You are on page 1of 9

Science of the Total Environment 615 (2018) 517–525

Contents lists available at ScienceDirect

Science of the Total Environment

journal homepage:

Irrigation and weed control alter soil microbiology and nutrient

availability in North Carolina Sandhill peach orchards
Yi Zhang a,c,⁎, Liangju Wang b,c, Yongge Yuan e, Jing Xu e, Cong Tu c, Connie Fisk d, Weijian Zhang f, Xin Chen e,
David Ritchie c, Shuijin Hu a,c,⁎
College of Resources & Environmental Sciences, Nanjing Agricultural University, Nanjing 210095, China
College of Horticulture, Nanjing Agricultural University, Nanjing 210095, China
Department of Entomology & Plant Pathology, North Carolina State University, NC 27695, USA
Department of Horticultural Science, North Carolina State University, NC 27695, USA
College of Life Sciences, Zhejiang University, Hangzhou 310029, China
Institute of Crop Science, Chinese Academy of Agricultural Sciences, Beijing 100081, China


• Irrigation and weed control effects were

assessed in peach Sandhill orchards.
• Irrigation increased AMF infection and
activities without altering AMF commu-
• Weed control reduced soil nutrient
availability, microbial biomass and ac-
• Weed control reduced mycorrhizal
spore density but not AMF infection
and community.

a r t i c l e i n f o a b s t r a c t

Article history: Orchard management practices such as weed control and irrigation are primarily aimed at maximizing fruit
Received 5 July 2017 yields and economic profits. However, the impact of these practices on soil fertility and soil microbiology is
Received in revised form 24 September 2017 often overlooked. We conducted a two-factor experimental manipulation of weed control by herbicide and trick-
Accepted 24 September 2017
le irrigation in a nutrient-poor peach (Prunus persica L. cv. Contender) orchard near Jackson Springs, North Car-
Available online 5 October 2017
olina. After three and eight years of treatments, an array of soil fertility parameters were examined, including soil
Editor: D. Barcelo pH, soil N, P and cation nutrients, microbial biomass and respiration, N mineralization, and presence of arbuscular
mycorrhizal fungi (AMF). Three general trends emerged: 1) irrigation significantly increased soil microbial bio-
Keywords: mass and activity, 2) infection rate of mycorrhizal fungi within roots were significantly higher under irrigation
Arbuscular mycorrhizal fungi (AMF) than non-irrigation treatments, but no significant difference in the AMF community composition was detected
Irrigation among treatments, 3) weed control through herbicides reduced soil organic matter, microbial biomass and activ-
Weed control ity, and mineral nutrients, but had no significant impacts on root mycorrhizal infection and AMF communities.
Peach orchard Weed-control treatments directly decreased availability of soil nutrients in year 8, especially soil extractable in-
Soil nutrients
organic N. Weed control also appears to have altered the soil nutrients via changes in soil microbes and altered
net N mineralization via changes in soil microbial biomass and activity. These results indicate that long-term
weed control using herbicides reduces soil fertility through reducing organic C inputs, nutrient retention and

⁎ Corresponding authors at: College of Resources & Environmental Sciences, Nanjing Agricultural University, Nanjing 210095, China.
E-mail addresses: (Y. Zhang), (S. Hu).
0048-9697/© 2017 Elsevier B.V. All rights reserved.
518 Y. Zhang et al. / Science of the Total Environment 615 (2018) 517–525

soil microbes. Together, these findings highlight the need for alternative practices such as winter legume cover
cropping that maintain and/or enhance organic inputs to sustain the soil fertility.
© 2017 Elsevier B.V. All rights reserved.

1. Introduction weed control and irrigation practices on soil microbial biomass and ac-
tivities as well as mycorrhizal colonization of peach roots and AMF com-
Effective weed and water management can critically affect the munity in the roots.
growth of fruit trees and development and productivity of young or-
chards in the southeastern US (Horton et al., 2004; Ritchie et al., 2. Materials and methods
2005). In this region, sandy soils, high precipitation and warm temper-
ature all facilitate rapid decomposition of organic residues, reducing the 2.1. The experimental location and design
nutrient reservoir and water retention (Sierra et al., 2001; Tu et al.,
2006a; Cross and Grace, 2010). However, irrigation is necessary in this The experiment was located at the Sandhills research station near
region as nutrient limitation and periodical drought often occurs. Also, Jackson Springs, North Carolina, located at 35°13′N and 79°41′W with
orchard floor vegetation can compete for water and nutrients, greatly an elevation of approximately 200 m. Average annual precipitation is
impacting fruit trees during the first few years after establishment about 1180 mm, and average monthly temperature ranges 9.9–21.6
(Buckelew, 2009). Therefore, young orchards may take longer to be a °C. The soil was a sedimentary Candor sand (sandy, kaolinitic, thermic
high-productive system if infested with weeds. Traditional manage- Grossarenic Kandiudults in the US Soil Taxonomy) with low fertility
ment regimes of weeds and water emphasize the net effects on short- (Buckelew, 2009) and a pH of ca. 5.0. The experiment was a 2 × 2 facto-
term tree growth and productivity (Forey et al., 2016), and limited at- rial design with weed-control by chemical herbicides and trickle irriga-
tention has been paid to the long-term consequences of irrigation and tion to the rhizosphere of trees as two treatment factors. Four
weed control on soil microbes, a major component of soil fertility, and experimental systems were thus formed, including no weed-control
also the long-term sustainability of peach orchards. plus irrigation (NWC-I), no weed-control plus no irrigation (NWC-NI),
Soil microbes are the primary agent of residue decomposition and weed-control by herbicides plus irrigation (WC-I), and weed-control
nutrient releases, and therefore critically affect plant growth by regulat- by herbicides plus no irrigation (WC-NI). The plot size was 21.9 m by
ing nutrient availability (Yao et al., 2005; Floch et al., 2009). They also 3.7 m with four plants per plot. Each treatment system was replicated
function as a temporary nutrient reservoir that retain soil N and influ- four times with a randomized block arrangement. Native weed species
ence plant productivity (van Der Heijden et al., 2008). In addition, across the field were mowed periodically to a height of 10–13 cm tall
many soil microbes have suppressive effects on soil pathogens and par- and the weed-control treatment was applied by using herbicides (see
asitic nematodes (Weller et al., 2002; Neher, 2010). Finally, some soil below for details).
microbes, particularly mycorrhizal fungi, form symbiotic associations
with peach trees (Gilmore, 1971). Arbuscular mycorrhizal fungi (AMF) 2.2. Peach tree establishment and management treatments
in general enhance the nutrient uptake of host plants (particularly P
and also N) and increase the tolerance and resistance of their host plants 2.2.1. Peach orchard establishment and fertilizer application
to environmental stresses such as drought (Gilmore, 1971; Augé, 2004; On Feb. 3, 2006, one-year-old grafted trees of peach (Prunus persica
Hodge and Fitter, 2010; Bowles et al., 2016; Yang et al., 2017; Zou et al., L. cv. Contender on Guardian rootstock) were planted. In the previous
2017). Wu et al. (2011) showed that AMF improved plant growth per- summer-fall, 4500 kg of chicken litter and 1125 kg of dolomitic lime
formance and nutrient acquisition of peach seedlings, but different per hectare were incorporated into the soil to raise the pH of ca. 5.8
AMF species led to different magnitudes of benefits. Some experiments (Buckelew, 2009). The trees were planted with a space of 5.5 m within
have shown that various orchard management practices (e.g. use of fer- row and 6.0 m between rows. On May 9, 2006, 280 g of compound fer-
tilizers or agrochemicals) may alter AMF communities and be detrimen- tilizer (a particular mix of fertilizers: urea, ammonium phosphate and
tal to AMF colonization of fruit plants, reducing AMF benefits (Ocampo muriate of potash, with N: P2O5: K2O = 10:10:10) per tree was applied.
and Barea, 1985; Jeffries et al., 2003; Baumgartner et al., 2005; Gosling et In 2007, fertilizer was surface-applied with 34 kg each of N and K2O
al., 2006; Druille et al., 2013; Van Geel et al., 2016; Turrini et al., 2017). ha− 1 in March, and 28.9 kg each of N, K2O and P2O5 ha− 1 in May. In
However, little is known about the effects of management practices on 2008, equal amounts of N and K2O were applied in two separate dress-
the biomass and activities of the soil microbial community and symbiot- ings of 45.4 and 22.7 kg ha−1 in April and June, respectively. Since 2009,
ic mycorrhizal fungi and their colonization of roots in peach orchards. all trees were fertilized uniformly with two applications of 44.8 kg each
In a long-term field experiment examining the impact of two major of N and K2O ha−1 in March, and 33.6 kg ha−1 again in May. Also, dolo-
management practices (weed control and irrigation) on peach tree mitic lime were incorporated into the soil every three or four years to
growth and productivity, we observed that peach leaves had premature neutralize the acidity so that soil pH is more suitable for peach plants.
discoloration and defoliation in the fall in both weed-control and irrigat-
ed system in the third year of the experiment (Buckelew, 2009; Fisk, 2.2.2. Weed control through herbicide applications
2013). We speculate that irrigation and weed control may have nega- The herbicides Chateau (flumioxazin, for PRE emergent control of
tively affected soil nutrient retention and plant health through altering weeds) at 440 mL ha−1 and Gramoxone Inteon (paraquat dichloride,
soil microbiology. More specifically, we hypothesized that 1) irrigation as a POST emergent burndown) at 3550 mL ha−1 were applied (plus
reduces nutrient retention through increasing leaching and plant up- nonionic surfactant) to maintain a 3.6 m weed-free strip under the
take, 2) weed control reduces soil organic carbon for microbes and mi- trees in the weed-control systems. The herbicides were tank-mixed to-
crobial nutrient retention, and 3) both irrigation and weed control gether and broadcast applied in the spring (March) and again when
suppress AMF infection of peach roots through reducing water stresses needed in the summer (usually early July). Other weeds like yellow nut-
and AMF spores in soil. Therefore, we collected soil samples in the fall of sedge were spot treated as needed with the herbicide Basagran (sodium
the 3rd year, and also in the growing season and the fall of the 8th year salt of bentazon). Therefore, the soil surface in these systems was free of
following orchard establishment. Our specific objectives were: (1) to as- vegetative ground cover throughout the growing season and without
sess the effects of irrigation and/or weed control by herbicides on nutri- plant residues other than defoliated peach leaves all years prior to col-
ent availability of peach orchard soils, (2) to determine the impact of lection of soil samples.
Y. Zhang et al. / Science of the Total Environment 615 (2018) 517–525 519

