Compositions and methods for crop-protection and uses thereof

Abstract

Disclosed herein are compositions for crop-protective compounds. These compositions are formed by functionalizing physically or chemically associating crop-protective compounds in chitosan nanoparticles. In a first aspect, the disclosure provides a composition comprising chitosan, and an insecticidal compound. In some embodiments, chitosan comprises between about 1.2 wt% and about 6 wt% of the composition. In some embodiments, the chitosan is about 1.7 wt% of the composition.

Description

Docket No.10860-10910-US COMPOSITIONS AND METHODS FOR CROP-PROTECTION AND USES THEREOF TECHNICAL FIELD

[0001] The invention relates to plant protection and specifically to physical and / or chemical association of compounds with chitosan for protecting plants from biotic stress and abiotic stress. BACKGROUND

[0002] Agricultural production across the globe is facing unprecedented challenges due to climate change, extreme weather, soil erosion, pathogen resistance and environmental pollution. Global population is projected to reach about 10 billion by 2050, thus the demand for food is expected to increase by 60%. One of the key challenges facing modern agriculture is the overuse or higher dosage of existing active ingredients due to their low effectiveness or due to higher / rapid environmental losses, leading to concerns such as resistance among target organisms, off-target impacts on human health, and ecological and environmental degradation. Therefore, regulatory pressure is increasing or limiting their use. SUMMARY

[0003] In aspects of the invention, the disclosure provides compositions for crop-protective compounds. These compositions are formed by physically and / or chemically associating crop- protective compounds on or within chitosan nanoparticles.

[0004] In a first aspect, the disclosure provides a composition comprising chitosan, and an insecticidal compound. The insecticidal compound may be any compound that has an insecticidal effect. In embodiments, the insecticidal compound comprises one or more of a spinosyn, 2- phenylethanol, Malathion, Parathion, Diazinon, Chlorpyrifos, Carbaryl, Methomyl, Aldicarb, Propoxur, Permethrin, Cypermethrin, Deltamethrin, Lambda-cyhalothrin, Imidacloprid, Clothianidin, Thiamethoxam, Acetamiprid, Dichlorodiphenyltrichloroethane (DDT), Lindane, Chlorane, Pyrethrins, Rotenone, Azadirachtin, Beauveria bassiana, Metarhizium anisopliae, Methoprene, Pyriproxfen, Fenoxycarb, Diflubenzuron, Sarin, Tabun, Soman, VX, Sulfur, Bacillus thuringiensis and Diatomaceous Earth. In some embodiments, the spinosyn comprises at least two chemically distinct spinosyns. In some embodiments, the at least two chemically distinct spinosyns are spinosyn A and spinosyn D. In some embodiments, the spinosyn comprises of Spinosad.

[0005] In a second aspect, the disclosure provides a composition comprising chitosan, and an herbicidal compound.Docket No.10860-10910-US

[0006] The herbicidal compound may be any compound that has an herbicidal effect. In embodiments, the herbicidal compound comprises one or more of a photosynthesis inhibitor, an auxin mimic, a microtubule assembly inhibitor, a cell membrane disruptor, or an amino acid synthesis inhibitor. In some of these embodiments, the amino acid synthesis inhibitor is glyphosate. In some of these embodiments, glyphosate containing potassium salt form and / or isopropylamine salt with or without ammonium sulphate.

[0007] In a third aspect, the disclosure provides a composition comprising chitosan, and a fungicidal compound.

[0008] The fungicidal compound may be any compound that has a fungicidal effect. In embodiments, the fungicidal compound comprises one or more of copper, chlorothalonil, azoxystrobin, prothioconazole, iprodione, or fludioxonil. In some embodiments, the copper comprises one or more of copper, copper nitrate, copper sulfate, or copper hydroxide.

[0009] In a fourth aspect, the disclosure provides a composition comprising chitosan, and a nematocidal compound.

[0010] The nematocidal compound may be any compound that has a nematocidal effect. In embodiments, the nematocidal compound comprises one or more of methyl bromide, 1,3- Dichloropropene, Dazomet, Aldicarb, Fenamiphos, Oxamyllus thuringiensis, Paecilomyces lilacinus, Pochonia chlamydosporia, Azadirachtin, nicotinamides, thiophenes, Terpenoids, Flavenoids, and Saponins.

[0011] In addition to physically or chemically associating crop-protective compounds, the chitosan nanoparticles are also useful for physically and / or chemically associating nutrient compounds such as fertilizers. In some embodiments, the nutrient compounds are nitrogen.

[0012] Further aspects and embodiments are provided in the foregoing drawings, detailed description and claims. BRIEF DESCRIPTION OF THE DRAWINGS

[0013] The following drawings are provided to illustrate certain embodiments described herein. The drawings are merely illustrative and are not intended to limit the scope of claimed inventions and are not intended to show every potential feature or embodiment of the claimed inventions. The drawings are not necessarily drawn to scale; in some instances, certain elements of the drawing may be enlarged with respect to other elements of the drawing for purposes of illustration.

[0014] Figures 1A.1B, and 1C are images of a copper fungicidal compound without chitosan taken with a Transmission Electron Microscope (TEM).Docket No.10860-10910-US

[0015] Figures 2A, 2B, and 2C are TEM images of the shell of chitosan nanoparticles without the copper fungicidal compound.

[0016] Figures 3A, 3B, and 3C are TEM images of copper fungicidal chitosan nanoparticles without a coupling enhancer.

[0017] Figures 4A, 4B, and 4C are TEM images of copper fungicidal chitosan nanoparticles with a coupling enhancer.

[0018] Figures 5A, 5B, and 5C are photos depicting the effect of different compositions on Palmer amaranth. 1) untreated control; 2) glyphosate and ammonium sulfate; 3) glyphosate, ammonium sulfate and 0.25 % chitosan; 4) glyphosate, ammonium sulfate and 0.5 % chitosan; 5) glyphosate, ammonium sulfate and 1.0 % chitosan; 6) ammonium sulfate and 0.25 % chitosan. Figure 5A is a photo taken 7 days after treatment. Figure 5B is a photo taken 14 days after treatment. Figure 5C is a photo taken 21 days after treatment.

[0019] Figures 6A, 6B, and 6C are photos depicting the effect of different compositions on Palmer amaranth. 1) untreated control; 2) glyphosate and ammonium sulfate; 3) glyphosate, ammonium sulfate and 0.25 % chitosan; 4) glyphosate, ammonium sulfate and 0.5 % chitosan; 5) glyphosate, ammonium sulfate and 1.0 % chitosan; 6) ammonium sulfate and 0.25 % chitosan. Figure 6A is a photo taken 7 days after treatment. Figure 6B is a photo taken 14 days after treatment. Figure 6C is a photo taken 21 days after treatment.

[0020] Figures 7A, 7B, and 7C are photos depicting the effect of different compositions on Common lambsquarters 1) untreated control; 2) glyphosate and ammonium sulfate; 3) glyphosate, ammonium sulfate and 0.25 % chitosan; 4) glyphosate, ammonium sulfate and 0.5 % chitosan; 5) glyphosate, ammonium sulfate and 1.0 % chitosan; 6) ammonium sulfate and 0.25 % chitosan. Figure 7A is a photo taken 7 days after treatment. Figure 7B is a photo taken 14 days after treatment. Figure 7C is a photo taken 21 days after treatment.

