Nanotechnology and its applications in agriculture.pptx

Nanotechnology in Agriculture revolutionizes farming by enhancing crop productivity, improving soil health, and reducing pesticide use. Nano-fertilizers and nano-pesticides ensure precise nutrient delivery, minimizing waste and environmental impact. This advanced technology paves the way for sustainable and efficient agricultural practices.

1. MBB 599 Master Seminar Department of Biotechnology Topic- Nanotechnology And its Applications in Agriculture Seminar Incharge- Dr. Karuna Dhiman By - NH 2023-24-M Sourabh

2. Content What is Nanotechnology 01 Classification of Nanoparticles 03 Methods of Synthesis of Nanoparticles 02 Applications of Nanoparticles in Agriculture 04 05 Challenges of Nanoparticles Case Studies 06

3. What is Nanotechnology ? Nanotechnology is the study, engineering, and use of materials and devices on the nanoscale, which generally ranges from 1 to 100 nanometers. Materials at this size often exhibit distinct physical, chemical, and biological features from their larger-scale counterparts. Feynman RP (1959). 20 nm 20 nm 50 nm 100 nm Fig 1- Silver Nanoparticles Zhang et al. (2016 )

4. Size Comparison Fig 2- A pictorial representation of matter ranging from (nm to mm) Neme et al. (2021) The word ‘Nano’ is a Greek prefix derived from the word dwarf, indicate a billionth. 1 (nm) is equal to 1 × 10 -9 m

5. What makes Nanoparticles so special ? When particles are sized between 1–100 nanometers , their properties differ significantly from larger particles due to quantum effects At this scale, scientists can tune material properties by altering particle size . Key properties like melting point, fluorescence, electrical conductivity, magnetic permeability, and chemical reactivity change at the nanoscale For instance, gold nanoparticles display colors ranging from red to purple , influenced by quantum effects that modify their interaction with light, unlike bulk gold.

6. Comparison of Bulk Gold and Gold Nanoparticles Property Bulk Gold Gold Nanoparticle Size Macroscopic size (visible to the naked eye) 1-100 nm Electrical Conductivity High conductivity Lower conductivity Melting point 1064 °C 850 ​°C. Chemical Reactivity Inert and stable Highly reactive Colour Shiny metallic yellow Varies based on size (red,purple )

7. Pioneers of Nanotechnology In 1959 , Richard Feynman ’s lecture " There's Plenty of Room at the Bottom " envisioned manipulating individual atoms . Direct atomic observation became possible in the 1980s with the invention of the scanning tunneling microscope (STM) by Gerd Binnig and Heinrich Rohrer (1981). The STM marked the beginning of nanotechnology as a distinct scientific field. Binnig and Rohrer received the 1986 Nobel Prize in Physics for their groundbreaking invention Richard Feynman Gerd Binnig Credit - Britannica Credit- https://commons.wikimedia.org/wiki/User:NIMSoffice Heinrich Rohrer Credit- Britannica

8. Synthesis of Nanoparticles Two type of approaches are used for the synthesis of nanoparticles Top down approach- Breakdown bulk material into smaller size particles Bottoms up approach- Growing nanoparticles from indiviual atom to aggregates Top-down approach Mechanical-physical process cm µm nm Coarse grinding Fine grinding Basic material Bottoms up approach nm µm Å Physiochemical processes Agglomerates Germ nanoparticles Atoms

9. Mechanical milling (Top- Down Approach) Nanoparticles are created during the shear action of grinding. The ultimate structure of the powder is defined by the deformation of particles that are captured between colliding balls during milling. A planetary ball mill is often used for mechanical milling. The energy transfer from a steel ball to a powder depends on the milling environment, duration, ball-to-powder mass ratio, size and number of balls, and rotational (vibration) speed. Neumeister et al. (2021) Fig 3- Mechanical milling synthesis

10. Laserablation Also called as photoablation this method involves exposing metal-containing precursors to a laser beam in order to create metal and metal oxide nanoparticles. The precursor is vaporized by the laser beam's extreme heat, and the vapor is quickly cooled to create NPs. By varying the laser intensity and precursor concentration, this technique may create NPs with a high degree of purity and crystallinity, and the particle size can be regulated. Barizão et al. (2021) Fig 4- Laser ablation

