Compositions and methods for agricultural inputs and their use

Abstract

This specification discloses compositions comprising chitosan and at least 1% (w / v) nitrogen. In certain cases, at least 50% of the nitrogen is in the chemical form of amines and / or nitrates and / or ammonia. This specification also discloses a method for producing chitosan nanoparticles by dissolving chitosan in acidic water in which water has been acidified with an acid selected from the group consisting of acetic acid, nitric acid, sulfuric acid, hydrochloric acid, and combinations thereof, and forming nanoparticles from the dissolved chitosan. Furthermore, treatment of plants or plant parts with the compositions disclosed herein is disclosed.

Description

Technical Field

[0001] The present invention relates to plant nutrition, and more specifically, to the assembly, encapsulation, functionalization, capture, and coating of agricultural inputs such as nutrients.

Background Art

[0002] Due to climate change, extreme weather, soil erosion, pathogen resistance, and environmental pollution, agricultural production worldwide is facing unprecedented challenges. The world's population is projected to reach approximately 10 billion by 2050, and food demand is expected to increase by 60%. The rate of increase in yields and the fertilizer (agricultural input) - to - yield ratio achieved by the Green Revolution has been continuously decreasing, and harmful climate change is expected to further limit crop productivity and nutritional value. To increase yields and improve nutritional value, plants require balanced nutrition in addition to water and sunlight. However, conventional agricultural inputs and their supply are very inefficient due to low use efficiency and environmental emissions. Therefore, there is a need for more effective formulations and compositions that can stimulate plant growth, mitigate the adverse effects of abiotic / biotic stresses, efficiently deliver active ingredients to plants in a timely and appropriate amount, and be released in response to signals emitted by plants.

[0003] Biomass - based renewable materials are being studied worldwide due to their importance for a circular green economy and sustainability. Chitin is the most abundant natural amino polysaccharide. Chitin is most abundantly present in crustaceans, insects, and fungi. Chitin and its derivatives are considered new functional biomaterials with high potential in interdisciplinary applications. For example, chitosan, a deacetylated derivative of chitin, is used in industries / sectors such as pharmaceuticals, textiles, food, energy, and water. So far, the translational applications of chitosan in agriculture have not been fully utilized.

Summary of the Invention

[0004] In a first embodiment, the disclosure provides a method for modifying chitosan for the formulation of chitosan-based nanoparticles.

[0005] In a second aspect, the present disclosure provides chitosan-based nanoparticles and methods for producing them. Chitosan-based nanoparticles may comprise nitrogen and / or nitrogen-rich compounds and / or mixtures of compounds, such as, but not limited to, amino acids including, potassium nitrate, calcium nitrate, magnesium nitrate, urea nitrate, ammonium urea nitrate, ammonium nitrate, ammonium hydroxide, ammonium sulfate, urea hydroxyapatite, arginine, cystine, histidine, leucine, lysine, methionine, threonine, tryptophan, tyrosine, valine, glutamine, glutamic acid, glycine, proline, taurine, aspartic acid, serine, and alanine.

[0006] In a third aspect, the disclosure provides a method for treating a plant or plant part. The method for treating a plant or plant part includes application with a composition comprising chitosan nanoparticles. The chitosan nanoparticles may be present in a solution, suspension, or formulation having a physical form that may be solid, liquid, or aerosol, and may be applied to any plant part of the plant, including seeds, leaves, stems, flowers, fruits, or roots, throughout the plant's life cycle. The method may include replacing a certain amount of conventional nitrogen fertilizer with a solution of chitosan nanoparticles.

[0007] In a fourth embodiment, chitosan nanoparticles may be used as part of a method for preparing a plant growth medium. For example, this method may involve treating the growth medium with a chitosan nanoparticle solution. The growth medium may be soil, peat, moss, woody residue, leaf mold, sawdust, bark, bagasse, rice hulls, sand, perlite, vermiculite, calcined clay, polystyrene, urea-formaldehyde resin, agar or agarose, and hydroponic solutions, in vitro and in vivo plant tissue culture media used for hybrid, genetically modified, or non-genetically modified seeds / plant varieties, as well as combinations thereof.

[0008] Further aspects and embodiments are provided in the drawings, detailed description, and claims described above.

[0009] The following drawings are provided to illustrate specific embodiments described herein. The drawings are illustrative only and are not intended to limit the scope of the claimed invention, nor are they intended to show all potential features or embodiments of the claimed invention. The drawings are not necessarily drawn to a fixed scale, and in some cases, for illustrative purposes, certain elements of the drawings may be enlarged relative to other elements of the drawings. [Brief explanation of the drawing]

[0010] [Figure 1] This is a diagram showing the morphological characteristics of liquid chitosan preparations (TgA-TgE) obtained by electron microscopy. [Figure 2] This is a diagram showing the morphological characteristics of liquid chitosan preparations (CTgA~CTgE) obtained by electron microscopy. [Figure 3] This figure shows the colloidal characterization of liquid chitosan formulations (TgA~E and CTgA~CTgE) obtained by dynamic light scattering, with reference to their hydrodynamic sizes. [Figure 4] This figure shows the colloidal characterization of liquid chitosan formulations (TgA~E and CTgA~CTgE) obtained by dynamic light scattering, with reference to their surface zeta potentials. [Figure 5] This table shows the pH and TKN of chitosan-based nanoparticle batches 1-7. [Figure 6] This is a UV wavelength analysis of chitosan-based nanoparticle batches 1-7. [Figure 7] This is an FTIR analysis of chitosan-based nanoparticles 1. [Figure 8] This is an FTIR analysis of chitosan-based nanoparticles 2. [Figure 9] This is an FTIR analysis of chitosan-based nanoparticles 3. [Figure 10] This is an FTIR analysis of chitosan-based nanoparticles 4. [Figure 11] This is an FTIR analysis of chitosan-based nanoparticles 5. [Figure 12] This is an FTIR analysis of chitosan-based nanoparticles 6. [Figure 13] This is an FTIR analysis of chitosan-based nanoparticles 7. [Figure 14] This graph shows the plant toxicity of chitosan-based nanoparticle formulations used in hydroponic systems. [Figure 15] This graph shows the vitality evaluation after application of a chitosan-based nanoparticle formulation used in hydroponic systems. [Figure 16] This graph shows the change in plant width after application of chitosan-based nanoparticle formulations used in hydroponic systems. [Figure 17] This graph shows the weight of shoots after application of a chitosan-based nanoparticle formulation used in hydroponic systems. [Figure 18] This graph shows the root weight after application of a chitosan-based nanoparticle formulation used in hydroponic systems. [Figure 19A] This graph shows the total harvest weight of corn after applying multiple different formulations as foliar treatment to different plots of corn. [Figure 19B] This graph shows the percentage of nitrogen present in leaf tissue samples at the V8 stage of corn growth. [Figure 20] This graph shows the effects of chitosan nanoparticle formulations when added to corn planted with nitrogen fertilizer according to standard farming methods, and when added to corn planted with nitrogen fertilizer according to standard farming methods to a soil nitrogen application of less than 40 pounds (18.14 kg). [Figure 21] This graph shows the effects of chitosan-based nanoparticle formulations containing glyphosate. [Figure 22] This graph shows the effects of chitosan-based nanoparticle formulations containing 2,4-D. [Modes for carrying out the invention]

[0011] The following description lists various aspects and embodiments of the invention disclosed herein. The specific embodiments are not intended to define the scope of the invention. Rather, the embodiments provide non-limiting examples of various compositions and methods within the scope of the claimed invention. This description should be read from the perspective of one of ordinary skill in the art. Thus, information well known to one of ordinary skill in the art is not necessarily included.

[0012] Definitions The following terms and phrases have the meanings set forth below unless otherwise defined herein. The present disclosure may employ other terms and phrases not explicitly defined herein. Such other terms and phrases shall have the meanings that would be understood by one of ordinary skill in the art within the context of the present disclosure. In some cases, a term or phrase may be defined in either the singular or plural form. In such cases, it is understood that the singular form may include the plural and vice versa, unless otherwise explicitly stated to the contrary.

[0013] As used herein, the singular forms "a," "an," and "the" include plural referents unless the context clearly dictates otherwise. For example, reference to "a substituent" includes a single substituent as well as two or more substituents.

[0014] As used herein, "for example," "for instance," "such as," or "including" means introducing examples to clarify a more general subject. Unless otherwise specifically stated, such examples are provided only as an aid to understanding the embodiments illustrated in the present disclosure and are not intended to limit in any way. Also, these phrases do not imply any kind of preference for the disclosed embodiments.

[0015] As used herein, “chitosan-based nanoparticles” means chitosan-containing particles in which nutrient-containing molecules are conjugated, bound, functionalized, encapsulated, or trapped within a chitosan polymer matrix via ionic, nonionic, non-covalent, or covalent intermolecular or intramolecular bonds. The nutrient in the nutrient-containing molecule may be nitrogen. The nutrient may be any primary macronutrient, secondary macronutrient, or micronutrient. Such nutrients include phosphorus (P), potassium (K), calcium (Ca), magnesium (Mg), sulfur (S), iron (Fe), zinc (Zn), copper (Cu), manganese (Mn), molybdenum (Mo), nickel (Ni), and boron (B), as well as combinations thereof.

[0016] Several formulations for chitosan-based nanoparticles and chitosan-based nanoparticle solutions are manufactured under the trademark alignN®.

[0017] As used herein, “associated with or linked to” means molecules that are bonded, conjugated, functionalized, captured, coated, encapsulated, or otherwise joined together. Such joining may be achieved by ionic, nonionic, noncovalent, or covalent bonds.

[0018] As used herein, “conventional nitrogen fertilizer” means any substance added to the soil or directly to plants that increases the availability of nitrogen to the plants. These fertilizers may be granular or liquid. Granular fertilizers are pellets that can be scattered around plants or spread over an entire field. Liquid fertilizers can be absorbed by plants when applied through an irrigation system or sprayed directly onto the leaves. Such fertilizers contain nitrogen in one of several forms, including ammonia, ammonium, urea, and nitrates.

[0019] As used herein, “plant toxicity” means any adverse effect on plant growth, physiology, or metabolism caused by a chemical substance, such as increased levels of fertilizers, herbicides, heavy metals, or nanoparticles. Common plant toxicity effects include altered plant metabolism, growth inhibition, or plant death. Altered plant metabolism and growth are the result of interference with physiological functions, such as photosynthesis, water and nutrient uptake, cell division, or seed germination.

[0020] Nutrient association and sustained release is one way to balance the nutritional requirements of plants. One such requirement is nitrogen in the form of amines, nitrates, ammonium, and ammonia. For example, the association of nitrogen compounds having chemical functional groups as amines, nitrates, ammonia, ammonium, or any one or all of these, including but not limited to amino acids such as potassium nitrate, calcium nitrate, magnesium nitrate, urea nitrate, ammonia urea nitrate, ammonium nitrate, ammonium hydroxide, ammonium sulfate, urea hydroxyapatite, arginine, cystine, histidine, leucine, lysine, methionine, threonine, tryptophan, tyrosine, valine, glutamic acid, glycine, proline, taurine, aspartic acid, serine, and alanine, as well as all possible combinations thereof, is a way to release an effective amount of nitrogen for plant growth. There are several advantages to using chitosan as a nitrogen association agent. The first advantage is that chitosan polymers are biologically safe and biodegradable. Chitosan polymers are bio-derived and have been used for immobilizing food enzymes and in biosensing applications. Chitosan can be extracted from fungi, shrimp, crabs, lobsters, or any combination of these chitosan sources. A second advantage compared to inorganic coatings such as sulfur coatings is that they do not crack. Cracking in coatings causes the coated nutrients to be released immediately upon contact with water. Immediate release of nutrients undermines the purpose of the coating, which is to release nutrients slowly. A third advantage of chitosan polymers is the overall health of the soil. Chitosan polymers contribute to reducing soil acidification. Soil acidification can adversely affect certain plant species. Coatings such as sulfur lower the soil pH. Furthermore, soil microorganisms are a crucial element of soil health. Soil microorganisms cannot metabolize synthetic polymers.

[0021] The embodiments include chitosan-based nanoparticles. In certain embodiments, the chitosan-based nanoparticles contain nitrogen in the form of amines and / or nitrates and / or ammonia and / or ammonium. Nitrogen can be associated, conjugated, attached, and linked by / within the chitosan-based nanoparticles. In certain embodiments, nitrogen is in the form of amines, nitrates, ammonium and / or ammonia, and may be selected potassium nitrate, calcium nitrate, magnesium nitrate, urea nitrate, ammonia urea nitrate, ammonium nitrate, ammonium hydroxide, ammonium sulfate, urea hydroxyapatite, amino acids such as arginine, cystine, histidine, leucine, lysine, methionine, threonine, tryptophan, tyrosine, valine, glutamine, glutamic acid, glycine, proline, taurine, aspartic acid, serine, alanine, and all combinations thereof.

