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Recent developments in nanotechnology transforming the agricultural sector: a transition

replete with opportunities

Dae-Young Kim1, Avinash Kadam2, Surendra Shinde1, Rijuta Ganesh Saratale2, Jayanta Patra2,

and Gajanan Ghodake1*

Department of Biological and Environmental Science, College of Life Science and
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Biotechnology, Dongguk University-Seoul, Ilsandong-gu, 10326, Goyang-si, Gyeonggi-do,

Republic of Korea
Research Institute of Biotechnology and Medical Converged Science, Dongguk University-

Seoul, Ilsandong-gu, 10326, Goyang-si, Gyeonggi-do, Republic of Korea

Corresponding author:

Gajanan Ghodake

Tel.: +82-31-961-5159

Fax: +82-31-961-5122



This article has been accepted for publication and undergone full peer review but has not
been through the copyediting, typesetting, pagination and proofreading process, which
may lead to differences between this version and the Version of Record. Please cite this
article as doi: 10.1002/jsfa.8749

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The applications and benefits of nanotechnology in agricultural sector have attracted
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considerable attention, particularly in the invention of unique nanopesticides and nanofertilizers.

The contemporary developments in nanotechnology are acknowledged and the most significant

opportunities awaiting in the agriculture sector from the recent scientific and technical literature

are addressed. This review discusses the significance of recent trends in nanomaterial-based

sensors available for the sustainable management of agricultural soil, as well as the role of

nanotechnology in detection and protection against plant pathogens, and for food quality and safety.

Novel nanosensors have been reported for primary applications in improving crop practices, food

quality, and packaging methods, thus will change the agricultural sector for potentially better and

healthier food products. Nanotechnology is well-known to play a significant role in the effective

management of phytopathogens, nutrient utilization, controlled release of pesticides, and fertilizers.

Research and scientific gaps to be overcome and fundamental questions have been addressed to

fuel active development and application of nanotechnology. Together, nanoscience,

nanoengineering, and nanotechnology, offer a plethora of opportunities, proving a viable

alternative in the agriculture and food processing sector, by providing a novel and advanced


Keywords: agriculture, nanotechnology, pathogen, pesticide, sensors, fertilizer.

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1 Introduction

The global population is projected to reach 8.5 billion by 2030 and the world will need to

produce at least 50% more food, to feed approximately 9 billion people by 2050, according to the

United Nations reports and the recent report.1 Conversely, biodiversity, agricultural estates, oceans,

forests, and other forms of natural resources, are being depleted at an unprecedented rate.2 Thus,

increasing agricultural productivity, and improving postharvest processing, and food processing

practices, which are vital to feed a growing population. For many decades, farmers have been

attempting to increase agricultural yields by using conventional fertilizers, pesticides, and hybrid

seeds. Global climate change is also causing a reduction in agricultural productivity, due to the

enormous industrial growth.3 Environmental impacts on agriculture can be assessed using agri-

environmental indicators to measure the adverse effects of cropping and farming, such as water

and soil pollution, soil erosion, and emission of greenhouse gases.4 Sequestration of organic carbon

in West African soils significantly increased maize yields by 24% in addition to this soil organic

C was increased by 14 to 26%.5

As a result of these ecological crises, the cost of essential foodstuffs and the inflation in

fresh food prices are volatile in nature, which will continue alarming the common population. The

social and nutritional implications of high food prices have wide-ranging impacts on well-being

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and health of millions of people is at stake, placing pressure on vulnerable households to follow
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an incomplete diet and consume less nutritious food.6, 7 Moreover, one-third of all child deaths

globally, are mainly due to malnourishment.8 The worlds food security will be at great risk, unless,

and until, we advance agricultural practices and manage our natural resources sustainably.9

Nanotechnology has the imminent potential to make a useful influence on several sectors,

agriculture, food processing, forestry, environmental problems, water industry, energy production,

and sustainable utilization of waste resources.10 In recent times, a wide range of nanotechnology

applications have been predicted in agriculture, thus attracting intensive research and development

practices at both academic and industrial levels is illustrated in Figure 1.11, 12 Nanotechnology has

tremendous opportunities particularly, in the postharvest processing of agricultural waste materials

including the production of nano-cellulose13, nanocomposites,14 and biochar.15 Some of the main

goals in agricultural nanotechnology requiring immediate attention are the development of novel

nanocomposites that are able to carry active agents such as nutrients, fertilizers, and pesticides.

The demonstration of real and practical advantages of nanoformulation systems, required design

of facile and scalable processes, risk, toxicity, and life-cycle assessment of the nanomaterials

(nanofertilizers and nanopesticides), and also simplicity in the regulations about the

commercialization of nanoproducts.16-18 Other achievements yet to be accomplished includes

climate-oriented farming systems, restoring infertile soil, and breeding new drought resistant crop

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species, to convey sustainable agricultural practices worldwide. Moreover, environmental
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sustainability and the principles of green chemistry need to be incorporated into nanotechnologies

at the source. Thus, despite the considerable advances in nanotechnology, many issues must still

be resolved, to create a significant impact in the agricultural sector.

The major production and economic losses in agricultural sector are caused by plant

diseases 19. For example, an estimated 2629% of soybean, wheat, and cotton losses worldwide,

are due to pests whereas the losses increases up to 31, 37 and 40%, in case of maize, rice, and

potatoes, respectively.20, 21 Hence, investments in developing advanced agricultural technologies

and their effective implementation with high priority is essential to boost the agriculture and food

processing sector. Finally, improving food processing, storage, and transport can also be

implemented to reduce the food losses and to improve food security.22 Nano- and bio-technology

includes the use of modern tools to facilitate genetic engineering crops programs, to improve food

security and to promote agricultural sustainability on a global scale.23 Thus, scientific

developments in nanotechnology have great potential to increase agricultural productivity, food

security, and processing capacity, while promoting social and economic equity.