11. Bottoms up approach Sol- gel process - This process involves dissolving the molecular precursor , typically a metal alkoxide are added to solvents such as alcohol and then converted it into a gel through of hydrolysis & Condensation . As the gel is moist and must be dried using suitable methods to synthesize specific nanoparticles. The sol-gel process is more widely used and applied in industry it also synthesizes extremely uniform composites with a purity level of 99.9% This technique can generate multiple types of nanoparticles at once Fig 5- Sol-Gel Process Bokov et al. (2021) M(OR) x​ +H 2​ O→M(OH) x​ +R−OH (Hydrolysis) M(OH) x​ +M(OH) x​ →M−O−M+H 2 O (Condensation)

12. Chemical Precipitaion Chemical precipitation is a method used to synthesize nanoparticles by creating a solid phase from a solution. It involves the reaction of two or more soluble precursors to form an insoluble product , which then precipitates out of the solution as nanoparticles One of the most frequently employed method Fig 6- Chemical precipitation synthesis of ZnS NPs Hamed et al. (2021)

13. Microwave assisted Synthesis Microwave-assisted synthesis has emerged as an efficient method for the synthesis of NPs due to its rapid heating and energy efficiency. Unlike conventional heating methods, microwaves heat the entire reaction mixture volumetrically , resulting in faster reaction rates, improved product quality, and sometimes unique morphologies . Can also be used as function for green synthesis of NPs. Fig 7- Microwave assisted synthesis of MoO 2 NPs Phalswal et al. (2021)

14. Green synthesis of Nanoparticles Fig 8- Green Synthesis (Plant Based ) Singh et al. (2023) Green Synthesis : A sustainable, eco-friendly method utilizing non-toxic chemicals, biological stabilizers, and reducing agents from plants and microbes. Plant Compounds' Role: Phytochemicals in plants synergize to create stable and efficient nanoparticles.

15. Classification of Nanomaterials Mekuye and Abera (2023) 2 nm 2nm-50 nm >50 nm (Mixed type) Carcinogenic, Mutagenic, Asthma genic and Reproductive toxin

16. Classification based on Structure Carbon-based nanomaterials which are made of carbon consist of five primary materials. Carbon nanotubes- are shaped like hollow cylinders with a diameter of 0.7 nm. Graphene- consists of hexagonal lattice structures arranged in a honeycomb pattern, composed of carbon atoms on a plane surface just around 1 nm thick. Fullerenes- are carbon structures, spherical in shape, with a diameter of 8.2 nm and are also known as Bucky balls . Carbon nano fibers- are carbon materials consisting of graphene sheets ranging from 50 to 200 nm in size. Carbon black- typically has a round shape and ranges in size from 20 to 70 nanometers in diameter. Carbon based nanomaterials are generally used to reinforce steel Fig 9- Carbon based Nanoparticles Mekuye and Abera (2023)

17. Organic Nanomaterials - Organic nanoparticles are nanosized particles predominantly made up of organic substances, typically consisting of carbon, hydrogen, nitrogen, and oxygen . They are extensively utilized in biomedical fields, especially in drug delivery and cancer treatment , because of their biocompatibility, biodegradability , and the capability to alter their surfaces. Organic nanoparticles can be produced from polymers, lipids, proteins, or alternative organic substances. Organic nanoparticles encompass Dendrimers - are synthetic polymers with extensive branching (under 15 nm) featuring layered structures comprising a central core, a central region, and numerous terminal groups, utilized as scaffolds for tissue repair. Liposomes- are vesicles made of phospholipids (50-100 nm) featuring an aqueous interior and a bilayer membrane structure similar to that of biological membranes . Micelles – comparable to liposomes but consist of a single layer and are smaller in size (5-100 nm) , and they are also utilized for drug delivery. Ferritin- commonly present in nature, has a hollow core with inner and outer diameters of 8 and 12 nm, respectively, allowing it to encapsulate up to 4500 iron atoms as ferric oxyhydroxide. They are typically utilized for drug delivery in cancer therapy.

