The dried soil sample was analyzed for
soil parameters according to British Standards Institute (BSI). Each soil sam-
ple was placed into three Petri dishes, which were 2 cm in depth and 2 cm in
diameter. The soil in the Petri dish was shaken and pressed gently before
leveling with a spatula. The soil samples were scanned in diffuse reflectance
mode using the same mobile, fiber type, vis–NIR spectrophotometer

(AgroSpec from Tec5 Technology for Spectroscopy, Germany) used during
the online measurement. A 100% white reference was used before scanning.
A total of 10 scans were collected from each container, and these were aver-
aged in one spectrum.
6.1.3.4 Spectra Pretreatment and Development of Calibration Models

Pretreatment of the soil spectra was conducted using Unscrambler 9.8 Soft-
ware (CAMO Inc., Oslo, Norway) following the methodology reported in
Mouazen et al. (2005). Calibration models for TN, OC, pH, P, K, Mg, Ca,
CEC, and MC were established by PLSR, based on one-leave-out
crossvalidation. Model performance was evaluated in terms of coefficient
of determination (R2), root-mean square error of prediction (RMSEP) in
calibration, and prediction and residual prediction deviation (RPD), which
is the SD divided by RMSEP. Viscarra Rossel et al. (2006) classified RPD
values as follows: RPD < 1.0 indicates very poor model/predictions; RPD
between 1.0 and 1.4 indicates poor model/predictions; RPD between 1.4
and 1.8 indicates fair model/predictions; RPD values between 1.8 and 2.0
indicate good model/predictions; RPD between 2.0 and 2.5 indicates very
good; and RPD > 2.5 indicates excellent model/predictions. Generally, a
good model prediction would have large values of R2 and RPD, and a small
value of RMSEP.
6.1.3.5 Soil Mapping

The online measured soil properties were subjected to semivariogram anal-
ysis in Vesper software (ACPA, Australia). After a satisfactory estimation of
semivariograms, these were applied in ArcGIS (Esri, USA) software to pre-
dict the variable qualities at unsampled sites to make a prediction map for
every soil property mentioned earlier.
6.1.3.6 Data Fusion and MZ Delineation

For every crop and soil feature, the interpolated map was transformed into a
common 5 m raster network in ArcGIS (Esri, USA) so as to help data syn-
thesis (Frogbrook and Oliver, 2007). The raster cells of the layers were
converted into this familiar network of points by pulling out the value at
the average of every raster square. The 5 m network size offered a balance
between sufficiently characterizing the spatial variability and sensible farm
management (e.g., farm equipment size and lag time).
Data fusion of soil and crop datasets were performed through a k-means
clustering algorithm (Hartigan and Wong, 1979) that employs the unscaled,
squared Euclidean distance, for distance computation. During this process,
an error decline of less than 5% was selected as the frontier. In order to iden-
tify MZs, the clustering production was introduced into ArcGIS (Esri, USA)
to help spatial analysis and visualization. Maps of grid-point class member-
ship were created so as to demonstrate the clustering effect. Possible MZs
were outlined by means of the “Draw” instrument in ArcGIS, to create
polygons about the borders of every category. Diverse MZ maps of the
research field were created (Fig. 5). Management region review offers
insight into the kind of the MZ formed. This consists of carrying out (1)
cluster study to find out the proportions of every parent qualities in every
category, (2) discriminate study to notice the mainly influential attributes,
and (3) yield analysis to explore whether any declines in yield discrepancy
for every delineation technique are statistically significant. Cluster analysis
was done through producing the graphs to signify values for every property
in every class by means of Statistical software (StatSoft Inc., USA). The
graphs were evaluated to locate the distinguishing features in every class
and the key dissimilarities between every class.
6.1.3.7 Site-Specific Fertilizer Applications

The application of fertilizer requires the delineation of practical MZ, the cal-
culation of appropriate application rates for each zone, and the production of
an application map. The higher resolution and greater number of input data
layers provided by the VR2 data produced more complex potential MZ than
VR1. As in VR1, the MZ map was delineated by hand using the drawing
tool in ArcGIS. Irregular cluster edges were smoothed and small areas were
merged with neighbor’s in order to delineate MZs, to which the pneumatic
fertilizer spreader (Kuhn Aero 2224’) can respond (Fig. 5).
The variation of fertility was found to be best described by five classes.
These formed the basis of the five application rates chosen with middle fer-
tility class assigned the rate recommended according to DEFRA RB209
guidelines (DEFRA, 2013) according to the crop and soil type. Higher
and lower steps of 12% were chosen for each of the remaining four classes
according to the fertility rank of the MZs (Table 2). The total N fertilizer
recommendation was divided into three applications over the growing sea-
son. This included a blanket early spring treatment of nitrate and sulfur mix.
The two further applications were of nitrate only. Both VR strategies
applied more N fertilizer to the least fertile areas and vice versa. For example,
Fig. 5 Comparison between traditional variable rate (VR1) based on interpolated soil properties (A) and innovative variable rate (VR2) based on
kriged maps of soil properties derived from the online visible and near-infrared (vis–NIR) sensor, normalized difference vegetation index (NDVI),
and yield data (B). Data fusion using clustering by K-means algorithm was used for management zone (MZ) delineation for nitrogen fertilizer.
a higher fertilizer rate (e.g., +12% compared to the middle fertility class) was
applied in poor fertility zones, and a lower rate (e.g., 12%) in rich zones.
6.2 Results and Discussion

6.2.1 Modeling and Soil Mapping
Prediction models were successfully created for the nine soil properties.
Validation was carried out by comparing the predicted values made by
the models with the chemical analysis of soil samples. The performance
of the calibration models is shown in Table 3. The results are in general
agreement with previous work (Kuang et al., 2012), i.e., properties with
a direct spectral response to the vis–NIR spectral range can be modeled
more successfully than those without. The calibration models were success-
fully applied to the spectra collected by the vis–NIR survey to produce pre-
diction points for the nine soil properties. According to Viscarra Rossel et al.
(2006) RPD classification guidance, the MC model provided excellent pre-
diction accuracy while the pH model provided very good quantitative pre-
dictions. All other models could be used to discriminate between low and
high values. These results are considered as sufficiently accurate for the pur-
poses of site-specific management.
The spatial variation of all soil properties were best described using spher-
ical semivarigram models. OK was performed on the predicted data points
Table 3 The Accuracy of Online Prediction of Moisture Content (MC),

Organic Carbon (OC), Total Nitrogen (TN), pH, Calcium (Ca), Cation
Exchange Capacity (CEC), Magnesium (Mg), Potassium (K), and
Phosphorous (P)
Online Prediction
Property RMSEP RPD R2
MC 1.10 (%) 4.40 0.93
OC 0.22 (%) 1.83 0.68
TN 0.02 (%) 1.91 0.75
pH 0.36 2.14 0.78
1
Ca 7.11 (mg kg ) 1.30 0.61
CEC 0.97 (cmol+/kg) 1.61 0.62
1
Mg 0.34 (mg kg ) 1.49 0.67
K n/a n/a n/a
1
P 0.60 (mg kg ) 1.50 0.60
based on the derived semivariogram models for all soil properties using
ArcGIS software (Esri, USA). The interpolated maps for all nine soil prop-
erties are shown in Fig. 5.
6.2.2 MZ Analysis
The traditional approach (VR1) produced three classes. However, the inno-
vative approach (VR2) produced five classes (Fig. 5). Class 5 was the most
fertile class, since it was associated with the highest yield among the remaining
four classes. It is worth noting that class 5 has the lowest concentrations of soil
properties. No clear differences in yield of the remaining four classes could be
observed, hence NDVI was considered for classifying the fertility of these clas-
ses. Class 2 had the highest NDVI, together with high levels of TN and P,
hence, it was considered as the second highest YP class. The lowest ranked
YP class was class 1 due to the lowest NDVI and low yield, together with
low P and K. Class 4 contained high levels of several influential soil properties
but low NDVI and historical yield. These factors ranked class 4 as the second
lowest fertility class. Although classes 1 and 4 have the highest concentration
of soil properties, their agronomic responses measured as NDVI and yield
were the poorest. The reason is the poor drainage system of areas of these
two classes, which negatively affect crop growth and yield, although they
are rich in soil nutrients. Good levels of MC, OC, and NDVI but low levels
of P, K, and MC rated class 3 as medium-fertility (YP) class. The final rank of
the four fertility classes from most fertile to the poorest class is Class 5 > Class
2 > Class 3 > Class 4 > Class 1 (Fig. 5).
6.2.3 Cost–Benefit Analysis

The cost–benefit analysis results of the studied field are shown in Tables 4
and 5. From the results obtained, it is possible to estimate the financial gain
the farmer would have made by employing VR over his normal UR appli-
cation for the whole field for this single year experiment (Table 5). The
innovative MZ delineation approach based on VR2 would have been more
profitable for this crop by £60 ha1 (or £1319 per 21.99 ha of the field area)
over the traditional UR approach and £34 ha1 (£946 per 21.99 ha of the
field area) over the traditional VR1 approach (Table 5). These margin
figures do not take the cost of surveys and equipment into account but it
is expected that a commercial vis–NIR survey would cost about £40 ha
to be carried out once every 4 years. Commercial precision agriculture com-
panies in the UK charge £22 ha1 for nutrient mapping once every 4 years
and £30 to include an ECa survey (figures are from Soyl Ltd., February 20,
Table 4 Amount of Fertilizer Applied and Yield Gained for the Three Nitrogen
Fertilization Approaches, Namely, Uniform Rate (UR), Traditional Variable Rate (VR1), and
Innovative Variable Rate (VR2)
Treatment
(5 Plots) Area (ha) Yield (t ha21) Yield (t) Fertilizer Input (t)
UR 2.983 5.023 14.990 2.246
VR1 2.981 5.021 14.967 2.091
a
VR2 2.990 5.174 15.475 2.235
Fertilizer Input
Per ha Area (ha) Yield (t ha21) (t ha21)
UR 21.99 5.023 0.752
VR1 21.99 5.021 0.701
a
VR2 21.99 5.174 0.747
a
Significant difference between treatments at P < 0.05 level.
Table 5 Margin Calculation Comparing the Three Nitrogen/Sulfur Fertilization