[0021] Figures 8A, 8B, and 8C are photos depicting the effect of different compositions on Common lambsquarters 1) untreated control; 2) glyphosate and ammonium sulfate; 3) glyphosate, ammonium sulfate and 0.25 % chitosan; 4) glyphosate, ammonium sulfate and 0.5 % chitosan; 5) glyphosate, ammonium sulfate and 1.0 % chitosan; 6) ammonium sulfate and 0.25 % chitosan. Figure 8A is a photo taken 7 days after treatment. Figure 8B is a photo taken 14 days after treatment. Figure 8C is a photo taken 21 days after treatment.

[0022] Figures 9A, 9B, and 9C are photos depicting the effect of different compositions on Green foxtail 1) untreated control; 2) glyphosate and ammonium sulfate; 3) glyphosate, ammonium sulfate and 0.25 % chitosan; 4) glyphosate, ammonium sulfate and 0.5 % chitosan; 5) glyphosate, ammonium sulfate and 1.0 % chitosan; 6) ammonium sulfate and 0.25 % chitosan. Figure 9A is a photoDocket No.10860-10910-US taken 7 days after treatment. Figure 9B is a photo taken 14 days after treatment. Figure 9C is a photo taken 21 days after treatment.

[0023] Figures 10A, 10B, and 10C are photos depicting the effect of different compositions on Green foxtail 1) untreated control; 2) glyphosate and ammonium sulfate; 3) glyphosate, ammonium sulfate and 0.25 % chitosan; 4) glyphosate, ammonium sulfate and 0.5 % chitosan; 5) glyphosate, ammonium sulfate and 1.0 % chitosan; 6) ammonium sulfate and 0.25 % chitosan. Figure 10A is a photo taken 7 days after treatment. Figure 10B is a photo taken 14 days after treatment. Figure 10C is a photo taken 21 days after treatment.

[0024] Figures 11A, 11B, and 11C are photos depicting the effect of different compositions on Velvetleaf 1) untreated control; 2) glyphosate and ammonium sulfate; 3) glyphosate, ammonium sulfate and 0.25 % chitosan; 4) glyphosate, ammonium sulfate and 0.5 % chitosan; 5) glyphosate, ammonium sulfate and 1.0 % chitosan; 6) ammonium sulfate and 0.25 % chitosan. Figure 11A is a photo taken 7 days after treatment. Figure 11B is a photo taken 14 days after treatment. Figure 11C is a photo taken 21 days after treatment.

[0025] Figures 12A, 12B, and 12C are photos depicting the effect of different compositions on Velvetleaf 1) untreated control; 2) glyphosate and ammonium sulfate; 3) glyphosate, ammonium sulfate and 0.25 % chitosan; 4) glyphosate, ammonium sulfate and 0.5 % chitosan; 5) glyphosate, ammonium sulfate and 1.0 % chitosan; 6) ammonium sulfate and 0.25 % chitosan. Figure 12A is a photo taken 7 days after treatment. Figure 12B is a photo taken 14 days after treatment. Figure 12C is a photo taken 21 days after treatment.

[0026] 13A is a photograph of a microtiter plate treated with chitosan formulation EXP-12960 which is a chitosan preparation with 2 wt% LMW Chitosan and 6 wt% glacial acetic acid along with a 6 wt% glacial acetic acid solution in combination with various amounts of fungicide azoxystrobin to assess what effect acetic acid had on fungal growth.

[0027] Figures 13B, 13C, 13D, 13E, and 13F are photographs of agar plates used for determining the minimum fungicidal concentration. Figure 14B shows fungal growth on an agar plate treated with chitosan formulation EXP-12982 and azoxystrobin. Figure 14C shows fungal growth on an agar plate treated with chitosan formulation EXP-12982 and pyraclostrobin. Figure 14D shows fungal growth on an agar plate treated with chitosan formulation EXP-12982 and propiconazole. Figure 14E shows fungal growth on an agar plate treated with chitosan formulation EXP-12982 and azoxystrobin. Figure 14F shows fungal growth on an agar plate treated with chitosan formulation EXP- 12982 and fluopyram.

[0028] Figures 14A, 14B, and 14C are graphs of the effectiveness of several compositions of commercial copper-based fungicides, chitosan copper nanoparticles, and commercial copper-basedDocket No.10860-10910-US fungicides combined with chitosan solutions against Xanthomonas citri at different time intervals such as pre-application, 3, 32, 62, 94, and 125 days after application.

[0029] Figures 15A and 15B are graphs depicting the effect of several compositions on insects. Figure 13A is for male Drosophila and figure 13B is for female Drosophila.

[0030] DETAILED DESCRIPTION

[0031] The following description recites various aspects and embodiments of the inventions disclosed herein. No particular embodiment is intended to define the scope of the invention. Rather, the embodiments provide non-limiting examples of various compositions, and methods that are included within the scope of the claimed inventions. The description is to be read from the perspective of one of ordinary skill in the art. Therefore, information that is well known to the ordinarily skilled artisan is not necessarily included. Definitions

[0032] The following terms and phrases have the meanings indicated below, unless otherwise provided herein. This disclosure may employ other terms and phrases not expressly defined herein. Such other terms and phrases shall have the meanings that they would possess within the context of this disclosure to those of ordinary skill in the art. In some instances, a term or phrase may be defined in the singular or plural. In such instances, it is understood that any term in the singular may include its plural counterpart and vice versa, unless expressly indicated to the contrary.

[0033] 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” encompasses a single substituent as well as two or more substituents, and the like.

[0034] As used herein, “for example,” “for instance,” “such as,” or “including” are meant to introduce examples that further clarify more general subject matter. Unless otherwise expressly indicated, such examples are provided only as an aid for understanding embodiments illustrated in the present disclosure and are not meant to be limiting in any fashion. Nor do these phrases indicate any kind of preference for the disclosed embodiment.

[0035] As used herein “crop protective compound” means compounds, chemical or biological agents, used to safeguard agricultural crops from damage caused by pests, diseases, weeds, and other harmful factors including those of biotic and abiotic stressors. These compounds include herbicides (to control weeds), insecticides (to manage insect pests), fungicides (to combat fungal infections), bactericides, nematicides, and other specialized agents.Docket No.10860-10910-US

[0036] As used herein “chitosan” means a natural polysaccharide derived from the deacetylation of chitin, which is found in the exoskeletons of crustaceans, insect cuticles, and fungal cell walls. It is composed of β-(1→4)-linked D-glucosamine and N-acetyl-D-glucosamine units. Chitosan exhibits unique physicochemical properties, including biocompatibility, biodegradability, non-toxicity, and a cationic nature. Its cationic nature allows it to interact with negatively charged biological molecules. Chitosan’s solubility depends on its degree of acetylation and molecular weight. It is soluble in acidic media due to protonation of its amine groups but has limited solubility in neutral or basic conditions. Chitosan may be further defined in terms of molecular weight. Ultra-low molecular weight, low molecular weight, medium molecular weight, and high molecular weight. Chitosan from these different groups may exhibit different properties. Throughout this disclosure ultra-low molecular weight chitosan is referred to as chitosan with a molecular weight of 20 kDa or lower. Throughout this disclosure low molecular weight chitosan is referred to as chitosan with a molecular weight of between about 100 kDa and 300 kDa. Throughout this disclosure high molecular weight chitosan is referred to as chitosan with a molecular weight of greater than 300 kDa.

[0037] A novel platform or formulation that enhances the effectiveness, and synergy of the active ingredients used for crop protection from various pathogens including bacteria, fungi, nematodes, insects, and non-intended growth of monocot-dicot plants often referred as herbs, would be beneficial. By leveraging the synergistic effects of engineered chitosan, this innovation aims to address these challenges head-on. The reduced dependence on high agrochemical input rates not only enhances cost- effectiveness for farmers but also aligns with global efforts to promote sustainable and environmentally friendly agricultural practices.