[0022] In certain embodiments, chitosan-based nanoparticles may contain at least 1, 5, 10, 15, 20, 25, 30, 35, 40, 45, 50, 55, 60, 65, 70, 75, 85, 90, or 95% (w / v) of nitrogen. In certain embodiments, at least 50, 55, 60, 65, 70, 75, 85, 90, or 95% of the nitrogen is in the chemical form of amines and / or nitrates, ammonium, and / or ammonia. In certain embodiments, chitosan-based nanoparticles may contain about 19.87% (w / v) of nitrogen. In certain embodiments, chitosan-based nanoparticles may contain about 15.64% (w / v) of nitrogen.

[0023] In certain embodiments, chitosan-based nanoparticles may contain at least 1, 5, 10, 15, 20, 25, 30, 35, 40, 45, 50, 55, 60, 65, 70, 75, 85, 90, or 95% (w / w) of nitrogen. In certain embodiments, at least 50, 55, 60, 65, 70, 75, 85, 90, or 95% of the nitrogen is in the chemical form of amines and / or nitrates and / or ammonia and / or ammonium. In certain embodiments, chitosan-based nanoparticles may contain about 16.15% (w / w) of nitrogen. In certain embodiments, chitosan-based nanoparticles may contain about 13.37% (w / w) of nitrogen.

[0024] In a particular embodiment, the chitosan-based nanoparticles may contain at least 5, 10, 15, 20, 25, 30, 35, or 40% (w / v) total nitrogen, at least 5, 10, 15, 20, 25, or 30% nitrogen derived from urea, and at least 1, 5, 10, 15, or 20% nitrogen derived from ammonium sulfate.

[0025] In a particular embodiment, the chitosan-based nanoparticles may contain: chitosan = 0.3%, total nitrogen (w / v) = 15.64%, total ammonium sulfate (including linker) = 19.06%, N in urea = 11.65%, N in AS = 3.99%, S in AS = 4.62%, sulfate = 13.84%, and potassium sorbate: 0.01%.

[0026] In a particular embodiment, the chitosan-based nanoparticles may contain at least 5, 10, 15, 20, 25, 30, 35, or 40% (w / v) total nitrogen, at least 5, 10, 15, 20, 25, or 30% nitrogen derived from urea, and at least 1, 5, 10, 15, or 20% nitrogen derived from ammonium sulfate.

[0027] In a particular embodiment, the chitosan-based nanoparticles may contain: chitosan = 0.2%, total nitrogen (w / v) = 19.87%, total ammonium sulfate (including linker) = 37.7%, N in urea = 11.96%, N in AS = 7.91%, S in AS = 9.1%, sulfate = 27.4%, and potassium sorbate: 0.01%.

[0028] In one embodiment, the process for forming chitosan-based nanoparticles begins with a chitosan polymer. The source of the chitosan can be fungi, shrimp, crab, lobster, or squid bone, or a mixture thereof. In certain embodiments, the chitosan has a degree of deacetylation (DDA): >80% and MW: <110 kDa. In certain embodiments, the DDA is >85%. The DDA may be 90%. The DDA may be >95%. In certain embodiments, DDA is approximately 60%, 61%, 62%, 63%, 64%, 65%, 66%, 67%, 68%, 69%, 70%, 71%, 72%, 73%, 74%, 75%, 76%, 77%, 78%, 79%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100%.

[0029] Before use in the formulation of nanoparticles, chitosan must be dissolved in solution. Dissolving medium: Acidic water with a pH in the range of 1 to 6.5, or with a pH of <1, <2, <3, <4, <5, <5.5. The acid used may be any acid. In certain embodiments, the acids used are combinations of acids containing one or more of the following: acetic acid, citric acid, lactic acid, malic acid, tartaric acid, formic acid, acetylsalicylic acid, oxalic acid, succinic acid, benzoic acid, folic acid, pyruvic acid, butyric acid, propionic acid, caproic acid, hydrochloric acid, sulfuric acid, nitric acid, phosphoric acid, hydrofluoric acid, perchloric acid, hydrobromic acid, hydroiodic acid, chloric acid, bromic acid, iodic acid, humic acid, fulvic acid, amino acids, nucleic acids (DNA and RNA), boric acid, chromic acid, cyanuric acid, hyaluronic acid, arsenic acid, and carboxylic acids, as well as hydrogen peroxide and carbon dioxide. To dissolve the chitosan, the acidic aqueous solution may be kept at a temperature of approximately 68 to 212 degrees Fahrenheit. In certain embodiments, the temperature is kept at approximately 68, 72, 82, 92, 201, 112, 122, 132, 142, 152, 162, 172, 182, 192, 202, or 212 degrees Fahrenheit. Throughout the entire reaction period, the solution may be stirred at rpm between approximately 50 and 2500 rpm. After the completion of the reaction (typically 0.3 to 4 hours), the solution was filtered through a 200-mesh (74 micron) mesh followed by a mesh of over 635 (<20 micron). An antimicrobial agent, such as potassium sorbate but not limited to it (0.1% v / v), is added to the filtered solution. In certain cases, a coupling enhancer, such as sodium tripolyphosphate and / or ammonium sulfate, is added to the solution before the addition of the antimicrobial agent, as described below.

[0030] In one embodiment, the reaction is carried out at a temperature of 68–250 degrees Fahrenheit in a specific solution prepared according to one and / or all of the procedures of Example 1. Throughout the entire reaction period, the solution was stirred at 400 rpm to homogenize the substances. After the completion of the reaction (typically 0.3–4 hours), the solution was filtered through a 200 mesh (74 microns) followed by a mesh of over 635 (<20 microns). The nitrogen % in the final solution was maintained between 0.1% and 100%, specifically 18–24%, using the compounds and / or mixtures of compounds described above.

[0031] In certain embodiments, chitosan-based nanoparticles may be formed in the presence of nitrogen in the form of amines and / or nitrates and / or ammonia and / or ammonium, and / or may be loaded with such nitrogen. Nitrogen may be at least partially associated by / within the chitosan-based nanoparticles. In certain embodiments, nitrogen in the form of amines, nitrates, ammonium, and / or ammonia may be selected amino acids such as potassium nitrate, calcium nitrate, magnesium nitrate, urea nitrate, ammonia urea nitrate, ammonium nitrate, ammonium sulfate, ammonium hydroxide, urea hydroxyapatite, arginine, cystine, histidine, leucine, lysine, methionine, threonine, tryptophan, tyrosine, valine, glutamine, glutamic acid, glycine, proline, taurine, aspartic acid, serine, alanine, and combinations thereof.

[0032] >95% of the particles may be spherical, nearly spherical, or small linear fragments with sizes ranging from 0.1 nm to 8 microns (measured by TEM). Polydispersion indexes ranged from 0.01 to 0.99, with particle sizes from 0.1 nm to 10 microns (measured by DLS). Surface zeta potentials ranged from +5 to 70 mV (measured by DLS).

[0033] Embodiments include methods for treating plants or plant parts with any of the compositions described herein. Plant parts may be any plant part, including but not limited to seeds, leaves, stems, flowers, fruits, or roots. Examples of plant part seeds include, but are not limited to, grains such as wheat, corn or maize, barley, rye, and oats; canola, cotton, eggplant, lettuce, sorghum, soybeans, rice, rapeseed, sugar beet, sugarcane, grapes, lentils, sunflowers, alfalfa, citrus pome fruits; drupes; nuts, peanuts; coffee; tea; strawberries; grass; and vegetables such as tomatoes, potatoes, cucumbers, and lettuce.

[0034] Embodiments of methods for treating plants or plant parts may include adding chitosan nanoparticles in conjunction with the application of conventional agricultural inputs, or replacing all or part of conventional agricultural inputs. Replacement of agricultural inputs with chitosan nanoparticles may replace 0% to 100% of the total amount of agricultural inputs with the chitosan nanoparticles described herein. In some embodiments, the percentage of agricultural input replaced is about 0, 1, 5, 10, 15, 20, 25, 30, 35, 40, 45, 50, 55, 60, 65, 70, 75, 80, 85, 90, 95, or 100. In some embodiments, the amount of nitrogen replaced is 25%. In some embodiments, the agricultural input replaced is nitrogen. In some embodiments, the percentage of nitrogen replaced is approximately 0, 1, 5, 10, 15, 20, 25, 30, 35, 40, 45, 50, 55, 60, 65, 70, 75, 80, 85, 90, 95, or 100. In some embodiments, the total nitrogen applied was reduced by approximately 25% by the addition of the chitosan nanoparticle formulation at statistically equal yields. In other words, the total amount of nitrogen used with low nitrogen and chitosan nanoparticles was 75% of the nitrogen used with high nitrogen at the same yield. Replacing other agricultural inputs with chitosan nanoparticles begins with removing a certain percentage of the other agricultural inputs. In some embodiments, the percentage of agricultural input removed is about 0, 1, 5, 10, 15, 20, 25, 30, 35, 40, 45, 50, 55, 60, 65, 70, 75, 80, 85, 90, 95, or 100 percent. In some embodiments, the agricultural input removed is nitrogen. In some embodiments, the percentage of nitrogen removed is about 0, 1, 5, 10, 15, 20, 25, 30, 35, 40, 45, 50, 55, 60, 65, 70, 75, 80, 85, 90, 95, or 100 percent. In addition to the removal of agricultural inputs such as nitrogen, the replacement of agricultural inputs such as nitrogen requires that a certain amount of nitrogen in chitosan nanoparticles be added to or applied to plants or plant parts.In some embodiments, the proportion of nitrogen in the added chitosan nanoparticles is about 0, 1, 2, 3, 4, 5, 5.5, 10, 15, 20, 25, 30, 35, 40, 45, or 50 of the amount of agricultural inputs such as nitrogen removed. In some embodiments, the proportion of added chitosan nanoparticles is about 5.5 percent of the amount of nitrogen removed.

[0035] The replacement of agricultural inputs with chitosan nanoparticles can replace 1% to 100% of the total amount of agricultural inputs with chitosan nanoparticles. In some embodiments, the agricultural inputs are added to a hydroponic system. In some embodiments, the percentage of agricultural inputs replaced is about 1, 5, 10, 15, 20, 25, 30, 35, 40, 45, 50, 55, 60, 65, 70, 75, 80, 85, 90, 95, or 100. In some embodiments, the amount of agricultural input replaced is 70%. In some embodiments, the agricultural input replaced is nitrogen. In some embodiments, the percentage of nitrogen replaced is about 1, 5, 10, 15, 20, 25, 30, 35, 40, 45, 50, 55, 60, 65, 70, 75, 80, 85, 90, 95, or 100. In some embodiments, the total nitrogen applied was reduced by approximately 70% with the addition of chitosan nanoparticle formulations at statistically equivalent biomass yields. In other words, the total amount of nitrogen used with low-nitrogen and chitosan nanoparticle formulations was 30% of the nitrogen used with high-nitrogen formulations yielding the same yield. Replacing other agricultural inputs with chitosan nanoparticles begins with removing a certain percentage of the other agricultural inputs. In some embodiments, the percentage of agricultural inputs removed is approximately 0, 1, 5, 10, 15, 20, 25, 30, 35, 40, 45, 50, 55, 60, 65, 70, 75, 80, 85, 90, 95, or 100 percent. In some embodiments, the agricultural input removed is nitrogen. In some embodiments, the percentage of nitrogen removed is about 0, 1, 5, 10, 15, 20, 25, 30, 35, 40, 45, 50, 55, 60, 65, 70, 75, 80, 85, 90, 95, or 100 percent. In some embodiments, the amount of nitrogen removed is 70 percent. In addition to the removal of agricultural inputs such as nitrogen, the replacement of agricultural inputs such as nitrogen requires that a certain amount of nitrogen in chitosan nanoparticles be added to or applied to the plant or plant part.In some embodiments, the proportion of nitrogen in the added chitosan nanoparticles is about 0, 1, 2, 3, 4, 5, 5.5, 10, 15, 20, 25, 30, 35, 40, 45, or 50 of the amount of agricultural inputs such as nitrogen removed. In some embodiments, the proportion of added chitosan nanoparticles is about 8.4 percent of the amount of nitrogen removed.

[0036] As described herein, the replacement of agricultural inputs such as nitrogen with chitosan nanoparticles replaces a larger amount of agricultural input such as nitrogen with a smaller amount of chitosan nanoparticles. Typically, in a field, 0 to 200 kg of nitrogen is applied to a cornfield. The reduction in nitrogen can be about 15 to 25 percent. In some embodiments, for example, 88.5 kg / acre is reduced by 18.14 kg for a 20% reduction. In a particular embodiment, for example, a cornfield conventionally fertilized by improving the soil with 68.04 kg / acre of nitrogen can be replaced by improving the soil with 49.90 kg / acre of nitrogen and spraying the plants with chitosan nanoparticles at a rate of 5 l / acre. This results in a 27% reduction in the total amount of nitrogen applied to the plants. In this embodiment, plants that have been sprayed with chitosan nanoparticles grow at the same rate as or better than plants that have received the full amount of nitrogen fertilizer.