2.2.3. Water management vigorously stirring to suspend spores and gently precipitating soil for
For the irrigated systems, two microsprinklers were fixed under about 2 min, the slurry water was decanted through 2 sieves (# 40
each tree, about 20 cm from either side of the trunk within the tree and # 200), where even small spores of AMF would be caught on the
row. Soil moisture was monitored twice a week from May through Au- # 200 sieve. This procedure was repeated a few times until the slurry
gust (after harvest) by Watermark™ granular matrix sensors. The sen- water was almost clear. Then, with a jet of water, the collected materials
sors were installed at 30 cm of soil depth and 46 cm from the trunk, on the #200 sieve were transferred into a test tube to a constant volume.
and were 76 to 91 cm from the microsprinklers. The microsprinklers, Ten milliliter aliquots of the spore extraction were placed in a petri dish
tree trunk and sensor were in a triangular arrangement. From May to to observe directly under a Nikon eclipse E600 microscope for spore
August, the more precipitation there was, the less irrigation was ap- abundance (Zarei et al., 2008).
plied, and the irrigated plots were watered when the soil moisture read-
ings were 20 kPa or above, as measured by the granular matrix sensors. 2.4.3. AMF community in roots
The irrigation range for peach trees was 20 to 60 kPa for sandy soils in AMF communities in the peach roots were analyzed by nested PCR-
the southeastern US (Taylor and Rieger, 2005), and we chose 20 kPa denaturing gradient gel electrophoresis (DGGE)-cloning-sequencing
as the threshold to ensure that soil moisture is not a limiting factor to method (Schechter and Bruns, 2008; Zhang et al., 2010) (Method A1).
peach plants. The deliver rate of the microsprinklers was 70 L per hour Phylogenetic analysis was carried out on the sequences obtained in
in a 6.0 m diameter around the tree. For each irrigation event, water this study. We conducted closest matches of sequences deposited in
was applied in the amount equivalent to a precipitation of 2.54 cm to the GenBank DNA database by using the BLAST and aligned sequences
ensure that a substantial amount of water moves down along the soil via the program Clustal X. A phylogenetic tree based on neighbor-join-
profile. This irrigation regime has been continuous since. ing (NJ) implemented in MEGA version 5.0 and the Kimura 2-Parameter
model was constructed. The NJ bootstrap values were calculated with
2.3. Soil and root sampling 1000 repetitions. Different sequence types or phylotypes were defined
as groups of closely related sequences, usually with a high level of boot-
Of the four peach plants per plot, the middle two were used for soil strap support in the phylogenetic analyses (higher than 80%) and with
sampling and the outer two on each end were buffer trees. Composite pairwise similarity (higher than 97%). Each sequence has been deposit-
soil samples were collected from the 0–15 cm layer under the outer ed in GenBank under accession numbers KY929306-KY929319.We
drip line of the tree canopy in late October of 2008 and 2013. Soil sam- compared the AMF community composition in roots of the four treat-
pling was intentionally set in the fall when active root growth and nutri- ments. We used the AMF phylotype presence/absence data based on
ent uptake occur. One composite sample consisting of 4 cores (5 cm DGGE banding patterns (Quantity one 4.6.2) and sequencing results.
diameter × 15 cm depth) was collected from the two center trees in The Bray–Curtis similarity matrix was used to perform non-metric mul-
each plot. Thus, four composite samples (one for each replicate) were tidimensional scaling (NMDS) in order to graphically visualize commu-
collected per treatment. For the no weed-control systems, the weeds nity composition patterns (Primer 7.0).
were removed from the sampling areas before coring in order to avoid
contamination of surface organic substrates. Soil samples (15 cm 2.5. Total soil organic C and organic matter
depth) were also collected in May 2013 during the fruiting season to
check whether soil N cycling and microbial parameters were signifi- Soil samples were air-dried and ground to a fine powder (b 100
cantly different from those obtained in the fall. All samples were imme- mesh). Total organic carbon content in soil was determined on a
diately sealed in plastic bags, stored in coolers and transported to the Perkin-Elmer 2400 CHNS/O elemental analyzer (Norwalk, Connecticut,
laboratory. The field-moist samples were sieved with a 2-mm screen USA), while the soil organic matter content was estimated from organic
and stored in sealed plastic bags at 4 °C for further analysis. Peach C by applying the van Bemmelen factor, 1.724 (Read and Ridgell, 1922;
roots on the sieve were collected and carefully washed with deionized Howard, 1966).
H2O for analysis of mycorrhizal colonization and AMF community.
2.6. Soil extractable carbon (SEC) and nitrogen (SEN)
2.4. Arbuscular mycorrhizal fungi (AMF)
Twenty grams (dry weight equivalent) of soil samples were shake-
2.4.1. Mycorrhizal infection determination extracted in 50 mL of 0.5 mol L−1 K2SO4 solution. After filtration, the
A subsample of peach roots was obtained from the mixed roots to concentration of carbon in the solutions was then determined using a
determine mycorrhizal colonization of roots, following the method de- total organic carbon (TOC) analyzer (TOC-5050A, Shimadzu Corpora-
scribed by Phillips and Hayman (1970). The fine roots with 0.5–1 mm tion, Kyoto, Japan) (Hu et al., 1997), while the concentrations of NO−
diameter were cut to 1 cm lengths and immerged in 10% KOH solution and NH+ 4 in the extracts were, respectively, determined on a Lachat
in filtering crucibles in plastic boxes. Then the boxes were put in a hot flow injection analyzer (Lachat Instruments, Milwaukee, Wisconsin,
water steamer for about 30 min until roots were cleaned. The roots USA) (Tu et al., 2006a).
were rinsed with tap water to remove KOH and then submerged in 1%
HCl solution for at least 1 h. After being removed from HCl solution, 2.7. Extractable macro nutrients in soil
the roots were stained in a trypan blue staining solution (containing
0.05% trypan blue, 50% glycerine, 25% lactic acid) and heated again in Twenty grams (dry weight equivalent) of soil samples were shake-
the steamer for 18 min. Finally, the roots were transferred into water extracted in 50 mL of 0.5 mol L−1 (NH4)2SO4 solution. After filtration,
to wash off the excess stain and the AMF infection was observed the concentrations of potassium, phosphate, calcium, magnesium in
under a Nikon SMZ-1B dissecting microscope (Nikon, Tokyo, Japan) the filtered extraction were measured by Perkin Elmer 2000 ICP-atomic
and pictures were taken under a Nikon eclipse E600 microscope with emission spectrometer (Norwalk, Connecticut, USA) (Kovács et al.,
SPOT v4.7 software (Diagnostic Instruments Inc., Sterling Heights, MI, 1998).
2.8. Soil microbial parameters
2.4.2. Abundance of AMF spores
A wet sieve method modified from Daniels and Skipper (1982) was 2.8.1. Soil microbial biomass
used to collect AMF spores. One hundred grams of soil sample was put Microbial biomass C (MBC) and microbial biomass N (MBN) were
into a 500 mL flask and extracted with 300 mL tap water. After assayed by the chloroform-fumigation extraction method (Ross, 1992;
520 Y. Zhang et al. / Science of the Total Environment 615 (2018) 517–525