In this context, we are motivated to report recent trends in nanotechnology and

nanomaterial-based systems that could provide sustainable assistance to manage the food supply

chain and precision farming. Nanoparticles, nanowires, nanotubes, and/or nanocrystals have

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unique thermal, electrical, optical, and surface chemistry properties.24 These materials are useful
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to establish excellent sensitivity toward different analytes, rapid response times, and low detection

limits, and multiplex detection sensors.25, 26 Thus, increasing the efficacy of agricultural practices

is possible by using modern applications that are involved in managing pests or diseases more


For instance, nanotechnology postulates various applications in improving plant protection

and nutrition (Figure 1). Various types of hydrogels and nanoclay-polymer composites have been

applied to increase the soil moisture content and to improve the water-holding capacity of soil

during the cultivation season.27 Also, fertilizer loaded superabsorbent polymer composites,

graphene quantum dots, and nanoclays, act as slow releasing agents and potential carriers to

manage mineral nutrient release and plant growth regulators.28 Engineered nanoparticles also have

significant advantage in improving crop protection and pesticide residual detection.12, 29 However,

nanoagrochemicals established for agricultural use is expensive in comparison to conventional

agrochemicals.30 Furthermore, comprehensive investigations are required to ensure the ease of use

of the nanoproduct to fulfill the safety parameters and have the ability to provide improved

alternatives to the traditional formulations.31

The uptake, bioaccumulation, and biotransformation of nanoparticles by plants and their

risks to food crops are not fully understood.9, 32 Consequently, the safety of nanoproducts in

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agriculture and food packaging, 33, 34 and the potential of adverse ecosystem responses and negative
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impacts on animal and human health from nanoparticles, remains clearly unknown. A previous

study recommends that the impact of nanomaterials on seed germination, root elongation, shoot

growth, and biomass may be not profound enough to evaluate their toxicity in agriculturally

important plant species.21 Hence, research into development of new methods are required to

address the specific properties of nanomaterials. Most of the recent nanotechnology research is

incomplete without the consideration of phytotoxicity and genotoxicity caused by nanoparticles.35,

Although detailed data about the risks associated with individual nanotechnologies is missing

due to an early stage of research and development and thus, guidance based on proactive measures

can be developed to control the risk factor.37, 38

2 Nanotechnology-based sensing opportunities in agriculture

Nanotechnology has accomplished a great deal in advancing a wide range of fields, from

electronics to energy, medicine, agriculture, food processing and more.9, 39, 40 Nanofabrication is a

pivotal component of modern technologies. Advances in materials science, device fabrication, and

manufacturing of nano-enabled products will facilitate the development of advanced

technologies.41 Automation should be implemented in agriculture to improve plant health,

agricultural yield, and quality.42 The properties of nanoscale products are tunable to regulate their

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size distribution, morphology, and composition, therefore, devices need to be engineered for
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specific uses.43 These physicochemical properties of nanoparticles have been actively used in

developing efficient biomedical, diagnostic, and therapeutic technologies.44 Many of these tools

have been proven by nanofabrication of chemosensors based on surface plasmon resonance,45

wireless sensor networks,46 electrochemistry,47 chromatography,48 electrophoresis chips,49 and

micro/nanofluidic techniques.50 Among the commercial nanotechnologies achieved, so far,

smartphones have gained huge market shares in various sectors, due to their portability, practicality,

easy operations, and convenience.51 Thus, advanced analytical tools are possible using different

types of nanomaterials, interfaces, and devices as well as their properties through systematic

interdisciplinary investigation and effective knowledge transfer.52 Current nanotechnology-

enabled sensors available on the market include optical sensors, electrical or electromagnetic

sensors, and electrochemical sensors.53, 54

Selective detection and efficient remediation of various persistent pesticides and

environmental pollutants is a major challenge. In particular, sensors developed for diagnostic, and

remediation purposes may allow on-site application with efficient monitoring in diverse

environmental conditions are more desirable. Therefore, achieving agricultural productivity in a

precision farming with plant growth and protection agents is ongoing challenge, while reducing

the use of hazardous agrochemicals.10 Modern agricultural management depends intensely on

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various sensing systems that provide precise information about crop conditions, soil quality,
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climate change, and weather conditions. The practical applications of electrochemical sensors are

being established for the direct measurement of soil chemistry and soil pH, and for monitoring
nutrient content in the soil. Hi-tech digital systems suitable for diverse soils and weather

conditions are in demand to assist growers. Detailed information about these factors is required

for the sensor technologies to define soil quality, fertilizers, and best treatment practices. Thus,

nanosystems must integrate sensors into the device, to improve the potential applications of

computation, communication, and power.55, 56 Nano-enabled sensors developed for monitoring the

ions, heat, nutrients, and pH could prevent overuse of water, fertilizers, and pesticides while

increasing the agricultural yield.57, 58

Additionally, facile, rapid, and cost-effective sensing of different target analytes was ably

monitored from the food contaminants (e.g., pesticide content, heavy metal traces, toxin content,

and food preservatives) to ensure food quality and safety.59, 60 Heavy metal contaminations in

agriculture soils pose threats to public health due to their toxicity and ability to accumulate. Thus,

the sensitive and selective biosensors methods for detecting heavy metals in trace amount and

developing various nano-adsorbents for the removal of heavy metal ions are largely required.61

The detection methodology based on the statistical algorithm and chemometrics method was

proposed for qualitative and quantitative analysis of heavy metals.62 Different colorimetric and

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electrochemical assays have been reported for the detection of alkali and alkaline earth metal
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ions,63-65 and heavy metal ions.66, 67
Thus, innovations in sensor technology by employing

nanomaterials continues to overcome industrial challenges for food and water safety. There are

many issues to be addressed before launching sensing tools, selectivity toward the target species,

reaching satisfactory results, dealing with the complex food medium, achieving ultrasensitive

limits of detection, and operating in versatile environments.56

The limited efforts are functional concerns to detect pesticide residues in vegetable and

fruit samples, even though the use of pesticides in agriculture has increased tremendously.

Detection of chemical residues in the fresh agricultural harvest will help growers and food

processors to safeguard and certify standards for organic foods and nutritional products.68

Analytical, laboratory-based methods, such as gas chromatography, high-performance liquid

chromatography, and mass spectroscopy used to identify and quantify residual pesticide contents

are not cost-effective and portable.69, 70 Enzymes, antibodies, cells, and DNA-based biosensors

have been industrialized for the detection of pesticide residues with higher sensitivity and accuracy

than the conventional methods.71, 72 However, accurate monitoring of minute residues of pesticide

in a large number of agricultural samples, without the involvement of complex pretreatment

procedure is encouraged to revolutionize the modern analytical methods with high selectivity,

rapidity, and sensitivity. The detection of organochlorine pesticides, such as aldrin, dieldrin,

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lindane, and -endosulfan, was reported by using surface-enhanced Raman spectroscopy (SERS)
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and optimization of the SERS-sensing substrate.73 Functionalization of the plasmonic nanometal

surfaces that exhibit interesting scattering, absorbance, and coupling properties can be used

effectively to form sensitive hot spots in SERS measurement of the pesticides.74-77 Nearly, all

sensing techniques invented could have an application in the agriculture and food processing.