[0037] In the second embodiment, the chitosan nanoparticle solution is used in a hydroponic system. Nutrient additives are added to the hydroponic solution. In a high-nitrogen environment, nitrogen is added to the hydroponic solution so that each plant receives an average of 105.50 mg of N, while in a low-nitrogen environment, plants receive an average of 24.99 mg of nitrogen per plant. In a low-nitrogen environment, plants also receive 6.73 + / - 0.1 mg of nitrogen per plant from the chitosan nanoparticles via foliar application. This represents a 76% removal of nitrogen in the hydroponics system, where 8.4% of the removed nitrogen is replaced by the chitosan nanoparticles via foliar application.

[0038] In certain embodiments, chitosan-based nanoparticles are used in conjunction with conventional nitrogen fertilizers. The chitosan-based nanoparticles are added on top of or together with the conventional nitrogen fertilizer. In one example, the addition of the chitosan-based nanoparticle composition increased the yield by 7%.

[0039] Several types of nitrogen agricultural inputs are used, and chitosan nanoparticles can replace any of them. Replacing all or part of agricultural inputs such as nitrogen with the composition, or chitosan nanoparticles, results in yields that are statistically equivalent to or greater than those produced with conventional agricultural inputs such as nitrogen. The composition, or chitosan nanoparticles, may be used in any type of growing environment. Growing environments include conventional agriculture where crops are grown in the ground, hydroponic systems, greenhouses, environmentally controlled agriculture (CEA), or aquaponic systems. In some embodiments, the composition, or chitosan nanoparticles...

[0040] Chitosan nanoparticles may be applied to plants in any way used to administer agricultural inputs such as nitrogen. Methods of application to crops include soil application, spraying (e.g., foliar application), and water infusion. Soil application is carried out by mixing chitosan nanoparticles into the soil using one or a combination of methods including spraying, top dressing, slow-release fertilization, drill application, lateral application, strip application, and pellet application. Foliar application involves scattering chitosan nanoparticles onto the leaves of growing plants. Some nutrients are readily absorbed by the leaves, especially when dissolved in water. Foliar application can be carried out via aerial application from handheld devices, backpack devices, sprayers, applicators, airplanes, or unmanned aerial vehicles (UAVs). Chitosan nanoparticles may also be applied through water infusion. This may be done via irrigation systems, drip irrigation systems, or sprinkler systems. Methods of application for hydroponic and / or aeroponic and / or geoponic systems include injection into liquid and / or gel-like growing media, or foliar application by spraying or misting systems.

[0041] Chitosan nanoparticles may be applied at any stage of soil preparation or plant growth. In some embodiments, chitosan nanoparticles may be applied at the germination stage. In some embodiments, chitosan nanoparticles may be applied to plant parts at any point during the vegetative growth stage of plant growth. In some embodiments, chitosan nanoparticles may be applied to plant parts during the reproductive stage of plant growth. In some embodiments, chitosan nanoparticles may be applied for seed coating. In some embodiments, chitosan nanoparticles are applied to the soil before planting. In some embodiments, chitosan nanoparticles are applied to the soil during the growth stage. In some embodiments, chitosan nanoparticles are applied to the soil during the maturation stage. In some embodiments, chitosan nanoparticles are applied to the soil during the dormancy stage. In some embodiments, chitosan nanoparticles are applied in any combination of those described above. In one embodiment, chitosan nanoparticles are applied at at least two different time points. In one embodiment, chitosan nanoparticles are applied at at least three different time points. In one embodiment, the chitosan nanoparticles are applied at least four different time points. In one embodiment, the chitosan nanoparticles are applied at least five different time points. In one embodiment, the chitosan nanoparticles are applied at least six different time points. Further embodiments include the use of the chitosan nanoparticles in combination with other agricultural inputs. Such agricultural inputs include primary macronutrients such as nitrogen, phosphorus, and potassium; secondary macronutrients such as calcium, magnesium, and sulfur; micronutrients such as iron, zinc, copper, manganese, and boron; herbicides; insecticides; fungicides; nematicides; plant growth regulators (PGRs) such as auxins, cytokinins, gibberellins, and ethylene; and seed treatments.

[0042] Further embodiments include methods for treating a plant growth medium with any of the compositions described herein. The plant growth medium may comprise one or more of the following: soil, peat, moss, woody residue, humus, sawdust, bark, bagasse, rice hulls, sand, perlite, vermiculite, calcined clay, polystyrene, urea-formaldehyde resin, agar, or agarose, in vitro and in vivo plant tissue culture / growth media, as well as hydroponic solutions, and substances used in aeroponic and geoponic systems. Methods for treating the plant growth medium may comprise soil conditioners, additions to solid media, or additions to liquid media as found in hydroponic systems.

[0043] The first embodiment provides a composition comprising chitosan and at least 1% (w / w) of nitrogen.

[0044] In the second embodiment, the composition of Embodiment 1 has about 11% (w / w) to about 25% (w / w) of nitrogen.

[0045] In a third embodiment, the composition of Embodiment 1 contains at least 50% of the nitrogen in the chemical form of an amine and / or nitrate and / or ammonia.

[0046] In a fourth embodiment, the composition of Embodiment 3 comprises chitosan nanoparticles and a nitrogen-rich compound and / or mixture of compounds selected from the group consisting of amino acids such as potassium nitrate, calcium nitrate, magnesium nitrate, urea nitrate, ammonium urea nitrate, ammonium nitrate, ammonium hydroxide, ammonium sulfate, urea hydroxyapatite, arginine, cystine, histidine, leucine, lysine, methionine, threonine, tryptophan, tyrosine, valine, glutamine, glutamic acid, glycine, proline, taurine, aspartic acid, serine, and alanine.

[0047] In the fifth embodiment, the composition of Embodiment 1 contains chitosan present in an amount of about 0.01% to about 4% (w / v).

[0048] In the sixth embodiment, the composition of Embodiment 1 and the coupling enhancer are added.

[0049] The composition according to Embodiment 6, wherein in the seventh embodiment, the coupling enhancer is sodium tripolyphosphate and / or ammonium sulfate.

[0050] The eighth embodiment is a method for producing chitosan nanoparticles. This method involves dissolving chitosan in acidic water, where the water is acidified with an acid selected from the group of acetic acid, nitric acid, sulfuric acid, hydrochloric acid, and combinations thereof. Nanoparticles are formed from the dissolved chitosan.

[0051] The method according to Embodiment 8, in the ninth embodiment, the coupling enhancer is added to the dissolved chitosan.

[0052] The method according to Embodiment 9, wherein in the tenth embodiment, the coupling enhancer is sodium tripolyphosphate and / or ammonium sulfate.

[0053] The method according to Embodiment 8, in an eleventh embodiment, a nitrogen-rich compound and / or mixture of compounds is selected from the group consisting of amino acids such as potassium nitrate, calcium nitrate, magnesium nitrate, urea nitrate, ammonium urea nitrate, ammonium nitrate, ammonium sulfate, urea hydroxyapatite, arginine, cystine, histidine, leucine, lysine, methionine, threonine, tryptophan, tyrosine, valine, glutamine, glutamic acid, glycine, proline, taurine, aspartic acid, serine, and alanine, and added to dissolved chitosan.

[0054] The method according to Embodiment 11, wherein in the twelfth embodiment, the nanoparticles comprise chitosan and a nitrogen-rich compound and / or a mixture of compounds.

[0055] The method according to Embodiment 12, in a thirteenth embodiment, wherein the nanoparticles contain at least 1% (w / w) nitrogen.

[0056] The method according to Embodiment 13, in a 14th embodiment, wherein nitrogen is present at a concentration of approximately 11% (w / w) to approximately 25% (w / w).

[0057] The method according to Embodiment 11, in a 15th embodiment, in which chitosan is present in an amount of about 0.01% to about 4% (w / v).

[0058] The sixteenth embodiment is a method for treating a plant or plant part, the method comprising treating the plant or plant part with a composition described in any one of embodiments 1 to 7.

[0059] The method according to embodiment 16, wherein in the 17th embodiment, the plant part is selected from the group consisting of plant seeds, leaves, stems, flowers, or roots.

[0060] In the 18th embodiment, the method of Embodiment 17 is carried out, wherein a predetermined amount of the composition according to any one of claims 1 to 7 is used in place of a predetermined amount of conventional fertilizer, resulting in an equivalent or better yield.

[0061] In the 19th embodiment, the method of Embodiment 18 is implemented, wherein a predetermined amount of the composition according to any one of claims 1 to 7 is about 250 mL to about 30 liters per acre applied by foliar spraying, and is used in place of about 0 kilograms to about 150 kilograms of nitrogen units per acre applied by conventional nitrogen fertilizers.

[0062] In the 20th embodiment, the method according to claim 19 is carried out, wherein a predetermined amount of the composition according to any one of claims 1 to 7 is about 5 liters per acre applied by foliar spray, and is used in place of about 18.14 kilograms of conventional nitrogen fertilizer per acre.

[0063] The 21st embodiment is a method for preparing a plant growth medium, comprising treating the growth medium with a composition described in any one of Embodiments 1 to 7.

[0064] The method according to Embodiment 18, in a 22nd embodiment, the growth medium is soil, peat, moss, woody residue, leaf mold, sawdust, bark, bagasse, rice hulls, sand, perlite, vermiculite, calcined clay, polystyrene, urea-formaldehyde resin, and hydroponic solution, as well as combinations thereof.

[0065] The method according to Embodiment 18, in the 23rd embodiment, wherein the proportion of the composition used is about 4 percent to about 12 percent of the conventional nitrogen fertilizer to be removed.

[0066] The method according to Embodiment 25, in the 24th embodiment, wherein the proportion of the composition used is about 8.4 percent of the conventional nitrogen fertilizer to be removed.

[0067] The method according to Embodiment 18, in the 25th embodiment, wherein the total nitrogen used is reduced by about 0 percent to about 80 percent.

[0068] The method according to embodiment 25, wherein the total nitrogen used is reduced by about 70 percent in the 26th embodiment.

[0069] The method according to claim 25, wherein, in the 27th embodiment, the total nitrogen used is reduced by about 76 percent.

[0070] In the 28th embodiment, chitosan-based nanoparticles are added together with a standard amount of conventional fertilizer.

[0071] In the 29th embodiment, the chitosan-based nanoparticles of embodiment 28 result in a yield improvement of approximately 0% to approximately 80%.

[0072] In the 30th embodiment, the chitosan-based nanoparticles of embodiment 29 result in an improvement in yield of approximately 7%.

[0073] Example 1 Preparation of chitosan using various acids [Acetic Acid] Sterile water or autoclaved water was filled into a tank with a stirring speed of 400 rpm. When the temperature reached 68-194 degrees Fahrenheit, chitosan flakes (0.1-5% w / v) with 80% but more than 75% DDA were added. The source of chitosan could be fungi, shrimp, crabs, or lobsters, or a mixture thereof. Furthermore, the chitosan variant could be α, β, or γ, or a mixture thereof. After immersing the flakes in water, 0.3-1.9% (v / v) acetic acid was added. The temperature and stirring were maintained throughout the reaction for up to 4 hours. Finally, when the flakes were no longer visible and the solution was formed, it was filtered through a 200-mesh filtration unit followed by a <635-mesh filtration unit. The solution temperature was cooled to 77 degrees Fahrenheit, and potassium sorbate solution (0.01-1% v / v) was added and mixed well with stirring.

[0074] [Acetic acid + NaTPP] Sterile water or autoclaved water was filled into a tank with a stirring speed of 400 rpm. When the temperature reached 68-194 degrees Fahrenheit, chitosan flakes (0.1-5% w / v) with 80% but more than 75% DDA were added. The source of chitosan could be fungi, shrimp, crabs, or lobsters, or a mixture thereof. Furthermore, the chitosan variant could be α, β, or γ, or a mixture thereof. After immersing the flakes in water, 0.3-1.9% (v / v) acetic acid was added. When the flakes were no longer visible and the mixture became a solution, sodium tripolyphosphate solution (0.001-1% v / v or v / w) was added. The temperature and stirring were maintained throughout the reaction for up to 4 hours. Finally, the solution was filtered through a 200-mesh filtration unit followed by a <635-mesh filtration unit. The solution was cooled to 77 degrees Fahrenheit, potassium sorbate solution (0.01-1% v / v) was added, and the mixture was thoroughly mixed using stirring.

[0075] [Acetic acid + ammonium sulfate] Sterile water or autoclaved water was filled into a tank with a stirring speed of 400 rpm. When the temperature reached 68-194 degrees Fahrenheit, chitosan flakes (0.1-5% w / v) with 80% but more than 75% DDA were added. The source of chitosan could be fungi, shrimp, crabs, or lobsters, or a mixture thereof. Furthermore, the chitosan variant could be α, β, or γ, or a mixture thereof. After immersing the flakes in water, 0.3-1.9% (v / v) acetic acid was added. When the flakes were no longer visible and the mixture became a solution, ammonium sulfate solution (0.001-1% v / v or v / w) was added. The temperature and stirring were maintained throughout the reaction for up to 4 hours. Finally, the solution was filtered through a 200-mesh filtration unit followed by a <635-mesh filtration unit. The solution was cooled to 77 degrees Fahrenheit, potassium sorbate solution (0.01-1% v / v) was added, and the mixture was thoroughly mixed using stirring.