[0038] Chitosan's ability to act as a synergistic platform for existing active ingredients in crop protection agents opens new avenues for sustainable agriculture. The integration of chitosan with conventional formulations creates a powerful synergy, amplifying the overall performance and efficacy of the applied agents. This synergy explored has the potential to revolutionize crop protection strategies by significantly reducing the required application rates of agrochemical inputs per acre.

[0039] Chitosan may be used as a crop protective compound on it own. Chitosan may be particularly effective as a fungicide. Certain molecular weights of chitosan may be more effective as a crop protective compound as compared to other molecular weights of chitosan.

[0040] Chitosan has a positive effect on mitigating health and environmental risks associated with conventional crop protection methods. The innovative use of chitosan not only enhances the performance of active ingredients but also contributes to the overall reduction of chemical residues in crops and the surrounding environment. As a result, this technology aligns with the growing demand for agricultural solutions that prioritize human health and environmental sustainability.Docket No.10860-10910-US

[0041] There are several benefits of the use of chitosan as a physical and / or chemical associating / embedding agent. The first benefit is that the chitosan polymer is bio-safe and biodegradable. The chitosan polymer has a biological origin and has been used for immobilization of enzymes for food, and in biosensing applications. Chitosan can be harvested from fungus, shrimp, crab, lobster, squid pens, insect exoskeletons, or any combination of these sources. A second benefit compared to inorganic coatings such as sulfur coatings is the absence of cracks. Cracks in the coating cause an immediate release of the coated nutrient upon contact with water. Immediate release of the nutrient defeats the purpose of the physical and / or chemical association, which is to slowly release the nutrient. A third benefit of the chitosan polymer coating is the overall health of the soil. The chitosan polymer results in lowered acidification of the soil. Acidification of the soil can be detrimental to specific plant species. Coatings such as sulfur lower the pH of the soil. Additionally, the soil microbes are an important part of soil health. Synthetic polymers cannot be metabolized by soil microbes.

[0042] The addition of chitosan enhances the overall activity of active ingredients used for crop protection due to action mechanisms. These action mechanisms include but are not limited to Adhesion and Penetration, Cuticle Penetration, Synergistic mode of action, and Reduced application rates. Chitosan, with its adhesive properties, may facilitate better adhesion of the fungicidal mixture to the plant surfaces. This improved adhesion increases the residence time of the active ingredients on the leaves, enhancing their ability to combat foliar diseases. Chitosan has been shown to enhance the penetration of active ingredients through the plant cuticle. In this example, it could assist active ingredients in penetrating the plant tissues more effectively, reaching the site of infection and providing better control over diseases. Chitosan may act synergistically with crop protection substances, augmenting their modes of action. For instance, azoxystrobin inhibits mitochondrial respiration, while chlorothalonil disrupts fungal cell membranes. Chitosan's presence may enhance these actions, creating a more robust defense against a broader spectrum of foliar pathogens. The synergistic effects enabled by chitosan might allow for a reduction in the application rates of crop protection chemicals. This reduction not only contributes to economic benefits for farmers but also aligns with sustainable agriculture practices by minimizing the total amount of chemical inputs.

[0043] In some embodiments, the crop protective compound is associated with a chitosan nanoparticle. To physically and / or chemically associate the crop-protective compound with the chitosan nanoparticle, the crop-protective compound must be held in place on or within the nanoparticle. The compound is generally held in place by ionic bonds or van der Waals force bonds. In some embodiments, a coupling enhancer assists in the formation of these bonds. In some embodiments, the coupling enhancer is sodium tripolyphosphate. In some embodiments, the coupling enhancer is an aldehyde or an anionic polymer. In embodiments where the coupling enhancer is an aldehyde, the aldehyde is glutaraldehyde or formaldehyde. In embodiments where the coupling enhancer is an anionic polymer, the anionic polymer is sodium alginate.Docket No.10860-10910-US

[0044] In some embodiments, the crop protective compound is part of a composition. In these embodiments, the composition includes chitosan. In these embodiments, the chitosan and crop protective compound may be in solution.

[0045] The composition comprises chitosan, and a crop protective compound. In certain embodiments, chitosan comprises between about 0.3 wt% and about 5.0 wt% of the composition. In some embodiments, chitosan comprises between about 1.2 wt% and about 2.2 wt% of the composition. In some embodiments, the chitosan is about 1.7 wt% of the composition. In embodiments, the wt% of the chitosan is about 0.3, 0.4, 0.5, 0.6, 0.7, 0.8, 0.9, 1.0, 1.1, 1.2, 1.3, 1.4, 1.5, 1.6, 1.7, 1.8, 1.9, 2.0, 2.1, 2.2, 2.3, 2.4, 2.5, 2.6, 2.7, 2.8, 2.9, 3.0, 3.1, 3.2, 3.3, 3.4, 3.5, 3.6, 3.7, 3.8, 3.9, 4.0, 4.1, 4.2, 4.3, 4.4, 4.5, 4.6, 4.7, 4.8, 4.9, or 5.0 wt%.

[0046] In some embodiments the amount of chitosan in a chitosan and crop protective composition is between about 20 ppm and about 2000 ppm. In embodiments, the amount of chitosan in a composition comprising chitosan and a crop protective compound is about 28.12, 56.25, 112.5, 225, 448, 896, or 1792 ppm. In embodiments, the amount of chitosan in a composition comprising chitosan and a crop protective compound is about 20, 30, 40, 50, 60, 70, 80, 90, 100, 110, 120, 130, 140, 150, 160, 170, 180, 190, 200, 210, 220, 230, 240, 250, 260, 270, 280, 290, 300, 310, 320, 330, 340, 350, 360, 370, 380, 390, 400, 410, 420, 430, 440, 450, 460, 470, 480, 490, 500, 510, 520, 530, 540, 550, 560, 570, 580, 590, 600, 610, 620, 630, 640, 650, 660, 670, 680, 690, 700, 710, 720, 730, 740, 750, 760, 770, 780, 790, 800, 810, 820, 830, 840, 850, 860, 870, 880, 890, 900, 910, 920, 930, 940, 950, 960, 970, 980, 990, 1000, 1010, 1020, 1030, 1040, 1050, 1060, 1070, 1080, 1090, 1100, 1110, 1120, 1130, 1140, 1150, 1160, 1170, 1180, 1190, 1200, 1210, 1220, 1230, 1240, 1250, 1260, 1270, 1280, 1290, 1300, 1310, 1320, 1330, 1340, 1350, 1360, 1370, 1380, 1390, 1400, 1410, 1420, 1430, 1440, 1450, 1460, 1470, 1480, 1490, 1500, 1510, 1520, 1530, 1540, 1550, 1560, 1570, 1580, 1590, 1600, 1610, 1620, 1630, 1640, 1650, 1660, 1670, 1680, 1690, 1700, 1710, 1720, 1730, 1740, 1750, 1760, 1770, 1780, 1790, 1800, 1810, 1820, 1830, 1840, 1850, 1860, 1870, 1880, 1890, 1900, 1910, 1920, 1930, 1940, 1950, 1960, 1970, 1980, 1990, or 2000 ppm.