Table 1
Results (actual P values) of the ANOVA for the effects of irrigation, weed control and year on soil physical, chemical and microbial properties in peach orchards.

Source of variation pH SOM SEC SEN P K Ca Mg AMF SPR SMR NNM MBC MBN

WC 0.003 b0.001 b0.001 b0.001 b0.001 b0.001 0.07 0.001 0.53 0.05 b0.001 0.001 0.03 b0.001
IR 0.219 0.37 0.80 0.005 b0.001 b0.001 0.94 0.70 b0.001 0.02 0.006 0.07 0.02 0.001
Year b0.001 0.03 b0.001 b0.001 b0.001 b0.001 0.94 0.18 0.70 b0.001 b0.001 0.001 b0.001 b0.001
WC ∗ IR 0.53 0.65 0.40 0.09 0.20 0.62 0.64 0.07 0.19 0.92 0.07 0.41 0.66 0.39
WC ∗ Year 0.05 0.82 b0.001 b0.001 0.53 0.98 0.74 0.49 0.37 0.57 0.09 0.28 0.04 0.02
IR ∗ Year 0.48 0.98 0.72 0.02 0.77 0.57 0.56 0.92 0.65 0.13 0.33 0.06 0.02 0.13
WC ∗ IR ∗ Year 0.60 0.36 0.47 0.13 0.71 0.92 0.61 0.83 0.96 0.43 0.30 0.25 0.20 0.22

WC, Weed control; IR, Irrigation. SOM, Soil organic matter; SEC, Soil extractable carbon; SEN, Soil extractable nitrogen; P, K, Ca, and Mg represent soil extractable phosphorous, potassium,
calcium and magnesium, respectively; AMF, Arbuscular mycorrhizal fungal infection rate of peach roots; SPR, Spore density; SMR, Soil microbial respiration; NNM, Net nitrogen mineral-
ization; MBC, Microbial biomass carbon; MBN, Microbial biomass nitrogen. Bold values indicate P b 0.05.

Vance et al., 1987). Twenty grams (dry weight equivalent) of soil were 2.9. Data analysis
fumigated with ethanol-free chloroform for 48 h. Both fumigated and
non-fumigated soils were extracted in 50 mL of 0.5 mol L−1 K2SO4 by Statistical analyses were performed using the SPSS 19.0 software
shaking for 30 min. The TOC analyzer was used to determine the organic (SPSS, Chicago, IL, USA). We used three-way ANOVA to test for main ef-
C (Corg) in the extracts. The MBC was calculated as follows: MBC = (Corg fects of weed control, irrigation, year, and their interactions on soil
in fumigated soil-Corg in non-fumigated soil) / kec, where, kec = 0.33, the physical, chemical and microbial properties. Subsequently, Tukey multi-
factor used here to convert the extracted organic C to MBC (Sparling and ple comparison test was further used when the interaction effects were
West, 1988). The concentration of N in the extract was determined on significant, while one-way ANOVA was performed to compare the main
the Lachat flow injection analyzer after digestion using alkaline persul- effects when the interaction was not significant.
fate oxidation (Cabrera and Beare, 1993). The MBN was calculated using Structural equation modeling (SEM) is a multivariate statistical
the equation: MBN = (total N in fumigated soil-total N in non-fumigat- modeling technique that test pathways and relationship among the ob-
ed soil)/ken, where, ken is 0.45, the factor used to convert the extracted served variables. We use SEM to analyze different hypothetical path-
organic N to MBN (Jenkinson, 1988). ways that may explain weed control and irrigation effects on soil
nutrients, net N mineralization, and linkages to AMF and soil microbial
responses (Fig. S1). Because similar trends of treatment effects were ob-
2.8.2. Soil microbial respiration (SMR)
served, we combined the data of Oct in 2008 and 2013 together and per-
Microbial activity was measured with the method modified from
formed the analysis. To facilitate our analysis, we classified all variables
Coleman et al. (1977). A soil sub-sample (equivalent to 20 g dry weight)
into eight effect and response functional groups before SEM models
was put into an Erlenmeyer flask, which was then placed in a 1 L Mason
were evaluated. These groups include (1) irrigation; (2) weed control;
jar, where 5 mL water were added to increase the air moisture, and a
(3) soil environment (soil pH and soil organic matter); (4) soil microbes
50 mL beaker containing 5 mL of 0.5 mol L−1 NaOH was suspended be-
(soil microbial biomass C and N); (5) soil nutrients (soil extractable C,
tween the inner wall of jar and the Erlenmeyer flask (Hu and van
soil extractable N, P and K, Ca, and Mg); (6) soil microbial respiration;
Bruggen, 1997). The jars were incubated at 25 °C in the dark immediate-
(7) soil net N mineralization; and (8) AMF (AMF infection of peach
ly after sealing. After 7 d incubation, the beaker was replaced with a bea-
root and soil AMF spore density). Principal components analysis (PCA)
ker containing fresh NaOH solution, and the jars were incubated for
was used to create multivariate functional indexes for each group with
additional 7 d. Adding 3–5 drops of phenolphthalein indicator, the CO2
multiple variables (Chen et al., 2013).
trapped in NaOH solution was titrated with 0.1 mol L−1 HCl. Microbial
respiration was estimated as mg CO2 kg−1 soil d−1 by averaging data
of 14 d (Tu et al., 2006a).
3. Results