[0076] [Nitric Acid + Acetic Acid] Sterile water or autoclaved water was filled into a tank with a stirring speed of 400 rpm. When the temperature reached 68-194 degrees Fahrenheit, chitosan flakes (0.1-5% w / v) with 80% DDA but more than 75% DDA were added. The source of chitosan could be fungi, shrimp, crabs, or lobsters, or a mixture thereof. Furthermore, the chitosan variant could be α, β, or γ, or a mixture thereof. After immersing the flakes in water, acetic acid and nitric acid were added in ratios of 1:1, 1:2, or 2:1, with a total v / v acid strength of 0.3%-1.9%. The temperature and stirring were maintained throughout the reaction for up to 4 hours. Finally, when the flakes were no longer visible and the solution was formed, it was filtered through a 200-mesh filtration unit followed by a <635-mesh filtration unit. The solution was cooled to 77 degrees Fahrenheit, potassium sorbate solution (1% v / v) was added, and the mixture was thoroughly mixed using stirring.

[0077] [Nitric acid + Acetic acid + NaTPP] Sterile water or autoclaved water was filled into a tank with a stirring speed of 400 rpm. When the temperature reached 68-194 degrees Fahrenheit, chitosan flakes (0.1-5% w / v) with 80% DDA but more than 75% DDA were added. The source of chitosan could be fungi, shrimp, crabs, or lobsters, or a mixture thereof. Furthermore, the chitosan variant could be α, β, or γ, or a mixture thereof. After immersing the flakes in water, acetic acid and nitric acid were added in ratios of 1:1, 1:2, or 2:1 with a total v / v acid strength of 0.3-1.9%. When the flakes were no longer visible and the solution was formed, sodium tripolyphosphate solution (0.001-1% v / v or v / w) was added. The temperature and stirring were maintained throughout the reaction for up to 4 hours. Finally, the solution was filtered through a 200-mesh filtration unit followed by a <635-mesh filtration unit. The solution temperature was cooled to 77 degrees Fahrenheit, potassium sorbate solution (0.01-1% v / v) was added, and the mixture was thoroughly mixed using stirring.

[0078] [Nitric acid + Acetic acid + Ammonium sulfate] Sterile water or autoclaved water was filled into a tank with a stirring speed of 400 rpm. When the temperature reached 68-194 degrees Fahrenheit, chitosan flakes (0.1-5% w / v) with 80% but more than 75% DDA were added. The source of chitosan could be fungi, shrimp, crabs, or lobsters, or a mixture thereof. Furthermore, the chitosan variant could be α, β, or γ, or a mixture thereof. After immersing the flakes in water, acetic acid and nitric acid were added in ratios of 1:1, 1:2, or 2:1 with a total v / v acid strength of 0.3-1.9%. When the flakes were no longer visible and the mixture was in solution, ammonium sulfate solution (0.001-1% v / v or v / w) was added. The temperature and stirring were maintained throughout the reaction for up to 4 hours. Finally, the solution was filtered through a 200-mesh filtration unit followed by a <635-mesh filtration unit. The solution temperature was cooled to 77 degrees Fahrenheit, potassium sorbate solution (0.01-1% v / v) was added, and the mixture was thoroughly mixed using stirring.

[0079] [Nitric acid + NaTPP] sterile water or autoclaved water was filled into a tank with a stirring speed of 400 rpm. When the temperature reached 68-194 degrees Fahrenheit, chitosan flakes (0.1-5% w / v) with 80% DDA but more than 75% DDA were added. The source of chitosan could be fungi, shrimp, crabs, or lobsters, or a mixture thereof. Furthermore, the chitosan variant could be α, β, or γ, or a mixture thereof. After immersing the flakes in water, 0.3-1.9% (v / v) nitric acid was added. When the flakes were no longer visible and the solution was formed, sodium tripolyphosphate solution (0.001-1% v / v or v / w) was added. The temperature and stirring were maintained throughout the reaction for up to 4 hours. Finally, the solution was filtered through a 200-mesh filtration unit followed by a <635-mesh filtration unit. The solution was cooled to 77 degrees Fahrenheit, potassium sorbate solution (0.01-1% v / v) was added, and the mixture was thoroughly mixed using stirring.

[0080] [Nitric acid + ammonium sulfate] Sterile water or autoclaved water was filled into a tank with a stirring speed of 400 rpm. When the temperature reached 68-194 degrees Fahrenheit, chitosan flakes (0.1-5% w / v) with 80% DDA but more than 75% DDA were added. The source of chitosan could be fungi, shrimp, crabs, or lobsters, or a mixture thereof. Furthermore, the chitosan variant could be α, β, or γ, or a mixture thereof. After immersing the flakes in water, 0.3-1.9% (v / v) nitric acid was added. When the flakes were no longer visible and the solution was formed, ammonium sulfate solution (0.001-1% v / v or v / w) was added. The temperature and stirring were maintained throughout the reaction for up to 4 hours. Finally, the solution was filtered through a 200-mesh filtration unit followed by a <635-mesh filtration unit. The solution was cooled to 77 degrees Fahrenheit, potassium sorbate solution (0.01-1% v / v) was added, and the mixture was thoroughly mixed using stirring.

[0081] [Nitric Acid] Sterile water or autoclaved water was filled into a tank with a stirring speed of 400 rpm. When the temperature reached 68-194 degrees Fahrenheit, chitosan flakes (0.1-5% w / v) with 80% but more than 75% DDA were added. The source of chitosan could be fungi, shrimp, crabs, or lobsters, or a mixture thereof. Furthermore, the chitosan variant could be α, β, or γ, or a mixture thereof. After immersing the flakes in water, 0.3-1.9% (v / v) nitric acid was added. The temperature and stirring were maintained throughout the entire reaction for up to 4 hours. Finally, when the flakes were no longer visible and the solution was formed, it was filtered through a 200-mesh filtration unit followed by a <635-mesh filtration unit. The solution temperature was cooled to 77 degrees Fahrenheit, and potassium sorbate solution (0.01-1% v / v) was added and mixed well with stirring.

[0082] [Hydrochloric acid] Sterile water or autoclaved water was filled into a tank with a stirring speed of 400 rpm. When the temperature reached 68-194 degrees Fahrenheit, chitosan flakes (0.1-5% w / v) with 80% but more than 75% DDA were added. The source of chitosan could be fungi, shrimp, crabs, or lobsters, or a mixture thereof. Furthermore, the chitosan variant could be α, β, or γ, or a mixture thereof. After immersing the flakes in water, 0.3-1.9% (v / v) hydrochloric acid was added. The temperature and stirring were maintained throughout the entire reaction for up to 4 hours. Finally, when the flakes were no longer visible and the solution was formed, it was filtered through a 200-mesh filtration unit followed by a <635-mesh filtration unit. The solution temperature was cooled to 77 degrees Fahrenheit, and potassium sorbate solution (0.01-1% v / v) was added and mixed well with stirring.

[0083] [Hydrochloric acid + NaTPP] Sterile water or autoclaved water was filled into a tank with a stirring speed of 400 rpm. When the temperature reached 68-194 degrees Fahrenheit, chitosan flakes (0.1-5% w / v) with 80% DDA but more than 75% DDA were added. The source of chitosan could be fungi, shrimp, crabs, or lobsters, or a mixture thereof. Furthermore, the chitosan variant could be α, β, or γ, or a mixture thereof. After immersing the flakes in water, 0.3-1.9% (v / v) hydrochloric acid was added. When the flakes were no longer visible and the mixture became a solution, sodium tripolyphosphate solution (0.001-1% v / v or v / w) was added. The temperature and stirring were maintained throughout the reaction for up to 4 hours. Finally, the solution was filtered through a 200-mesh filtration unit followed by a <635-mesh filtration unit. The solution was cooled to 77 degrees Fahrenheit, potassium sorbate solution (0.01-1% v / v) was added, and the mixture was thoroughly mixed using stirring.

[0084] [Hydrochloric acid + ammonium sulfate] Sterile water or autoclaved water was filled into a tank with a stirring speed of 400 rpm. When the temperature reached 68-194 degrees Fahrenheit, chitosan flakes (0.1-5% w / v) with 80% but more than 75% DDA were added. The source of chitosan could be fungi, shrimp, crabs, or lobsters, or a mixture thereof. Furthermore, the chitosan variant could be α, β, or γ, or a mixture thereof. After immersing the flakes in water, 0.3-1.9% (v / v) hydrochloric acid was added. When the flakes were no longer visible and the mixture became a solution, ammonium sulfate solution (0.001-1% v / v or v / w) was added. The temperature and stirring were maintained throughout the reaction for up to 4 hours. Finally, the solution was filtered through a 200-mesh filtration unit followed by a <635-mesh filtration unit. The solution was cooled to 77 degrees Fahrenheit, potassium sorbate solution (0.01-1% v / v) was added, and the mixture was thoroughly mixed using stirring.

[0085] [Sulfuric acid] Sterile water or autoclaved water was filled into a tank with a stirring speed of 400 rpm. When the temperature reached 68-194 degrees Fahrenheit, chitosan flakes (0.1-5% w / v) with 80% DDA but more than 75% DDA were added. The source of chitosan could be fungi, shrimp, crabs, or lobsters, or a mixture thereof. Furthermore, the chitosan variant could be α, β, or γ, or a mixture thereof. After immersing the flakes in water, 0.3-1.9% (v / v) sulfuric acid was added. The temperature and stirring were maintained throughout the reaction for up to 4 hours. Finally, when the flakes were no longer visible and the solution was formed, it was filtered through a 200-mesh filtration unit followed by a <635-mesh filtration unit. The solution temperature was cooled to 77 degrees Fahrenheit, and potassium sorbate solution (0.01-1% v / v) was added and mixed well with stirring.

[0086] [Sulfuric acid + NaTPP] Sterile water or autoclaved water was filled into a tank with a stirring speed of 400 rpm. When the temperature reached 68-194 degrees Fahrenheit, chitosan flakes (0.1-5% w / v) with 80% DDA but more than 75% DDA were added. The source of chitosan could be fungi, shrimp, crabs, or lobsters, or a mixture thereof. Furthermore, the chitosan variant could be α, β, or γ, or a mixture thereof. After immersing the flakes in water, 0.3-1.9% (v / v) sulfuric acid was added. When the flakes were no longer visible and the mixture became a solution, sodium tripolyphosphate solution (0.001-1% v / v or v / w) was added. The temperature and stirring were maintained throughout the reaction for up to 4 hours. Finally, the solution was filtered through a 200-mesh filtration unit followed by a <635-mesh filtration unit. The solution was cooled to 77 degrees Fahrenheit, potassium sorbate solution (0.01-1% v / v) was added, and the mixture was thoroughly mixed using stirring.

[0087] [Sulfuric acid + ammonium sulfate] Sterile water or autoclaved water was filled into a tank with a stirring speed of 400 rpm. When the temperature reached 68-194 degrees Fahrenheit, chitosan flakes (0.1-5% w / v) with 80% DDA but more than 75% DDA were added. The source of chitosan could be fungi, shrimp, crabs, or lobsters, or a mixture thereof. Furthermore, the chitosan variant could be α, β, or γ, or a mixture thereof. After immersing the flakes in water, 0.3-1.9% (v / v) sulfuric acid was added. When the flakes were no longer visible and the mixture became a solution, ammonium sulfate solution (0.001-1% v / v or v / w) was added. The temperature and stirring were maintained throughout the reaction for up to 4 hours. Finally, the solution was filtered through a 200-mesh filtration unit followed by a <635-mesh filtration unit. The solution was cooled to 77 degrees Fahrenheit, potassium sorbate solution (0.01-1% v / v) was added, and the mixture was thoroughly mixed using stirring.

[0088] [Nitric acid + Acetic acid + Hydrochloric acid + Sulfuric acid] Sterile water or autoclaved water was filled into a tank with a stirring speed of 400 rpm. When the temperature reached 68-194 degrees Fahrenheit, chitosan flakes (0.1-5% w / v) with 80% DDA but more than 75% DDA were added. The source of chitosan could be fungi, shrimp, crabs, or lobsters, or a mixture thereof. Furthermore, the chitosan variant could be α, β, or γ, or a mixture thereof. After immersing the flakes in water, acetic acid, nitric acid, hydrochloric acid, and sulfuric acid were added in ratios of 1:1:0:0, 1:0:1:0, 1:0:0:1, 1:1:1:0, 1:1:2:0, 1:1:1:1, and 1:1:1:2, with a total v / v acid strength of 0.3%-1.9%. Temperature and stirring were maintained throughout the reaction for up to 4 hours. Finally, once the flakes were no longer visible and the mixture was in solution, it was filtered through a 200-mesh filtration unit followed by a <635-mesh filtration unit. The solution temperature was cooled to 77°F, and potassium sorbate solution (1% v / v) was added and thoroughly mixed with stirring.