Approaches, Namely, Uniform Rate (UR), Traditional Variable Rate (VR1), and Innovative
Variable Rate (VR2)
Treatment Input Cost Yield Price (£)
(5 Plots) Fertilizer (£) (t ha21) Margin (£)
UR 831 5846 5015
VR1 774 5837 5063
VR2 827 6035 5208
Input Cost Yield Price (£)
Per ha Fertilizer (£) (t ha21) Margin (£)
UR 278 1960 1682
VR1 259 1958 1699
VR2 276 2018 1742
Cost was £0.37 kg1 for nitrate fertilizer product and £390 t1 the selling price for the oil seed rape
(OSR) crop.
2013, www.soyl.com). In-season nitrate fertilizer application based on crop

canopy measurement using the Yara N-sensor would be an additional
cost in Soyl quotations. Taking the online survey cost (£10 per year) into
account, the innovative VR2 N fertilization would lead to a net profit
to the farmer of £50 ha1, by mainly increasing the OSR overall yield
by 3% and marginal reductions in fertilizer use, when compared to UR
N fertilization. The ANOVA analysis indicated that the differences in yield

between the three treatments are significant at 5% confidence (P < 0.05)
level (Table 4). The significant increase in overall OSR yield with VR2
might be attributed to the improvement of MZ delineation with the new
approach adopted in this study, which better matches the actual
within-field variation in soil fertility. This enables the grower to apply fer-
tilizer according to need and where it will be used, rather than being lost into
the environment.
Regarding the amount of nitrogen in form of nitrate applied, the tradi-
tional VR1 consumed the smallest amount. VR2 consumed 5 kg ha1
(100 kg/22 ha field) less nitrate compared to that of the UR (Table 4).
Although the VR1 approach used much less fertilizer than the UR or
VR2 and matched the yield of UR, the VR2 yield increase was large enough
to offset the extra fertilizer used. It should be noted that the classification
interval in VR1 was greater than VR2. These encouraging results suggest
recommending the fusion of data on crop and soil properties collected with
a proximal crop sensor and online soil sensor, respectively, for the delinea-
tion of MZs for VR N fertilization.
7. CONCLUSIONS
This literature review has identified several research gaps in MZ delin-
eation and highlighted how technical advances have the potential to
improve the performance of fertility-based MZ for VR nitrogen fertilizer
application.
It is clear that measuring a single soil or crop property is insufficient to
inform VRAs. MZ created by a suite of properties are more likely to char-
acterize the complex spatial variation of soil fertility since it is unlikely that all
the yield-limiting factors are known beforehand. Indeed, fertility in different
parts of the same field may be influenced by different factors.
Soil sampling and subsequent laboratory soil chemical analysis on its own
are currently economically unfeasible for effective characterization of soil
property spatial variation. New research should make use of current sensor
systems, which can gather data at sufficiently high density to characterize
small-scale variations now known to be present in majority of fields.
Our investigation demonstrated that different integration pathways have
been applied in various contexts to map/model variations in soil properties.
For example, measurement of soil OC may use a different technology and
data mining than for soil texture, microbial biomass phosphorus, soil mois-
ture, salinity, or soil type. Our knowledge and understanding of which
“sensor soil/environmental data integration pathway method/model

approach” works best, or could be effectively applied to solve a specific
problem, is still limited, and is dependent on the required level of accuracy,
technical, and scientific expertise and know-how required to fuse and model
available sources of data.
On the positive side, multisensor and data fusion approaches provide a
broad source of information valuable for the improvement of precision
farming practices for maximizing crop yield at minimized input costs and
environmental impacts. In this context, the selected case study of delineation
of MZs for VR N fertilization based on the fusion of high-resolution data on
soil and crop properties showed high agronomic, environmental, and eco-
nomic potential. Results allowed the following conclusions to be drawn: the
online vis–NIR sensor platform can be successfully used to acquire high res-
olution, field-wide information on a range of influential soil properties
linked to soil fertility. It would be prohibitively expensive to achieve similar
results using traditional laboratory analysis of soil samples. The large number
of data points enabled advanced geostatistical methods (e.g., kriging) to be
employed to derive maps with a more complex variation in soil properties
and fertility than traditional methods.
While a significant amount of recent research and commercially available
services make use of ECa data from sensor surveys to characterize soil tex-
tural variations, this approach is not suitable for informing VRA of nitrogen
fertilizer. A suitable alternative may be to use data from crop growth mea-
surements, taken during the growing season, as part of the MZ delineation
technique. The cost of such data from satellite or UAVs is almost certain to
become more affordable in the future.
It is worth noting that, the majority of fertility-based MZ delineation tech-
niques in the literature, their effectiveness was not assessed by measuring per-
formance in actual VRAs. Rather, statistical assessments, such as yield variance
reduction in the MZ, were commonly used. While this may be understand-
able due to budget and time restrictions, any new MZ delineation approach
needs to be assessed against both a UR application and an established method
for VRA in a commercial setting. The ability of any new technique to provide
economic and/or environmental benefits will decide whether it is adopted
by growers or not. The cost–benefit analysis of the study case on VR
N fertilization presented in this paper confirmed that the innovative MZ
delineation approach based on fusion of high sampling resolution on soil
and crop to be more profitable as compared to the traditional UR and VR
approaches, with overall net margin of £50 and £34, respectively. This was
attributed mainly to increase in yield of OSR in one cropping season.
The literature indicates that in research which involves a comparison of

UR and VR-MZ performance, the experimental design has normally
included all the MZs delineated by the respective techniques and employed
randomized replicates. It is recommended in future work to follow this
approach, as literature confirms this to provide robust data allowing science
base comparison between different MZs approaches.
MZs are a tool which can improve decisions in precision agriculture by
better management of variation. Technological advances in MZ delinea-
tion provide more fine-grained information about soil constituents and
their complex interactions. Data gathered about these interactions and
the make-up of the zones themselves could provide a valuable resource
of agricultural informatics for optimized soil and crop management in
the future. It is increasingly important to manage soil in the most econom-
ical manner which has to guarantee minimizing the environmental
impacts simultaneously. The traditional management approach of treating
fields as homogenous units has been shown to result in increasing environ-
mental impacts by excessive use of agrochemicals, while reductions in
crop yield were reported. Management of within-field variability by
MZ approaches based on high sampling resolution of data on soil and crop
is the future for improving land productivity, while reducing the environ-
mental impacts. MZ can increase AE by reducing waste. In addition, crop
yield can be more uniform in yield and quality. This consistency is much
welcomed by companies involved in the food supply chain, such as
supermarkets.
Crops are grown in a wide variety of soils, landscapes, and climates.
There is therefore no standard MZ delineation technique for soil fertility,
which can be universally applied; MZ delineation should be case specific.
The key research challenges are: (1) to include the most influential
yield-limiting factors in the creation of MZ, (2) to measure effectively
these properties at a scale which is sufficient to characterize the spatial var-
iation, (3) to create zones using methods which maximize between-zone
variation yet minimize within-zone variation, and (4) to reduce the num-
ber of MZs to a number that is cost effective and practical for the grower to
implement. At present, it is technically possible to produce very small MZ
from fine-scale survey data (e.g., <2 m sampling) but they are impractical
to implement, as there are technical restriction regarding the size and speed
of response of the current agricultural machinery. The current farm
machinery cannot change application rates speedily enough when in
motion. An alternative, stop/start application process would be uneco-
nomic due to the time taken.
Future research into MZ delineation techniques should favor practical