18. Organic nanoparticles are used for drug delivery sytems and ligands are attached to it which are specific to bio receptors that helps in targeting of specific sites and delievery of drug (In a scenario for treatment of breast cancer ( HER- 2) Human Epidermal Growth Factor Receptor 2 can be targeted and Antibody- Trastuzumab can be used a ligand .) Fig 10 - a) Dendrimer b) Liposomes c) Micelles d) Ferritin Mekuye and Abera (2023) Lipid bilayer (50-100 nm) Single lipid layer (5-100 nm) Hollow core (8-12 nm) Scafold structure (<15 nm)

19. Classification based on No. of Dimensions Nanomaterials are categorized into four types according to their size dimensions: 0D, 1D, 2D, and 3D . 0D- These nanomaterials feature all three dimensions (x, y, and z) within the nanoscale range (>10 nm) . QDs and fullerenes serve as examples of zero-dimensional nanomaterials. Due to this confinement, these particles show no significant structure in any direction, rendering them effectively "point-like"(Spherical) regarding dimensions. 1D- In this class have two of their three dimensions (x, y) in the nanoscale range, but one dimension of the nanostructure is outside the non-metric range (>10 nm) . 1D nanomaterials, such as nanofibers, nanotubes, nanohorns, nano rods, thin films, and nanowires , are examples of needle-shaped nanomaterials . 2D- They possess plate-like structures that extend in two dimensions beyond the nanometer scale , while 1D (x) is within the nanoscale (ranging from 1 to 100 nm) . Coatings and thin-film multilayers, nanosheets or nano walls serve as examples of 2D nanomaterials. 3D- Nanomaterials that extend beyond the nanoscale in any dimension or range of dimensions. All aspects of a 3D material are beyond the nanometer scale or exceed 100 nm . Fig 11- A) OD B) 1D C) 2D D) 3D Mekuye and Abera (2023)

20. Applications of Nanotechnology in Agriculture

21. Quantum dots as a Nanosensor A quantum dot is a small semiconductor crystal ranging in size from 1 to 10 nm , made up of metals like Ag, Cd, Hg, Ln, P, Pb, Se, Te, and Zn . These brightly fluorescent quantum dots emit intense fluorescence in various colors when UV ray is projected on them. These tiny metal or semiconductor containers contain a specific quantity of electrons and have a variety of potential applications in electronics and photonics. Quantum dots are emerging as a new type of fluorescent nanosensors for detecting pathogens. Quantum dots have unique optical and electronic properties such as adjustable light emission based on size, increased signal intensity, resilience to photo bleaching , and the ability to emit light of specific colour under the influence of UV-light . The most commonly used QD system is the CdSe semiconductor core coated with ZnS outer shell . Alexei Ekimov , Louis Brus and Moungi Bawendi were awarded Noble prize in chemistry in the year 2023 for synthesis of “Quantum Dots”. (Will be explained extensively in Case study 2 )

22. Structure of Quantum Dot 1 to 10 nm UV ray Fluorescence Outer Shell Inner core Spherical in shape

23. Nanofertilizers Nanofertilizers are materials engineered at the nanoscale (1-100 nanometers) that provide essential nutrients to plants more efficiently compared to traditional fertilizers and a lso called as Smart fertilizers . Conventional fertilizers must be used in high quantities because their absorption efficiency is low. Nanofertilizers release nutrients slowly, potentially boosting nutrient utilization without causing any negative side effects. These nanofertilizers are designed to release nutrients gradually for a long period and minimize nutrient loss, ensuring environmental safety. Babu et al. (2022) Fig 12- Conventional fertilizers Vs Nanofertilizers

24. Why choose Nanofertilizers To Save the water bodies from eutrophication . Excess Nitrogen (>10mg/L) in water bodies due nitrogen runoff in water body causes Blue Baby syndrome(Methemoglobinemia or Cyanosis) . It causes a baby's skin to appear blue or purple in infants causing hinderance in the ability of haemoglobin to absorb oxygen. Cost effective as conventional fertilizers are more costly and have to be bought in bulk . Soil Degradation is caused by use of excessive fertilizers. It has been observed in a study conducted by IFFCO that use of nano N have halved the amount of urea and increased the grain yield and economic returns in wheat. (Kumar et al. 2020) Neem encapsulated urea not only works as a slow release nitrogen fertilizer but it also has beneficial properties of neem, particularly its pesticidal and antimicrobial effects .