[0089] [Nitric acid + Acetic acid + Hydrochloric acid + Sulfuric acid + NaTPP] Sterile water or autoclaved water was filled into a tank with a stirring speed of 400 rpm. When the temperature reached 68-194 degrees Fahrenheit, chitosan flakes (0.1-5% w / v) with 80% DDA but more than 75% DDA were added. The source of chitosan could be fungi, shrimp, crabs, or lobsters, or a mixture thereof. Furthermore, the chitosan variant could be α, β, or γ, or a mixture thereof. After immersing the flakes in water, acetic acid, nitric acid, hydrochloric acid, and sulfuric acid were added in ratios of 1:1:0:0, 1:0:1:0, 1:0:0:1, 1:1:1:0, 1:1:2:0, 1:1:1:1, and 1:1:1:2, with a total v / v acid strength of 0.3%-1.9%. Once the flakes were no longer visible and the mixture was in solution, sodium tripolyphosphate solution (0.001–1% v / v or v / w) was added. Temperature and stirring were maintained throughout the reaction for up to 4 hours. Finally, the solution was filtered through a 200-mesh filtration unit followed by a <635-mesh filtration unit. The solution temperature was cooled to 77°F, potassium sorbate solution (0.01–1% v / v) was added, and the mixture was thoroughly mixed using stirring.

[0090] [Nitric acid + Acetic acid + Hydrochloric acid + Sulfuric acid + Ammonium sulfate] Sterile water or autoclaved water was filled into a tank with a stirring speed of 400 rpm. When the temperature reached 68-194 degrees Fahrenheit, chitosan flakes (0.1-5% w / v) with 80% DDA but more than 75% DDA were added. The source of chitosan could be fungi, shrimp, crabs, or lobsters, or a mixture thereof. Furthermore, the chitosan variant could be α, β, or γ, or a mixture thereof. After immersing the flakes in water, acetic acid, nitric acid, hydrochloric acid, and sulfuric acid were added in ratios of 1:1:0:0, 1:0:1:0, 1:0:0:1, 1:1:1:0, 1:1:2:0, 1:1:1:1, and 1:1:1:2, with a total v / v acid strength of 0.3%-1.9%. Once the flakes were no longer visible and the mixture was in solution, ammonium sulfate solution (0.001–1% v / v or v / w) was added. Temperature and stirring were maintained throughout the reaction for up to 4 hours. Finally, the solution was filtered through a 200-mesh filtration unit followed by a <635-mesh filtration unit. The solution temperature was cooled to 77°F, potassium sorbate solution (0.01–1% v / v) was added, and the mixture was thoroughly mixed using stirring.

[0091] Example 2 Various chitosan solutions were prepared using chitosan from two different sources. These sources are designated as Tg and CTg. Using Tg and CTg, chitosan-based nanoparticles were generated using the procedure identified above, yielding the following products. Tg source chitosan-based nanoparticles prepared with TgA=acetic acid. TgB = Tg source chitosan-based nanoparticles prepared with acetate and sodium tripolyphosphate. TgC = Tg source chitosan-based nanoparticles prepared with acetic acid and nitric acid. TgD = Chitosan-based nanoparticles prepared with acetic acid, nitric acid, and sodium tripolyphosphate. TgE = Tg source chitosan-based nanoparticles prepared with nitrate and sodium tripolyphosphate. CTgA = CTg source chitosan-based nanoparticles prepared with acetic acid. CTgB = CTg source chitosan-based nanoparticles prepared with acetate and sodium tripolyphosphate. CTgC = CTg source chitosan-based nanoparticles prepared with acetic acid and nitric acid. CTgD = CTg source chitosan-based nanoparticles prepared with acetic acid, nitric acid, and sodium tripolyphosphate. CTgE = CTg source chitosan-based nanoparticles prepared with nitrate and sodium tripolyphosphate.

[0092] For physicochemical characterization, formulations and intermediates were characterized. Physical morphology (shape and size) was characterized using transmission or scanning electron microscopy (Figures 1 and 2). In the resulting solution for Tg-source chitosan, >95% of particles were spherical, nearly spherical, or small linear fragments with sizes ranging from 0.1 nm to 5 microns. More specifically, the desired size obtained and utilized for further formulation development was less than 0.5 microns and was used for further formulation development and / or other agricultural applications for crop growth, stimulation, germination, and trait improvement. Similarly, the resulting solution for CTg-source chitosan had a spherical physical structure, with sizes ranging from 0.1 nm to 8 microns. More specifically, the obtained sizes of 0.1 nm to 200 nm were used as selection parameters for supplying to crops for growth, stimulation, germination, and trait improvement.

[0093] The dynamic stability and hydrodynamic size in the solution were characterized using dynamic light scattering (DLS), and the nanoparticles were subjected to dynamic light scattering (DLS) and analyzed for Z-mean, intensity, number, and polydispersity (PDI) (Figure 3). The resulting solutions ranged from 0.1 nm to 10 microns with a polydispersity index of 0.01 to 0.99. Furthermore, the surface zeta potential of the resulting solutions was characterized by DLS (Figure 4), which ranged from +5 to 70 mV. The main determinants of the zeta potential were the chitosan concentration and pH of the solution. Higher chitosan concentrations and lower pH resulted in higher zeta potentials.

[0094] Example 3 The ability of various chitosan-based nanoparticles produced in Examples 1 and 2 to load nitrogen and / or mixtures of nitrogen-rich compounds and / or compounds, such as, but not limited to, amino acids including, potassium nitrate, calcium nitrate, magnesium nitrate, urea nitrate, urea, ammonium urea nitrate, ammonium nitrate, ammonium sulfate, ammonium hydroxide, urea hydroxyapatite, arginine, cystine, histidine, leucine, lysine, methionine, threonine, tryptophan, tyrosine, valine, glutamine, glutamic acid, glycine, proline, taurine, aspartic acid, serine, and alanine, was tested. Nutrient association and sustained release are one method for balancing the nutritional requirements of plants. Several solutions were prepared and evaluated.

[0095] Preparation of chitosan solution Preparation of Solution A: Chitosan solution in acetic acid Step 1. Add 150 ml of water to a 250 ml beaker. Step 2. Turn on a top-head stirrer at 300 RPM at room temperature. The stirrer was kept on continuously throughout the process until it indicated stop. Step 3. Three minutes after Step 2, add 8 grams of chitosan flakes. Step 4. Seven minutes after Step 3, add 8 ml of 99.7% glacial acetic acid. Step 5. After the flakes have completely dissolved (17 minutes after Step 4), add 50 ml of water. Step 6. Ten minutes after Step 5, stop the stirrer. Step 7. Filter the solution through a 200 mesh filter. The total volume of the solution was 210 ml, and the concentration was 104.335 g / 100 ml. The calculated percentage of chitosan in the solution is 3.8% (w / v). The pH of the solution is 3.88.

[0096] Preparation of Solution B: Chitosan solution in HCl Step 1. Add 150 ml of water to a 250 ml beaker. Step 2. Turn on a top-head stirrer at 300 RPM at room temperature. The stirrer was kept on continuously throughout the process until it indicated stop. Step 3. Add 8 grams of chitosan flakes 3 minutes after Step 2. Step 4. Add 8 ml of 35.4% HCl 7 minutes after Step 3. Step 5. Add 50 ml of water after the flakes have completely dissolved (85 minutes after Step 4). Step 6. Stop the stirrer 26 minutes after Step 5. Step 7. Filter the solution through a 200 mesh size filter. The total volume of the solution in mL was 222.096 mL at 105.37 g / 100 mL. The calculated percentage of CS in the solution was 3.6% (w / v), and the pH of the solution was 0.66.

[0097] Preparation of solution C: Chitosan solution in nitric acid Step 1. Add 800 ml of water to a 2000 ml beaker. Step 2. Turn on a top-head stirrer at 600 RPM at room temperature. The stirrer was kept on continuously throughout the process until it indicated stop. Step 3. Two minutes after Step 2, add 20 grams of chitosan flakes. Step 4. Five minutes after Step 3, add 20 ml of 69.71% nitric acid. Step 5. After the flakes have completely dissolved (110 minutes after Step 4), add 100 ml of water. Step 6. Fourteen minutes after Step 5, stop the stirrer. Step 7. Ten minutes after Step 6, add 80 ml of water to make the solution 1000 ml. Step 8. Filter the solution through a 200 mesh size filter. The total volume of the solution in mL was 1000 mL with a weight of 100.686 grams / 100 mL. The calculated percentage of CS in the solution is 2% (w / v), and the pH of the solution is 0.84.

[0098] Preparation of Solution D: Chitosan solution in acetic acid Step 1. Add 800 ml of water to a 2000 ml beaker. Step 2. Turn on a top-head stirrer at 600 RPM at room temperature. The stirrer was kept on continuously throughout the process until it indicated stop. Step 3. 4 minutes after Step 2, add 20 grams of chitosan flakes. Step 4. 10 minutes after Step 3, add 20 ml of 99.7% glacial acetic acid. Step 5. After the flakes have completely dissolved (10 minutes after Step 4). Step 6. 17 minutes after Step 5, turn off the stirrer. Step 7. 10 minutes after Step 6, add 180 ml of water to make the solution 1000 ml. Step 8. Filter the solution through a 200 mesh size filter. The total volume of the solution in mL was 1000 ml at 100.472 grams / 100 mL. The calculated percentage of CS in the solution was 2% (w / v), and the pH of the solution was 3.91.

[0099] Preparation of solution E: Chitosan solution in H2SO4 Since chitosan did not dissolve in the solution, the H2SO4 solution was ineffective and effectively ended the experiment, as the H2SO4 solution was not used for any association formation.

[0100] Preparation of solution F: Chitosan solution in HCl Step 1. Add 800 ml of water to a 2000 ml beaker. Step 2. Turn on a top-head stirrer at 600 RPM at room temperature. The stirrer was kept on continuously throughout the process until it indicated stop. Step 3. Two minutes after Step 2, add 20 grams of chitosan flakes. Step 4. Six minutes after Step 3, add 20 ml of 35.4% HCl. Step 5. After the flakes have completely dissolved (44 minutes after Step 4), add 100 ml of water. Step 6. Twelve minutes after Step 5, stop the stirrer. Step 7. Ten minutes after Step 6, add 80 ml of water to make 1000 ml and mix thoroughly. Step 8. Filter the solution through a 200 mesh size filter. The total volume of the solution in mL was 1000 mL at 100.328 grams / 100 mL. The calculated percentage of CS in the solution was 2% (w / v), and the pH of the solution was 1.09.

[0101] Preparation of a coupling enhancer - Ammonium sulfate (AS) solution Step 1. Place 0.200 grams of ammonium sulfate into a 50 mL volumetric flask. Step 2. Add 30 mL of water. Step 3. Vortex the solution until the resulting solution becomes clear. Step 4. Add 20 mL of water to make a volume of 50 mL. The total volume of the solution in mL was 50 mL; 49.764 g. The calculated percentage of AS in the solution was 0.4% w / v. The calculated percentage of N in the solution was 0.0856% w / v. The calculated percentage of S in the solution was 0.0980% w / v. The pH of the solution was 5.75.

[0102] Preparation of a coupling enhancer - Trisodium polyphosphate (TPP) solution Step 1. Place 0.200 grams into a 50 mL volumetric flask. Step 2. Add 30 mL of water. Step 3. Vortex the solution until the resulting solution becomes clear. Step 4. Add 20 mL of water to make a volume of 50 mL. The total volume of the solution in mL was 50 mL:49.821 g. The calculated percentage of TP in the solution was 0.4% w / v. The calculated percentage of P in the solution was 0.101%. The pH of the solution was 9.76.

[0103] Preparations of combined urea Chitosan-based nanoparticles #1 Step 1. Prepare a 2000 mL beaker. Step 2. Add 175 mL of Solution A. Step 3. Start the overhead stirrer and continue the process until it indicates stop. Step 4. Five minutes after Step 3, add 100 mL of water. Step 5. Six minutes after Step 4, add 100 g of urea. Step 6. Thirteen minutes after Step 5, add 100 g of urea. Step 7. One minute after Step 6, add 50 mL of water. Step 8. Eight minutes after Step 7, add 50 mL of water. Step 9. Ten minutes after Step 8, add 135 g of urea. Step 10. Nine minutes after Step 9, add 175 mL of Solution B. Step 11. Eighteen minutes after Step 10, add 25 mL of water. Step 12. Fifty-two minutes after Step 11, add 10 mL of ammonium sulfate solution (0.1 g in 10 mL, i.e., 1% w / v). Step 12. Fifteen minutes after Step 12, stop the stirrer. Dilute this solution to 1000 mL using 125 mL of water. Step 13. Filter the solution through a 200 mesh size filter. Step 14. The solution is ready for further use / QC test analysis and is stored in a 1 L bottle. This method results in the association of the nutrient chitosan, and the solution has the following composition: Urea: 43.5% (w / v), N% w / v: 20.01 (calculated); 18.24% (analyzed by TKN), Ammonium sulfate: 0.01 (w / v), Chitosan: 1.29% (w / v), Acetic acid: 0.7% (v / v), HCl: 0.7% (v / v), pH: 3.67. Weight (grams / 100 mL): 110.832 grams

[0104] Chitosan-based nanoparticles #2 Step 1. Prepare a 2000 mL beaker. Step 2. Add 200 mL of solution D. Step 3. Start the overhead stirrer and continue the process until it indicates stop. Step 4. Three minutes after step 3, add 200 mL of distilled water. Step 5. Five minutes after step 4, add 100 g of urea. Step 6. Seven minutes after step 5, add 100 g of urea. Step 7. Five minutes after step 6, add 100 g of urea. Step 8. Eight minutes after step 7, add 135 g of urea. Step 9. Ten minutes after step 8, add 100 g of solution F. Step 10. Thirteen minutes after step 9, add 100 g of solution F. Step 11. Nineteen minutes after step 10, add 75 mL of water. Step 12. Five minutes after step 11, add 25 mL of ammonium sulfate solution (0.1 g in 25 mL of water, i.e., 0.4% w / v). Step 13. 15 minutes after Step 12, the stirrer was stopped. The solution was 1 L. Step 14. Filter the solution through a 200 mesh size filter. Step 15. The solution is ready for further use / QC test analysis and is stored in a 1 L bottle. An example of this method yields the association of nutrients with chitosan, and the solution has urea: 43.5% (w / v), N% w / v: 20.01 (calculated); 18.05 (analyzed by TKN), ammonium sulfate: 0.01 (w / v), chitosan: 0.8% (w / v), acetic acid: 0.4% (v / v), HCl acid: 0.4% (v / v), pH: 4, and weight (grams / 100 mL): 110.622 grams.