and affordable systems which make use of the latest sensor technology
and advanced geostatistical techniques. Growers need to be encouraged
to adopt MZ and precision farming as “normal practice” so that the envi-
ronment and agricultural industry worldwide can benefit from increased
efficiency and a reduction in wasted inputs.
ACKNOWLEDGMENTS
Authors acknowledge the financial support received through Tru-Nject project (No. 36428-
267209), which was jointly sponsored by Innovate UK and Biotechnology and Biological
Sciences Research Council (BBSRC). The corresponding author acknowledge the FWO
funded Odysseus Project (No. G0F9216N).
REFERENCES
Abdu, H., Robinson, D.A., Seyfried, M., Jones, S.B., 2008. Geophysical imaging of water-
shed subsurface patterns and prediction of soil texture and water holding capacity. Water
Resour. Res. 44, W00D18.
Adamchuk, V., Hummel, J., Morgan, M., Upadhyaya, S., 2004. On-the-go soil sensors for
precision agriculture. Comput. Electron. Agric. 44, 71–91.
Adamchuk, V.I., Viscarra Rossel, R.a., Marx, D.B., Samal, A.K., 2011. Using targeted sam-
pling to process multivariate soil sensing data. Geoderma 163, 63–73.
Adhikari, K., Carre, F., Toth, G., 2009. Site Specific Land Management General Concepts and
Applications. European Commission, Luxembourg. http://dx.doi.org/10.2788/32619.
Al-Asadi, R.A., Mouazen, A.M., 2014. Combining frequency domain reflectometry and vis-
ible and near infrared spectroscopy for assessment of soil bulk density. Soil Tillage Res.
135, 60–70.
Ancev, T., Whelan, B., McBratney, A., Stafford, J.V., 2005. Evaluating the benefits from
precision agriculture: the economics of meeting traceability requirements and environ-
mental targets. In: Precision Agriculture ’05. Papers Presented at the 5th European Con-
ference on Precision Agriculture, Uppsala, Sweden. Wageningen Academic Publisher,
pp. 985–992.
Arnó, J., Martı́nez Casasnovas, J.A., Ribes Dasi, M., Rosell, J.R., 2009. Review. Precision
viticulture. Research topics, challenges and opportunities in site-specific vineyard man-
agement. Span. J. Agric. Res. 7, 779–790.
Baudron, F., Giller, K.E., 2014. Agriculture and nature: trouble and strife? Biol. Conserv.
170, 232–245. http://dx.doi.org/10.1016/j.biocon.2013.12.009.
Boufassa, A., Armstrong, M., 1989. Comparison between different kriging estimators. Math.
Geol. 21 (3), 331–345.
Brock, A., Brouder, S.M., Blumhoff, G., Hofmann, B.S., 2005. Defining yield-based man-
agement zones for corn–soybean rotations. Agron. J. 97, 1115–1128.
Brus, D.J., Heuvelink, G.B.M., 2007. Optimization of sample patterns for universal kriging
of environmental variables. Geoderma 138, 86–95. http://dx.doi.org/10.1016/j.
geoderma.2006.10.016.
Burgess, T.M., Webster, R., 1980. Optimal interpolation and isarithmic mapping of soil
properties: I. The semivariogram and punctual kriging. J. Soil Sci. 31, 315–331.
Burrough, P.A., McDonnell, R.A., 1998. Principles of Geographical Information Systems.
Oxford University Press, Oxford. 333 pp.
Castrignanò, A., Maiorana, M., Fornaro, F., Lopez, N., 2002. 3D spatial variability of soil
strength and its change over time in a durum wheat field in Southern Italy. Soil Tillage
Res. 65, 95–108.
Castrignanò, A., Wong, M.T.F., Stelluti, M., De Benedetto, D., Sollitto, D., 2012. Use of
EMI, gamma-ray emission and GPS height as multi-sensor data for soil characterisation.
Geoderma 175–176, 78–89. http://dx.doi.org/10.1016/j.geoderma.2012.01.013.
Chang, D., Zhang, J., Zhu, L., Ge, S.-H., Li, P.-Y., Liu, G.-S., 2014. Delineation of man-
agement zones using an active canopy sensor for a tobacco field. Comput. Electron.
Agric. 109, 172–178.
Chen, F., Kissel, D.E., West, L.T., Adkins, W., Clark, R., Rickman, D., Luvall, J.C., 2004.
Field scale mapping of surface soil clay concentration. Precis. Agric. 5, 7e26.
Christy, C.D., 2008. Real-time measurement of soil attributes using on-the-go near infrared
reflectance spectroscopy. Comput. Electron. Agric. 61, 10–19.
Corwin, D.L., Rhoades, J.D., 1982. An improved technique for determining soil electrical
conductivity-depth relations from above-ground electromagnetic measurements. Soil
Sci. Soc. Am. J. 46, 517.
De Benedetto, D., Castrignano, A., Sollitto, D., Modugno, F., Buttafuoco, G., Papa, G.L.,
2012. Integrating geophysical and geostatistical techniques to map the spatial variation of
clay. Geoderma 171–172, 53–63.
De Benedetto, D., Castrignano, A., Diacono, M., Rinaldi, M., Ruggieri, S.,
Tamborrino, R., 2013a. Field partition by proximal and remote sensing data fusion.
Biosyst. Eng. 114, 372–383.
De Benedetto, D., Castrignan, A., Rinaldi, M., Ruggieri, S., Santoro, F., Figorito, B.,
Gualano, S., Diacono, M., Tamborrino, R., 2013b. An approach for delineating homo-
geneous zones by using multi-sensor data. Geoderma 199, 117–127. http://dx.doi.org/
10.1016/j.geoderma.2012.08.028.
Department for Environment Food and Rural Affairs (DEFRA), 2013. Fertiliser Manual
RB209. TSO (The Stationery Office), Norwich, UK.
de Gruijter, J.J., McBratney, A.B., Taylor, J., 2010. Sampling for high-resolution soil map-
ping. In: Viscarra Rossel, R.A., McBratney, A.B., Minasny, B. (Eds.), Proximal Soil
Sensing. Springer, Dordrecht, Netherlands, pp. 3–14.
de Jong, E., Ballantyne, A.K., Cameron, D.R., Read, D.W.L., 1979. Measurement of appar-
ent electrical conductivity of soils by an electromagnetic induction probe to aid salinity
surveys. Soil Sci. Soc. Am. J. 43, 810.
Dickson, B.L., Scott, K.M., 1997. Interpretation of aerial gammaray surveys: adding the geo-
chemical factors. AGSO J. Aust. Geol. Geophys. 17, 187–200.
Diggle, P.J., Ribeiro Jr., P.J., 2007. Model-Based Geostatistics. Springer, New York, p. 228.
Diggle, P.J., Tawn, J.A., Moyeed, R.A., 1998. Model-based geostatistics. Appl. Stat. 47 (3),
299–350.
Dragovic, S., Onjia, A., 2006. Classification of soil samples according to their geographic
origin using gamma-ray spectrometry and principal component analysis. J. Environ.
Radioact. 89, 150–158.
Du, Q., Chang, N.-B., Yang, C., Srilakshmi, K.R., 2008. Combination of multispectral
remote sensing, variable rate technology and environmental modeling for citrus pest
management. J. Environ. Manage. 86, 14–26.
Duffera, M., White, J.G., Weisz, R., 2007. Spatial variability of southeastern U.S. coastal
plain soil physical properties: implications for site-specific management. Geoderma
137, 327–339.
Fleming, K.L., Westfall, D.G., Wiens, D.W., Brodahl, M.C., 2000a. Evaluating farmer
defined management zone maps for variable rate fertilizer application. Precis. Agric.
2, 201–215.
Fleming K.L., Westfall D.G., Collins F., 2000b. Evaluating management zone technology
and grid soil sampling for variable rate nitrogen application. In: Robert, P.C.,
Rust, R.H., Larson, W.E. (Eds.), Proceedings of the 5th International Conference on
Precision Agriculture, Minneapolis, MN, 16 -19 July 2000. ASA, CSSA, and SSSA,
Madison, WI.
Fleming, K.L., Heermann, D.F., Westfall, D.G., 2004. Evaluating soil color with farmer input
and apparent soil electrical conductivity for management zone delineation. Agron. J.
96, 1581–1587.
Foley, J.a., Ramankutty, N., Brauman, K.a., Cassidy, E.S., Gerber, J.S., Johnston, M.,
Mueller, N.D., O’Connell, C., Ray, D.K., West, P.C., Balzer, C., Bennett, E.M.,
Carpenter, S.R., Hill, J., Monfreda, C., Polasky, S., Rockstr€ om, J., Sheehan, J.,
Siebert, S., Tilman, D., Zaks, D.P.M., 2011. Solutions for a cultivated planet. Nature
478, 337–342.
Fraisse, C.W., Sudduth, K.A., Kitchen, N.R., 2001. Delineation of site-specific management
zones by unsupervised classification of topographic attributes and soil electrical conduc-
tivity. Am. Soc. Agric. Eng. 44, 155–166.
Franzen, D.W., Hopkins, D.H., Sweeney, M.D., Ulmer, M.K., Halvorson, A.D., 2002.
Evaluation of soil survey scale for zone development of site-specific nitrogen manage-
ment. Agron. J. 94, 381–389.
Frogbrook, Z.L., Oliver, M.a., 2007. Identifying management zones in agricultural fields using
spatially constrained classification of soil and ancillary data. Soil Use Manag. 23, 40–51.
Fu, Q., Wang, Z., Jiang, Q., 2010. Delineating soil nutrient management zones based on
fuzzy clustering optimized by PSO. Math. Comput. Model. 51, 1299–1305.
Gago, J., Douthe, C., Coopman, R.E., Gallego, P.P., Ribas-Carbo, M., Flexas, J.,
Escalona, J., Medrano, H., 2015. UAVs challenge to assess water stress for sustainable
agriculture. Agric. Water Manag. 153, 9–19.
Galagedara, L.W., Parkin, G.W., Redman, J.D., von Berktold, P., Endres, A.L., 2005. Field
studies of the GPR ground wave method for estimating soil water content during irri-
gation and drainage. J. Hydrol. 301, 182–197.
Galambošová, J., Rataj, V., Prokeinová, R., Prešinská, J., 2014. Determining the manage-
ment zones with hierarchic and non-hierarchic clustering methods. Res. Agr. Eng.
60, S44–S51.
Gaus, I., Kinniburgh, D.G., Talbot, J.C., Webster, R., 2003. Geostatistical analysis of arsenic
concentration in groundwater in Bangladesh using disjunctive kriging. Environ. Geol.
44 (8), 939–948.
Gish, T.J., Dulaney, W.P., Kung, K.-J.S., Daughtry, C.S.T., Doolittle, J.A., Miller, P.T.,
2002. Evaluating use of ground-penetrating radar for identifying subsurface flow path-
ways. Soil Sci. Soc. Am. J. 66, 1620. http://dx.doi.org/10.2136/sssaj2002.1620.
Godwin, R.J., Wood, G.A., Taylor, J.C., Knight, S.M., Welsh, J.P., 2003. Precision farming
of cereal crops: a review of a six year experiment to develop management guidelines.
Biosyst. Eng. 84, 375–391.
Gomez, C., Viscarra Rossel, R.A., McBratney, A.B., 2008. Soil organic carbon prediction by
hyperspectral remote sensing and field vis-NIR spectroscopy: an Australian case study.
Geoderma 146, 403–411.
Goovaerts, P., 1992. Factorial kriging analysis: a useful tool for exploring the structure of
multivariate spatial soil information. J. Soil Sci. 43, 597–619.
Goovaerts, P., 1997. Geostatistics for Natural Resources Evaluation. Oxford University
Press, New York. 483 pp.
Gregory, A.F., Horwood, J.L., 1961. A Laboratory Study of Gamma Ray Spectra at the Sur-
face of Rocks: Mines Branch Research Report R.85. Department of Mines and
Technical Surveys, Ottawa.
Griffin, T.W., Lowenberg-DeBoer, J., Lambert, D.M., Peone, J., Payne, T.,
Daberkow, S.G., 2004. Adoption, profitability, and making better use of precision farm-
ing data. Purdue University, Department of Agricultural Economics, Staff Papers, West
Lafayette, IN. http://ageconsearch.umn.edu/handle/2861. accessed 13.07.2016.
Grote, K., Hubbard, S., Rubin, Y., 2003. Field-scale estimation of volumetric water content
using ground-penetrating radar ground wave techniques. Water Resour. Res. 39 (1321),
1–13.
Grote, K., Anger, C., Kelly, B., Hubbard, S., Rubin, Y., 2010. Characterization of soil water
content variability and soil texture using GPR groundwave techniques. J. Environ. Eng.
Geophys. 15, 93–110.
Grunwald, S., Vasques, G.M., Rivero, R.G., 2015. Fusion of soil and remote sensing data to
model soil properties. Adv. Agron. 131, 1–109. http://dx.doi.org/10.1016/bs.
agron.2014.12.004.
Guastaferro, F., Castrignanò, A., De Benedetto, D., Sollitto, D., Troccoli, A., Cafarelli, B.,
2010. A comparison of different algorithms for the delineation of management zones.
Precis. Agric. 11, 600–620.
Haghverdi, A., Leib, B.G., Washington-Allen, R.A., Ayers, P.D., Buschermohle, M.J.,
2015. Perspectives on delineating management zones for variable rate irrigation. Com-
put. Electron. Agric. 117, 154–167.
Al Hagrey, A., M€ uller, 2000. GPR study of pore water content and salinity in sand. Geophys.
Prospect. 48, 63–85. http://dx.doi.org/10.1046/j.1365-2478.2000.00180.x.
Halcro, G., Corstanje, R., Mouazen, A.M., 2013. Site-specific land management of
cereal crops based on management zone delineation by proximal soil sensing. In:
Stafford, J. (Ed.), Precision Agriculture 2013, Proceedings of the 10th European Con-
ference on Precision Agriculture. Wageningen Academic Publishers, Wageningen,
pp. 475–481.
Hartigan, J.a., Wong, M.a., 1979. A K-means clustering algorithm. J. R. Stat. Soc.
28, 100–108. http://dx.doi.org/10.2307/2346830.
Heijting, S., de Bruin, S., Bregt, A.K., 2011. The arable farmer as the assessor of within-field
soil variation. Precis. Agric. 12, 488–507. http://dx.doi.org/10.1007/s11119-010-9197-y.
Hengari, G.M., Hall, C.R., Kozusko, T.J., Bostater, C.R., 2013. Use of ground
penetrating radar for determination of water table depth and subsurface soil characteris-
tics at Kennedy Space Center. In: Michel, U., Civco, D.L., Schulz, K., Ehlers, M.,
Nikolakopoulos, K.G. (Eds.), SPIE Remote Sensing. International Society for Optics
and Photonics, Bellingham, WA, USA, p. 889318.
Hornung, A., Khosla, R., Reich, R., Inman, D., Westfall, D.G., 2006. Comparison of
site-specific management zones, soil-color-based and yield-based. Agron. J.
98, 407–415.
Hossain, M.B., Lamb, D.W., Lockwood, P.V., Frazier, P., 2010. EM38 for volumetric soil
water content estimation in the root-zone of deep vertosol soils. Comput. Electron.
Agric. 74, 100–109.
Hu, K., Li, B., Lu, Y., Zhang, F., 2004. Comparison of various spatial interpolation methods
for non-stationary regional soil mercury content. Environ. Sci. 25 (3), 132–137.
Huang, J., Lark, R.M., Robinson, D.A., Lebron, I., Keith, A.M., Rawlins, B., Tye, A.,
Kuras, O., Raines, M., Triantafilis, J., 2014. Scope to predict soil properties at
within-field scale from small samples using proximally sensed -ray spectrometer and
EM induction data. Geoderma 232–234, 69–80.
Huisman, J.A., Hubbard, S.S., Redman, J.D., Annan, A.P., 2003. Measuring soil water con-
tent with ground penetrating radar. Vadose Zo. J. 2, 476. http://dx.doi.org/10.2136/
vzj2003.4760.
Hyyti€ainen, K., Niemi, J.K., Koikkalainen, K., Palosuo, T., Salo, T., 2011. Adaptive opti-
mization of crop production and nitrogen leaching abatement under yield uncertainty.
Agric. Syst. 104, 634–644.
Inman, D., Khosla, R., Reich, R., Westfall, D.G., 2008. Normalized difference vegetation
index and soil color-based management zones in irrigated maize. Agron. J. 100, 60–66.
Isaaks, E.H., Srivastava, R.M., 1989. An Introduction to Applied Geostatistics. Oxford
University Press, New York, p. 561.
Johnson, C.K., Doran, J.W., Duke, H.R., Wienhold, B.J., Eskridge, K.M., Shanahan, J.F.,
2001. Field-scale electrical conductivity mapping for delineating soil condition. Soil Sci.
Soc. Am. J. 65, 1829–1837.
Johnson, C.K., Mortensen, D.A., Wienhold, B.J., Shanahan, J.F., Doran, J.W., 2003.
Site-specific management zones based on soil electrical conductivity in a semiarid
cropping system. Agron. J. 95, 303–315.
Jonard, F., Mahmoudzadeh, M., Roisin, C., Weiherm€ uller, L., Andre, F., Minet, J.,
Vereecken, H., Lambot, S., 2013. Characterization of tillage effects on the spatial var-
iation of soil properties using ground-penetrating radar and electromagnetic induc-
tion. Geoderma 207–208, 310–322. http://dx.doi.org/10.1016/j.geoderma.2013.
05.024.
Journel, A.G., Rossi, M.E., 1989. When do we need a trend model in kriging? Math. Geol.
21, 715–738.
Kaivosoja, J., Jackenkroll, M., Linkolehto, R., Weis, M., Gerhards, R., 2014. Automatic
control of farming operations based on spatial web services. Comput. Electron. Agric.
100, 110–115.
Khaleghi, B., Khamis, A., Karray, F.O., Razavi, S.N., 2013. Multisensor data fusion: a
review of the state-of-the-art. Inf. Fusion 14, 28–44. http://dx.doi.org/10.1016/
j.inffus.2011.08.001.
Khosla, R., Alley, M.M., 1999. Soil-specific nitrogen management on Mid-atlantic coastal
plain soils. Better Crop. 83, 6–7.
Khosla, R., Fleming, K., Delgado, J.A., Shaver, T.M., Westfall, D.G., 2002. Use of
site-specific management zones to improve nitrogen management for precision agricul-
ture. J. Soil Water Conserv. 57, 513–518.
Khosla, R., Inman, D., Westfall, D.G., Reich, R.M., Frasier, M., Mzuku, M., Koch, B.,
Hornung, A., 2008. A synthesis of multi-disciplinary research in precision agriculture:
site-specific management zones in the semi-arid western Great Plains of the USA. Precis.
Agric. 9, 85–100.
Khosla, R., Westfall, D.G., Reich, R.M., Mahal, J.S., Gangloff, W.J., 2010. Spatial variation
and site-specific management zones. In: Oliver, M.A. (Ed.), Geostatistical Applications
for Precision Agriculture. Springer, Dordrecht, Netherlands, pp. 195–219.
King, J.A., Lark, R.M., Wheeler, H.C., Park, W., Bradley, R.I., Mayr, T.R., 2005. Mapping
potential crop management zones within fields : use of yield-map series and patterns of
soil physical properties identified by electromagnetic induction sensing. Precis. Agric.
6, 167–181.
Kitanidis, P.K., 1997. Introduction to Geostatistics: Applications in Hydrogeology.
Cambridge University Press, UK, p. 249.
Kitchen, N.R., Sudduth, K.A., Myers, D.B., Drummond, S.T., Hong, S.Y., 2005. Delin-
eating productivity zones on claypan soil fields using apparent soil electrical conductivity.
Comput. Electron. Agric. 46, 285–308.
Koch, B., Khosla, R., Frasier, W.M., Westfall, D.G., Inman, D., 2004. Economic feasibility
of variable-rate nitrogen application utilizing site-speicific management zones. Agron. J.
96, 1572–1580.
Kodaira, M., Shibusawa, S., 2013. Using a mobile real-time soil visible-near infrared sensor
for high resolution soil property mapping. Geoderma 199, 64–79.
Kohonen, T., 1995. Self-Organizing Maps. Springer, Berlin.
Kuang, B., Mouazen, A.M., 2013. Effect of spiking strategy and ratio on calibration of
on-line visible and near infrared soil sensor for measurement in European farms. Soil Till-
age Res. 128, 125–136.
Kuang, B., Mahmood, H.S., Quraishi, M.Z., Hoogmoed, W.B., Mouazen, A.M., van
Henten, E.J., 2012. Sensing soil properties in the laboratory, in situ, and on-line.
A review. Adv. Agron. 114, 155–223.
Kuang, B., Tekin, Y., Mouazen, A.M., 2015. Comparison between artificial neural network
and partial least squares for on-line visible and near infrared spectroscopy measurement of
soil organic carbon, pH and clay content. Soil Tillage Res. 146, 243–252.
Kweon, G., Lund, E., Maxton, C., 2013. Soil organic matter and cation-exchange capacity
sensing with on-the-go electrical conductivity and optical sensors. Geoderma
199, 80–89.
Laliberte, A.S., Rango, A., 2011. Image processing and classification procedures for analysis
of sub-decimeter imagery acquired with an unmanned aircraft over arid rangelands.
GISci. Remote Sens. 48, 4–23.
Lamb, J.A., Dowdy, R.H., Anderson, J.L., Rehm, G.W., 1997. Spatial and temporal stability
of corn grain yields. JPA 10, 410.
Lamb, D.W., Frazier, P., Adams, P., 2008. Improving pathways to adoption: putting the
right P’s in precision agriculture. Comput. Electron. Agric. 61, 4–9.
Lambot, S., Slob, E.C., Van, B., Den, I., Stockbroeckx, B., Vanclooster, M., 2004. Modeling
of ground-penetrating radar for accurate characterization of subsurface electric proper-
ties. IEEE Trans. Geosci. Remote Sens. 42, 2555–2568.
Li, Y., Shi, Z., Li, F., Li, H.-Y., 2007. Delineation of site-specific management zones using
fuzzy clustering analysis in a coastal saline land. Comput. Electron. Agric. 56, 174–186.
Li, Y., Shi, Z., Wu, C., Li, H., Li, F., 2008. Determination of potential management
zones from soil electrical conductivity, yield and crop data. J. Zhejiang Univ. Sci. B
9, 68–76.
Link, J., Senner, D., Claupein, W., 2013. Developing and evaluating an aerial sensor platform
(ASP) to collect multispectral data for deriving management decisions in precision farm-
ing. Comput. Electron. Agric. 94, 20–28.
Lopez-Granados, F., Jurado-Exposito, M., Pena-Barragan, J.M., Garcia-Torres, L., 2005.
Using geostatistical and remote sensing approaches for mapping soil properties. Eur. J.
Agron. 23, 279–289.
Lunt, I.A., Hubbard, S.S., Rubin, Y., 2005. Soil moisture content estimation using
ground-penetrating radar reflection data. J. Hydrol. 307, 254–269.
MacMillan, R.A., Pettapiece, W.W., Nolan, S.C., Goddard, T.W., 2000. A generic proce-
dure for automatically segmenting landforms into landform elements using DEMs,
heuristic rules and fuzzy logic. Fuzzy Sets Syst. 113, 81–109.
Magalhães, P.S.G., Sanches, G.M., Molin, J.P., Portz, G., Amaral, L., Kolln, O.T.,
Duft, G.D., 2013. Potential solution to contribute for sugar cane spatial variability man-
agement. In: EFITA-WCCA-CIGR Conference “Sustainable Agriculture through ICT
Innovation”, Turin, Italy, 24–27 June 2013.
Mahmood, H.S., Hoogmoed, W.B., van Henten, E.J., 2012. Sensor data fusion to predict
multiple soil properties. Precis. Agric. 13, 628–645.
Marı́n-González, O., Kuang, B., Quraishi, M.Z., Munóz-Garcı́a, M.Á., Mouazen, A.M.,
2013. On-line measurement of soil properties without direct spectral response in near
infrared spectral range. Soil Tillage Res. 132, 21–29.
Matheron, G., 1969. Le krigeage universel. Cahiers du Centre de Morphologie
Mathematique vol. 1 Ecole des Mines de Paris, Fontainebleau.
Mcbratney, A.B., Odeh, I.O.A., 1997. Application of fuzzy sets in soil science: fuzzy logic,
fuzzy measurements and fuzzy decisions. Geoderma 77, 85–113.
McBratney, A.B., Webster, R., 1983. Optimal interpolation and isarithmic mapping of soil
properties. V: Co-regionalization and multiple sampling strategies. Eur. J. Soil Sci.
34, 137–162.
McBratney, A.B., Hart, G.A., McGarry, D., 1991. The use of region partitioning to improve
the representation of geostatistically mapped soil attributes. J. Soil Sci. 42, 513–532.
McBratney, A.B., Whelan, B., Ancev, T., Bouma, J., 2005. Future directions of precision
agriculture. Precis. Agric. 6, 7–23.
Meirvenne, M.V., Islam, M.M., Smedt, P.D., Meerschman, E., Van De Vijver, E., Saey, T.,
2013. Key variables for the identification of soil management classes in the aeolian land-
scapes of northewest Europe. Geoderma 199, 99–105.
Metternicht, G., 1996. Detecting and Monitoring Land Degradation Features and Processes
in the Cochabamba Valleys, Bolivia. A Synergistic Approach. ITC Enschede, The
Netherlands.
Metternicht, G., Zinck, J., 2003. Remote sensing of soil salinity: potentials and constraints.
Remote Sens. Environ. 85, 1–20.
Meul, M., Van Meirvenne, M., 2003. Kriging soil texture under different types of non-
stationarity. Geoderma 112, 217–233.
Milne, A.E., Webster, R., Ginsburg, D., Kindred, D., 2012. Spatial multivariate classification
of an arable field into compact management zones based on past crop yields. Comput.
Electron. Agric. 80, 17–30.
Minasny, B., McBratney, A.B., 2002. FuzME version 3.0, Australian Centre for Precision
Agriculture. McMillan Building A05, The University of Sydney, NSW 2006. http://
sydney.edu.au/agriculture/pal/software/fuzme.shtml (accessed 13.07.2016).
Moral, F.J., Terrón, J.M., da Silva, J.R.M., 2010. Delineation of management zones using
mobile measurements of soil apparent electrical conductivity and multivariate
geostatistical techniques. Soil Tillage Res. 106, 335–343.
Moral, F.J., Terrón, J.M., Rebollo, F.J., 2011. Site-specific management zones based on the
Rasch model and geostatistical techniques. Comput. Electron. Agric. 75, 223–230.
Mouazen, A.M., 2006. Soil Sensing Device. In: International Publication, Published Under
the Patent Cooperation Treaty (PCT). World Intellectual Property Organization,
International Bureau, International Publication Number; W02006/015463; PCT/BE
2005/000129; G01N21/00GO1N21/00.
Mouazen, A.M., Kuang, B., 2016. On-line visible and near infrared spectroscopy for in-field
phosphorous management. Soil Tillage Res. 155, 471–477.
Mouazen, A.M., De Baerdemaeker, J., Ramon, H., 2005. Towards development of on-line
soil moisture content sensor using a fibre-type NIR spectrophotometer. Soil Tillage Res.
80, 171–183. http://dx.doi.org/10.1016/j.still.2004.03.022.
Mouazen, A.M., Maleki, M.R., De Baerdemaeker, J., Ramon, H., 2007. On-line measure-
ment of some selected soil properties using a VIS–NIR sensor. Soil Tillage Res. 93, 13–27.
Mouazen, A.M., Kuang, B., De Baerdemaeker, J., Ramon, H., 2010. Comparison among
principal component, partial least squares and back propagation neural network analyses
for accuracy of measurement of selected soil properties with visible and near infrared
spectroscopy. Geoderma 158, 23–31.
Mouazen, A.M., Alhwaimel, S.A., Kuang, B., Waine, T., 2014. Multiple on-line soil sensors
and data fusion approach for delineation of water holding capacity zones for site specific
irrigation. Soil Tillage Res. 143, 95–105.
Moyeed, R.A., Papritz, A., 2002. An empirical comparison of kriging methods for nonlinear
spatial point prediction. Math. Geol. 34 (4), 365–386.
Mulla, D.J., Bhatti, A.U., Hammond, M.W., Benson, J.A., 1992. A comparison of winter
wheat yield and quality under uniform versus spatially variable fertilizer management.
Agric. Ecosyst. Environ. 38, 301–311.
Mzuku, M., Khosla, R., Reich, R., Inman, D., Smith, F., MacDonald, L., 2005. Spatial var-
iability of measured soil properties across site-specific management zones. Soil Sci. Soc.
Am. J. 69, 1572–1579.
National Research Council, 1997. Precision Agriculture in the 21st Century: Geospatial and
Information Technologies in Crop Management. National Academies Press,
Washington, DC.
Nolan, S.C., Goddard, T.W., Lohstraeter, G., Coen, G.M., Robert, P.C., Rust, R.H.,
Larson, W.E., 2000. Assessing managements units on rolling topography.
In: Proceedings of the 5th International Conference on Precision Agriculture, Bloom-