[0047] The molecular weight of the chitosan in a crop protective composition may affect the release of the physically and / or chemically associated compounds. In embodiments, the chitosan has a molecular weight between about 0.5 kDa and about 900 kDa. In embodiments, the chitosan has a molecular weight of about 0.5, 0.6, 0.7, 0.8, 0.9, 1.0, 1.1, 1.2, 1.3, 1.4, 1.5, 1.6, 1.8, 1.9, 2.0, 2.1, 2.2, 2.3, 2.4, 2.5, 2.6, 2.7, 2.8, 2.9, 3.0, 3.1, 3.2, 3.3, 3.4, 3.5, 3.6, 3.7, 3.8, 3.9, 4.0, 4.1, 4.2, 4.3, 4.4, 4.5, 4.6, 4.7, 4.8, 4.9, 5.0, 10, 15, 20, 25, 30, 35, 40, 45, 50, 55, 60, 65, 70, 75, 80, 85, 90, 95, 100, 105, 110, 115, 120, 125, 130, 135, 140, 145, 150, 155, 160, 165, 170, 175, 180, 185, 190, 195, 200, 205, 210, 215, 220, 225, 230, 235, 240, 245, 250, 255, 260, 265, 270, 275, 280, 285, 290, 295, 300, 305, 310, 315, 320, 325, 330, 335, 340, 345, 350, 355, 360, 365, 370, 375, 380, 385, 390, 395, 400, 405, 410, 415, 420, 425, 430, 435, 440, 445, 450, 455, 460, 465, 470, 475, 480, 485, 490, 495, 500, 505,Docket No.10860-10910-US 510, 515, 520, 525, 530, 535, 540, 545, 550, 555, 560, 565, 570, 575, 580, 585, 590, 595, 600, 605, 610, 615, 620, 625, 630, 635, 640, 645, 650, 655, 660, 665, 670, 675, 680, 685, 690, 695, 700, 705, 710, 715, 720, 725, 730, 735, 740, 745, 750, 755, 760, 765, 770, 775, 780, 785, 790, 795, 800, 805, 810, 815, 820, 825, 830, 835, 840, 845, 850, 855, 860, 865, 870, 875, 880, 885, 890, 895, 900.

[0048] In embodiments, the chitosan in the composition comprises at least two different molecular weight classifications. In embodiments, the chitosan is one of ULMW chitosan, LMW chitosan, or HMW chitosan. In embodiments, the chitosan in the composition comprises at least three different molecular weights. In embodiments, the chitosan is one of ULMW chitosan, LMW chitosan, or HMW chitosan.

[0049] The formulation of chitosan and crop protective compound compositions utilizes chitosan dissolved in acid. Additionally, the formulation of chitosan nanoparticles utilizes chitosan dissolved in acid, therefore the composition requires an acid in the composition. The acid may be any one or more of 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.

[0050] In one embodiment, the physical and / or chemical association process begins with the chitosan polymer. The source of chitosan can be fungus, shrimp, crab or lobster or a mixture of them. In certain embodiments the chitosan has a Degree of DeAcetylation (DDA): >80%, MW: <110 KDa. In particular embodiments the DDA is > 85%. The DDA may be 90%. The DDA bay be > 95%. In particular embodiments the DDA is about 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%.

[0051] Prior to usage for the formulation of crop protective chitosan-based nanoparticles, or in the formulation of nanoparticles, the chitosan must be dissolved in a solution. Dissolution medium: acidic water having pH ranges between 1 and 5.5, or having a pH < 1, < 2, < 3, < 4, < 5, < 5.5. The acid used may be any acid. In particular embodiments the acid used is one of or a combination of the 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. To dissolve the chitosan, the aqueous acidic solution may be kept at temperature between about 68 and about 212 degrees F. In particularDocket No.10860-10910-US embodiments, the temperature is kept at about 68, 72, 82, 92, 201, 112, 122, 132, 142, 152, 162, 172, 182, 192, 202, or 212 degrees F. During the entire reaction period the solution may be kept stirring at rpms between about 50 and about 2500 rpm. After completion of the reaction, typically between 0.3 and 4 hours, the solution was filtered through 200 mesh (74 micron) followed by more than 635 mesh (<20 micron). In the filtered solution, antimicrobial agents such as but not limited to potassium sorbate are added (0.1% v / v). In certain cases, as described below a linker such as sodium tripolyphosphate and / or ammonium sulfate is added in the solution before the addition of antimicrobial agents.

[0052] Embodiments of chitosan-based crop protective compounds include chitosan formulations in solution, chitosan powder, chitosan-based salts, and chitosan-based nanoparticles. In embodiments, the chitosan powder and chitosan-based salts are water soluble. In certain embodiments the chitosan-based nanoparticles comprise crop protective compounds. The crop protective compounds are physically and / or chemically associated by / in the chitosan-based nanoparticles. The crop protective compounds include compounds to protect crop plants from pathogens and pests including but not limited to insects, fungus, non-crop plants, and nematodes.

[0053] > 95% particles were in the spherical, nearly spherical shape or small linear fragments having size between 0.1 nm and 8 microns. (By TEM measurement). 0.1 nm and 10 microns with polydispersity index between 0.01 to 0.99 (by DLS measurement). Surface zeta potential ranged between +5 and 70 mV (measured by DLS). N%: 18-24 Application onto the seed surface, mixing in the soil, exposing root and / or exposing leaves. The application rate ranged between 0.01% to 50% v / w or v / v.

[0054] In certain embodiments, the chitosan-based nanoparticles may comprise at least 1, 5, 10, 15, 20, 25, 30, 35, 40, 45, 50, 55, 60, 65, 70, 75, 85, 90, or 95% (wt%) active ingredient. The active ingredient may be any insecticide, fungicide, herbicide, bactericide, or nematicide. Many of the options for these active ingredients are listed below.

[0055] Embodiments include insecticides in suspension or in solution in a composition with chitosan or physically and / or chemically associated on or within chitosan-based nanoparticles. Insecticides may be naturally occurring compounds or artificially created compounds. They may repel insects or may kill insects. In particular embodiments, the insecticide may be any one or more of a spinosyn, a phenylethanol, Malathion, Parathion, Diazinon, Chlorpyrifos, Carbaryl, Methomyl, Aldicarb, Propoxur, Permethrin, Cypermethrin, Deltamethrin, Lambda-cyhalothrin, Imidacloprid, Clothianidin, Thiamethoxam, Acetamiprid, Dichlorodiphenyltrichloroethane (DDT), Lindane, Chlorane, Pyrethrins, Rotenone, Azadirachtin, Bacillus thuringiensis Beauveria bassiana, Metarhizium anisopliae, Methoprene, Pyriproxfen, Fenoxycarb, Diflubenzuron, Sarin, Tabun, Soman, VX, Sulfur, and Diatomaceous Earth. In more particular embodiments, the phenylethanol is 2-phenylethanol. In more particular embodiments, the spinosyn may be a combination of multiple chemically distinctDocket No.10860-10910-US spinosyns. In these embodiments, the chemically distinct spinosyns may be spinosyn A and spinosyn D, the combination of spinosyn A and spinosyn D is often referred to as Spinosad.