[0105] Chitosan-based nanoparticles #3 Step 1. Prepare a 2000 mL beaker. Step 2. Add 200 mL of water. Step 3. Start the overhead stirrer and continue the process until it indicates stop. Step 4. Two minutes after Step 3, add 200 mL of Solution D. Step 5. Five minutes after Step 4, add 100 g of urea. Step 6. Three minutes after Step 5, add 100 g of urea. Step 7. Six minutes after Step 6, add 100 g of urea. Step 8. Eleven minutes after Step 7, add 135 g of urea. Step 9. Ten minutes after Step 8, add 200 mL of Solution C. Step 10. Fifty minutes after Step 9, add ammonium sulfate solution (0.1 g in 25 mL of water, i.e., 0.4% w / v). Step 11. Thirty-five minutes after Step 10, stop the stirrer. Add 60 mL of water to make a total solution of 1000 mL. Step 12. Filter the solution through a 200 mesh filter. Step 13. The solution is ready for further use / QC test analysis and is stored in a 1L bottle. An example of this method yields the association of nutrients with chitosan, and the solution has urea: 43.5% (w / v), N% w / v: 20.1 (calculated); 18.17 (analyzed by TKN), ammonium sulfate: 0.01% (w / v), chitosan: 0.8% (w / v), acetic acid: 0.4 (v / v), nitric acid: 0.4 (v / v), pH: 3.22, and weight (grams / 100mL): 110.622 grams.

[0106] Chitosan-based nanoparticles #4 Step 1. Prepare a 1000 mL beaker. Step 2. Add 300 mL of water. Step 3. Start the magnetic stirrer and continue the process until it indicates stop. Step 4. 7 minutes after Step 3, add 100 g of urea. Step 5. 4 minutes after Step 4, add 100 g of urea. Step 6. 2 minutes after Step 5, add 100 g of urea. Step 7. 4 minutes after Step 6, add 135 g of urea. Step 8. 15 minutes after Step 7 (the solution was not completely dissolved, as semicrystals appeared at the bottom of the beaker), stop the stirrer. Step 9. After Step 8, prepare another 2 L beaker, add 200 mL of solution C, and start the overhead stirrer. Step 10. 2 minutes after Step 9, transfer the semi-dissolved urea solution (prepared in Step 7) to solution C. Step 11. 3 minutes after Step 10, add 100 mL of water. Step 12. 17 minutes after Step 11, add 50 mL of water. Step 13. 8 minutes after Step 12, add ammonium sulfate solution (0.1 g in 25 mL of water, i.e., 0.4% w / v). Step 14. 25 minutes after Step 13, stop the stirrer. Add 25 mL of water to make 1000 mL of solution. Step 15. Filter the solution through a 200 mesh filter. Step 16. The solution is ready for further use / QC test analysis and is stored in a 1 L bottle. An example of this method yielded a chitosan association of nutrients, and the solution had the following composition: urea: 43.5% (w / v), N% w / v: 20.01 (calculated); 18.42 (analyzed by TKN), ammonium sulfate: 0.01 (w / v), chitosan: 0.4% (w / v), nitric acid: 0.4% (v / v), pH: 2.87, and weight (grams / 100mL): 110.586 grams.

[0107] Chitosan-based nanoparticles #5 Step 1. Prepare a 2000 mL beaker. Step 2. Add 200 mL of solution D. Step 3. Start the head stirrer and continue the process until it indicates stop. Step 4. 2 minutes after step 3, add 200 mL of water. Step 5. 4 minutes after step 4, add 100 g of urea. Step 6. 4 minutes after step 5, add 100 g of urea. Step 7. 12 minutes after step 6, add 100 g of urea. Step 8. 11 minutes after step 7, add 135 g of urea. Step 9. 13 minutes after step 8, add 200 mL of solution C. Step 10. 25 minutes after step 9, add TPP solution (0.1 g of TPP in 25 mL of water, i.e., 0.4% w / v). Step 11. 20 minutes after step 10, stop the stirrer. Add 80 mL of water to this solution to make 1000 mL. Step 12. Filter the solution through 200 mesh size filter paper. An example of this method yielded a chitosan association of nutrients, and the solution had the following composition: urea: 43.5% (w / v), N% w / v: 20.01 (calculated), 18.06 (analyzed by TKN), TPP: 0.01% (w / v), chitosan: 0.8% (w / v), acetic acid: 0.4% (v / v), nitric acid: 0.4% (v / v), pH: 3.14, and weight (grams / 100mL): 110.726 grams.

[0108] Chitosan-based nanoparticles #6 Step 1. Take a 1000 mL beaker. Step 2. Add 400 mL of water. Step 3. Start a magnetic (bottom-head) stirrer and continue the process until it indicates stop. Step 4. Five minutes after Step 3, add 100 g of urea. Step 5. Nine minutes after Step 4, add 100 g of urea. Step 6. Four minutes after Step 5, add 100 g of urea. Step 7. Twelve minutes after Step 6, add 135 g of urea. Step 8. Thirty minutes after Step 7 (the solution was not completely dissolved and crystals had appeared at the bottom of the beaker), stop the stirrer. Step 9. Prepare another 2 L beaker and add 200 mL of solution C. Start an overhead stirrer. Step 10. Four minutes after Step 9, transfer the semi-dissolved urea solution (prepared in Step 7) to solution C. Step 11. Step 12. One minute after Step 10, add 50 mL of water. Step 13. 15 minutes after Step 11, add TPP solution (0.1 g in 25 mL of water, i.e., 0.4% w / v). Step 14. 20 minutes after Step 12, stop the stirrer. Step 15. Add 25 mL of water to make 1000 mL of solution. Step 16. Filter the solution through 200 mesh size filter paper. This example of the method yields chitosan association of nutrients, and the solution has urea: 43.5% (w / v), N% w / v: 20.01 (calculated); 17.9 (analyzed by TKN), TPP: 0.01% (w / v), chitosan: 0.4% (w / v), nitrate: 0.4% (v / v), pH: 2.88, weight (grams / 100 mL): 110.506 g.

[0109] Chitosan-based nanoparticles #7 Step 1. Take a 1000 mL beaker. Step 2. Add 400 mL of water. Step 3. Start with a magnetic stirrer and continue the process until it indicates stop. Step 4. Five minutes after step 3, add 100 g of urea. Step 5. Ten minutes after step 4, add 100 g of urea. Step 6. Twenty minutes after step 5, add 100 g of urea. Step 7. Ten minutes after step 6, add 135 g of urea. Step 8. Three minutes after step 7 (the solution was not completely dissolved, as semicrystals appeared at the bottom of the beaker), stop the stirrer. Step 9. Prepare another 2 L beaker, add 200 mL of solution D, and start an overhead stirrer. Step 10. Two minutes after step 9, transfer the semi-dissolved urea solution (prepared in step 8) to solution D. Step 11. Three minutes after step 10, add 25 mL of water. Step 12. Eleven minutes after Step 11, add ammonium sulfate solution (0.1 g in 25 mL of water, i.e., 0.4% w / v). Step 13. Twenty-six minutes after Step 12, stop the stirrer and add 50 mL of water to make the solution 1000 mL. Step 14. Filter the solution through 200 mesh size filter paper. This example of the method yields the association of nutrients with chitosan, and the solution has urea: 43.5% (w / v), N% w / v: 20.01 (calculated); 18.19 (analyzed by TKN), ammonium sulfate: 0.01% (w / v), chitosan: 0.4% (w / v), acetic acid: 0.4% (v / v), pH: 4.76, and weight (grams / 100 mL): 110.29 g.

[0110] Coupling agents include glutaraldehyde, genipin, epichlorohydrin, tripolyphosphate or sodium tripolyphosphate, sodium hexametaphosphate, and polyphosphates. Sulfates such as ammonium sulfate, dextran sulfate, ethylenediamine, tartaric acid, urea, and sodium trimetaphosphate are chemical agents or compounds that promote the association of chitosan molecules, resulting in the formation of a three-dimensional network or structure, and can be used with or in combination with chitosan and chitosan-containing nitrogen nanoparticles.

[0111] Stabilizers: Potential auxiliaries / stabilizers include: Surfactants: Nonionic surfactants (e.g., alkyl polyglucosides), anionic surfactants (e.g., alkyl sulfonates), cationic surfactants (e.g., alkylamines); Emulsifiers: Polyethylene glycol (PEG) derivatives, sorbitan esters (e.g., Tween series); Spreaders / stickers: Organosilicone surfactants, fatty acid spreaders (e.g., methylated seed oil); Penetrants: Crop oil concentrates, methylated seed oil. Buffers: Ammonium sulfate, potassium dihydrogen phosphate; Compatibilizers: Polyvinyl alcohol, polyacrylic acid; Antifoamers: Silicone-based antifoamers, polyethylene glycol-based antifoamers; pH adjusters: Ammonium hydroxide, phosphoric acid, sodium hydroxide, citric acid; Thickeners: Guar gum, acacia gum, xanthan gum; Humectants: Glycerol. Propylene glycol can be used with chitosan solution, chitosan nanoparticles, and combinations of chitosan and nitrogen to enhance stability and improve delivery to plant parts.

[0112] pH analysis (Figure 5) was performed according to the standard protocol for drinking water described by the U.S. EPA. Total Kjeldahl nitrogen (TKN) analysis (Figure 5) was performed according to U.S. EPA Water Administration Method 1688. UV-Vis analysis (Figure 6) was performed according to OPPTS830.7050. FTIR analysis (Figures 7-13) was performed using an Agilent-Cary 630 according to its manual.

[0113] Several formulations were manufactured using the method described above. Tables 1 and 2 show the concentrations of each component in the composition or chitosan-based nanoparticles. Tables 1 and 2 also list the physical properties of each chitosan-based nanoparticle, including EC, density, viscosity, zeta potential, and particle size. [Table 1-1] [Table 1-2] [Table 1-3] [Table 1-4] [Table 1-5] [Table 1-6] [Table 2-1] [Table 2-2] [Table 2-3]

[0114] Example 4: Application and use of chitosan and / or chitosan / nitrogen nanoparticles Chitosan and / or chitosan-based nanoparticle products or mixtures of products obtained as a result of the above examples were used for multidimensional effects on plants, such as growth, protection, and nutrition. Multidimensional effects on plants were obtained by applying the products obtained as a result of the above examples to fertilizer granules, seed surfaces, soil, soil, roots, and / or leaves. Application rates ranged from 0.01% to 50% v / w or v / v. Furthermore, the products obtained as a result of the above examples were used to be mixed with other pesticides to function as growth promoters, symbiotics, nutrients, pathogens, or protective agents against undesirable plants in order to obtain synergistic effects.

[0115] The multidimensional effects on plants were studied in several experiments. The experiments used either a 2% or 3% chitosan formulation.

[0116] The 2% chitosan preparation AN1136 (Tables 1 and 2) was manufactured according to the following procedure.