ington, Minnesota, USA, 16–19 July, 2000. American Society of Agronomy, pp. 1–12.
Odeh, I.O., McBratney, A.B., Chittleborough, D.J., 1995. Further results on prediction of
soil properties from terrain attributes: heterotopic cokriging and regression-kriging.
Geoderma 67, 215–226.
Oliver, M.A., Webster, R., McGrath, S.P., 1996. Disjunctive kriging for environmental
management. Environmetrics 7, 333–358.
Ortega, R.A., Santibáñez, O.A., 2007. Determination of management zones in corn (Zea
mays L.) based on soil fertility. Comput. Electron. Agric. 58, 49–59.
Panda, S., Sahu, S., Jena, P., Chattopadhyay, S., 2012. Comparing fuzzy-C means and
K-means clustering techniques: a comprehensive study. In: Wyld, D.C., Zizka, J.,
Nagamalai, D. (Eds.), Proceedings of 2nd International Conference on Computer Sci-
ence, Engineering & Applications. In: Advances in Intelligent and Soft Computing
(AISC), vol. 166, pp. 451–460.
Pantazi, X.E., Moshou, D., Mouazen, A.M., Alexandridis, T., Kuang, B., 2015. Data fusion
of proximal soil sensing and remote crop sensing for the delineation of management
zones in arable crop precision farming. In: CEUR Workshop Proceedings.
CEUR-WS, pp. 765–776.
Peralta, N.R., Costa, J.L., 2013. Delineation of management zones with soil apparent elec-
trical conductivity to improve nutrient management. Comput. Electron. Agric.
99, 218e226.
Peralta, N.R., Costa, J.L., Balzarini, M., Castro Franco, M., Córdoba, M., Bullock, D., 2015.
Delineation of management zones to improve nitrogen management of wheat. Comput.
Electron. Agric. 110, 103–113.
Petersen, H., Fleige, H., Rabbel, W., Horn, R., 2005. Applicability of geophysical pros-
pecting methods for mapping of soil compaction and variability of soil texture on farm
land. J. Plant Nutr. Soil Sci. 168, 68–79.
Phillips, J.D., 2001. Divergent evolution and the spatial structure of soil landscape variability.
Catena 43, 101–113.
Pierpaoli, E., Carli, G., Pignatti, E., Canavari, M., 2013. Drivers of precision agriculture
technologies adoption: a literature review. Procedia Technol. 8, 61–69.
Pike, R.J., Evans, I.S., Hengl, T., 2009. Geomorphometry—concepts, software, applications.
Dev. Soil Sci., 33, pp. 3–30.
Pracilio, G., Adams, M.L., Smettem, K.R.J., Harper, R.J., 2006. Determination of spatial
distribution patterns of clay and plant available potassium contents in surface soils at
the farm scale using high resolution gamma ray spectrometry. Plant Soil 282, 67–82.
Primicerio, J., Di Gennaro, S.F., Fiorillo, E., Genesio, L., Lugato, E., Matese, A.,
Vaccari, F.P., 2012. A flexible unmanned aerial vehicle for precision agriculture. Precis.
Agric. 13, 517–523.
Quinlan, J., 1993. Programs for Machine Learning, C4.5: Programs for Machine Learning.
Morgan Kaufmann Publishers Inc, San Francisco, CA.
Quraishi, M.Z., Mouazen, A.M., 2013. Calibration of an on-line sensor for measurement of
topsoil bulk density in all soil textures. Soil Tillage Res. 126, 219–228.
Rampant, P., Abuzar, M., 2004. Geophysical tools and digital elevation models : tools for
understanding crop yield and soil variability. In: SuperSoil 2004 3rd Aust. New Zeal.
Soils Conf. 5–9 December 2004, Univ. Sydney, Australia.
Rhoades, J.D., Corwin, D.L., 1981. Determining soil electrical conductivity-depth relations
using an inductive electromagnetic soil conductivity meter1. Soil Sci. Soc. Am. J.
45, 255.
Rodrigues, F.A., Bramley, R.G.V., Gobbett, D.L., 2015. Proximal soil sensing for precision
agriculture: simultaneous use of electromagnetic induction and gamma radiometrics in
contrasting soils. Geoderma 243–244, 183–195.
Rodriguez, H.G., Popp, J., Gbur, E., Chaubey, I., 2011. Environmental and economic
impacts of reducing total phosphorous runoff in an agricultural watershed. Agric. Syst.
104, 623–633.
Rokhmana, C.A., 2015. The potential of UAV-based remote sensing for supporting preci-
sion agriculture in Indonesia. Procedia Environ. Sci. 24, 245–253.
Ruß, G., Kruse, R., 2011. Advances in data mining. Applications and Theoretical Aspects,
Industrial Conference on Data Mining—Workshops, Lecture Notes in Computer
Science. Springer, Berlin, Heidelberg.
Saey, T., Meirvenne, M.V., Vermeersch, H., Ameloot, N., Cockx, L., 2009. A pedotransfer
function to evaluate the soil profile textural heterogeneity using proximally sensed appar-
ent electrical conductivity. Geoderma 150, 389–395.
Sarangi, S.K., Maji, B., Singh, S., Sharma, D.K., Burman, D., Mandal, S., Singh, U.S.,
Ismail, A.M., Haefele, S.M., 2015. Using improved variety and management enhances
rice productivity in stagnant flood-affected tropical coastal zones. F. Crop. Res.
190, 70–81. http://dx.doi.org/10.1016/j.fcr.2015.10.024.
Schepers, A.R., Shanahan, J.F., Liebig, M.a., Schepers, J.S., Johnson, S.H., Luchiari, A.,
2004. Appropriateness of management zones for characterizing spatial variability of soil
properties and irrigated corn yields across years. Agron. J. 96, 195–203.
Serbin, G., Or, D., 2003. Near-surface water content measurements using horn antenna
radar: Methodology and overview. Vadose Zone J. 2, 500–510.
Shaddad, S.M., Madrau, S., Castrignanò, A., Mouazen, A.M., 2015. Data fusion techniques
for delineation of site-specific management zones in a field in UK. Precis. Agric. 17 (2),
200–217.
Shibusawa, S., Made Anom, S.W., Sato, H.P., Sasao, A., 2001. Soil mapping using the 512
real-time soil spectrometer. In: Gerenier, G., Blackmore, S. (Eds.), In: ECPA 2001,
vol. 2. agro Montpellier, Montpellier, France, pp. 485–490.
Srinivasan, 2006. Handbook of Precision Agriculture: Principles and Applications. Food
Products Press, New York, NY. 708 p.
Stafford, J.V., 2000. Implementing precision agriculture in the 21st century. J. Agric. Eng.
Res. 76, 267–275.
Stenberg, B., Viscarra Rossel, R.A., Mouazen, A.M., Wetterlind, J., 2010. Visible and near
infrared spectroscopy in soil science. Adv. Agron. 107, 163–215.
Sudduth, K.A., Kitchen, N.R., Hughes, D.F., Drummond, S.T., 1995. Electromagnetic
induction sensing as an indicator of productivity on claypan soils. In: Robert, P.C.,
et al. (Eds.), Proc. Int. Conf. on Site- Specific Management for Agricultural Systems,
2nd, Minneapolis, MN. 27–30 March 1994. ASA, CSSA, and SSSA, Madison, WI.
Sumfleth, K., Duttmann, R., 2008. Prediction of soil property distribution in paddy soil land-
scapes using terrain data and satellite information as indicators. Ecol. Indic. 8, 485–501.
Sun, Y., Druecker, H., Hartung, E., Hueging, H., Cheng, Q., Zeng, Q., Sheng, W., Lin, J.,
Roller, O., Paetzold, S., Schulze Lammers, P., 2011. Map-based investigation of soil physical
conditions and crop yield using diverse sensor techniques. Soil Tillage Res. 112, 149e158.
Suzuki, R., Shimodaira, H., 2006. Pvclust: an R package for assessing the uncertainty in hier-
archical clustering. Bioinformatics 22, 1540–1542.
Tagarakis, A., Liakos, V., Fountas, S., Koundouras, S., Gemtos, T.A., 2013. Management
zones delineation using fuzzy clustering techniques in grapevines. Precis. Agric. 14, 18–39.
Taylor, M.J., Smettem, K., Pracilio, G., Verboom, W., 2002. Relationships between soil
properties and high-resolution radiometrics, central eastern Wheatbelt, western Austra-
lia. Explor. Geophys. 33, 95–102.
Taylor, J.A., Short, M., McBratney, A.B., Wilson, J., 2010. Comparing the ability of mul-
tiple soil sensors to predict soil properties in a scottish potato production system.
In: Viscarra Rossel, R.A., McBratney, A.B., Minasny, B. (Eds.), Proximal Soil Sensing,
Progress in Soil Science 1. Springer Netherlands, Dordrecht.
Tekin, A.B., 2010. Variable rate fertilizer application in Turkish wheat agriculture : eco-
nomic assessment. Afr. J. Agric. Res. 5, 647–652.
van Alphen, B.J., Stoorvogel, J.J., 1999. A methodology to define management units in sup-
port of an integrated, model-based approach to precision agriculture. In: Precision Agri-
culture. ACSESS Publications. American Society of Agronomy, Crop Science Society of
America, Soil Science Society of America, Madison, WI, pp. 1267–1278.
Van Meirvenne, M., Van De Vijver, E., Vandenhaute, L., Seuntjens, P., 2014. Investigating
soil pollution with the aid of EMI and GPR measurements. In: Proc. 15th Int. Conf. Gr.
Penetrating Radar, pp. 1006–1010.
Viscarra Rossel, R.A., McGlynn, R.N., McBratney, A.B., 2006. Determining the compo-
sition of mineral-organic mixes using UV–vis–NIR diffuse reflectance spectroscopy.
Geoderma 137, 70–82. http://dx.doi.org/10.1016/j.geoderma.2006.07.004.
Viscarra Rossel, R.A., Taylor, H.J., McBratney, A.B., 2007. Multivariate calibration of
hyperspectralg-ray energy spectra for proximal soil sensing. Eur. J. Soil Sci. 58, 343–353.
Viscarra Rossel, R.A., McBratney, A.B., Minasny, B. (Eds.), 2010a. Proximal soil sensing,
Progress in Soil Science Series. Springer-Verlag, New York.
Viscarra Rossel, R.A., Rizzo, R., Demattê, J.a.M., Behrens, T., 2010b. Spatial modeling of a
soil fertility index using visible–near-infrared spectra and terrain attributes. Soil Sci. Soc.
Am. J. 74 (4), 1293–1300.
Vrindts, E., Mouazen, A.M., Reyniers, M., Maertens, K., Maleki, M.R., Ramon, H., De
Baerdemaeker, J., 2005. Management zones based on correlation between soil compac-
tion, yield and crop data. Biosyst. Eng. 92, 419–428.
Wackernagel, H., 1988. Geostatistical techniques for interpreting multivariate spatial infor-
mation. In: Chung, C.F. (Ed.), Quantitative Analysis of Mineral and Energy Resources.
D. Reidel Publishing Company, Dordrecht, Holland.
Wackernagel, H., 1995. Multivariate Geostatistics. Springer, Berlin.
Wackernagel, H., 2003. Multivariate Geostatistics: An Introduction with Applications.
Springer, Berlin. 387 pp.
Walthall, C.L., Gish, T.J., Daughtry, C.S.T., Dulaney, W.P., Kung, K.-J.S., McCarty, G.,
Timlin, D., Angier, J.T., Buss, P., Houser, P.R., 2001. An innovative approach for
locating and evaluating subsurface pathways for nitrogen loss. Sci. World J. 1, 223–229.
Wang, Z.M., Song, K.S., Zhang, B., Liu, D.W., Li, X.Y., Ren, C.Y., Zhang, S.M., Luo, L.,
Zhang, C.H., 2009. Spatial variability and affecting factors of soil nutrients in croplands
of Northeast China: a case study in Dehui County. Plant Soil Environ. 55, 110–120.
Webester, R., 1994. The development of pedometerics. Geoderma 62, 1–5.
Webster, R., Oliver, M.A., 2007. Geostatistics for Environmental Scientists Statistics in Prac-
tice. John Wiley & Sons Ltd, The Atrium, England.
Welsh, J., Wood, G., Godwin, R., Taylor, J., Earl, R., Blackmore, S., Knight, S., 2003.
Developing strategies for spatially variable nitrogen application in cereals, part I: Winter
Barley. Biosyst. Eng. 84, 481–494.
Wetterlind, J., Stenberg, B., S€oderstr€om, M., 2008. The use of near infrared (NIR) spectros-
copy to improve soil mapping at the farm scale. Precis. Agric. 9, 57–69.
Whelan, B., McBratney, A., 2003. Definition and interpretation of potential management
zones in Australia. In: Proc. 11th Aust. Agron. Conf. 2–6.
Williams, B., Baker, G., 1982. An electromagnetic induction technique for reconnaissance
surveys of soil salinity hazards. Aust. J. Soil. Res. 20 (2), 107–118.
Yao, R.-J., Yang, J.-S., Zhang, T.-J., Gao, P., Wang, X.-P., Hong, L.-Z., Wang, M.-W.,
2014. Determination of site-specific management zones using soil physico-chemical
properties and crop yields in coastal reclaimed farmland. Geoderma 232–234, 381–393.
Zadeh, L.A., 1965. Fuzzy sets. Inf. Control 8 (3), 338–353.
Zhang, C., Kovacs, J.M., 2012. The application of small unmanned aerial systems for pre-
cision agriculture: a review. Precis. Agric. 13, 693–712.
INDEX
Note: Page numbers followed by “f ” indicate figures, and “t” indicate tables.
A Cross-polarization (CP), 67
Aegilops tauschii, 176–177 Cross-polarization magic angle spinning
Alfisols, 8–9 (CPMAS), 66–67
Algaenan Cutan
characteristics, 108–109 identification, 104
isolation, 109 structure of, 104–105, 104f
PyGC/MS, 109–110 thermochemolysis, 104–105
structure of, 108–109, 109f Cutin, 103
Application maps, 220–221
B D
Bacteran, 110 Database and drought stress
Biochar, 111–113 composition
Black carbon (BC), 111–113 classification of, 147–149,
Bloch decay (BD). See Direct polarization 148–149t
(DP) distribution of experiments, 147–149,
147f
C construction
Cellulose, 94–95, 94f criteria for, 145–147
Char, 111–113 process and meta-analysis, 145–147,
Chemical shift anisotropy (CSA), 68 146f
13
C nuclei NMR spectroscopy Data fusion
advantages, 66–67 advantages, 208
CP and spin diffusion, 67 ECa measurement
CPMAS, 66–67 clustering algorithms, GIS, 211
CSA, 68 EC shallow (ECSH) and deep (ECDP)
DD, 69 measurements, 211–212
dipolar and quadrupolar interactions, 70 EMI technique, 210–211
disadvantages, 66–67 regression kriging technique, 213
DP, 69 terrain elevation and soil depth,
MAS, 69 211–212
paramagnetic species, 70 GPR, 215–217
ramped amplitude CPMAS, 67–68 γ-ray spectrometer, 214–215
SSB, 68 location of, 208, 209f
Cokriging, 187 multisensor and
Collocated cokriging (CCK), 187–188 GPR data, 219
Comprehensive multiphase (CMP) NMR proximal sensor fusion, 218–219
spectroscopy remote and proximal sensors fusion,
applications of, 70–71 219–220
carboxylates and aromatic components, remote sensor (RS) fusion, 220
86–87 NIR region, 217
CPMAS, 85–86, 85f online vis–NIR spectroscopy sensors,
DMSO, 86 217–218
247
248 Index
Data fusion (Continued ) spatial scales, agricultural systems, 28, 28t