2.8.3. Net nitrogen mineralization (NNM) 3.1. Effects of weed and water managements on soil physical and chemical
Net nitrogen mineralization was determined using the method de- properties and nutrient availability
scribed by Hart et al. (1994). Soil sub-samples (equivalent to 20 g dry
weight) were weighted into an Erlenmeyer flask, which were placed Soil pH was significantly higher under the weed-control than the no
in 1 L Mason jar containing 5 mL water in order to maintain air moisture. weed control treatments (Tables 1 and 2a). The soil organic matter
After the soils were incubated in the dark at 25 °C for 4 weeks, NH+
4 and (SOM) content at the experimental site was low, ranging from 7.7 to
NO− 3 in the soils were extracted in 50 mL of 0.5 mol L
K2SO4 solution 11.6 g kg−1 in 2008. However, weed-control treatment significantly de-
and determined with the Lachat flow injection analyzer. Net mineral- creased SOM content whereas irrigation had no effect in both 2008 and
ized N in soil was the difference between K2SO4-extractable inorganic 2013 (Tables 2a & 2b). The SOM content was averagely 27.6% higher in
N contents before and after incubation. the no weed-control than in the weed-control systems after eight years

Table 2a
Effects of weed control on soil physical, chemical and microbial properties in peach orchards.

System Soil pH SOM AMF Spore SMR NNM Extractable nutrients

(g kg−1) (%) (100 g−1) (mg CO2 kg−1 d−1) (mg N kg−1 d−1) (mg kg−1 soil)

P K Ca Mg
NWC 5.3b 11.6a 76.3a 655a 15.6a 0.12a 0.504a 69.4a 163.1a 47.2a
WC 5.5a 8.7b 68.8a 456a 10.4b 0.06b 0.422b 57.7b 141.0a 38.7b

NWC, No weed control and WC, Weed control. SOM, soil organic matter; AMF, Arbuscular mycorrhizal fungal infection rate of peach roots; SMR, Soil microbial respiration; NNM, Net ni-
trogen mineralization.
All data are means of four replicates in 2008 and 2013 across the irrigation system (n = 16). Different letters indicate significant differences between NWC and WC treatments (P b 0.05).
Y. Zhang et al. / Science of the Total Environment 615 (2018) 517–525 521

Table 2b
Effects of irrigation on soil physical, chemical and microbial properties in peach orchards.

System Soil pH SOM AMF Spore SMR NNM Extractable nutrients

(g kg−1) (%) (100 g−1) (mg CO2 kg−1 d−1) (mg N kg−1 d−1) (mg kg−1 soil)

P K Ca Mg
IR 5.5a 9.9a 82.3a 680a 14.6a 0.10a 0.438b 57.4a 148.2a 43.4a
NIR 5.4a 10.5a 62.8b 430b 11.3b 0.08a 0.488a 69.6b 155.9a 42.3a

IR, Irrigation and NIR, No irrigation. SOM, soil organic matter; AMF, Arbuscular mycorrhizal fungal infection rate of peach roots; SPR, Spore density; SMR, Soil microbial respiration; NNM,
Net nitrogen mineralization.
All data are means of four replicates in 2008 and 2013 across the weed control system (n = 16). Different letters indicate significant differences between NIR and IR treatments (P b

of management. Irrigation and weed-control treatments also resulted in soil microbial respiration (Tables 2a and 2b and S1). Microbial biomass
a significant decline in the availability of soil P and K in both sampling C were significantly higher in no weed-control than the weed-control
years (Tables 2a & 2b). Available Ca did not show significant difference systems (Table 3a) and in irrigated than non-irrigated systems (Tables
among systems. Magnesium availability showed a significant reduction 3b and S1b) in both sampling years. Microbial biomass C and N in the
in the weed control systems, as compared to the no weed-control sys- no weed-control treatments were on average 18.5% and 42.5% higher
tems for both sampling dates (P b 0.05, Table 2a). Also, weed-control than the weed-control systems on Oct 2013, respectively.
treatments significantly reduced soil extractable C by 36.9% as com-
pared to no weed-control treatments (Table 3a). Soil extractable N
3.5. Treatment effects observed at different sampling times
(SEN) was low and no significant difference was observed among differ-
ent systems in 2008 but significantly increased by 2013 (P b 0.01, Tables
Similar trends of treatment effects were observed and year effects of
3a & 3b). However, weed-control treatments significantly reduced (P b
most variables were significant (Table 1). Also, we found similar trend
0.01) soil extractable N by 2013 (October), eight years after the orchard
of weed control and irrigation effect on soil N cycling and some soil mi-
establishment (Table 3a). In addition, weed-control treatments signifi-
crobial parameters in May_2013 (Table S1) and Oct_2013 (Tables 3a
cantly reduced soil net N mineralization (Tables 1 and 2a), in consistent
and 3b). Soil extractable N (Tables 3a and 3b), and microbial biomass
with the change in soil N availability.
(Tables 3a and 3b) and activities (Tables 2a and 2b) were very low in
2008, but increased multi-fold by 2013. Mycorrhizal infection rates of
3.2. Arbuscular mycorrhizal fungi: root infection and spore density in soil peach roots sampled in October were very similar between 2008 and
2013. Morphologies of mycorrhizal hyphae in roots were also similar
Weed-control treatments did not affect AMF colonization of peach in both 2008 (Fig. 1) and 2013 (pictures not shown). However, AMF
roots and mycorrhizal spore density (SPR) (Tables 2a and S1a). Howev- spores in soil significantly decreased in all treatments from 2008 to
er, irrigation significantly increased AMF infection and spore density in 2013 (P b 0.01).
the fall of both years (P b 0.01, Tables 1 and 2b). Greater hyphal density
of mycorrhizal fungi and vesicles within roots was also observed in the
3.6. Pathways determining soil nutrients and net N mineralization
irrigation than in the non-irrigation systems (Fig. 1: a vs. b and c vs. d).

Most variables examined were correlated with one another, making

3.3. AMF community in roots our data set appropriate for SEM analysis (Table S3). Direct paths in our
model from weed control directly to soil environment (organic matter
Non-metric multidimensional scaling (NMDS) analysis of AMF com- and soil pH), soil nutrients (soil extractable C, N and P) and AMF (AMF
munities in peach roots revealed no apparent trends of change (Fig. 2). infection of peach root and soil AMF spore density) were observed
There were 14 AMF phylotypes in root samples from the 4 treatments (Fig. 3). Model results suggest 1) Soil Environment and Soil Microbes
(Table S2). Based on the ANOSIM, a multivariate statistical measure, pathways affected soil microbial respiration, 2) Soil Microbes pathway
showed that AMF communities in root samples did not differed also affected the soil nutrients, and 3) both Soil Microbes and Soil Micro-
among the treatments (R = 0.25, P = 0.17). bial Respiration pathways affected net N mineralization. Surprisingly, ir-
rigation only directly altered AMF, and appear to have had no direct
3.4. Microbial biomass C, N and respiration effect on the soil nutrients and net N mineralization.
Soil Environment and Soil Microbes pathways together explained
Weed-control and irrigation treatments significantly impacted the 93% of the total variance in soil microbial respiration (Fig. 3). Changes
size of the soil microbial communities (Tables 3a and 3b and S1) and in the soil microbes were mainly related to changes in soil environment

Table 3a Table 3b
Effects of weed control on soil extractable carbon and nitrogen, microbial properties in Effects of irrigation on soil extractable carbon and nitrogen, and microbial properties in
peach orchards in years 2008 and 2013. peach orchards in years 2008 and 2013.