25. Serial No. Nanomaterials Crop Species Conc. Used Mode of Application Duration of Treatments Responses 1 ZnO Cyamopsis tetragonoloba 10 mg/L Foliar spray 4–6 weeks Increa sed biomass , accumulation, nutrient concentrationand enhancements to growth physiology 2 ZnO and Fe 3 O 4 Moringa peregrina 30, 60, and 90 mg/L Foliar application Watered every 3 days Salinity levels reduced the growth parameters significantly 3 Zn, Fe and NPK Cicer arietinum 20 L/plot at each stage Foliar application First at 4–6 leaf stage, second at 30 days, third during pod filling Significant increase in both biological and seed output 4 Zn and B Punica granatum 0, 60, and 120 mg ZnL−1 Foliar application Once every season, one week before first full bloom Increases in pomegranate fruit yield 5 Al 2 O 3 Solanum lycopersicum 400 mg/L Foliar application 20 days Effectively counteracted Fusarium as a biocontrol agent 6 ZnO Coffea arabica 15 mg/L Foliar application 40–45 days Increase of net photosynthesis and increased biomass production Nongbet et al. 2020 Positive Impacts of Nanoparticles on crops

26. Nanoparticles a possible solution for Post Harvest Losses Food loss from harvest to retail accounts for 13.2% globally , while food waste at retail and consumer levels makes up 17% . Collectively, approximately 30% of the food that is produced does not end up reaching its destination the stomach of a humans (FAO 2021) . India country still faces an estimated annual loss of (USD 18.5 billion) due to post-harvest losses of crops and agri-allied produce is projected for the year s 2020 to 2022 by the NABCONS study in 2022 . Nanoparticles are used as edible coating in which agricultural produce can be dipped as they safeguard by preventing dehydration , inhibiting respiration , enhancing texture, preserving volatile aromas , and limiting microbial growth . Edible nano-coatings applied to various foods act as a barrier for gas and moisture transfer, while also imparting flavors, colors, enzymes, antioxidants, and agents that resist browning , potentially prolonging the lifespan of processed foods.

27. How Nanopartilces extends Shelf life Chitosan-facilitated nano-silica coating enhanced the physical, chemical, and biological quality of Longan fruit at room temperature better than other methods by effectively creating an exceptional semi-permeable layer which reduced the gas exchange rate . (Neme et al. 2021) Pt/DMS(Platinum/-loaded dendritic mesoporous silica) was utilized as an ethylene scavenger to prolong the shelf life of Musa nana banana, featuring distinct central-radial pore formations . (Wei et al. 2023)

28. Nano-Pesticides in Agriculture Nano-pesticides are formulations that consist of engineered nanoparticles and have biocidal properties. Nanopesticides comes in different forms having specific properties Lipid-containing nanopesticides can increase longevity and durability. Metal-organic nanopesticides frameworks have the ability to slowly release pesticides due to their meticulously arranged pore structure. Nanopesticides are designed by encapsulating the active ingredient and adding ligands specific to pest receptors, enhancing precision and minimising risk of affecting non target organisms

29. Advantages of Nanopesticides Increased solubility in water Penetration of cuticle in insects More environmental friendly then convenional pesticides. Host specific Improved stability due to encapsulation Slow releasing An et al. 2022 Fig 13 - Use of nanopesticides

30. Use of Nanopesticides in Agriculture Nanopesticide Active Ingredient Type Target pest use Advantage Refrence Nano-encapsulated neem Azadirachtin (neem extract) Insecticide Aphids, whiteflies and caterpillars Controlled release, prolonged activity Pascoli et al. (2019) Nano copper (MV-Cu) (FQ-Cu) Copper nanoparticles Bactericide P. syringae Angular leaf spot of Cucurbits Ebrahim et al. (2018) Nano-pyrethroids Pyrethroid nano particle Insecticide M osquitoes, Reduced toxicity, targeted pest contro l Kah et al . ( 2013 )