sensor sampling watershed
calibration sampling, 208–209 challenges, 30–31
EMI, 209–210 degradation patterns, 31
GPR, 209–210 Revised Universal Soil Loss
Diffusion-editing (DE), 1H NMR, 66 Equation (RUSLE2) model, 31
Digital elevation model (DEM), 200–201 Elevation maps, 200–201
Dipolar and quadrupolar interactions, 70
Dipolar dephasing (DD), 69 F
Direct polarization (DP), 69 Factorial kriging (FK), 186
Disjunctive kriging (DK), 188 Farmer-defined MZs
Drought stress agricultural efficiency (AE), 198
climate change and evaluation of, 197
biomass allocation, 143 N application strategies, 198
CO2 concentration, 143–144 soil color-based management zone
wheat growth, 142–143 (SCMZ), 199
database spatial variability, 198–199
composition, 147–149, 147f, yield-based MZ (YBMZ), 199
148–149t
construction, 145–147, 146f G
hexaploid (6n) wheat, 141–142 Glomalin, 110–111
International Maize and Wheat Ground penetration radar (GPR)
Improvement Center (CIMMYT), multisensor and, 219
141–142 PSS, 215–217
meta-analysis, 144–145 sensor sampling, 209–210
physiological traits, 156–160
ploidy wheat H
limited gene pool, 168–169 Hemicellulose, 95–96
phenological stages, 166–167 Heteronuclear correlation experiments, 73
plant architecture, biomass allocation, Heteronuclear multiple quantum
and physiological traits, 156–160, coherence (HMQC) technique, 73,
157f 89–90, 90f
root environment and type, 160–166, Heteronuclear single-quantum coherence
161f, 164f (HSQC), 73, 91–92, 91f
root pruning, 167–168 High-resolution magic angle spinning
water content, 167–168 (HRMAS) 1H NMR spectroscopy,
yield and yield components, 151–156, 70–71
152f, 153–155t Homonuclear Hartmann–Hahn
sources of variation, 149–150 spectroscopy (HOHAHA).
statistical analyses, 150–151 See Total correlation spectroscopy
synthetic 6n wheat, 141–142 (TOCSY)
tetraploid (4n) wheat, 142 Humin
wheat domestication, 140–141, 141f acidified DMSO, 115
aliphatic hydrocarbon, 115
E alkyl hydrocarbon, dominance of, 119
Ecosystem services and soil clay fraction, 120–121
field, 28–30, 29f climate and vegetation, effects of,
landscape, 30 119–120
Index 249
CMP-NMR black carbon, char/biochar, 111–113