[0056] Embodiments include fungicides in suspension or in solution in a composition with chitosan or physically and / or chemically associated on or within chitosan-based nanoparticles. Fungicides may be naturally occurring compounds or artificially created compounds or any combination thereof. In embodiments, the fungicide includes copper. The copper in such embodiments may be in various forms including but not limited to copper, copper nitrate, copper sulfate, and copper hydroxide. In other embodiments, the fungicide may be one or more ofAzoxystrobin, Pyraclostrobin, Trifloxystrobin, Fluoxastobin, Picoxystrobin, Kresoxim methyl, Epoxiconazole, tebuconazole, Propiconazole, Cyproconazole, dienconazole, metconazole, Boscalid, Carboxin, Flutoanil, Sedaxane, Fluopyram, Penflufen, Fluxaapyroxad, Fludioxonil, Cymoxanil, Forety-al, Propamocarb, Mandipropamid, Cyazofamid, Famoxadone, Metafenone, Isoprothiolane, Validymycin, Fenhexamid, Pyroquilon, Carpropamid, Fenamidone, Fluopocolide, Harpin, Oxolinic acid, Aluminum sulfate, Bentonite clay, chitosan, Copper Octanoate, Cuprous Oxide, Peptides, Phosphoric Acids, Potassium Bicarbonate, Potassium iodide, Potassium phosphite, Potassium silicate, Potassium thiocyanate, Sodium bicarbonate, Sulfur, Tribasic copper sulphate, Hydrogen Peroxide, Chenopodium quinoa saponins, citric acid, Clove Oil, Coconut Oil, Esquisetum arvense, Eugenol, Garlic Oil, Gu-lutathione, Lamarin, Melaleuca, Peppermint Oil, Phospholipids, Propolis extract, Propylene glycol, Reynoutria, Rosemary Oil, Saponins, Seaweed Extract, Sodium lautyl, Soybean Oil, Sucrose esters, Terpenese, Thyme Oil, Natamycin, Agrobacterium, Bacillus amyloliquefaciens, Bacillus subtilllis, Bacillus pumilus, Pantoea,Streptomyces lydicus, Aspergillus flavus, Aureobasidium, Clavulanic acid, Conothyrium, Phliebiopsis gigantea, Pseudomonas fluorescens, Pythium oligandrum, Saccharomyces, Tricoderma harzianum, Tricoderma longibratum, Tricoderma spp, Tricoderma Virens, Gliocladium, Polyoxin D Zinc Salt, Bacteriophage, Metschnikowia, Mancozeb, Thiram, Propineb, Prothioconazole. Probenazole, Tricyclazole, Prochloraz, Copper, Sulphur, Fentin, Oxine Copper, Chlorothalonil, Captan, Folpet, Metaxyl, Benalaxyl, Oxadixyl, Carbendazim, Thiophanate, Thiabendazole, Benomyl, Fuberidazole, Spiroxamine, dimethomorph, Fenpropidin, Fenproprimorph, Cypodinil, Pyrimethanil, Mepanipyrim, Fluazinam, Dithianon, Pencycuron, Iminoctadine, Tolylfuanid, Iprodione, Procymidone, Vincllozolin, Fenarimol, Bupirimate, Triforine, Ferimzone, and Pyrifenox. Embodiments include herbicides in solution in a composition with chitosan or physically and / or chemically associated on or within chitosan-based nanoparticles. In embodiments, the herbicidal compound may be one or more of a photosynthesis inhibitor, an auxin mimic, a microtubule assembly inhibitor, a cell membrane disruptor, or an amino acid synthesis inhibitor. In embodiments, the herbicide may inhibit enzymatic activity in plants. In embodiments, the herbicide is glyphosate, atrazine, 2,4-D, metolachlor, pendimethalin,trifluralin, dicamba, imazethapyr, mesotrione, or sethoxydim. In particularDocket No.10860-10910-US embodiments, the herbicide is glyphosate. In some of those embodiments, the glyphosate is glyphosate K.

[0057] Embodiments include nematicides in suspension or in solution in a composition with chitosan or physically and / or chemically associated on or within chitosan-based nanoparticles. In embodiments the nematicide may be one or more of methyl bromide, 1,3-Dichloropropene, Dazomet, Aldicarb, Fenamiphos, Oxamyl, Bacillus thuringiensis, Paecilomyces lilacinus, Pochonia chlamydosporia, Azadirachtin, nicotinamides, thiophenes, Terpenoids, Flavenoids, Saponins Burkholderia spp., bacillus fermis, bacillus spp., pasteuria nishizawae, fluopyram, bacillus amyloiquefaciens, pyriflumetofen, and chromobacterium subtsugae.

[0058] Embodiments include bactericides in in suspension or in solution in a composition with chitosan or physically and / or chemically associated on or within chitosan-based nanoparticles. In embodiments, the bactericide may be a copper-based bactericide. The copper-based bactericide may be one or more of copper sulfate, copper hydroxide, copper oxide, copper oxychloride, copper octanoate, or copper (II) sulfate pentahydrate. In embodiments, the bactericide may be streptomycin sulfate, oxytetracycline, kasugamycin, or other bactericides.

[0059] In embodiments, the chitosan and crop protective compounds have synergistic effects. The synergistic effects of the chitosan in solution in a composition with a crop protective compound or on or within a chitosan-based nanoparticle with an associated crop protective compound show improvement over the effects of the crop protective compound alone. In embodiments, the chitosan nanoparticle and associated crop protective compound composition improves the effect by between about 1.0 % and about 50 % as compared to the crop protective compound alone. In embodiments, the chitosan nanoparticle and associated crop protective compound composition improves the effect by about 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, and 50 % as compared to the crop protective compound alone.

[0060] In embodiments, the chitosan and crop protective compounds have synergistic effects. The synergistic effects of the chitosan in solution in a composition with a crop protective compound or on or within a chitosan-based nanoparticle associated with a crop protective compound enable the amount of crop protective compound to be reduced and show the same effect as a standard treatment of the crop protective compound alone. In embodiment, the amount of the crop protective compound is reduced by between about 5.0 and about 60 wt%. In embodiments, amount of crop protective compound used in the chitosan nanoparticle and associated crop protective compound composition is reduced by about 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, and 60 wt% as compared to the standard use of the crop protective compound alone.Docket No.10860-10910-US

[0061] In addition to combining chitosan with active ingredients for crop protection, chitosan may also be used with nutritional compounds to assist in providing the nutritional needs of plants. Physical and / or chemical association of nutrients on or within a chitosan molecule enables the slow release of nutrients. The slow release of nutrients is one method for balancing the nutritional needs of plants. One such nutritional need is nitrogen, which for example, may occur in the form of amine, nitrate and ammonia.

[0062] In embodiments it is desirable to combine chitosan in solution in a composition with a crop protective compound or physically and / or chemically associate nitrogen in or on the chitosan- based nanoparticles along with the crop-protective particles. In embodiments, the nitrogen is in the form of amine and / or nitrate and / or ammonia. The nitrogen may be physically and / or chemically associated by / in the chitosan-based nanoparticles. In certain embodiments, the chitosan nanoparticles may be formed in the presence of and or loaded with nitrogen in the form of amine and / or nitrate and / or ammonia. The nitrogen may be physically and / or chemically associated by / in the chitosan-based nanoparticles. In particular embodiments, the nitrogen in the form of amine, nitrate, and / or ammonia may be selected potassium nitrate, calcium nitrate, magnesium nitrate, urea nitrate, urea ammonia nitrate, ammonium nitrate, 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 combination thereof.

[0063] In certain embodiments, the chitosan-based nanoparticles may comprise at least 1, 5, 10, 15, 20, 25, 30, 35, 40, 45, 50, 55, 60, 65, 70, 75, 85, 90, or 95% (wt%) nitrogen. In particular embodiments, at least 50, 55, 60, 65, 70, 75, 85, 90, or 95% of the nitrogen is in the chemical form of the amine and / or nitrate and / or ammonia.

[0064] In one embodiment, the reaction is carried out with specific solutions prepared in accordance with one and / or all of the procedures listed above at the temperature between 68 and 250 degrees F. During the entire reaction period the solution was kept stirring at 400 rpm for uniformity of the substances. After completion of the reaction, typically between 0.3 and 4 hours, the solution was filtered through 200 mesh (74 micron) followed by more than 635 mesh (<20 micron). The nitrogen % in the final solution was maintained between 0.1% and 100%, specifically between 18-24% using the compound and / or mixture of compounds described above.