According to the organic principles, monitoring of the pesticide content and the impact of food

processing on the food quality seems of high relevance.78 The use of nanoparticles in sensors and

software are being designed to determine salinity and water requirements in the field while

increasing the yield and saving water.79 Although nanosensors can assist farmers in maintaining

precise control of their crops, research efforts to develop the nanosensors will be vital to aid

decision-making in crop protection, monitor nutrient profiles, recommend pesticide dose, and

enable real-time monitoring of soil parameters (residual pesticides, nutrients, pH and soil

humidity).80 The use of real-time data for decision-making is significantly important in the

agriculture sector and in the environmental monitoring.81

Detection of foodborne pathogens and prevention of food poisoning diseases aids to

guarantee food quality and safety across the food chain.82 Laboratory-based disease identification

methods, such as polymerase chain reaction (PCR), fluorescence in-situ hybridization, enzyme-

linked immunosorbent assay, and flow cytometry, is being currently used in agriculture.83-85

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However, the most promising alternative is the application of real-time quantitative polymerase
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chain reaction (Q-PCR), which offers a specific, sensitive and rapid method for the diagnosis of

plant pathogens in water, soil, air and plant samples. The most common disadvantage of Q-PCR

is the complex and time-consuming sample preparation, and the lack of necessary facilities in rural

areas.86-88 Modern methods based on nucleic acids, biosensors, and immunological assays, allow
rapid detection of pathogens, although they possess certain disadvantages including low

selectivity and sensitivity as compared to the nanosensor methods.90 Thus, the design of smart

sensors in precision farming will influence the agricultural productivity by providing accurate

information to make accurate decisions related to the agricultural problems.

The diagnosis of various types of plant diseases is essential before significant damage

occurs to the crop through extensive pathogen dispersal. Thus, farmers require cost-effective, light-

weight, safe, and compatible sensor technologies.91, 92 The ability to sense and detect pathogens

and biomolecules will be improved radically by current developments in nanoscience and

nanotechnology.93 Nanotechnology has the ability to revolutionize the agriculture sector by

detecting plant pathogens and enabling their efficient treatment, via developing smart delivery

systems and best management strategies.94 Thus, pathogen detection, disease diagnosis, and

pesticide residual examination will be possible using nanotechnology-based sensors.

Nanoparticles and quantum dots, used in nanotechnology devices, have shown potential to increase

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sensitivity, specificity, and speed for the detection of specific biomarkers and pathogens.95 Surface
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functionalization of nanoparticles, using biomolecules allows creating specificity needed to

identify a particular pathogen.96 However, efficient application of nanomaterials and nano-enabled

devices to provide desirable technical support, holds a grand investigation at the joint of

agricultural yield and safety.56, 97 The ability of nanosensors to detect plant pathogens and disease

is in the early stage, but would fulfill the demand for food and also reduce the food price inflation.95,

3 Nanotechnology-based opportunities in crop protection

The rapid expansion of nanotechnology has founded a global scientific revolution in the

twenty-first century; driven by multiple factors, including the interdisciplinary nature of the field,

nanoscale phenomena, and increased interest between academics and enterprises.99

Nanotechnology applications are already being explored in biomedicine, diagnosis, and

antibacterial treatments, however, their implementation in crop protection is relatively recent.

Specifically, in agriculture, nanotechnological innovations are of the prime importance with regard

to addressing global challenges such as population growth, climate change, toxicity of

agochemicals, and the limited availability of minerals. Some nanopesticides have already arrived

in the market, while, for example, controlled-release agrochemicals is still under development and

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may take several years before they are commercialized. The excessive application of pesticides
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should be replaced by nanoemulsion, nanocomposite, and nanoparticle formulations that are safe,

effective, and environmentally sustainable in the killing of agricultural pests and pathogens. The

development of nanoformulations of agrochemicals for applying fertilizers (discussed in section

4) and pesticides is one aspect of the agriculture sector that is receiving increasing interest.100, 101

The huge usage of pesticides has grown potential to pollute the environment and pose a

hazardous danger to living beings. In green revolution, pesticides and chemical fertilizers have

been practiced markedly, thus loss of soil biodiversity, ground water contamination, resistance

evolved in pathogen and pests as well.102 Nanotechnology is becoming progressively significant

for delivering pesticides for their safe applications. The application of nanotechnology in

agriculture aims in particular to reduce the dose of plant protection agents, minimize nutrient loss,

identify plant pathogens, detect pesticide residue, and increase yields.103 A nanopesticide is a

formulation having nanomaterials as an active ingredient, employed to enhance the efficacy of

plant protection products.104 Figure 2 presents various type of nanomaterials for the development

of modern applications include, nanoemulsions, nanocapsules, carbon nanotubes, graphene oxide,

zinc oxide, copper-gold nanoparticles, iron nanoparticles, silver nanoparticles, and gold

nanoparticles.105-107 Particular emphasis is placed on formulations based on biodegradable

nanocomposite materials prepared from biopolymers, which offer an opportunity to replace

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petroleum by-products, addressing significant concerns regarding pollution and sustainability.108,
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Biogenic nanosilica and nanofibrillated cellulose based nanocomposites were reported for slow-

release of tebuconazole with the intention of biocidal purpose and reduction of the initial costs.110

Most of the recent biopolymer nanocomposites have been developed based on renewable resources

such as chitin, starch, and cellulose.111-114 For instance, starch-based nano-composites are widely

examined due to their nontoxic nature and advantage in controlled release of agrochemicals and

biostimulants.115 Such nano-enabled methods are particularly important to enhance the

effectiveness of hydrophobic pesticides and to reduce the negative impact of pesticides on the

environment and health.116

The application of nanomaterials in pesticide industry is the relatively new field and

designing delivery mechanisms are in the early stages of growth. There are many techniques

emerging as a novel antimicrobial, comprising use of those numerous organic and inorganic

products in coordination complexes, composite materials and antimicrobial treatments.117

Nanoencapsulated pesticide formulation could help to reduce phytotoxicity of the active

ingredients, thus, enhanced solubility and stability need to be demonstrated, by protecting

pesticides in nanocapsules.118 Challenges associated with the development of encapsulated

pesticides are ensuring excellent reproducibility, selectivity, specificity, and its controlled