(mg kg−1 soil) (mg kg−1 soil) (mg C kg−1) (mg N kg−1) (mg kg−1 soil) (mg kg−1 soil) (mg C kg−1) (mg N kg−1)

2008_OCT NWC 24.1aa 0.28a 47.0a 9.1a 2008_OCT IR 18.9aa 0.24a 46.3a 6.65a
WC 17.6b 0.31a 38.4b 6.3a NIR 22.8a 0.34a 39.1b 8.75a
2013_OCT NWC 83.4A 3.47A 154A 17.7A 2013_OCT IR 60.2A 3.14A 155.9A 16.6A
WC 27.6B 1.78B 130B 12.4B NIR 54.9A 2.11A 128.8B 13.5A

NWC, No weed control; WC, Weed control; SEC, Soil extractable carbon; SEN, Soil extract- IR, Irrigation and NIR, No irrigation; SEC, Soil extractable carbon; SEN, Soil extractable ni-
able nitrogen; MBC, Microbial biomass carbon; MBN, Microbial biomass nitrogen. trogen; MBC, Microbial biomass carbon; MBN, Microbial biomass nitrogen.
a a
All data are means of four replicates across the irrigation system (n = 8). Different low All data are means of four replicates across the weed control system (n = 8). Different
case and capital case letters indicate significant differences between NWC and WC treat- low case and capital case letters indicate significant differences between NWC and WC
ments for 2008 and 2013, respectively (one-way ANOVA, P b 0.05). treatments for 2008 and 2013, respectively (one-way ANOVA, P b 0.05).
522 Y. Zhang et al. / Science of the Total Environment 615 (2018) 517–525

Fig. 1. Irrigation and weed control affected arbuscular mycorrhizal (AM) fungal fungi of peach roots. a: non weed-control with irrigation, b: non weed-control without irrigation, c: weed-
control with irrigation, and d: weed-control without irrigation. The peach roots was pretreated and stained according to the method described by Phillips and Hayman (1970), and AMF
infection pictures were taken under a Nikon eclipse E600 microscope with the computer software. Pictures a, b and c were taken by magnifying 40 times, while d was 100 times.

(r2 = 84%). Changes in soil microbes and soil microbial respiration ex- studies (Andersen et al., 2013; Forey et al., 2016). Results from our ex-
plained about 59% of the total variance in soil net N mineralization. periment showed that microbial biomass and activity, and mycorrhizal
The changes in soil nutrients could be attributed largely to changes in colonization of peach roots were higher in 2008 and continued to be
weed control and soil microbes. Both weed control and irrigation higher in 2013 in the irrigated than the non-irrigated plots. A previous
could explain about 60% of the total variance in AMF. report of tree growth and nutrient status from the same experiment
showed that three year old peach trees grew 14% larger (expressed by
4. Discussion Tree Cross-Sectional Area), produced 18% heavier individual fruit and
yielded 45% more total fruit harvest under irrigation than no irrigation
Weed and water management regimes are traditionally aimed at in- (Buckelew, 2009). Soil microbes control the decomposition and nutri-
creasing tree growth and productivity in orchards, but their long term ent release from organic materials and their biomass functions as a la-
effects on the soil microbes and soil nutrient status are often overlooked bile nutrient reservoir for plant growth (Dalal, 1998; Friedel et al.,
since leaf analysis has frequently been undertaken in these types of 2001; Sugihara et al., 2010). Over the short term, higher microbial

Fig. 2. Non-metric multidimensional scaling (NMDS) analysis of AMF communities in peach roots from the four treatments. NWC-I: non weed-control with irrigation, NWC-NI: non weed-
control without irrigation, WC-I: weed-control with irrigation, and WC-NI: weed-control without irrigation.
Y. Zhang et al. / Science of the Total Environment 615 (2018) 517–525 523

Fig. 3. Structural equation model analysis (SEM) examining the effects of irrigation (IR) and weed control (WC) on the soil nutrients and net N mineralization via pathways of soil
resources, soil microbes and AMF. Square boxes indicate variables included in the model: irrigation (IR) and weed control (WC), soil environment includes soil pH and organic matter
(OM), soil microbes include microbial biomass C (MBC) and microbial biomass N (MBN), soil nutrients include soil extractale N (SEN) and extractable P (P), soil net N mineralization
(NNM), soil microbial respiration, and AMF includes AMF infection of peach root and soil AMF spore density. Results of model fitting: χ2 = 2.858, P = 0.414, AIC = 84.858. Solid and
dashed arrows indicate significant positive and negative effects (P b 0.05), respectively; the pathways without significant effects did not show (P N 0.05). r2 values associated with
response variables indicate the proportion of variation explained by relationships with other variables. Values associated with arrows represent standardized path coefficients.

biomass and activity in irrigated systems will provide more nutrients for Vesicules and arbuscules are the important structure for nutrient ex-
the growth and production of peach trees shown by the positive rela- change (Smith and Read, 2010) and significantly higher infection
tionship between microbial biomass (and activity) and net mineraliz- (Table 2b) and density of vesicules (Fig. 1) in our irrigated systems fur-
able N (Molina et al., 2016). Enhanced N mineralization by stimulated ther indicate that AMF may more actively take up and transfer nutrients.
microbial biomass and activity has also been observed in the present Also, other studies have showed that nutrient inputs such as P and N
study (Fig. 3) and previous experiments. For example, in organic tomato often suppressed AMF (Treseder, 2004; Collins and Foster, 2009) and
systems, the net mineralizable N was significantly positively correlated our results of significantly lower AMF spore densities in 2013 than in
with the increased microbial biomass and activity (Tu et al., 2006b) and 2008 were consistent with these observations. A decrease in spore den-
the enhanced N mineralization may substantially result from the turn- sity may lead to changes in AM fungal diversity and community, and the
over of microbial biomass itself (Bonde et al., 1988). Together, higher symbiosis functionality, considering that spores represent an important
microbial biomass (Table 3b) and activity (Tables 2a and 2b), as well source of infective propagules of AMF in soil (Smith and Read, 2010).
as higher extractable (Table 3b) and mineralizable N (Table 2b) in the Yet, the mycorrhizal infection rate of peach roots did not change with
irrigated plots suggest that enhanced plant growth and leaf residues time, with relatively similar levels of infection in October in both sam-
may be the major factor enhancing nutrient retention (Buckelew, pling years. The implications of these results are unknown. One possibil-
2009). ity is that perennial plant roots present in the soil provide more
The results of irrigation effects on mycorrhizal infection of peach opportunities for AM fungal colonization over time (Alguacil et al.,
roots and AMF spore densities were particularly interesting and directly 2012). Also, AMF likely infected the peach roots when the peach tree
contradicted our hypothesis. Because periodical drought frequently oc- was very young and the initial community dominates the composition
curs in the sandy soil at the experiment site and weed-control treat- of the AMF in roots as the host plant may be unable to discriminate a
ment causes fewer host plants for mycorrhizal fungi, we expected that specific species in roots (Kiers and Van Der Heijden, 2006).
mycorrhizal colonization of peach roots would be highest in the non-ir- Weed-control treatment has been shown to enhance the growth
rigation, no weed-control systems. Contrary to this expectation, mycor- and production of peach trees in the first few years (Buckelew, 2009),
rhizal colonization of peach roots was significantly higher in irrigation likely due to reduced competition for nutrients and water. However,
than non-irrigation treatments while there was no difference between our data showing lower microbial biomass N and soil nutrients by
the non-weed and weed-control treatments (Tables 2a and 2b). Some 2013 (Tables 2a, 2b, 3a and 3b) indicated that weed control systems re-
studies suggest that herbicides can negatively affect the establishment duced ecosystem nutrient retention that occurs in weed biomass, soil
and function of AMF (Baumgartner et al., 2005; Druille et al., 2013), organic matter and soil microbial biomass, facilitating nutrient losses
but these effects are highly variable depending on the herbicide type through leaching and runoff (Gómez et al., 2009). Also, the model re-
and application rate, the host plant, and soil nutrient status (Smith et sults of the SEM analysis support the hypothesis that weed control pri-
al., 1981; Alguacil et al., 2012; Njeru et al., 2015). In our experiment, her- marily reduce soil nutrients and N cycling via altering biomass and
bicide treatment did not significantly affect peach AMF colonization and activities of soil microbes (Fig. 3). Since equal amounts of manures
community (Table 2a and Fig. 2). We postulate that the AMF communi- and fertilizers were applied to all treatments, the higher levels of soil
ty in soil at the early stage of orchard establishment may be the domi- nutrients remaining in the non-weed control (NWC) systems suggest
nant factor that controls the AMF community composition in roots, a combination of increased nutrient cycling in the NWC treatments
but that carbohydrate supply from host plants dominates the infection and greater nutrient leaching losses in the weed control treatments.
rate over time. Irrigation facilitates plant growth, prompting plants to Furthermore, soil microbes typically are C-limited (Smith and Paul,
enhance allocation of carbohydrates below ground for scavenging nutri- 1990), and reduced organic C in soils from weed control systems likely
ents (Haynes and Gower, 1995; Hu et al., 1997; Jia et al., 2010). contributed to lower microbial biomass (Fig. 3) and fewer carbon
524 Y. Zhang et al. / Science of the Total Environment 615 (2018) 517–525