[0065] Embodiments include methods of treating a plant or pant part with any of the compositions described herein. The plant part may be any plant part including, but not limited to a seed, leaf, stem, flower, or root of a plant. Examples species of plant parts include, but are not limited to cereals such as wheat, barley, rye, oat; canola, cotton, eggplant, lettuce, sorghum, soybean, rice, oil seed rape, sugar beet, sugarcane, grapes, lentils, sunflowers, alfalfa, pome fruits; stone fruits; peanuts; coffee; tea; strawberries; turf; vegetables, such as tomatoes, potatoes, cucurbits and lettuce.Docket No.10860-10910-US

[0066] One example of the synergistic potential of chitosan in combination with an existing active ingredient is the partnership between chitosan and copper-based fungicides, such as copper hydroxide or copper sulfate, in the realm of crop protection. Copper-based fungicides have long been utilized to control various fungal diseases in crops. However, they often face challenges related to adherence, persistence, and overall efficacy. Chitosan, with its unique properties, can address these limitations and enhance the effectiveness of copper-based fungicides.

[0067] When chitosan is combined with copper-based fungicides, a synergistic interaction occurs. Chitosan acts as an adjuvant, improving the adhesion of copper particles to plant surfaces and enhancing their penetration into plant tissues. This increased adherence and penetration lead to more sustained and efficient control of fungal pathogens. Chitosan assists in forming a protective barrier on the grapevine leaves, promoting better retention and even distribution of copper ions. This synergy enhances the fungicidal activity against downy mildew, providing a more robust defense mechanism for the grape crop. The chitosan-copper combination not only improves the fungicidal efficacy but also allows for a reduction in the amount of copper required for effective disease control. This reduction is significant in addressing concerns related to the environmental impact of copper accumulation in soils and the potential development of copper-resistant strains of fungi. Furthermore, the chitosan-copper synergy contributes to a sustained release of copper ions, extending the duration of protection against fungal infections. This prolonged activity reduces the frequency of fungicide applications, offering economic benefits to farmers while minimizing the environmental footprint associated with frequent chemical treatments.

[0068] Further embodiments include methods of treating a plant growth medium with any of the compositions described herein. Plant growth medium may comprise one or more of soil, peat, moss, wood residue, leaf mold, sawdust, bark, bagasse, rice hull, sand, perlite, vermiculite, calcinated clay, polystyrene, urea formaldehyde resins, and hydroponic liquids.

[0069] In a first embodiment, a composition comprises chitosan, and a fungicidal compound.

[0070] A second embodiment comprises the composition of embodiment 1, wherein chitosan is present in the composition in an amount sufficient to enhance the antifungal activity of the fungicidal compound.

[0071] A third embodiment comprises the composition of embodiment 1, wherein the chitosan and the fungicidal compound, together, have synergistic antifungal activity.

[0072] A fourth embodiment comprises the composition of any of embodiments 1-3, wherein the chitosan and the antifungal compound are the same.Docket No.10860-10910-US

[0073] A fifth embodiment comprises the composition of any of embodiments 1-4, wherein the chitosan comprises ultra-low molecular weight, low molecular weight, or high molecular weight chitosan.

[0074] A sixth embodiment comprises the composition of any of embodiments 1-5, wherein the majority of the chitosan in the composition by weight is ultra-low molecular weight, low molecular weight, or high molecular weight chitosan.

[0075] A seventh embodiment comprises the composition of any of embodiments 1-6, wherein the composition comprises at least two of ultra-low molecular weight, low molecular weight, and high molecular weight chitosan.

[0076] An eighth embodiment comprises the composition of embodiment 5, wherein the ultra- low molecular weight chitosan has a molecular weight between about 2 kDa and about 10 kDa.

[0077] A ninth embodiment comprises the composition of embodiment 5, wherein the ultra- low molecular weight chitosan has a molecular weight between about 3 kDa and about 6 kDa.

[0078] A tenth embodiment comprises the composition of any of embodiments 1-9, wherein the fungicidal compound comprises one or more of copper, Azoxystrobin, Pyraclostrobin, Trifloxystrobin, Fluoxastobin, Picoxystrobin, Kresoxim methyl, Epoxiconazole, tebuconazole, Propiconazole, Cyproconazole, dienconazole, metconazole, Boscalid, Carboxin, Flutoanil, Sedaxane, Fluopyram, Penflufen, Fluxaapyroxad, Fludioxonil, Cymoxanil, Forety-al, Propamocarb, Mandipropamid, Cyazofamid, Famoxadone, Metafenone, Isoprothiolane, Validymycin, Fenhexamid, Pyroquilon, Carpropamid, Fenamidone, Fluopocolide, Harpin, Oxolinic acid, Aluminum sulfate, Bentonite clay, chitosan, Copper Octanoate, Cuprous Oxide, Peptides, Phosphoric Acids, Potassium Bicarbonate, Potassium iodide, Potassium phosphite, Potassium silicate, Potassium thiocyanate, Sodium bicarbonate, Sulfur, Tribasic copper sulphate, Hydrogen Peroxide, Chenopodium quinoa saponins, citric acid, Clove Oil, Coconut Oil, Esquisetum arvense, Eugenol, Garlic Oil, Gu-lutathione, Lamarin, Melaleuca, Peppermint Oil, Phospholipids, Propolis extract, Propylene glycol, Reynoutria , Rosemary Oil, Saponins, Seaweed Extract, Sodium lautyl, Soybean Oil, Sucrose esters, Terpenese, Thyme Oil, Natamycin, Agrobacterium, Bacillus amyloliquefaciens, Bacillus subtilllis, Bacillus pumilus, Pantoea, Streptomyces lydicus, Aspergillus flavus, Aureobasidium, Clavulanic acid, Conothyrium, Phliebiopsis gigantea, Pseudomonas fluorescens, Pythium oligandrum, Saccharomyces, Tricoderma harzianum, Tricoderma longibratum, Tricoderma spp, Tricoderma Virens, Gliocladium, Polyoxin D Zinc Salt, Bacteriophage, Metschnikowia, Mancozeb, Thiram, Propineb, Prothioconazole. Probenazole, Tricyclazole, Prochloraz, Copper, Sulphur, Fentin, Oxine Copper, Chlorothalonil, Captan, Folpet, Metaxyl, Benalaxyl, Oxadixyl, Carbendazim, Thiophanate, Thiabendazole, Benomyl, Fuberidazole, Spiroxamine, dimethomorph, Fenpropidin, Fenproprimorph, Cypodinil, Pyrimethanil,Docket No.10860-10910-US Mepanipyrim, Fluazinam, Dithianon, Pencycuron, Iminoctadine, Tolylfuanid, Iprodione, Procymidone, Vincllozolin, Fenarimol, Bupirimate, Triforine, Ferimzone, and Pyrifenox.

[0079] An eleventh embodiment comprises the composition of embodiment 10, wherein the copper comprises one or more of copper, copper nitrate, copper sulfate, or copper hydroxide.

[0080] A twelfth embodiment comprises the composition of any of the preceding embodiments, wherein the chitosan comprises between about 1.2 wt% and about 2.2 wt% of the composition.

[0081] A thirteenth embodiment comprises the composition of any of the preceding embodiments, wherein the chitosan comprises about 1.7 wt% of the composition.

[0082] A fourteenth embodiment comprises the composition of any of the preceding embodiments, further comprises an acid.

[0083] A fifteenth embodiment comprises the composition of embodiment 14, wherein the acid comprises one or more of 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.