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Besides, it is important to prove the ineffectiveness of the pesticide by leaching, and
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pesticide drainage in soil, and groundwater, which could cause potential life-threatening
consequences . The development of nanodevices, smart delivery systems, and targeted

nanocarriers for controlled release of pesticides and fertilizers have a wide scope. Nonetheless,

several challenges must be addressed to prove the efficacy of nanotechnology-based plant

protection agents against bacterial, fungal, and viral pathogens. Nanoencapsulated pesticide

formulations are useful to reduce the dose of pesticides while achieving successful pest removal

with an environmentally friendly approach 118. However, there is need of sufficient development

in nanoencapsulation, in terms of cost-benefits and effectiveness to enhance the applications of

nanoencapsulated pesticides. In recent lab scale experiments, it has been identified that modern

nano-fertilizers can improve agricultural yield by increasing the rate of seed germination,

photosynthetic activity, seedling growth, nitrogen fixation, and carbohydrate-protein metabolism

101, 121

Moreover, the rising threat of resistance in plant pathogenic fungi is a major challenge for

the agriculture sector. Hence, the need to explore novel and cost-effective antimicrobial bactericide

and fungicide agents.122, 123 Particularly, a variety of nanomaterials have been used in developing

novel antimicrobial agents for agricultural disease control includes, graphene oxide, titanum oxide,

carbon-based nanomaterials, copper nanoparticles, and silver nanoparticles.124 Recent studies

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showed the antibacterial potential of Taraxacum officinale mediated AgNPs against Xanthomonas
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axonopodis and Pseudomonas syringae. Nanoformulation with standard antibiotics mainly

tetracycline showed higher antibacterial potential against antibiotic resistant phytopathogens.125

Both the Ag- and Cu-based monometallic graphene composites were reported to suppress bacterial

growth, in particular, Ag-based graphene composite showed higher antibacterial activity as

compared to the Cu-based graphene composite.126 Recently, several important multi-drug resistant

bacteria have been successfully treated by using the antibacterial activity of reduced graphene-

silver nanocomposite.127 The development of recyclable and synergistic graphene oxide

nanocomposite was reported by using both silver nanoparticles and iron oxide nanoparticles as a

novel multifunctional antibacterial material.128

Numerous studies have confirmed that metal nanoparticles are effective against plant

pathogens, pests and insects and, therefore, the topic is deserving of extended discussion. Effective

control of plant pathogens is challenging because their populations vary according to time, space,

and genotype. Evolved resistance in plant pathogens is the most important issue in current plant

protection practices.19 Bacterial pathogens often entrench and destroy the crop at the time

symptoms are recognized.129 Pathogenic bacteria cause many serious diseases of crops. For

example, bacterial blight disease of pomegranate fruits, caused by Xanthomonas axonopodis pv.

Punicae, is a serious issue in several countries.130, 131 Furthermore, bacterial pathogens can become

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systemic in a plants vascular tissue system, making it unfeasible to eliminate all the pathogens,
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simply by applying the pesticide to the plant surface.132 Metal nanoparticles, such as silver, have

been comprehensively investigated against agriculturally important plant pathogens, using in-vitro

experiments, to reveal their inhibitory mechanism and plant protection applications.133, 134

Consequently, silver-based commercial products have been extensively manufactured for their

bactericidal applications against a wide range of microorganisms.135, 136 Silver nanoparticles act as

antibacterial agents by releasing silver ions into the intracellular matrix of pathogens, affecting the

metabolism of the plant, in a similar way to chemical-based crop protection agents.137

The existing antifungal drugs are well known for their fate, toxicity and environmental

pollution, thus, new formulations need to be discovered and industrialized to sustain the plant

protection practices30 Aguilar-Mndez showed that silver nanoparticles significantly delayed, the

growth of Colletotrichum gloesporioides, which causes anthracnose in a wide range of fruits in a

dose-dependent manner.138 Silver nanoparticles have also demonstrated broad-spectrum antifungal

activity against wood-degrading fungal pathogens, such as Gloeophyllum abietinum,

Gloeophyllum trabeum, Chaetomium globosum, and Phanerochaete sordida.139 For instance,

many Alternaria and Fusarium species are commonly reported as the causal agents of fungal

disease outbreak in diverse plant species.140 The spores and hyphae of Fusarium graminearum

were damaged successfully using a graphene oxide-silver nanoparticle composite through

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releasing chemical reactive oxygen species and physical injury.141 Both silver ions and
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nanoparticles have shown a significant inhibitory effect on colony formation of spores and disease

progression of the cereal pathogenic fungi Bipolaris, which causes spot blotch disease, and

Magnaporthe grisea.142 Fluconazole antibiotic was in presence of the silver nanoparticles and the

nanoparticles evaluated for their ability to enhance the antifungal activity of fluconazole against

diverse fungal cultures, including Phoma herbarum, Phoma glomerata, Trichoderma sp.,

Fusarium semitectum, and Candida albicans.143 Although, no significant enhancement of activity

was found against P. herbarum and F. semitectum, the combination of antibiotic and silver

nanoparticles displayed maximum inhibition against C. albicans, followed by P. glomerata and

Trichoderma sp. Silver nanoparticles significantly lower the respiration of mycelium and increase

mycelial membrane permeability. Thus, ultrastructure analysis revealed many alterations, such as

cell collapse, expansion, and apoptosis of mycelium cells.144, 145

The carbon nanoscrolls composed of graphene oxides and silver nanoparticles identified

for local antifungal therapy with enhanced and prolonged antifungal activity.146 Graphene oxide-

silver nanocomposite was investigated for its antifungal activity against Fusarium graminearum,

the results showed that the inhibition efficiency was increased ~ 3- and 7-fold over the pure silver

nanoparticles and graphene oxide.141 Zinc nanoparticles are also well-known for their excellent

antifungal activity against Penicillium expansum, and Botrytis cinerea biocompatibility with the