applications of, 70–71 cellulose, 94–95, 94f
carboxylates and aromatic components, cutan, 104–105, 104f
86–87 cutin, 103
CPMAS, 85–86, 85f glomalin, 110–111
DMSO, 86 hemicelluloses, 95–96
composition of, 60–61 latex materials, 108
cuticular components, 116–117 lignin, 96–98, 96–97f
definition, 48–49 lipids, 100–102
DE 1H NMR spectra melanins, 111
albumin, 84–85, 84f suberan, 107
methyl and methylene functionalities, suberin, 105–107, 106f
81–83, 83f tannins, 98–100, 99f
spectral regions, 81, 82f radiocarbon studies, 120
from denaturation, 58–59 solid-state 13C NMR, 114–116
2D NMR Spectroscopy, 87–93 SOM
extraction and fractionation alkali insoluble fraction, 50
carbohydrate content, 77 carbon, terrestrial reservoir of, 118
CPMAS 13C, 76–77, 76f contents of, 49
DD spectra, 79, 80f humic acids (HAs) and fulvic acids
DMSO treatments, 75 (FAs), 50
IHSS Mollisol soil standard, 77, 78f humified materials, 50
VACP 13C, 75, 75f microorganism activity, 49–50
vs. humic substances (HSs), 48–49 resistance to biological degradation, 50
hydrophobic species, 120–121 resistant plant materials, 118–119
incubation experiments, 120 soil structure, 118
isolation water retention by, 117
artifacts, 59 structure of, 60
DMSO and H2SO4, 64–65 theories for genesis
HA-like materials (HALMs), 62–63 acid filtrates, 52–53
inorganic mineral matter, 61–62 Browning reaction, 54
methylisobutyl ketone (MIBK), 62–63 enzymatic catalytic synthesis, 57
organic solvents, 63 HAs and FAs, 57–58
solvent systems, 62 ligno-casein complex, 54
urea, 64 Maillard reaction, 54–55
lack of solubility, 114 melanoidins and caramel products, 55
NMR spectroscopy methylglyoxal glycine products, 55
CMP, 70–71 polycyclic aromatic compounds, 56–57
13
C nuclei, 66–70 proteins and encrusting substances,
2D, 71–74 interaction, 54
DE, 1H NMR, 66 soil formation and soil fertility, 52
electromagnetic (EM) radiation, 65 sources of, 53
free-induced decay (FID), 65
organic carbon (OC), 50–51 I
partitioning approach, 114 Innovative variable-rate (VR2) MZ
plant and microbial sources delineation
algaenan, 108–110, 109f applications, 228–230, 229f
bacteran, 110 classification, 231
250 Index
Innovative variable-rate (VR2) MZ fatty acids, 100