31. Challenges of Nanoparticles Nanoparticles can enter the human body through the lung, intestinal tract, or skin and have the potential to be harmful to the brain, trigger lung inflammation , and lead to cardiac issues . Silver nanoparticles (AgNP) can contaminate water bodies as they can lead to toxicity in marine creatures like fish and algae.. TiO 2 NPs cause DNA Damage . Zn-NP have the ability to cause harm to plant cells, resulting in hindered growth , decreased photosynthesis, and changed gene expression in plants. The toxicity of nanoparticles depends upon the characteristics of the particles such as concentration, pH, size, and shape , all of which impact the reactivity of the nanoparticles . Solutions Using green approach for synthesis of nanoparticles. Surface coating of nanoparticles to decreases its bioavailability by (PEG,PVP and Natural membranes). Controlled exposure and proper handling.

32. Case Studies

33. Case study 1 Nanofertilizers IFFCO conducted 730 field demonstrations in various districts of Uttar Pradesh , demonstrating that using Nano-N can cut the amount of urea applied by farmers in half to supply nitrogen to their crops. The crop yields achieved with various treatments were anlayzed with respect farmers typical fertilizer practice (FFP). Applying 2 sprays of Nano-N on standing crops resulted in higher yields compared to FFP in the majority of the crops evaluated in this study. In addition to this, the impact of Nano-Zn and Nano-Cu was also assessed. Kumar et al. (2020)

34. A total of 730 demonstrations on 12 crops resulted in a successful harvest. The planting of the crops took place in November and December of 2019 using 5 treatments , as outlined in Table. Nanofertilizers namely, Nano-N , Nano-Zn and Nano-Cu had nutrient concentrations of 25000 , 5000 and 2000 mg L -1 , respectively. 4 mL of these liquid fertilizers were added in 1L of water and for one acre 500 mL of nanofertilizers were added to 125 L of water and sprayed as per treatments detailed in Table The initial spraying took place after complete germination. Second application was applied after 10-15 days following the initial spray or 5 weeks after complete germination. Crops were harvested at full maturity, and yield data was collected based on net plot area harvested. Fig- Treatments and Nanofertilzers used Kumar et al. (2020)

35. Mean effects of nanofertilizers on grain yield of Wheat (out of the 12 crops) Grain yield and economic yield T2 [(FFP-50% N) + 2 sprays of of Nano-N] was the highest (4,779 kg ha -1 ). with additional increase of 425 kg ha -1 over FFP, giving 9.76% increase. The economic return over FFP was also highest under T2 of Rs. 8,182 per ha -1 ). Conclusion - The yield and economic return was increased along with 50 % reduction of Conventional Nitrogen fertilizer with usage Treatment 2 Fig 14- Grain Yield and Economic Return Kumar et al. (2020) Kumar et al. (2020)

36. Case study 2 Nanoparticles as Biosensors Safarnejad et al (2017) developed (QD) Quantum dot based nanosensors to detect Tristeza Disease in Citrus. The most prevalent techniques employed for detecting CTV involve enzyme-linked immunosorbent test (ELISA) and polymerase chain reaction (PCR). Above mentioned methods have limitations such as lengthy and demanding of time and unable to identify minimal levels of pathogens which is essential for identifying the illness in the initial phases of the infection QDs were used as a biosensor due to its fluorosence, brightness, stability, size-tunable light emission, and resistance to photo-bleaching Safarnejad et al. (2017)

37. How does Nanosensors work ? A nanosensor is a sensing instrument that has at least one dimension measuring 100 nm , utilized to collect information at the nanoscale and transform it into analyzable data . Main Subunits of a Nanosensors Receptor- Intercepts the incoming signal/analyte Transducer- Converts the incoming signal into information Analyte- substance to be indentified Output Device- Gives graphical or a simple visual indication 2 types of QD based nanosensors were synthesized and compaired ( Cd-Te based) C admium Telluride Quantum Dot particle in the following case.