[0084] A sixteenth embodiment comprises the composition of any of the preceding embodiments, further comprises a coupling enhancer.

[0085] A seventeenth embodiment comprises the composition of embodiment 16, wherein the coupling enhancer is sodium tripolyphosphate.

[0086] A eighteenth embodiment comprises the composition of embodiment 17, wherein the fungicidal compound is comprised in a nanoparticle.

[0087] An nineteenth embodiment comprises the composition of any of the preceding embodiments, wherein the fungicidal activity of the composition is increased by between about 5% and about 50% as compared to the same fungicide not in a composition with chitosan.

[0088] A twentieth embodiment comprises the composition of any of the preceding embodiments, wherein the composition comprises between about 10 wt% and about 40 wt% nitrogen.

[0089] A twenty-first embodiment comprises the composition of embodiment 19, wherein the composition comprises about 20 wt% nitrogen.

[0090] A twenty-second embodiment comprises the composition of embodiment 20, wherein at least 50% of the nitrogen is in the chemical form of amine and / or nitrate and / or ammonia.Docket No.10860-10910-US

[0091] A twenty-third embodiment comprises the composition of any of the preceding embodiments, wherein the composition has antifungal activity against pythium irregulare.

[0092] In a twenty-fourth embodiment, a composition comprises: chitosan, and an insecticidal compound.

[0093] A twenty-fifth embodiment comprises the composition of embodiment 24, wherein the chitosan is present in the composition in an amount sufficient to enhance the insecticidal activity of the insecticidal compound.

[0094] A twenty-sixth embodiment comprises the composition of embodiment 24, wherein the chitosan and the insecticidal compound, together have synergistic insecticidal activity.

[0095] A twenty-seventh embodiment comprises the composition of any of embodiments 24- 26, wherein the chitosan comprises ultra-low molecular weight, low molecular weight, or high molecular weight chitosan.

[0096] A twenty-eighth embodiment comprises the composition of embodiments 24-27, wherein the majority of the chitosan in the composition by weight is ultra-low molecular weight, low molecular weight, or high molecular weight chitosan.

[0097] A twenty-ninth embodiment comprises the composition of embodiment 24, wherein the chitosan comprises between about 1.2 wt% and about 2.2 wt% of the composition.

[0098] A thirtieth embodiment comprises the composition of embodiment 29, wherein the chitosan is about 1.7 wt% of the composition.