[0117] First, pour 200 ml of water into a 500 ml beaker, ensuring accurate measurement. Switch on a magnetic stirrer, set to room temperature (26.4°C) and programmed to operate at 300 revolutions per minute (RPM). Maintain continuous stirring throughout the procedure until further instructions. Stir the water for 5 minutes to establish a uniform environment in the beaker. After this period, carefully add 5 grams of chitosan flakes to the stirring water. After adding the chitosan flakes, stir for a further 15 minutes to promote their dispersion and interaction with water molecules. At this point, add 5 ml of 37% hydrochloric acid (HCl) to the solution. Carefully observe the solution and allow sufficient time (approximately 60 minutes) for the chitosan flakes to dissolve completely in the liquid medium, ensuring thorough mixing and dissolution. Once complete dissolution is achieved, add an additional 40 ml of water to the solution and maintain the stirring process to ensure homogeneity. Continue stirring the solution with the newly added water for a further 15 minutes to ensure proper mixing and equilibrium. After the stirring period, stop the magnetic stirrer. Add 5 mL of water to adjust the solution volume to 250 mL to compensate for any volume loss during the process. Ensure thorough mixing to obtain a homogeneous solution. Filter the solution using a 200-mesh (74-micron) filter to remove any residual particulate matter or impurities, allowing only the liquid portion to pass through while capturing any solid residue. [Table 3]

[0118] Prepare the nitrogen precursor solution. First, select a 2000 mL beaker and ensure it is clean, dry, and ready for experimental use. Carefully measure 490 mL of water using a graduated cylinder and pour it into the selected beaker. Transfer the prepared beaker containing the measured water to an overhead stirrer apparatus. Ensure the apparatus is properly set up and calibrated and set the stirring speed to 300 revolutions per minute (RPM). Furthermore, ensure that the room temperature is maintained at 25°C, providing the optimal environment for the experiment. After stirring the water for 5 minutes, gradually introduce 260 grams of nitrogen source, in this case urea. Slowly add the urea to the water while stirring, ensuring that it is uniformly dispersed and dissolved in the solution. After adding the urea, continue stirring the solution for a further 120 minutes, giving sufficient time for the urea to completely dissolve and dissolve in the water. After the urea has dissolved, add 377 grams of ammonium sulfate to the solution. Similar to the addition of urea, gradually introduce the ammonium sulfate, ensuring that it is uniformly dissolved and dispersed in the solution. After adding the ammonium sulfate, stir the solution for a further 120 minutes, then carefully observe the solution to determine if the ammonium sulfate has completely dissolved. Confirm complete mixing and dissolution by checking for any remaining solid particles in the solution. Once dissolution is confirmed, stop the overhead stirrer and cease the stirring process. Ensure the stirrer is completely off to prevent unnecessary stirring of the solution. Measure the final volume of the solution using a graduated cylinder or volumetric flask. An ideal volume is approximately 900 ml, indicating the success of the experiment and accurate preparation of the solution. [Table 4]

[0119] Once the precursor chitosan and nitrogen solution are prepared, a chitosan-based nanoparticle solution is prepared.

[0120] Select a clean, dry 2000 mL beaker to serve as the container for the experiment. Carefully measure 900 mL of nitrogen precursor solution using accurate measuring instruments such as a 1000 mL graduated cylinder. Pour this measured volume of solution into the prepared beaker, ensuring accuracy of the measurement to obtain consistent results. Once the nitrogen precursor solution is added to the beaker, transfer the beaker to an overhead stirrer apparatus. Ensure the apparatus is properly set up and calibrated, and set the stirring speed to 300 revolutions per minute (RPM). Maintain ambient room temperature to provide the best environment for the experiment. After stirring the solution for 30 minutes, then add 100 mL of 2% chitosan solution to the stirred nitrogen precursor solution. Slowly introduce the chitosan solution into the stirring solution to ensure complete mixing and uniform dispersion in the beaker. After adding the chitosan solution, continue stirring the solution for a further 3 hours to allow sufficient time for any reactions or interactions between the components of the solution to occur. After this period, stop the overhead stirrer to end the stirring process. To determine the final volume of the solution, measure it using appropriate measuring instruments to ensure accuracy of the measurement. Ideally, the measured volume should be approximately 1000 mL, indicating the success of the experiment and the accurate preparation of the solution. Proceed to filtering the solution using a 200-mesh filtration unit. This filtration process takes approximately 18 seconds to complete and effectively removes larger particles or impurities present in the solution. Following the initial filtration, repeat the filtration process using a 5-micron filtration unit. This secondary filtration step further purifies the solution, removing smaller particles and ensuring a higher level of purity. The filtration process with the 5-micron unit takes approximately 20 seconds to complete. Once the solution is filtered, transfer it to a suitable storage container, such as a high-density polyethylene (HDPE) bottle, ensuring it is tightly sealed to prevent contamination. Store the filtered solution at a temperature of 20°C until further use. [Table 5] [Table 6]

[0121] The 3% chitosan preparation EZ2241293 (Tables 1 and 2) was manufactured according to the following procedure.

[0122] The preparation begins with the preparation of the intermediate solution. This starts with the preparation of the N-source stock solution (w / w). First, to ensure accurate measurement, carefully prepare a 2000 ml glass beaker by placing it on a weighing machine. Pay attention to the initial weight. Precisely pour 400 grams of water into the beaker to confirm the accuracy of the measurement. After adding the water, activate the overhead stirrer and set it to a constant rotation speed of 300 RPM. Ensure that the temperature of the solution remains stable at 25°C throughout this process. Gradually add 435 grams of urea to the stirring solution, taking care to maintain a consistent mixing rate. Maintain a stirring speed of 300 RPM and a temperature of 25°C and allow the solution to mix completely for 3 hours. This ensures that the urea is completely dissolved in the solution. Once the stirring process is complete, carefully stop the overhead stirrer. To determine the final weight of the solution, weigh the beaker containing the solution and note any change from the initial weight. The final weight is recorded, which is 833 grams, indicating that the urea has been successfully mixed into the solution. Next, the solution is filtered through a 200-mesh mesh to purify it. This step ensures the removal of any impurities or undissolved particles. The composition (w / w) of this intermediate solution is urea: 52.09% and nitrogen: 23.96%. Table 3 lists the properties of the solution. [Table 7]

[0123] The second step is the preparation of the ammonium sulfate stock solution (w / w). First, select a dry, clean 1000 mL beaker and ensure that it is completely free of residues or contaminants. Carefully place the beaker on the weighing scale to prepare for accurate measurement. With precision, carefully pour 290 grams of water into the clean beaker to ensure accurate measurement. After adding the water, start the overhead stirrer and set it to rotate at a constant speed of 300 RPM. Maintain a stable temperature of 25°C throughout this process to ensure optimal conditions for mixing. Introduce 210 grams of ammonium sulfate into the stirring solution and mix thoroughly to ensure complete dispersion. Maintain a stirring speed of 300 RPM and a temperature of 25°C and allow the solution to mix harmoniously for 3 hours. After the stirring cycle is complete, carefully remove the overhead stirrer and bring the mixing process to a controlled halt. Using a weighing scale, weigh the beaker containing the solution and note any deviation from the initial weight. The recorded final weight will be a total weight of 498.09 grams, reflecting the successful incorporation of ammonium sulfate. To further purify the solution, the filtration process is carried out using a 200-mesh mesh. This step serves to remove any impurities or undissolved particles. The composition (w / w) of the ammonium stock solution is ammonium sulfate: 42%, sulfur: 10.18%, nitrogen: 8.90%, sulfate: 30.5%. [Table 8]

[0124] The preparation of chitosan-based nanoparticles EZ2244810 proceeds as follows, using the stock solution prepared above. First, carefully weigh a 1000 mL glass beaker. Add 486.32 grams of urea-derived N source to the beaker. Carefully introduce approximately 0.7 mL of hydrochloric acid (HCl) to adjust the pH to the desired level of 3.57, and mix thoroughly to ensure uniformity of the pH distribution. Similarly, prepare another 1000 mL glass beaker by tare the weighing machine to ensure accurate measurements for the subsequent steps. Add 453.48 grams of ammonium sulfate-derived N source to this prepared beaker, maintaining accuracy throughout the process. Prepare for the addition of 0.1 grams of potassium sorbate by tare the weighing machine using another weighing paper to ensure accurate measurements of each component. While maintaining measurement accuracy, prepare another 20 mL glass beaker by tare the measuring instrument and adding 7.5 grams of water. Weigh the 2000 mL glass beaker, tare the measuring instrument, and add 30 grams of Zale-2 solution from the stock. Place the beaker under an overhead stirrer and set to rotate at 300 RPM, maintaining a constant temperature of 25°C. After stirring for 15 minutes, introduce 486.32 grams of N source from urea into the solution and mix gently to ensure uniform dispersion. After 30 minutes, add 453.48 grams of N source from ammonium sulfate into the solution and maintain the stirring process for uniform mixing. Then, after another 30 minutes, introduce 0.1 grams of solid potassium sorbate into the solution and mix thoroughly to ensure homogeneity. After another 5 minutes, add 30 grams of water to the solution and maintain the stirring process to ensure complete dissolution. Continue stirring the solution for a total of 3 hours, giving sufficient time for all components to completely dissolve and interact. Once the stirring cycle is complete, carefully stop the stirrer and bring the mixing process to a controlled halt. Measure the final weight of the solution, which should be 991 grams, taking into account an additional 8.42 grams for solution transfer losses. Next, filter the solution using a 200-mesh filtration unit with a filtration time of approximately 10 seconds to ensure the removal of any impurities or undissolved particles.The solution is further purified by filtering it through a 5-micron filtration unit for approximately 13 seconds, increasing its purity and clarity. After filtration, the solution is weighed again to determine the final weight, which should be approximately 982.5 grams, taking into account an additional 9 grams of solution loss during filtration. The purified solution is stored in a 1000 mL HDPE bottle, ensuring proper sealing to maintain its integrity until further use. [Table 9]

[0125] The final composition (w / w) of chitosan-based nanoparticles EZ2241293 is as follows: chitosan = 0.3%, total nitrogen (w / v) = 15.64%, total ammonium sulfate (including linker) = 19.06%, N in urea = 11.65%, N in AS = 3.99%, S in AS = 4.62%, sulfate = 13.84%, potassium sorbate: 0.01%, and concentrated hydrochloric acid (used to maintain the pH of the urea solution): 700 microliters.

[0126] The first experiment was conducted in a hydroponic environment. Lettuce was grown in a hydroponic system, and the effect of chitosan-based nanoparticles EZ2241293 (see Tables 1 and 2) on the lettuce was evaluated.

[0127] The experiment included five test parameters. In the positive control, high nitrogen (105.50 mg of nitrogen per plant) was added to the growth medium and water was applied as a foliar spray. In the negative control, low nitrogen (24.99 mg of nitrogen per plant) was added to the growth medium and water was applied as a foliar spray. The water application used in these two control groups was to eliminate the possibility that water applied to the foliar surface could affect the plants. A commercially available control was used to simulate a foliar spray as currently available, consisting of low nitrogen (24.99 mg of applied nitrogen per plant) and urea foliar spray added to the growth medium. A second commercially available control was used to simulate another foliar spray as currently available, consisting of low nitrogen (24.99 mg of applied nitrogen per plant) and urea and ammonium nitrate foliar spray added to the growth medium. The formulation tested was a chitosan-based nanoparticle containing 15.65 wt% nitrogen and 0.3 wt% chitosan, with a nitrogen-to-chitosan ratio of 0.019. Chitosan-based nanoparticles were applied as a foliar spray, and nitrogen was applied at a concentration of 6.73 + / - 0.1 per plant.

[0128] Chitosan nanoparticles have a positive effect on plants, resulting in increased plant width, shoot weight, vigor, root weight, and reduced plant toxicity. Foliar spraying often has adverse effects on plants because the substances in the spray can damage or burn the leaves. Chitosan nanoparticles have the lowest plant toxicity scores, statistically similar to positive controls of high nitrogen in hydroponic media and negative controls of low nitrogen in hydroponics (Figure 14). In contrast, foliar spraying of urea and ammonium sulfate has a statistically high plant toxicity rating. The low plant toxicity rating of chitosan nanoparticles demonstrates that foliar spraying with chitosan nanoparticles causes less damage to plants than foliar spraying with urea and ammonium sulfate. The effect of nitrogen on plants can be measured in several ways. In hydroponic studies using lettuce, plant vigor is one parameter that indicates the effect of nitrogen and chitosan nanoparticles (Figure 15). Plants exhibit greater vigor with higher levels of nitrogen in the hydroponic medium than with lower levels of nitrogen in the hydroponic medium. The addition of chitosan nanoparticles bridges the gap between high and low levels of nitrogen in the hydroponic medium. In other words, the vigor of lettuce receiving chitosan nanoparticles is statistically equivalent to that of lettuce receiving high levels of nitrogen in the hydroponic medium. Plant width is another parameter that identifies the effect of chitosan nanoparticles (Figure 16). Plants exhibit wider width with higher levels of nitrogen in the hydroponic medium than with lower levels of nitrogen in the hydroponic medium. The addition of chitosan nanoparticles bridges the gap between high and low levels of nitrogen in the hydroponic medium. In other words, the width of lettuce receiving chitosan nanoparticles is statistically equivalent to that of lettuce receiving high levels of nitrogen in the hydroponic medium. Commercial controls were comparable to negative controls (Figure 17). Plants exhibit greater shoot weight with higher levels of nitrogen in the hydroponic medium than with lower levels of nitrogen in the hydroponic medium. The addition of chitosan nanoparticles bridges the gap between high and low levels of nitrogen in the hydroponic medium. In other words, the shoot weight of lettuce treated with chitosan nanoparticles is statistically equivalent to that of lettuce treated with high levels of nitrogen in the hydroponic medium. The commercial control was comparable to the negative control.Another parameter for measuring the effect of nitrogen is root weight. Roots grow in search of nitrogen and other nutrients (Figure 18). High and low nitrogen in hydroponic media are statistically equivalent. Chitosan nanoparticles resulted in an increase in lettuce root weight, indicating that lettuce plants treated with chitosan nanoparticles are beneficial to the plants.