sources for soil microbes (Lister et al., 2004; Huang et al., 2013; Andersen, L., Kühn, B.F., Bertelsen, M., Bruus, M., Larsen, S.E., Strandberg, M., 2013. Alter-
natives to herbicides in an apple orchard, effects on yield, earthworms and plant di-
Gómez-Muñoz et al., 2014; Wan et al., 2014). Therefore, orchard floor versity. Agric. Ecosyst. Environ. 172, 1–5.
management can critically affect soil microbial biomass, activities and Augé, R.M., 2004. Arbuscular mycorrhizae and soil/plant water relations. Can. J. Soil Sci.
even composition (Yao et al., 2005; Tu et al., 2006b; Liu et al., 2015). 84, 373–381.
Baumgartner, K., Smith, R.F., Bettiga, L., 2005. Weed control and cover crop management
In addition, significant lower AMF spore densities in weed-control affect mycorrhizal colonization of grapevine roots and arbuscular mycorrhizal fungal
soils than the no weed-controls (Fig. 1) suggest that herbicides may spore populations in a California vineyard. Mycorrhiza 15, 111–119.
have reduced AMF density and diversity through reducing AMF host Bonde, T.A., Schnürer, J., Rosswall, T., 1988. Microbial biomass as a fraction of potentially
mineralizable nitrogen in soils from long-term field experiments. Soil Biol. Biochem.
plants, resulting in a decrease in the capacity of plants to take up nutri- 20, 447–452.
ents and lower soil fertility (Jeffries et al., 2003). Bowles, T.M., Barrios-Masias, F.H., Carlisle, E.A., Cavagnaro, T.R., Jackson, L.E., 2016. Effects
Consistently higher nutrient availability and microbial biomass and of arbuscular mycorrhizae on tomato yield, nutrient uptake, water relations, and soil
carbon dynamics under deficit irrigation in field conditions. Sci. Total Environ. 566,
activity in the irrigated but no-weed control treatment (Tables 2a, 2b,
3a and 3b) suggest that nutrient retention through plant-microbe sys- Buckelew, J.K., 2009. Orchard Floor Management in Young Peach [Prunus persica (L.)
tems play a major role. Moderate weed growth provides C sources for Batsch.]: Effects of Irrigation, Vegetation-free Width, and Certain Pre Herbicides. (Dis-
microbes and promotes soil nutrient retention (Welch et al., 2016), par- sertation). North Carolina State University.
Cabrera, M.L., Beare, M.H., 1993. Alkaline persulfate oxidation for determining total nitro-
ticularly in areas with heavy precipitation. Increasing surface coverage gen in microbial biomass extracts. Soil Sci. Soc. Am. J. 57, 1007–1012.
by allowing weed growth has been proposed in apple (Andersen et al., Chen, D., Lan, Z., Bai, X., Grace, J.B., Bai, Y., 2013. Evidence that acidification-induced de-
2013; Neilsen et al., 2014), grape (Pardini et al., 2002) and olive clines in plant diversity and productivity are mediated by changes in below-ground
communities and soil properties in a semi-arid steppe. J. Ecol. 101, 1322–1334.
(Gómez et al., 2009). In particular, Gómez et al. (2009) showed that Coleman, D.C., Anderson, R.V., Cole, C.V., Elliott, E.T., Woods, L., Campion, M.K., 1977. Tro-
weed elimination with herbicides led to the largest soil loss in young phic interactions in soils as they affect energy and nutrient dynamics. IV. Flows of
olive groves in southern Spain when most of the soil is unprotected by metabolic and biomass carbon. Microb. Ecol. 4, 373–380.
Collins, C.D., Foster, B.L., 2009. Community-level consequences of mycorrhizae depend on
the small olive canopy. Practices that enhance nutrient inputs and re- phosphorus availability. Ecology 90, 2567–2576.
duce water runoff and leaching should, therefore, be incorporated into Cross, A., Grace, J., 2010. The effect of warming on the CO2 emissions of fresh and old or-
management regimes. Because of the warm temperature, significant ganic soil from under a Sitka spruce plantation. Geoderma 157, 126–132.
Dalal, R.C., 1998. Soil microbial biomass-what do the numbers really mean? Aust. J. Exp.
microbial decomposition occurs in this region in the winter when Agric. 38, 649–665.
peach trees are dormant (Grisi et al., 1998), suggesting the need to re- Daniels, B.A., Skipper, H.A., 1982. Methods for the recovery and quantitative estimation of
duce nutrient losses in winter. Winter cover cropping that incorporates propagules from soil. In: Schenck, N.C. (Ed.), Methods and Principles of Mycorrhizal
Research. Phytopathological, St Paul, Minn, USA, pp. 29–35.
legume plants may deserve the most attention.
Druille, M., Cabello, M.N., Omacini, M., Golluscio, R.A., 2013. Glyphosate reduces spore viabil-
ity and root colonization of arbuscular mycorrhizal fungi. Appl. Soil Ecol. 64, 99–103.
5. Conclusions Fisk, C.L., 2013. Effect of orchard management practices on peach tree growth, yield, and
soil ecology. (Dissertation). North Carolina State University.
Floch, C., Capowiez, Y., Criquet, S., 2009. Enzyme activities in apple orchard
Our results showed that irrigation enhanced soil microbial biomass agroecosystems: how are they affected by management strategy and soil properties.