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plants . Detailed investigation of the nanopesticides includes behavior, inhibitory properties,
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and the potential in suppressing the plant pathogens in field trials will help to establish a regulatory

framework for their commercialization. A new method for the preparation of nanohexaconazole

was reported using polyethyleneglycol-400 (PEG) and it was found that nano form of

hexaconazole were efficient fungicidal agent in comparison to that of conventional

hexaconazole.148 Sodium alginate-chitosan matrix is reported for formulation of nanoetofenprox

through entrapment having potential for effective management of Spodoptera litura.149 In-vitro

the insecticidal activity of pyridalyl nanosuspension was reported against tomato shoot and fruit

borer Helicoverpa armigera, showing that insecticide load to the environment can be reduced

significantly in comparison to the commercial formulations.150

Colloidal copper has been used since 1931 in a fungicide product known as Bouisol, to

control fungal diseases in grapes and fruit trees. Applications of copper nanoparticle/polymer

composites as antifungal agents have been proposed in previous reports.151 Copper nanoparticles

showed a strong antifungal effect against agriculturally important fungal phytopathogens, like

Curvularia lunata, Alternaria alternate, Fusarium oxysporum, and Phoma destructive.152, 153

However, there must be clarity about whether the copper is present in an ionic or nanoparticulate

form, as this can impact the control of several bacterial and fungal pathogens.154, 155 Moreover, the

health consequences of consuming residual copper ions and colloid forms of copper from

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agricultural produce are unknown.156, 157 Various metal-based nanomaterials can be possibly used
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for plant disease management using efficient crop protection practices, particularly, to avoid the

agricultural losses posed by fungal and bacterial pathogens. Metal nanoparticles could be used to

inhibit the pathogen in multiple ways and in different growth stages. Thus, nanopesticides

containing metal nanoparticles, could be useful in killing various phytopathogens, eliminating the

opportunity to develop resistance to the pathogens. However, comprehensive antifungal studies

are required to develop new disease management strategies using metal nanoparticles that are less

harmful to the plant and the environment.158 In order to establish the potential use of antifungal

and antibacterial nanoparticles in control of plant diseases, a detailed understanding of the

antifungal activity of plant pathogens and the accomplishment of application strategies to increase

the effectiveness in the disease suppression is required. Nanotechnology systems can detect

diseases in both plants and animals, thus have potential to improve food quality by delivering better

plant nutrients. The great hope lies in the nanotechnology innovations that fills shortened food

supply by finding ways to improve the health of agricultural crops.

4 Nanotechnology-based opportunities in fertilizers

Increasing agricultural yields are required to feed a growing world population. Effective

strategies to improve agricultural practices are urgently needed, with limited use of important

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mineral resources, such as phosphorus and potassium. Scientific research, and publications and
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patents on nanomaterials with promising agricultural applications, for plant protection and

fertilization, began several decades ago, however, very few products containing nanoproducts are

present in the market.159 Nanoproducts called soil-enhancers have been developed by technology-

oriented companies, to improve water distribution, storage and saving in the agricultural sector.

Water-holding polymers, gels, nanoclays and nanozeolites,160, 161 have been used to improve the

water-quality and -holding capacity of the soil. Organic polymers, inorganic metals, metal oxide

nanomaterials, and carbon nanotubes have also been used to reduce the time and cost of soil

remediation treatment.162, 163

The uses of nanotechnology in remediation of water, includes

nanomaterials for water filtration and catalysis and disinfection.164, 165 Nanotechnology has the

huge possibilities in contributing to the long-term water quality assessment, and the use of

advanced filtration nanomaterials that enables sustainable desalinization, water reuse, and

recycling.166 To address scarcity of water resources and low rainfall, an effective treatment of a

wastewater is recommended by using advanced wastewater treatment technologies. The excessive

application of mineral fertilizers and pesticides is causing enormous pollution and serious health

issues. Thus, better release mechanism and targeted delivery of minerals and nanopesticides that

shows better activity against broad-spectrum pests with excellent protection efficiency that is

desirable and sustainable in comparison with conventional pesticides.167 In particular,

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implementation of novel and improved sensors to detect organic and inorganic contaminants in the
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trace quantity from the soil and water samples are not yet achieved magnificently. Interest in

nutrient accumulation and its release is desirable to increase agricultural yield by improving the

nutritional quality of plants as foods and feeds.168 Micronutrients packaged in nanoparticles have

profound effects on uptake of micronutrients and fertilizer use efficiency relative to conventional

salts and ions.169 Agronomic fortification with micronutrients was successfully established in a

greenhouse experiments with composite of three micronutrient nanoparticles (ZnO, B2O3, and

CuO) to mitigate drought stress in soybean.170

Precision agriculture, in particular, is documented to reduce applications of pesticides,

optimize nutrient management, and increase agricultural yields. The long-held belief that excess

application of fertilizers can enhance crop yields has seriously impacted the soil quality and soil

contamination. The farming community has realized that there is an inadequate amount of nitrogen

in the soil, thus, most farmers are boosting the nitrogen levels in the soil to achieve optimum

growth of their crops. The dependence on synthetic nitrogen fertilizer dramatically increased after

the discovery of the HaberBosch process, in 1913, because nitrogen is a key nutrient source in

agriculture for protein production.171 Dr. Carlos Monreal from Carleton University, stated that for

every $100 a farmer puts into the soil, only $30 get utilized by the crop plants due to limited

nitrogen use efficiency by crops.171 Conventional fertilizers, such as urea and NPK (nitrogen-

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phosphate-potassium), can only fulfill the nitrogen requirement for a short time because excess
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nitrogen is lost in the gaseous or ionic form through leaching, denitrification, and volatilization.

Nitrate not absorbed by the plant roots is washed away in runoff or leaches into the soil along with

water. Moreover, sustainable agricultural practices, energy conservation, and resource utilization

do not allow excessive use of pure fertilizers.172 Nitrogen fertilizer losses may cause environmental

problems, such as air and groundwater pollution through ammonia volatilization, denitrification,

and leaching.173 Nano urea successfully increased the agronomic efficiency by 44.5 % and the

grain yield by 10.2 %, with modern nitrogen fertilization in comparison to the normal urea.174

Nanoparticle-based fertilizers are proposed to address excess fertilizer application, by

releasing the required nutrient on demand and preventing premature loss. However, although

various aspects of nanotechnology in agriculture have been investigated by many research groups,

no specific strategies to address the problem of loss of nitrogen fertilizer have yet been

investigated.175, 176 The main factor hindering research and development activities in agricultural

nanotechnology is the low returns in the agricultural sector. Conversely, nano-enabled commercial

products have only been achieved in laboratory scale research, due to high production costs.