delineation (Continued ) hydrocarbons and, 101–102
crop growth and yield measurements, 226 lipases, 101
data fusion and, 227–228 lipoproteins, 101
laboratory chemical and optical analyses, long-term stabilization, 102
226–227 mass spectrometry methods, 102
online sensor measurement and soil waxes, 100–101
sampling, 226
soil mapping, 227
spectra pretreatment and development, M
calibration models, 227 Magic angle spinning (MAS), 69
Inverse distance weighting (IDW) Management zone (MZ) delineation
vs. kriging, 182–183 application (prescription) maps, 220–221
spatial dependence, 182 classification, effectiveness of, 191–197,
192–195t
K cost–benefit analysis
Kriging comparitive analysis, 234
CCK, 187–188 fertilizer applied and financial gain,
cokriging, 187 231–233, 232t
definition, 182 nitrogen/sulfur fertilization
DK, 188 approaches, 231–233, 232t
FK, 186 SSMZ–constant yield goal
vs. IDW, 182–183 (SSMZ–CYG), 222–223
KED, 186–187 SSMZ–variable yield goal
MBK, 188 (SSMZ–VYG), 222–223
OK, 184–185 VRA, 223
PCK, 187 VR1 vs. VR2, 233
PK, 188 crop coverage
SK, 183–184 ECa data, 205–206
SKlm, 186 NDVI, 205–206
UK, 185–186 remote sensing data, 207
variogram model, 183 UAVs, 207
Kriging with an external drift (KED), VRA, 205
186–187 definition, 177–178
Kriging with a trend (KT), 185–186 environmental challenges, 176
farmer knowledge
L agricultural efficiency (AE), 198
Latex materials, 108 evaluation of, 197
Lignin N application strategies, 198
chemical stability and resistance, 98 soil color-based management zone
degradation, 98 (SCMZ), 199
from gymnosperms, 96–98, 97f spatial variability, 198–199
phenylpropenoic alcohols, yield-basedMZ (YBMZ), 199
96–98, 96f final MZ, 191
Lipids geomorphology, 200–201
aliphatic hydrocarbons, 102 homogeneous areas
characteristics, 100 hierarchical clustering, 189
eukaryotic, 100 k-means clustering algorithm, 189
Index 251
principal component analysis (PCA), Duck End Farm and study Horn End
189 Field, 223, 224f
soil fertility index (SFI), 189–190 22 ha study area, 223, 224f
SOM, 190 uniform-rate (UR) fertilization,
IDW 223–224, 225t
vs. kriging, 182–183 VR1, 225, 225t
spatial dependence, 182 VR2, 226–230, 229f
innovative approach (VR2), 231 soil class, 203
kriging soil-limiting factors, 178
CCK, 187–188 spatial interpolation methods, 181
cokriging, 187 traditional soil chemical analyses
definition, 182 fuzzy clustering analysis, 202
DK, 188 limitation, 201
FK, 186 PCA, 201–202
vs. IDW, 182–183 site-specific MZ (SSMZ), 202
KED, 186–187 UR and VR strips, 201
MBK, 188 VR1, 231
OK, 184–185 VRA
PCK, 187 cost–benefit analysis, 223
PK, 188 economic benefits, 222–223
SK, 183–184 SSMZ–constant yield goal
SKlm, 186 (SSMZ–CYG), 222–223
UK, 185–186 SSMZ–variable yield goal
variogram model, 183 (SSMZ–VYG), 222–223
modeling and soil mapping, 230–231, whole-field approach, 177
230t within-field variability, 235
multisensor and data fusion approaches, yield maps, 204
234 Mapping properties, MZ delineation
optimal number of classes, 190–191 IDW
precision agriculture/precision farming, vs. kriging, 182–183
176–178 spatial dependence, 182
PSS and data fusion kriging
advantages, 208 CCK, 187–188
ECa measurement, 210–213 cokriging, 187
GPR, 215–217 definition, 182
γ-ray spectrometer, 214–215 DK, 188
location of, 208, 209f FK, 186
multisensor and, 218–220 vs. IDW, 182–183
NIR region, 217 KED, 186–187
online vis–NIR spectroscopy sensors, MBK, 188
217–218 OK, 184–185
sensor sampling, 208–210 PCK, 187
research challenges, 235 PK, 188
sensing technologies, 178 SK, 183–184
site-specific management practice, SKlm, 186
179–180, 180f UK, 185–186
site-specific N fertilizer application, oil variogram model, 183
seed rape (OSR) spatial interpolation methods, 181
252 Index
Melanins, 111 DMSO treatments, 75

Model-based kriging (MBK), 188 IHSS Mollisol soil standard, 77, 78f
Mollisols, 8–9 VACP 13C, 75, 75f
free-induced decay (FID), 65
N Nuclear Overhauser effect spectroscopy
Nuclear magnetic resonance (NMR) (NOESY), 72–73, 92–93, 93f
spectroscopy Nutrient cycling, 5–7
CMP
applications of, 70–71 O
carboxylates and aromatic components, Oil seed rape (OSR), site-specific
86–87 N fertilizer application
CPMAS, 85–86, 85f Duck End Farm and study Horn End
DMSO, 86 Field, 223, 224f
13
C nuclei 22 ha study area, 223, 224f
advantages, 66–67 uniform-rate (UR) fertilization, 223–224,
CP and spin diffusion, 67 225t
CPMAS, 66–67 VR1, 225, 225t
CSA, 68 VR2
DD, 69 applications, 228–230, 229f
dipolar and quadrupolar Interactions, crop growth and yield measurements,
70 226
disadvantages, 66–67 data fusion and MZ delineation,
DP, 69 227–228
MAS, 69 laboratory chemical and optical
paramagnetic species, 70 analyses, 226–227
ramped amplitude CPMAS, 67–68 online sensor measurement and soil
SSB, 68 sampling, 226
2D soil mapping, 227
applications of, 74 spectra pretreatment and development,
bond interactions, 72 calibration models, 227
F1 and F2 frequencies, 71–72 Ordinary kriging (OK), 184–185
heteronuclear correlation experiments,
73 P
HMQC, 89–90, 90f Paramagnetic species, 70
HSQC, 91–92, 91f Ploidy wheat and drought stress
NOESY, 72–73, 92–93, 93f biomass allocation, 159
TOCSY, 72, 87–89, 88–89f effect of, 157–158, 157f
DE, 1H NMR, 66 leaf water potential (LWP), 157–158
1D 1H DE spectra limited gene pool, 168–169
albumin, 84–85, 84f phenological stages, 166–167
methyl and methylene functionalities, plant architecture, 156–157
81–83, 83f primary and high-order tiller, 160
spectral regions, 81, 82f relative water content (RWC), 157–158
electromagnetic (EM) radiation, 65 root environment and wheat type
extraction and fractionation in pots/field, 160–163, 161–162f
carbohydrate content, 77 spring and winter, 163–166, 164–165f
CPMAS 13C, 76–77, 76f root pruning, 167–168
DD spectra, 79, 80f water content, 167–168
Index 253
yield and yield components 22 ha study area, 223, 224f