and activities, mycorrhizal infection of peach roots and mycorrhizal fun- Soil Biol. Biochem. 41, 61–68.
Forey, O., Metay, A., Wery, J., 2016. Differential effect of regulated deficit irrigation on
gal spore densities, and soil N availability in a North Carolina Sandhill growth and photosynthesis in young peach trees intercropped with grass. Eur.
peach ecosystem. Weed control through herbicide applications, howev- J. Agron. 81, 106–116.
er, reduced soil organic C, microbial biomass and activities as well as the Friedel, J.K., Gabel, D., Stahr, K., 2001. Nitrogen pools and turnover in arable soils under
different durations of organic farming: II: source- and -sink function of the soil micro-
labile nutrient pools. These results indicate that soil microbes in this
bial biomass or competition with growing plants? J. Plant Nutr. Soil Sci. 164, 421–429.
highly sandy system are very sensitive to alterations in C availability Gilmore, A.E., 1971. Influence of endotrophic mycorrhizae on growth of peach seedlings.
and moisture. These findings suggest that enhancement of organic in- Am. Soc. Hort. Sci. J. 96, 35.
puts such as through planting winter legume cover crops and/or reduc- Gómez, J.A., Sobrinho, T.A., Giráldez, J.V., Fereres, E., 2009. Soil management effects on
runoff, erosion and soil properties in an olive grove of Southern Spain. Soil Tillage
ing the rate of herbicide applications may be needed to maintain and Res. 102, 5–13.
increase soil fertility and microbial activities for sustainable peach pro- Gómez-Muñoz, B., Hatch, D.J., Bol, R., García-Ruiz, R., 2014. Nutrient dynamics during de-
duction in the southeastern United States. composition of the residues from a sown legume or ruderal plant cover in an olive oil
orchard. Agric. Ecosyst. Environ. 184, 115–123.
Gosling, P., Hodge, A., Goodlass, G., Bending, G.D., 2006. Arbuscular mycorrhizal fungi and
Acknowledgments organic farming. Agric. Ecosyst. Environ. 113, 17–35.
Grisi, B., Grace, C., Brookes, P.C., Benedetti, A., Dell'Abate, M.T., 1998. Temperature effects
on organic matter and microbial biomass dynamics in temperate and tropical soils.
The study was supported in part by the Fundamental Research Soil Biol. Biochem. 30, 1309–1315.
Funds for Central Universities (KJQN201743) of China, and USDA grants Hart, S.C., Stark, J.M., Davidson, E.A., Firestone, M.K., 1994. Nitrogen mineralization, immo-
bilization, and nitrification. In: Weaver, R.W., Bottomley, S., Bezdicek, P., Smith, D.,
(USDA_2004-51101-02213, USDA_2007-34381-18625, USDA_2009-
Tabatabai, S., Wollum, A. (Eds.), Methods of Soil Analysis. Part 2: Microbiol Biochem
35101-05351). YZ and LJW each were partially supported by a fellow- Properties. Soil Sci Soc Am Inc, Madison, WI, pp. 985–1018.
ship from China Scholarship Council (CSC) when they were at North Haynes, B.E., Gower, S.T., 1995. Belowground carbon allocation in unfertilized and fertil-
Carolina State University. The authors would like to thank Yi Wang, ized red pine plantations in northern Wisconsin. Tree Physiol. 15, 317–325.
Hodge, A., Fitter, A.H., 2010. Substantial nitrogen acquisition by arbuscular mycorrhizal
Liang Wang, and Lei Cheng for assistance with this project. We express fungi from organic material has implications for N cycling. Proc. Natl. Acad. Sci. U. S.
our appreciation to the staff of the N.C. Agricultural Research Service’s A. 107, 13754–13759.
Sandhills research station for their management and maintenance of Horton, D.L., Brannen, P., Schnabel, G., Mitchem, W., Taylor, K., Layne, D., Bellinger, B.,
Guillebeau, P., 2004. Crop Profile for Peaches in Georgia and South Carolina. http://
the research systems.
Howard, P.J.A., 1966. The carbon-organic matter factor in various soil types. Oikos 15,
Appendix A. Supplementary data 229–236.
Hu, S., van Bruggen, A.H.C., 1997. Microbial dynamics associated with multiphasic decom-
position of 14C-labeled cellulose in soil. Microb. Ecol. 33, 134–143.
Supplementary data to this article can be found online at https://doi. Hu, S.J., Coleman, D.C., Carroll, C.R., et al., 1997. Labile soil carbon pools in subtropical for-
org/10.1016/j.scitotenv.2017.09.265. est and agricultural ecosystems as influenced by management practices and vegeta-
tion types. Agric. Ecosyst. Environ. 65, 69–78.
Huang, S., Pan, X., Sun, Y., Zhang, Y., Hang, X., Yu, X., Zhang, W., 2013. Effects of long-term
References fertilization on the weed growth and community composition in a double-rice eco-
system during the fallow period. Weed Biol. Manag. 13, 10–18.
Alguacil, M.M., Torrecillas, E., Roldán, A., Díaz, G., Torres, M.P., 2012. Perennial plant Jeffries, P., Gianinazzi, S., Perotto, S., Turnau, K., Barea, J.M., 2003. The contribution of
species from semiarid gypsum soils support higher AMF diversity in roots than the arbuscular mycorrhizal fungi in sustainable maintenance of plant health and soil fer-
annual Bromus rubens. Soil Biol. Biochem. 49, 132–138. tility. Biol. Fertil. Soils 37:1–16.
Y. Zhang et al. / Science of the Total Environment 615 (2018) 517–525 525