[0129] Several growth characteristics showed that replacing nitrogen with smaller-volume chitosan nanoparticles resulted in biomass yields at least equal to those of the high-nitrogen control group. This means that the plants benefited from the small amount of nitrogen associated with chitosan nanoparticles as much as they benefited from the high nitrogen. High nitrogen provided 105.50 mg of nitrogen per plant. Low nitrogen provided 24.99 mg of nitrogen per plant. Chitosan nanoparticles added 6.73 + / - 0.1 mg of nitrogen per plant. Therefore, chitosan nanoparticles and low hydroponic nitrogen together provided 31.72 mg of nitrogen to the plants, representing a 70% reduction in the total amount of nitrogen applied to each plant. In other words, 80.51 mg of removed nitrogen was replaced with 6.73 + / - 0.1 mg of nitrogen, or 8.4% of the nitrogen removed from high nitrogen was replaced with nitrogen in chitosan nanoparticles with an equivalent benefit.

[0130] The second experiment was a field study using chitosan-based nanoparticles as a foliar spray. This study was a small-scale plot replicate study conducted in Florida during the off-season.

[0131] Corn was sown in plots improved with either a high nitrogen content (68.04 kg / acre nitrogen) or a low nitrogen content (49.90 kg / acre nitrogen). High nitrogen (68.04 kg / acre nitrogen) and low nitrogen (49.90 kg / acre nitrogen) control groups were established. Three different chitosan-based nanoparticle formulations were applied to different plots, creating high-nitrogen and low-nitrogen plots for each chitosan-based nanoparticle formulation (Figure 19A). The chitosan-based nanoparticle formulations were AN1132 = 0.2% chitosan, 12% N (urea-derived), and 8% N (ammonium sulfate-derived). LMW chitosan was used.

[0132] The formulation was sprayed onto the plot at a rate of 5 l / acre. This resulted in approximately 1 kg of nitrogen per acre. Therefore, the 18.14 kg / acre of nitrogen removed from high nitrogen to low nitrogen was replaced by 1 kg / acre in the low nitrogen setting. The yield for at least a portion of the chitosan nanoparticle formulation was at least statistically equal to the yield in the high nitrogen setting. Therefore, adding the chitosan nanoparticle formulation with a statistically equal yield reduced the nitrogen by approximately 25% from high nitrogen to low nitrogen. In other words, the total amount of nitrogen used in the low nitrogen setting with chitosan nanoparticles was approximately 75% of the nitrogen used in the high nitrogen setting that yielded the same yield. Furthermore, the nitrogen in the chitosan nanoparticle formulation replaced approximately 5.5% of the nitrogen removed from the low nitrogen setting. Adding chitosan nanoparticles AN1132 to the low nitrogen setting resulted in higher nitrogen levels in the V8 stage leaves compared to the high nitrogen or low nitrogen control groups (Figure 19B).

[0133] The third experiment involved the addition of chitosan nanoparticles to corn fields fertilized with nitrogen according to generally accepted standard fertilization methods. The tests were conducted in Nebraska, Kansas, and Ohio. Plot sizes were approximately 10 x 45 feet, and each treatment was repeated five times. Test parameters included a control group with no nitrogen fertilizer or chitosan nanoparticle composition added; a standard nitrogen plot, which was fertilized with nitrogen according to agricultural standards; a standard nitrogen + AN1113 chitosan nanoparticle solution applied at 5 liters / acre at two locations during growth; a low nitrogen plot with 18.14 kg / acre of nitrogen added below standard nitrogen; and a low nitrogen + AN1113 chitosan nanoparticle solution applied at 5 liters / acre at two locations during growth. Throughout the tests, the total nitrogen applied in the chitosan nanoparticle + low nitrogen scenarios was 15–25% less nitrogen than grower standards.

[0134] Nitrogen was applied according to the treatment definitions in Table 10. For the standard nitrogen rate, the minimum acceptable value was used to represent the nitrogen rate that is representative of the standard practice of local growers relative to the average yield potential. [Table 10]

[0135] Adding chitosan nanoparticles to standard nitrogen increased yield by 7% (Figure 20). Adding chitosan nanoparticles to low nitrogen yield resulted in a yield equivalent to that of standard nitrogen.

[0136] Chitosan-based nanoparticle formulations may be used in combination with other agricultural inputs, for example, with herbicides. Experiments were conducted with several herbicides to confirm any synergistic and antagonistic effects. Experiments were conducted with two herbicides, glyphosate and 2,4-D. Glyphosate did not show statistical evidence of antagonistic activity when used with the chitosan-based nanoparticle solution (Figure 21). In the glyphosate experiments, seven different crops were tested: wheat, corn, rye, mustard, sunflower, melon, and beans. Each crop was tested under untreated conditions, glyphosate conditions, and chitosan-based nanoparticle solution / glyphosate combination conditions. There was statistical separation between the untreated condition and glyphosate, and between the untreated condition and the chitosan-based nanoparticle solution / glyphosate combination condition. There was no statistical separation between the glyphosate condition and the chitosan-based nanoparticle solution / glyphosate combination condition.

[0137] When 2,4-D was used with a chitosan-based nanoparticle solution, no statistical evidence of antagonistic activity was observed (Figure 22). The 2,4-D experiments involved testing four different crops: mustard, sunflower, melon, and bean. Under untreated conditions, 2,4-D was tested. D Each crop was tested under different conditions, including the combined use of a 2,4-D-chitosan nanoparticle solution. Statistical separation was observed between the untreated condition and the 2,4-D condition, and between the untreated condition and the combined use of the chitosan nanoparticle solution and 2,4-D condition. No statistical separation was observed between the 2,4-D condition and the combined use of the chitosan nanoparticle solution and 2,4-D condition.

[0138] This invention is described with reference to various specific and preferred embodiments and techniques. Nevertheless, it should be understood that many modifications and changes can be made while remaining within the spirit and scope of the invention.

Claims

1. A composition comprising chitosan and at least 1% (w / w) of nitrogen.

2. The composition according to claim 1, wherein the nitrogen is present in an amount of about 11% (w / w) to about 25% (w / w).

3. The composition according to claim 1, wherein at least 50% of the nitrogen is in the chemical form of an amine and / or nitrate and / or ammonia.

4. The composition according to claim 3, wherein the composition comprises chitosan nanoparticles and a nitrogen-rich compound and / or mixture of compounds selected from the group consisting of amino acids such as potassium nitrate, calcium nitrate, magnesium nitrate, urea nitrate, ammonium urea nitrate, ammonium nitrate, ammonium sulfate, ammonium hydroxide, urea hydroxyapatite, arginine, cystine, histidine, leucine, lysine, methionine, threonine, tryptophan, tyrosine, valine, glutamine, glutamic acid, glycine, proline, taurine, aspartic acid, serine, and alanine.

5. The composition according to claim 1, wherein the chitosan is present in an amount of about 0.01% to about 4% (w / v).

6. The composition according to claim 1, further comprising a coupling enhancer.

7. The composition according to claim 6, wherein the coupling enhancer is sodium tripolyphosphate and / or ammonium sulfate.

8. A method for producing chitosan nanoparticles, A method comprising: dissolving chitosan in acidic water, wherein the water is acidified with an acid selected from the group of acids including acetic acid, citric acid, lactic acid, malic acid, tartaric acid, formic acid, acetylsalicylic acid, oxalic acid, succinic acid, benzoic acid, folic acid, pyruvic acid, butyric acid, propionic acid, caproic acid, hydrochloric acid, sulfuric acid, nitric acid, phosphoric acid, hydrofluoric acid, perchloric acid, hydrobromic acid, hydroiodic acid, chloric acid, bromic acid, iodic acid, humic acid, fulvic acid, amino acids, nucleic acids (DNA and RNA), boric acid, chromic acid, cyanuric acid, hyaluronic acid, arsenic acid, and carboxylic acids, and combinations thereof; and forming nanoparticles from the dissolved chitosan.

9. The method according to claim 8, wherein a coupling enhancer is added to the dissolved chitosan.

10. The method according to claim 9, wherein the coupling enhancer is glutaraldehyde, genipin, epichlorohydrin, tripolyphosphate or sodium tripolyphosphate, sodium hexametaphosphate, polyphosphate, sulfates such as ammonium sulfate and dextran sulfate, ethylenediamine, tartaric acid, urea, sodium trimetaphosphate, etc.

11. The method according to claim 8, wherein an auxiliary agent is added to improve the stability of chitosan.

12. Auxiliary agents include surfactants: nonionic surfactants (e.g., alkyl polyglucosides), anionic surfactants (e.g., alkyl sulfonates), cationic surfactants (e.g., alkylamines); emulsifiers: polyethylene glycol (PEG) derivatives, sorbitan esters (e.g., Tween series); spreaders / stickers: organosilicone surfactants, fatty acid-based spreaders (e.g., methylated seed oils); penetrating agents: crop oil concentrates, methylated seed oils. The method according to claim 11, wherein the buffering agent is ammonium sulfate, phosphoric acid, sodium acetate, potassium dihydrogen phosphate; the compatibilizer is polyvinyl alcohol, polyacrylic acid; the defoaming agent is a silicone-based defoaming agent, a polyethylene glycol-based defoaming agent; the pH adjuster is ammonium hydroxide, citric acid, hydrochloric acid, sodium hydroxide; the thickener is guar gum, xanthan gum, acacia gum; and the humectant is glycerol, propylene glycol.

13. The method according to claim 8, wherein a nitrogen-rich compound and / or mixture of compounds is selected from the group consisting of potassium nitrate, calcium nitrate, magnesium nitrate, urea nitrate, ammonium urea nitrate, ammonium hydroxide, ammonium nitrate, ammonium sulfate, urea hydroxyapatite, arginine, cystine, histidine, leucine, lysine, methionine, threonine, tryptophan, tyrosine, valine, glutamine, glutamic acid, glycine, proline, taurine, aspartic acid, serine, and alanine, and is added to the dissolved chitosan.

14. The method according to claim 13, wherein the nanoparticles comprise chitosan and a nitrogen-rich compound and / or a mixture of compounds.

15. The method according to claim 14, wherein the nanoparticles contain at least 1% (w / w) of nitrogen.

16. The method according to claim 15, wherein the nitrogen is present in an amount of about 11% (w / w) to about 25% (w / w).

17. The method according to claim 13, wherein the chitosan is present in an amount of about 0.01% to about 4% (w / v).

18. A method for treating a plant or plant part, comprising treating the plant or plant part with a composition according to any one of claims 1 to 7.

19. The method according to claim 18, wherein the plant part is selected from the group consisting of plant seeds, leaves, stems, flowers, or roots.

20. The method according to claim 19, wherein a predetermined amount of the composition according to any one of claims 1 to 7 is used in place of a certain proportion of a conventional nitrogen fertilizer to produce an equivalent or better yield.

21. The method according to claim 20, wherein the predetermined amount of the composition according to any one of claims 1 to 7 is about 0.250 liters to about 30 liters per acre applied by foliar spraying, and is used in place of about 0 kilograms to about 150 kilograms of nitrogen units per acre applied by conventional nitrogen fertilizers.

22. The method according to claim 21, wherein the predetermined amount of the composition according to any one of claims 1 to 7 is about 5 liters per acre applied by foliar spraying using a device capable of producing droplet sizes of 0.0001 to 2 mm, and is used in place of about 18.14 kilograms of nitrogen units per acre applied by conventional nitrogen fertilizers.

23. A method for preparing a plant growth medium, comprising treating the growth medium with a composition according to any one of claims 1 to 7.

24. The method according to claim 23, wherein the growth medium is soil, peat, moss, woody residue, leaf mold, sawdust, bark, bagasse, rice hulls, sand, perlite, vermiculite, calcined clay, polystyrene, urea-formaldehyde resin, agar, or agarose, in vitro and in vivo plant tissue culture / growth media, and hydroponic solutions, aeroponic and geoponic media, and combinations thereof.

25. The method according to claim 18, wherein the proportion of the composition used is about 2 percent to about 7 percent of the conventional nitrogen fertilizer to be removed.

26. The method according to claim 25, wherein the proportion of the composition used is about 5.4 percent of the conventional nitrogen fertilizer to be removed.

27. The method according to claim 18, wherein the total nitrogen used is reduced by about 50 percent to about 90 percent.

28. The method according to claim 27, wherein the total nitrogen used is reduced by approximately 70 percent.

29. A method for treating plant parts, comprising treating the plant or plant parts with a composition according to any one of claims 1 to 7, together with a standard amount of conventional fertilizer.

30. The method according to claim 29, wherein treatment with the composition results in an improvement in yield of about 0% to about 80%.

31. The method according to claim 29, wherein treatment with the composition results in an improvement in yield of about 7%.

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