Nonetheless, the high sorption capacity and high aspect ratio of nanoparticles are suitable for the

controlled release of micronutrients to the target site. Consequently, nanofertilizer delivery

systems have the prospective in increasing nutrient efficacy and crop productivity, through

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enhancing photosynthetic activity, nitrogen metabolism, and protein synthesis.101 Development of
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nanoporous materials, nanopolymers, and nanocomposites that can immobilize nutrients and

release nutrients into the soil have the advantage to prevent loss of nutrients from soil and also

mitigates eutrophication.177 . In this regard, predominantly nanoparticles resulting from

biopolymers are attractive such as proteins, chitosan, and the starch having biocompatible

properties and low impact on the human health and the environment. There is a huge prospective

for nanotechnology in agriculture, however, many issues are still to be fixed, such as increasing

the yield of nanotechnology processes to lower the cost and completing the risk assessment trials.

Conventional formulations of pesticides and fertilizers can contaminate the environment,

thus there is a need for controlled-release formulations of agrochemicals using hydrophilic,

biodegradable and sustainable polysaccharide biopolymers.178 The design of different soft

nanomaterials from renewable feedstocks (biomass) is gaining tremendous attention for their

unique properties and safer applications.179 The soft materials having nanometer scale such as

polymers, gels, biopolymers, membranes, nanocomposites, and biomimetic materials are

considered as soft nanomaterials. Harvesting, processing and producing ligno-cellulosic biomass

from agriculture waste are desirable for commercialization of cellulose based nanocomposites.

Soft nanomaterials have several applications in our daily life; including cosmetics, adhesives,

foams, gels, biomedical sciences, food packaging, and agricultural formulations.180-183 Polymer-

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coated fertilizers have the potential to minimize agriculture inputs during the growth period and
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their utilization should not create harm to the environment and crop plants.184 A urea-

hydroxyapatite nanocomposite (6:1 by weight) has been developed to maintain yield, while

reducing the amount of urea applied to the soil, by reducing its solubility, providing a platform for

its slow release compared to pure urea.185 Chitosan has also attracted great attention due to its

ability to form complexes and its suitability to develop controlled release of active compounds,

which, together has allowed a major advances in the agricultural, pharma, and biomedical

industries.183 Applications of nanotechnology as plant nutritional mediator in the form of modern

nanofertilizers are referred to as controlled release fertilizers (CRFs). CRFs designed for

controlling nutrient release behaviors that rely on thermal post-treatment parameters, can enhance

the nutrient use efficiency of crops.186 Yet, soil temperature-dependent CRFs need to be

comprehensively monitored with different crops to obtain reliable data regarding release

characteristics and duration of nutrient release. An increase in temperature, from 20 to 40 C, was

shown to differentially increase the rate of release of various nutrients from a polymer-coated CRF,

such that nitrogen release into the water was rapid in comparison to potassium, whereas phosphate

was significantly slower.187 Shaviv et al. (2003) developed a statistical model, as an effective

agronomic tool, to monitor the release of nutrients from CRF granules. Besides CRFs, slow release

fertilizers (SRFs), for the slow release of nutrients have also been demonstrated.188 SRFs consist

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of urea coated with water-insoluble polymers that contain the fertilizer, thus, the runoff can be
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avoided. SRFs aim to supply nutrients required for the optimal growth of crop plants, thereby,

enhancing nutrient efficiency, lowering labor costs, and reducing negative side effects.184, 189

A semi-arid regions, often experiences severe drought conditions. Despite enough rainfall,

the quality of soils allows low water retention capacity, rapid evaporation, and more insolation

may result in diffusion of water into a bottomless layer of soil. In this report, we summarize the

application of soil conditioners derived from the polymer, hydrogels, and nanoclays to achieve

effective water holding capacity. Water and nutrients conservation is important to safeguard these

limited resources after inputs to agriculture and essential to be used sensibly with sophisticated

efficiency. For these motives, a series of nanoclaypolymer composite used as a superabsorbent

of nutrient carriers and water holder were reported.190 Multifunctional hydrogel materials prepared

from nanoclay, agar, -carrageenan, and konjac glucomannan were reported for their water barrier

properties such as water contact angle, water vapor permeability, water vapor uptake, and water

uptake efficiency.191 In addition to this value added products from agriculture and industrial waste,

in particular biochar prepared by slow pyrolysis has also shown great potential to improve soil

conditions. Biochar produced from orange peel, sugar cane waste, coconut shells, palm oil waste,

sugarcane bagasse, water hyacinth etc. are applicable for conditioning the soil. Recently, results

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reported to explain the polarity of the biochars, and excellent hydrophilicity can be increased by
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tuning the micropores that could hold and retain the water.192

5 Nanotechnology-based opportunities in postharvest processing

The most recent literature on the application of nanotechnology in agriculture

emphasize nanofertilizers, nanopesticides, nanosensors, crop protection, and pollution control.

However, a wide scope lies in renewable agricultural waste, which can be processed effectively to

produce raw materials and final nanoproducts for the development of several different applications.

Agriculture waste materials have attracted attention as a potential source for substitution of fossil

fuel resources. Numerous postharvest technologies are yet to be implemented efficiently to

generate nanocelluloses, nanocomposites, and biochars from agricultural wastes. Abundant

lignocellulosic biomass is obtained from agricultural waste and is a renewable resource that has

great hope for the preparation of various functional nanomaterials.193 Nanocomposites based on

biopolymers, such as starch, proteins, and carbohydrates, are safer and sustainable compared to

chemical-based composite materials. Rice and wheat husks are byproducts of the harvest that can

be used as raw materials for the fabrication of renewable energy, high-quality nanosilica and

biochar, for generating other value-added products.194 Implementing the mass production of

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nanosilica using nanotechnology and biochar production systems, can alleviate the growing
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concerns about disposal of agricultural waste.195, 196

The most important factor is that the use of waste lignocellulosic waste biomass does

not compete with the food chain and fodder industry for the synthesis of green, safer, and

sustainable nanomaterials (Figure 3). Nanotechnology can be used to transform waste cotton fibers

and cellulose into value-added products, using electrospinning.197 The nano-micro fibers produced,

can be used as high-performance absorbents to develop targeted applications in fertilizers or

pesticides.198, 199 The fascinating characteristics of nanofibres, such as porosity, high surface-to-

volume ratio, and safety makes nanofibres as a suitable candidates for developing variety of

applications.200 Cellulose, a linear polysaccharide consisting of several hundred to many thousands

of (14)-linked-D-glucose monomers, is the most abundant natural biopolymer as shown in

Figure 3. Cellulose is the backbone of tree trunks and branches, and it is the main component in

the paper and cardboard industry. Large amounts of cellulosic- containing biomass, available from

renewable agricultural waste, need to be utilized efficiently for production and application of

natural fibers and nanocellulose.201, 202 Nanometer-scale cellulose fibrils are very hydrophilic and

look like jelly, called aerogels.203, 204 Nanometer-sized cellulose materials offer a plethora of

applications and their properties are vastly distinct from the bulk cellulose of the same material.