between-Group Heterogeneity (QB), uniform-rate (UR) fertilization, 223–224,
151–152, 153–155t 225t
grain number, 152–156 VR1, 225, 225t
preanthesis stem reserves, 152–156 VR2
thousand kernel weight (TKW), applications, 228–230, 229f
151–152 crop growth and yield measurements,
yield losses, 158–159 226
Prescription maps, 220–221 data fusion and MZ delineation,
Principal component kriging (PCK), 187 227–228
Probability kriging (PK), 188 laboratory chemical and optical
Proximal soil sensors (PSS) analyses, 226–227
advantages, 208 online sensor measurement and soil
ECa measurement sampling, 226
clustering algorithms, GIS, 211 soil mapping, 227
EC shallow (ECSH) and deep (ECDP) spectra pretreatment and development,
measurements, 211–212 calibration models, 227
EMI technique, 210–211 SK with varying local means (SKlm), 186
regression kriging technique, 213 Soil
terrain elevation and soil depth, Alfisols, 8–9
211–212 challenges
GPR, 215–217 awareness of, 36
γ-ray spectrometer, 214–215 food, energy, water nexus, 35–36
location of, 208, 209f reversing soil degradation, 34–35
multisensor and soil carbon, 34–35
GPR data, 219 cropping system, 33–34
proximal sensor fusion, 218–219 degradation
remote and proximal sensors fusion, global evaluation, 10
219–220 global land degradation assessments, 10
remote sensor (RS) fusion, 220 impact of, 2–3
NIR region, 217 types and extent of, 10–12, 11f
online vis–NIR spectroscopy sensors, dryland and irrigated soils, 9–10
217–218 ecosystem services, 4–5
sensor sampling ecosystem services and
calibration sampling, 208–209 field, 28–30, 29f
EMI, 209–210 landscape, 30
GPR, 209–210 spatial scales, agricultural systems, 28,
28t
R watershed, 30–31
Ramped amplitude CPMAS, 67–68 erosion
climate change, extreme rainfall and
S heat, 14–18, 15f
Simple kriging (SK), 183–184 growing food demand, 19–21, 20f
Single-pulse experiment (SPE). See Direct improvment of, 26–28
polarization (DP) linkage of, 21–26
Site-specific N fertilizer application, OSR organic matter content, 12
Duck End Farm and study Horn End quantity of, 12
Field, 223, 224f soil renewal, 13
254 Index
Soil (Continued ) improvment of

urban expansion, 18–19 rehabilitation, 26–27
food security, 32–33 SOM and plant-available water,
functionality of 27–28
aggradation process, 7–8, 8f organic matter content, 12
environmental and societal issues, 5–7 quantity of, 12
organic carbon, 7–8 soil renewal, 13
properties, 5, 6f urban expansion, 18–19
Mollisols, 8–9 water and soil, linkage between
precipitation, 4 deficit irrigation, 23–24
production requirements, 3–4 nutrients and photosynthates
role of, 2–3, 3f transmission, 23
soil aggregate stability, 33–34 pumping, irrigation water, 23
SOM and soil properties, 33–34, 33f water use efficiency (WUE), 23–24
water availability, 4 Soil organic matter (SOM)
Soil degradation alkali insoluble fraction, 50
global evaluation, 10 carbon, terrestrial reservoir of, 118
global land degradation assessments, 10 contents of, 49
impact of, 2–3 homogeneous areas, MZ delineation, 190
types and extent of, 10–12, 11f humic acids (HAs) and fulvic acids (FAs),
Soil erosion 50
climate change, extreme rainfall and heat humified materials, 50
crop productivity, 17–18 microorganism activity, 49–50
management practices, 17–18 plant-available water and, 27–28
multiobjective optimization approach, resistance to biological degradation, 50
14–15 resistant plant materials, 118–119
negative environmental impacts, soil properties and, 33–34, 33f
14–15 soil structure, 118
projected changes, average annual water retention by, 117
precipitation, 15–16, 15f Spin diffusion, 67
temperature and precipitation, Spinning side bands (SSB), 68
combinations, 15–16 Suberan
energy and soil, linkage between characteristics, 107
biofuel, 25–26 saponification processes, 107
correlation coefficient, 24–25 Suberin
direct and indirect uses, 25 composition of, 105–106
Energy Independence and Security Act extraction, 107
(EISA), 26 model for, 105–106, 106f
food and soil, linkage between solid-state NMRstudies, 106
characteristics, soil pore space, 22
plant growth, 21–22 T
source of, 21 Tannins
thermal properties, 22–23 characteristics, 98
growing food demand condensed tannins, structures of, 99, 99f
components, 20–21 gallotannins, structures of, 99, 99f
food security, 19–20 hydrolyzable tannins, 99–100
increased productivity and sustainable monomer units, 98–99, 99f
intensification, 20–21, 20f split peak, 100
Index 255
Total correlation spectroscopy (TOCSY) NOESY, 72–73

applications of TOCSY, 72
aromatic resonances, 87
hydrocarbon compositions, 88–89, 89f U
Mollisol soil, 87, 88f Universal kriging (UK), 185–186
2D NMR, 72
Traditional variable-rate (VR1) MZ V
delineation, 225, 225t Variable-rate application (VRA)
Triticum species, 2–3, 3f application (prescription) maps, 220–221
Two dimensional (2D) NMR spectroscopy cost–benefit analysis, 223
applications of economic benefits, 222–223
HMQC, 89–90, 90f SSMZ–constant yield goal
HSQC, 91–92, 91f (SSMZ–CYG), 222–223
NOESY, 92–93, 93f SSMZ–variable yield goal (SSMZ–VYG),
TOCSY, 87–89, 88–89f 222–223
bond interactions, 72
F1 and F2 frequencies, 71–72 Y
heteronuclear correlation experiments, 73 Yield maps, 204

(Advances in Agronomy Volume 143) Donald L. Sparks (Eds.) - Academic Press (2017) PDF

Hochgeladen von

Dokumentinformationen

Originaltitel

Copyright

Verfügbare Formate

Dieses Dokument teilen

Dokument teilen oder einbetten

Freigabeoptionen

Stufen Sie dieses Dokument als nützlich ein?

Sind diese Inhalte unangemessen?

Copyright:

Verfügbare Formate

(Advances in Agronomy Volume 143) Donald L. Sparks (Eds.) - Academic Press (2017) PDF

Hochgeladen von

Copyright:

Verfügbare Formate

ADVANCES IN AGRONOMY

KATE M. SCOW ALFRED E. HARTEMINK

Emeritus Advisory Board Members

EUGENE J. KAMPRATH PAUL M. BERTSCH

First edition 2017

Copyright © 2017 Elsevier Inc. All rights reserved.

For information on all Academic Press publications

Publisher: Zoe Kruze

Soil: The Forgotten Piece of the

Advances in Agronomy, Volume 143 # 2017 Elsevier Inc. 1

supporting role for crop production; this is particularly concerning as

2. SOIL—A MISSING NEXUS COMPONENT

Food, energy, water security

Soil organic carbon

Soil aggradation climb

Fig. 3 Process of soil aggradation to restore soil functions.

2.2 Current State of Soils

fertilizers or manures if physically and economically available, with the

2.3 Soil Degradation

Typology of degradation Intervention options

Type 1—High degradation trend Rehabilitate if economically feasible; mitigate

Type 2—Moderate degradation trend Introduce measure to mitigate degradation

Type 3—Stable land, slightly Preventive interventions

Type 4—Improving lands Reinforcement of enabling

heavily on human judgement. GLASOD estimates soil erosion is responsible

2.4 Soil Erosion

2.4.1 Climate Change, Especially Extreme Rainfall and Heat

climate, management, and soils information together with simulation

minimum temperatures on the physiology of organisms reveals a disruption

2.4.2 Urban Expansion—Covering of Productive Soils

1982). The changes in the landscape as suggested by Ajmone-Marsan et al.

2.4.3 Growing Food Demand Placing Increased Pressure on the Soil

resources ability to provide the soil functions, especially, water, nutrients,

2.4.4 Linkage of Soils to Food

spatially and temporally heterogeneous, involve a large diversity of organ-

2.4.5 Linkage of Soils to Water

2.4.6 Linkage of Soils to Energy

soil degradation. Optimizing crop production on a land resource base will

drought (Wright and Wimberly, 2012), and on the environmental impacts

2.4.7 Improving Soils

change adaptation. Practices to maintain productive soils are generally well

3. SCALE OF THE INTERFACE BETWEEN SOILS AND

Table 1 Overview of Ecosystem Services at Different Spatial Scales of Agricultural

availability is the major factor affecting crop production (Nazarifar et al.,

4. IMPLICATIONS OF SOIL AS A COMPONENT OF THE

residue removal and enhancement of soil will require a reduction in the

• Understanding the response of different soils to different management

5.2 Soil as a Component in the Food, Energy, Water Nexus

agricultural problems rather than an agronomic approach (Lal, 2016). That

Humin: Its Composition and

Advances in Agronomy, Volume 143 # 2017 Elsevier Inc. 47

5.14 Summary of Section 5 113

thought to lie mainly in properties such as higher molecular weight and

2. HISTORICAL PERSPECTIVES OF HUMIC SUBSTANCES

as dung” (translation by Dakyns, 1897). In De Re Rustica, Lucius Columella

2.1 Selected Theories for the Genesis of Humic Substances

(4) that microbial enzymatic processes are involved in the decomposition

That stimulated interest in the abiotic synthesis of humic substances from

However, based on the reasoning of Burdon (2001), acceptable evidence

there is no convincing evidence to support Haworth’s structural hypothesis

2.2 Humin in the Historical Context

he viewed as irreversible. These concepts were reasonable assumptions based

Based on the studies of: Preston and Ripmeester (1982), Saiz-Jimenez