38. The FRET based Nanobiosensor ( Contains Ab - QD and CP - Rd ) (Antibodies and Coat protein were synthesized by injecting Antigens in New Zealand white rabbit) In this case Cd-Te QD acts as a Transducer & Donor Fluorophore Antibody IgA acts as a Bio receptor Analyte is coat protein Rhodamine 123 is used as a fluoroscent molecule Flurometer- Used as Input device Working- The QD were conjugated with antibodies and Complex of protein coated- Rhodopsin then excited by a light with excitation of light at 390 nm wavelength value of emission obtained is 522 nm in QDs and due to FRET absorption is observecd at QD with emission of 580 nm in Rhodamine. FRET (Förster Resonance Energy Transfer) - It is the phenomenon in which energy is transfered from Donor Fluorophore (QD) to Acceptor Fluorophore (Rd). Ab - QD - Antibody conjugated to Quantom Dot CP - Rd - Coat protein conjugated to Rhodamine Fig 15- FRET based Nanobiosensor of Ab- QD and CP-Rd Safarnejad et al. (2017)

39. Working of FRET based Quantum Dots Ab-QD = > 3500 AU and 522 nm (Base line) Ab-QD + CP-Rd = 1500 AU and 580 nm Light is emitted by Ab -QD is 522nm when excited at 390 nm singularly. but when CP - Rd is also conjugated it causes a Quenching effect which decreases the emission intensity from 3500 AU to 1500 AU and Increase in the wave light emission from 522 nm to 580 nm is caused due to FRET. Safarnejad et al. (2017) 3500 AU 1500 AU

40. The FRET based Nanobiosensor Graphical Representation In the initial method for identifying CTV , the FRET- based nanobiosensor was employed. The rise in wave light emitted from 390 nm to 522nm and t he Emission Intensity decreased from 5700 AU to 2200 AU is attributed to the separation of the CP-Rd from the Ab-QD in reaction and when antigens (CP) gets bind at receptor increase in intensity is observed. The excitation occurs through a light with 390 nm wavelength and peak emission value acquired at 522 nm . The emission intensity of the Ab-QDs by themselves was assessed at approximately 5700 AU ; nonetheless, following the sepearation of Rd-CP and addition of CP value exhibited incline at 4900 AU. 5700 AU Increase in light intensity due separation of CP-Rd and binding of CP Safarnejad et al. (2017) Safarnejad et al. (2017) Fig 16- FRET-based mechanism for detection of infected CTV plant Decreased in light intensity

41. The Non FRET based Nanobiosensor Graphical Representation In the second method for identifying CTV , the Non FRET- based nanobiosensor was employed. The rise in wave light emitted from 390 nm to 522nm and t he Emission Intensity decreased. In this context, the identification relied primarily on the clustering of Ab-QD particles through the inclusion of antigens (CP) The excitation occurs through a light with 390 nm wavelength and peak emission value acquired at 522 nm of the Ab-QDs The non-FRET method relies on antigen-induced aggregation of QD-conjugates, resulting in increased fluorescence. 5700 AU 2200 AU Safarnejad et al. (2017) Safarnejad et al. (2017) Fig 17- Non FRET-based mechanism for detection of infected CTV plant

42. Conclusion- Both the FRET based and Non FRET Based Nano Biosenoser was able to detect the Citrus Tristeza Virus (CTV). The comparative analysis of two above described approaches showed that the non-FRET based method is more sensitive for detection of CTV-CP in the solution. The synthesized nanobiosensor d emonstrated full accuracy in identifying the infected samples containing the CTV particles .

43. Conclusion Th is document explores nanotechnology's foundational concepts, focusing on nanoparticle synthesis, classification, and their unique properties arising from nanoscale phenomena. Applications in agriculture include innovations such as nanofertilizers, nano-coatings for post-harvest preservation, and nanopesticides that improve crop yield and environmental safety. Challenges of nanoparticles, including potential toxicity and environmental impacts, are also discussed alongside sustainable solutions like green synthesis. Case studies highlight practical uses, such as enhanced crop yields with nanofertilizers and biosensors for early disease detection, emphasizing nanotechnology's transformative potential in agriculture

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