Tough cellulose hydrogels obtained by a solvent exchange method was reported for their stability

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under changing temperature, pH, and solvents.205. The complete nucleotide sequence of
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Gluconacetobacter hansenii strain NQ5 (ATCC 53582), an efficient producer of bacterial

cellulose, has been elucidated with the aim of contributing to the understanding of the molecular

mechanisms involved in cellulose biosynthesis.206 The nano-fibrillar like network structures of

holocellulose aerogels, with high surface area and high adsorption property, indicating potential

application in environmentally friendly adsorption materials, were synthesized using tulip tree and

an aqueous alkali hydroxide-urea solution.207 Nanocellulose obtained from natural materials have

gained much courtesy for the evolution of modern medical tools due to its excellent properties,

unique surface chemistry, biocompatibility, and biodegradability.208

Silica is an inorganic element abundantly present in the rice husk, different processes have

been developed in the recent time for extracting amorphous silica having high-surface area.209 The

rice husk is an excellent source of micro- and nanosilica, simply an alkaline hydrogen peroxide

with controlled reaction pH can be used to eliminate of lignin bonds while preserving cell wall and

most of the silica.210 A small nanosilica particles powder having high purity, and high surface area

was prepared from the rice husk by using alkali extraction and acid precipitation technique.211

Nano-silica has been explored to control insect pests, preparation of silica nanoparticles conjugated

with validamycin has been reported by Liu et al., 2006 for effective use and controlled delivery of

water soluble pesticide.212 Nanoscale silicon combined with methyltrimethoxysilane was

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identified as a plant growth regulator, mildew disease controlling nano-fungicide as well as a
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conjugating and dispersing agent.35

The biochar enterprise was inspired by the discovery of the layer of black soil terra preta,

made by dumping waste charcoal at the sites of ancient settlements in the Amazon forest.213, 214

According to local farmers in the Amazon, productivity on the terra preta is much higher than

surrounding soils.215, 216 Biochar can be produced from almost all kinds of agricultural waste and

from all organic matter rich in carbon and hydrogen. Many biomass-based countries, such as India,

Canada, USA, and Ghana, have huge potential to address global warming by transforming their

agricultural biomass residues into biochar production. Literature reports highlight biochar as a

valuable product to mitigate global climate change and also suggest its vast potential in agriculture,

construction, water purification, and environmental remediation.217, 218 Use of water contaminated

by ammonium ions for irrigation purposes have huge risks to the agricultural productivity and soil

fertility. A recent report has evaluated the potential of biochar as an excellent adsorbent to

remediate an ammonium ions from the water samples.219

A recent report suggests that the global estimate of biochar requirement already exceeds

gigatons (approximately, 0.6 0.1 109 tons per year), thus, there is a huge gap in the availability

of waste biomass and production of biochar.220 Recently it was stated that 1 gigaton per year of

carbon emission is equivalent to about 12% of current anthropogenic carbon emission contributed

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from the industrial chimney and automobiles.217 Biochar goals are achievable by utilizing
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agricultural, municipal and industrial biomass, for biochar production in a sustainable manner

without endangering food security, habitat, and forest resources.220 If this scale of biomass

transformation into biochar is achieved by co-operation between countries worldwide, it may be

possible to address the consequences that have emerged from global atmospheric carbon emissions.

Biochar has been produced from various types of feedstocks such as agricultural waste, forestry

residues, wood industry waste, sugar cane industry waste, organic materials of municipal waste,

and also from animal manures.221 Biochar production and processing, based on a particular market,

and their application has been under certain assumptions because the commercial scale processes

have not yet been completely established. Biochar had the greatest influence on the overall growth

of lettuce as shown in Figure 4, thus can provide an opportunity for a soil management system as

an amendment agent after establishing proper recommendations.222 The rice husk biochar used as

an alternative substrate for production of leafy vegetables in hydroponic technology.223 However,

according to the applicability of each type of biomass as feedstock, different physical

characteristics, chemical compositions, porosity of structures, as well as economic, and logistic

factors are influenced by the overall process.224, 225 The density and porosity of biochar restrict its

mobility in the environment, it has impacted on the soil hydrologic cycle and suitable for the

ecological niche for soil microorganisms.226

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Biochar materials produced in controlling thermal decomposition processes have great
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potential in soil remediation and waste disposal management. Biochar is a carbon-rich, porous

material that is stable for thousands of years and produced by the thermal decomposition process,

called pyrolysis, of biomass in a reactor at 350700 C.227, 228 There is no significant distinction

between biochar and activated carbon from the structural viewpoint, considering both are

amorphous carbon, with an abundant porosity in the macroscale range. The potential of biochar-

amended soils based on the surface functionality, and porosity of biochar and soil water retention

features are interesting.229 Scanning electron microscope images of biochar revealed that the

pyrolytic processes releases volatile organics, hemicellulose and lignin, thus porosity results from

the shrinkage, merging, and cracking of the pores.230 Biochar processed in ultrasonic bath

treatment allows to prepare carbon nanoparticles in demineralized aqueous media, subsequently,

atomic force microscopy was used to characterize the graphene sheets prepared from the biochar

materials by the same method.231

The use of biochar having macroporous structures (as shown in Figure 4) is a powerful tool

to battle climate change and to conserve water and nutritions, by sequestering atmospheric carbon

in the agricultural soil. Thus, providing a great opportunity for the processing of agricultural waste

materials and other organic waste into renewable energy.232 The chemical composition, physical

structures, and surface properties of biochar can vary greatly with the type of biomass; thus, the

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utilization of different biomass as a potential feedstock depending on various factors, such as
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moisture content, composition, and ash content.233, 234 The surface functionality and porosity of

biochar are useful to apply directly for the synthesis of many functional materials useful in

catalysis, remediation, and adsorption. Application of biochar to raise soil fertility and to improve

nutrient efficacy, seems promising in agriculture to increase crop yields and has a high prospective

in mitigating the climate.235 Biochar production goals can be achieved by making biochar

production economically feasible and commercial scale operations of energetically autonomous

pyrolysis plants worldwide. Thus, there is the need for comprehensive utilization and investigation,

to reduce carbon emissions and sustain soil productivity by solving the emerging problems.