Jenkinson, D.S., 1988. Determination of microbial biomass carbon and nitrogen in soil. In: Sugihara, S., Funakawa, S., Kilasara, M., Kosaki, T., 2010. Dynamics of microbial biomass ni-
Wilson, J.R. (Ed.), Advances in Nitrogen Cycling in Agricultural Ecosystems. CAB Inter- trogen in relation to plant nitrogen uptake during the crop growth period in a dry
national, Wallingford, UK, pp. 368–386. tropical cropland in Tanzania. Soil Sci. Plant Nutr. 56, 105–114.
Jia, S.X., Wang, Z.Q., Li, X.P., Sun, Y., Zhang, X., Liang, A., 2010. N fertilization affects on soil Taylor, K.C., Rieger, M., 2005. Irrigation scheduling. Southeastern Peach Growers' Hand-
respiration, microbial biomass and root respiration in Larix gmelinii and Fraxinus book. Cooperative Extension Service, University of Georgia College of Agricultural
mandshurica plantations in China. Plant Soil 333, 325–336. and Environmental Sciences.
Kiers, E.T., Van Der Heijden, M.G.A., 2006. Mutualistic stability in the arbuscular mycorrhi- Treseder, K.K., 2004. A meta-analysis of mycorrhizal responses to nitrogen, phosphorus,
zal symbiosis: exploring hypotheses of evolutionary cooperation. Ecology 87, and atmospheric CO2 in field studies. New Phytol. 164, 347–355.
1627–1636. Tu, C., Booker, F.L., Watson, D.M., Chen, X., Rufty, T.W., Shi, W., Hu, S., 2006a. Mycorrhizal
Kovács, B., Daniel, P., Gyori, Z., Loch, J., Prokisch, J., 1998. Studies on parameters of induc- mediation of plant N acquisition and residue decomposition: impact of mineral N in-
tively coupled plasma spectrometer. Commun. Soil Sci. Plant Anal. 29, 2035–2054. puts. Glob. Chang. Biol. 12, 793–803.
Lister, T.W., Burger, J.A., Patterson, S.C., 2004. Role of vegetation in mitigating soil quality Tu, C., Ristaino, J.B., Hu, S.J., 2006b. Soil microbial biomass and activity in organic tomato
impacted by forest harvesting. Soil Sci. Soc. Am. J. 68 (1), 263–271. farming systems: effects of organic inputs and straw mulching. Soil Biol. Biochem. 38,
Liu, R.J., Sheng, P.P., Hui, H.B., Lin, Q., Chen, Y.L., 2015. Integrating irrigation management 247–255.
for improved grain yield of winter wheat and rhizosphere AM fungal diversity in a Turrini, A., Agnolucci, M., Palla, M., Tomé, E., Tagliavini, M., Scandellari, F., Giovannetti, M.,
semi-arid cropping system. Agric. Syst. 132, 167–173. 2017. Species diversity and community composition of native arbuscular mycorrhizal
Molina, A.J., Josa, R., Mas, M.T., Verdú, A.M.C., Llorens, P., Aranda, X., Savé, R., Biel, C., 2016. fungi in apple roots are affected by site and orchard management. Appl. Soil Ecol. 116,
The role of soil characteristics, soil tillage and drip irrigation in the timber production 42–54.
of a wild cherry orchard under Mediterranean conditions. Eur. J. Agron. 72, 20–27. Van Der Heijden, M.G., Bardgett, R.D., Van Straalen, N.M., 2008. The unseen majority: soil
Neher, D.A., 2010. Ecology of plant and free-living nematodes in natural and agricultural microbes as drivers of plant diversity and productivity in terrestrial ecosystems. Ecol.
soil. Annu. Rev. Phytopathol. 48, 371–394. Lett. 11 (3), 296–310.
Neilsen, G., Forge, T., Angers, D., Neilsen, D., Hogue, E., 2014. Suitable orchard floor man- Van Geel, M., De Beenhouwer, M., Ceulemans, T., Caes, K., Ceustermans, A., Bylemans, D.,
agement strategies in organic apple orchards that augment soil organic matter and Gomand, A., Lievens, B., Honnay, O., 2016. Application of slow-release phosphorus
maintain tree performance. Plant Soil 378, 325–335. fertilizers increases arbuscular mycorrhizal fungal diversity in the roots of apple
Njeru, E.M., Avio, L., Bocci, G., Sbrana, C., Turrini, A., Bàrberi, P., Giovannetti, M., Oehl, F., trees. Plant Soil 402, 291–301.
2015. Contrasting effects of cover crops on ‘hot spot’ arbuscular mycorrhizal fungal Vance, E.D., Brookes, P.C., Jenkinson, D.S., 1987. An extraction method for measuring soil
communities in organic tomato. Biol. Fertil. Soils 51, 151–166. microbial biomass C. Soil Biol. Biochem. 19, 703–707.
Ocampo, J.A., Barea, J.M., 1985. Effect of carbamate herbicides on VA mycorrhizal infection Wan, N.F., Ji, X.Y., Gu, X.J., Jiang, J.X., Wu, J.H., Li, B., 2014. Ecological engineering of ground
and plant growth. Plant Soil 85 (3), 375–383. cover vegetation promotes biocontrol services in peach orchards. Ecol. Eng. 64,
Pardini, A., Faiello, C., Longhi, F., Mancuso, S., Snowball, R., 2002. Cover crop species and 62–65.
their management in vineyards and olive groves. Adv. Hortic. Sci. 16, 225–234. Welch, R.Y., Behnke, G.D., Davis, A.S., Masiunas, J., Villamil, M.B., 2016. Using cover crops
Phillips, J.M., Hayman, D.S., 1970. Improved procedures for clearing roots and staining in headlands of organic grain farms: effects on soil properties, weeds and crop yields.
parasitic and vesicular-arbuscular mycorrhizal fungi for rapid assessment of infection. Agric. Ecosyst. Environ. 216, 322–332.
Trans. Br. Mycol. Soc. 55, 158–161. Weller, D.M., Raaijmakers, J.M., Gardener, B.B.M., Thomashow, L.S., 2002. Microbial popu-
Read, J.W., Ridgell, R.H., 1922. On the use of the conventional carbon factor in estimating lations responsible for specific soil suppressiveness to plant pathogens 1. Annu. Rev.
soil organic matter. Soil Sci. 13, 1–6. Phytopathol. 40, 309–348.
Ritchie, D.F., Parker, M.L., Parker, K.A., Parker, J.R., Parker, W.E., 2005. Crop Profile for Wu, Q.S., Li, G.H., Zou, Y.N., 2011. Roles of arbuscular mycorrhizal fungi on growth and nu-
Peaches in North Carolina. trient acquisition of peach (Prunus persica L. Batsch) seedlings. J. Anim. Plant Sci. 21,
ncpeaches.html. 746–750.
Ross, R.J., 1992. Influence of sieve mesh size on estimates of microbial carbon and nitro- Yang, H., Zhang, Q., Koide, R.T., Hoeksema, J.D., Tang, J., Bian, X., Hu, S., Chen, X., 2017. Tax-
gen by fumigation extraction procedures in soils under pasture. Soil Biol. Biochem. onomic resolution is a determinant of biodiversity effects in arbuscular mycorrhizal
24, 343–350. fungal communities. J. Ecol. 105, 219–228.
Schechter, S.P., Bruns, T.D., 2008. Serpentine and non-serpentine ecotypes of Collinsia Yao, S.R., Merwin, I.A., Bird, G.W., Abawi, G.S., Thies, J.E., 2005. Orchard floor management
sparsiflora associate with distinct arbuscular mycorrhizal fungal assemblages. Mol. practices that maintain vegetative or biomass groundcover stimulate soil microbial
Ecol. 17, 3198–3210. activity and alter soil microbial community composition. Plant Soil 271, 377–389.
Sierra, J., Fontaine, S., Desfontaines, L., 2001. Factors controlling N mineralization, nitrifica- Zarei, M., Saleh-Rastin, N., Jouzani, G.S., Savaghebi, G., Buscot, F., 2008. Arbuscular mycor-
tion, and nitrogen losses in an Oxisol amended with sewage sludge. Aust. J. Soil Res. rhizal abundance in contaminated soils around a zinc and lead deposit. Eur. J. Soil Boil
39, 519–534. 44, 381–391.
Smith, J.L., Paul, E.A., 1990. The significance of soil microbial biomass estimations. In: Zhang, Q., Yang, R., Tang, J., Yang, H., Hu, S., Chen, X., 2010. Positive feedback between my-
Bollag, J.M., Stotzky, G. (Eds.), Soil Biochemistry. Marcel Dekker, Inc., New York, NY, corrhizal fungi and plants influences plant invasion success and resistance to inva-
pp. 357–396. sion. PLoS One 5, e12380.
Smith, S.E., Read, D.J., 2010. Mycorrhizal Symbiosis [M]. Academic press. Zou, Y.N., Wang, P., Liu, C.Y., Ni, Q.D., Zhang, D.J., Wu, Q.S., 2017. Mycorrhizal trifoliate or-
Smith, T.F., Noack, A.J., Cosh, S.M., 1981. The effect of some herbicides on vesicular- ange has greater root adaptation of morphology and phytohormones in response to
arbuscular endophyte abundance in the soil and on infection of host roots. Pestic. drought stress. Sci Rep 7:41134.
Sci. 12, 91–97.
Sparling, G.P., West, A.W., 1988. A direct extraction method to estimate soil microbial C:
calibration in situ using microbial respiration and 14C labelled cells. Soil Biol. Biochem.
20, 337–343.