6 Nanotechnology risk assessment

The number of patent applications from the nanotechnology sector has increased

approximately ten-fold over the last two decades, signifying a great potential in nanotechnology-

enabled commercial products and applications.236, 237 However, the main factors that are limiting

the development of nanotechnology in agriculture and applications are less investments in research

infrastructure,35 low returns from the agriculture sector, high-cost of production, and inefficient

technology transfer in the agriculture field.12, 238 Agricultural production and food products are the

most sensitive fields to apply nanotechnology. The accumulation and uptake of newly engineered

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nanomaterials by plants as shown in Figure 5, may adulterate food chain and causes risk to the
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human health and the environment.239 Terms like, nanoemulsion, nanocomposite, nanopesticide,

and nanofertilizer have been widely used, however, these nanoproducts have different meanings

according to their application. Moreover, the nano term does not designate specific nanoproducts,

regarding their composition, safety, and significance of the agricultural practices.238

The rise of nanotechnology-enabled consumer products in the marketplace has raised a

number of ethical and social concerns, regarding health and environmental safety.240 Thus, the

public is demanding proper labeling and safety standards on packages from all applicants of

nanotechnology products, to identify health and environmental concerns of nano-enabled

products.241 Mukhopadhyay, 2014 reports greater role of nanotechnology in agriculture and food

processing, to guard security of the livelihoods, public approval of nanoproducts, and smart human

resource teams for the advanced nanotechnology research.242 However, these sectors are gateways

of food chain contamination via foliar spray of nanopesticides and soil applications of

nanofertilizers, as illustrated in Figure 5. Risk analysis of nanomaterials in the food and agriculture

sectors, has not yet been comprehensively performed.

The interactions of nanomaterials and plants is of great concern, the key routes of exposure

of nanomaterials to the plants are the accumulation and entrance in the food chain as the plants

closely involve interaction with water, soil, and the atmosphere.243 Thus, the further research is

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required to understand the interactions of nanomaterials with the plants that can enable the rapid
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evolution nanotechnology by establishing biocompatible nanomaterials and uses. It is evident that

the risk assessment approaches and assumptions used in the conventional chemical pesticides may

be inappropriate for nanoagrochemicals.244, 245 Considering the uptake, bioavailability, and toxicity

of nanopesticides depends on the particle number, particle stability, and particle size distribution,

here is a prompt endorsement to develop specific risk assessment guidelines for nanomaterials.100

The safe use of nanoproducts in agriculture, risk assessment factors, and understanding the fate,

interactions, and toxicity of nanoparticles have been proposed to secure food supply chain.246

The regulation of nanoagrochemicals should be based on the scientific evidence regarding

the efficiency, safety, and benefits from the whole formulation (i.e., active ingredients and

formulation base), and not merely on the basis of core size of the active individual nanoparticles.

The importance of combining and adapting different approaches is emphasized, to achieve a

comprehensive assessment of the risks associated with nanoproducts for the development of safe

pesticides from nanoparticle formulations.100 However, R and D funding is focused on the positive

environmental applications of the nanotechnology and is limited to basic knowledge/research,

nanoenvironmental research, and for the potential risk assessment of the nanotechnology.247

The nano-enabled concept has huge potential to serve agriculture and food processing

sectors by building collective experience from all the cross-disciplinary sectors (e.g.,

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nanotechnology, biotechnology, food science), and will be significantly valuable to fuel the growth
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of safe and sustainable agrochemicals. Millions of dollars being spent by both government and

industry to apply nanotechnologies in areas such as agricultural production, food processing, and

packaging.242, 248 According to the current developments and public perception of nanotechnology,
in the near future, nanotech innovations will be the most important tool in the modern

agriculture and agri-food processing industry.12 Nanotechnology also promises creating new nano-

industrial products to reduce pesticide need, improve plant growth, and livestock care. It is

expected that the nanoproducts are prepared to transform the food industry by conveying

revolution in the agricultural yield, food processing, food packaging, and quality of food. The right

time arrived to stare curiosity toward nano-engineered food products, the potential consumer

benefits curiosity, and risk associated to health and environment.

7 Conclusions

In summary, this review discussed potential nanotechnology applications and opportunities

in the agricultural sector. Literature reports suggest that nanotechnology will have significant and

long-term impacts on the agriculture and food processing industries. The potential implications of

nanomaterials and nano-enabled systems are yet to be established to advance sustainable progress

in agriculture and the environment. The current scenario of challenges associated with agriculture,

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food security, and climate change, is the source of improvement through constant exploration of
Accepted Article
science and engineering involved in nanotechnology. A major contribution of nanotechnology is

predictable to emerge from the promising results of nanopesticides is the use of nanoparticles

encapsulation and stabilization of crop protection agents, which will increase agricultural



This research was supported by National Research Foundation South Korea under the

project number 2017R1C1B-5017360. This work was partly supported by Dongguk University-

Seoul, Korea Research Fund 2016-2017.


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Figure Legends:

Figure: 1 Different applications of the nanotechnology projected in the agricultural sector.

Figure: 2 Different types of nanomaterials promoted in the development of agricultural

applications as labeled in respective images. Graphene oxide, gold nanoparticles, zinc

nanoparticles, and silver nanoparticles are cited in the text. TEM image of carbon nanotubes

This article is protected by copyright. All rights reserved.

(photo credit: Leonhardt, Leibniz Institute, Germany), TEM image of iron oxide magnetic
Accepted Article
nanoparticles (image source-, Canada), gold-copper hybrid nanoparticles

(Image credit-Zhichuan Xu, China), vesical image credit- Esposito University of Ferrara, Italy.

Figure: 3 Preparation of cellulose nano crystal by acidic hydrolysis of waste biomass (source of

wood image- Geo Arizona Antevs Biomes, Source of cell wall image- thing link, Source of TEM

Image-Cellulose Lab, Canada).

Figure: 4 Electron micrograph of biochar (image source- biochar and biochar mediated

growth enhancement of the crop plants (image source-cool planet energy systems, Colorado,

United States).

Figure: 5 Nanopesticides and nanofertilizers can pose the threat to the food chain and food

processing industry due to uptake, translocation, and accumulation.

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Figure: 1

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Figure: 2

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Figure: 3

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Figure: 4

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Figure: 5

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