[0099] A thirty-first embodiment comprises the composition of embodiment 24, wherein the chitosan has a molecular weight between about 4 kDa and about 500 kDa. [000100] A thirty-second embodiment comprises the composition of embodiment 31, wherein the chitosan has a molecular weight between about 20 kDa and about 380 kDa. [000101] A thirty-third embodiment comprises the composition of embodiment 32, wherein the chitosan has a molecular weight between about 50 kDa and about 300 kDa. [000102] A thirty-fourth embodiment comprises the composition of embodiment 33, wherein the chitosan has a molecular weight between about 100 kDa and about 250. [000103] A thirty-fifth embodiment comprises the composition of embodiment 34, wherein the chitosan has a molecular weight between about 150 kDa and about 200 kDa. [000104] A thirty-sixth embodiment comprises the composition of embodiment 24, further comprises an acid.Docket No.10860-10910-US [000105] A thirty-seventh embodiment comprises the composition of embodiment 36, wherein the acid comprises one or more of 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. [000106] A thirty-eighth embodiment comprises the composition of embodiment 24, further comprises a coupling enhancer. [000107] A thirty-ninth embodiment comprises the composition of embodiment 38, wherein the coupling enhancer is sodium tripolyphosphate. [000108] A fortieth embodiment comprises the composition of any of embodiments 24-39, wherein the insecticidal compound comprises one or more of a spinosyn, a phenylethanol, Malathion, Parathion, Diazinon, Chlorpyrifos, Carbaryl, Methomyl, Aldicarb, Propoxur, Permethrin, Cypermethrin, Deltamethrin, Bacillus thuringiensis Lambda-cyhalothrin, Imidacloprid, Clothianidin, Thiamethoxam, Acetamiprid, Dichlorodiphenyltrichloroethane (DDT), Lindane, Chlorane, Pyrethrins, Rotenone, Azadirachtin, Beauveria bassiana, Metarhizium anisopliae, Methoprene, Pyriproxfen, Fenoxycarb, Diflubenzuron, Sarin, Tabun, Soman, VX, Sulfur, and Diatomaceous Earth.. [000109] A forty-first embodiment comprises the composition of embodiment 40, wherein the spinosyn comprises at least two chemically distinct spinosyns. [000110] A forty-second embodiment comprises the composition of embodiment 41, wherein the at least two chemically distinct spinosyns are spinosyn A and spinosyn D. [000111] A forty-third embodiment comprises the composition of embodiment 24, wherein the composition comprises between about 10 wt% and about 30 wt% nitrogen. [000112] A forty-fourth embodiment comprises the composition of embodiment 24, wherein the composition comprises about 20 wt% nitrogen. [000113] A forty-fifth embodiment comprises the composition of embodiment 44, wherein at least 50% of the nitrogen is in the chemical form of amine and / or nitrate and / or ammonia. [000114] In a forty-sixth embodiment, a composition comprises: chitosan and an herbicidal compound. [000115] A forty-seventh embodiment comprises: the composition of embodiment 46, wherein the chitosan comprises between about 1.2 wt% and about 2.2 wt% of the composition.Docket No.10860-10910-US [000116] A forty-eighth embodiment comprises: the composition of embodiment 47, wherein the chitosan is about 1.7 wt% of the composition. [000117] A forty-ninth embodiment comprises: the composition of embodiment 47, wherein the chitosan has a molecular weight between about 4 kDa and about 60 kDa. [000118] A fiftieth embodiment comprises: the composition of embodiment 49, wherein the chitosan has a molecular weight between about 4 kDa and about 16 kDa. [000119] A fifty-first embodiment comprises: the composition of embodiment 50, wherein the chitosan has a molecular weight between about 6 kDa and about 12 kDa. [000120] A fifty-second embodiment comprises: the composition of embodiment 51, wherein the chitosan has a molecular weight of about 8 kDa. [000121] A fifty-third embodiment comprises: the composition of embodiment 49, wherein the chitosan has a molecular weight between about 20 kDa and about 60 kDa. [000122] A fifty-fourth embodiment comprises: the composition of embodiment 53, wherein the chitosan has a molecular weight between about 30 kDa and about 50 kDa. [000123] A fifty-fifth embodiment comprises: the composition of embodiment 54, wherein the chitosan has a molecular weight of about 40 kDa. [000124] A fifty-sixth embodiment comprises: the composition of embodiment 46, further comprises an acid. [000125] A fifty-seventh embodiment comprises: the composition of embodiment 56, wherein the acid comprises one or more of 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. [000126] A fifty-eighth embodiment comprises: the composition of embodiment 46, further comprises a coupling enhancer. [000127] A fifty-ninth embodiment comprises: the composition of embodiment 58, wherein the coupling enhancer is sodium tripolyphosphate. [000128] A sixtieth embodiment comprises: the composition of any of embodiments 46-59 wherein the herbicidal compound comprises one or more of a photosynthesis inhibitor, an auxin mimic, a microtubule assembly inhibitor, a cell membrane disruptor, or an amino acid synthesis inhibitor.Docket No.10860-10910-US [000129] A sixty-first embodiment comprises: the composition of embodiment 60, wherein the amino acid synthesis inhibitor is glyphosate. [000130] A sixty-second embodiment comprises: the composition of any of embodiments 46- 61, wherein the composition comprises between about 10 wt% and about 40 wt% nitrogen. [000131] A sixty-third embodiment comprises: the composition of embodiment 62, wherein the composition comprises about 20 wt% nitrogen. [000132] A sixty-fourth embodiment comprises: the composition of embodiment 63, wherein at least 50% of the nitrogen is in the chemical form of amine and / or nitrate and / or ammonia. [000133] In a sixty-fifth embodiment, a composition comprises: chitosan, and a nematocidal compound. [000134] A sixty-sixth embodiment comprises: the composition of embodiment 65, wherein the chitosan comprises between about 1.2 wt% and about 2.2 wt% of the composition. [000135] A sixty-seventh embodiment comprises: the composition of embodiment 66, wherein the chitosan is about 1.7 wt% of the composition. [000136] A sixty-eighth embodiment comprises: the composition of embodiment 65, wherein the chitosan has a molecular weight between about 4 kDa and about 60 kDa. [000137] A sixty-ninth embodiment comprises: the composition of embodiment 68, wherein the chitosan has a molecular weight between about 4 kDa and about 16 kDa. [000138] A seventieth embodiment comprises: the composition of embodiment 69, wherein the chitosan has a molecular weight between about 6 kDa and about 12 kDa. [000139] A seventy-first embodiment comprises: the composition of embodiment 70, wherein the chitosan has a molecular weight of about 8 kDa. [000140] A seventy-second embodiment comprises: the composition of embodiment 65, wherein the chitosan has a molecular weight between about 20 kDa and about 60 kDa. [000141] A seventy-third embodiment comprises: the composition of embodiment 72, wherein the chitosan has a molecular weight between about 30 kDa and about 50 kDa. [000142] A seventy-fourth embodiment comprises: the composition of embodiment 73, wherein the chitosan has a molecular weight of about 40 kDa. [000143] A seventy-fifth embodiment comprises: the composition of embodiment 65, further comprises an acid. [000144] A seventy-sixth embodiment comprises: the composition of embodiment 75, wherein the acid comprises one or more of Acetic acid, Citric acid, Lactic acid, Malic acid, Tartaric acid,Docket No.10860-10910-US 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. [000145] A seventy-seventh embodiment comprises: the composition of embodiment 65, further comprises a coupling enhancer. [000146] A seventy-eighth embodiment comprises: the composition of embodiment 77, wherein the coupling enhancer is sodium tripolyphosphate. [000147] A seventy-ninth embodiment comprises: the composition of any of embodiments 65- 78, wherein the nematocidal compound comprises one or more of methyl bromide, 1,3- Dichloropropene, Dazomet, Aldicarb, Fenamiphos, Oxamyl, Bacillus thuringiensis, Paecilomyces lilacinus, Pochonia chlamydosporia, Azadirachtin, nicotinamides, thiophenes, Terpenoids, Flavenoids, Saponins Burkholderia spp., bacillus fermis, bacillus spp., pasteuria nishizawae, fluopyram, bacillus amyloiquefaciens, pyriflumetofen, and chromobacterium subtsugae. [000148] An eightieth embodiment comprises: the composition of any of embodiments 65-79, the composition comprises between about 10 wt% and about 40 wt% nitrogen. [000149] An eighty-first embodiment comprises: the composition of embodiment 80, wherein the composition comprises about 20 wt% nitrogen. [000150] An eighty-second embodiment comprises: the composition of embodiment 81, wherein at least 50% of the nitrogen is in the chemical form of amine and / or nitrate and / or ammonia. [000151] In an eighty-third embodiment a composition comprises any two or more of the compositions of groups A, B, C, and D, wherein group A comprises the compositions of embodiments 1-23, wherein group b comprises the compositions of embodiments 24-45, wherein group C comprises the compositions of embodiments 46-64, and wherein group D comprises the compositions of embodiments 65-82. [000152] An eighty-fourth embodiment comprises: a method of producing crop-protective chitosan nanoparticles, the method comprises: dissolving chitosan in acidic water, where the water is made acidic by an acid selected from the group of 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 HumicDocket No.10860-10910-US acid, Fulvic acid, Amino acids , Nucleic acids (DNA and RNA), Boric acid, Chromic acid, Cyanuric acid, Hyaluronic acid, Arsenic acid and Carboxylic acids; dissolving a crop-protective compound in the acidic water; and forming nanoparticles from the dissolved chitosan and the crop protective compound. [000153] An eighty-fifth comprises: the method of embodiment 84, wherein the chitosan comprises between about 1.2 wt% and about 2.2 wt% of the composition. [000154] An eighty-sixth embodiment comprises: the method of embodiment 85, wherein the chitosan is about 1.7 wt% of the composition. [000155] An eighty-seventh embodiment comprises: the method of embodiment 84, wherein the chitosan has a molecular weight between about 4 kDa and about 160 kDa. [000156] An eighty-eighth embodiment comprises: the method of embodiment 87, wherein the chitosan has a molecular weight between about 4 kDa and about 16 kDa. [000157] An eighty-ninth embodiment comprises: the method of embodiment 88, wherein the chitosan has a molecular weight between about 6 kDa and about 12 kDa. [000158] A ninetieth embodiment comprises: the method of embodiment 89, wherein the chitosan has a molecular weight about 8 kDa. [000159] A ninety-first embodiment comprises: the method of embodiment 87, wherein the chitosan has a molecular weight between about 20 kDa and about 50 kDa. [000160] A ninety-second embodiment comprises: the method of embodiment 91, wherein the chitosan has a molecular weight between about 25 kDa and about 40 kDa. [000161] A ninety-third embodiment comprises: the method of embodiment 92, wherein the chitosan has a molecular weight of about 32 kDa. [000162] A ninety-fourth embodiment comprises: the method of embodiment 93, wherein the chitosan has a molecular weight between about 60 kDa and about 100 kDa. [000163] A ninety-ninth embodiment comprises: the method of embodiment 87, wherein the chitosan has a molecular weight between about 75 kDa and about 85 kDa. [000164] A one hundredth embodiment comprises: the method of embodiment 98, wherein the chitosan has a molecular weight of about 78 kDa. [000165] A one hundred and first embodiment comprises: the method of embodiment 87, wherein chitosan has a molecular weight between about 120 kDa and about 160 kDa. [000166] A one hundred and second embodiment comprises: the method of embodiment 101, wherein chitosan has a molecular weight between about 130 kDa and about 150 kDa.Docket No.10860-10910-US [000167] A one hundred and third embodiment comprises: the method of embodiment102, wherein the chitosan has a molecular weight of about 141 kDa. [000168] A one hundred and fourth embodiment comprises: the method of embodiment 83, further comprises a coupling enhancer. [000169] A one hundred and fifth embodiment comprises: the method of embodiment 104, wherein the coupling enhancer is sodium tripolyphosphate. [000170] A one hundred and sixth embodiment comprises: the method of any of embodiments 83-105, wherein the crop-protective compound is an insecticidal compound. [000171] A one hundred and seventh embodiment comprises: the method of embodiment 106, wherein the insecticidal compound comprises one or more of a spinosyn, a phenylethanol, Malathion, Parathion, Diazinon, Chlorpyrifos, Carbaryl, Methomyl, Aldicarb, Propoxur, Permethrin, Cypermethrin, Deltamethrin, Lambda-cyhalothrin, Imidacloprid, Clothianidin, Thiamethoxam, Acetamiprid, Dichlorodiphenyltrichloroethane (DDT), Lindane, Chlorane, Pyrethrins, Rotenone, Azadirachtin, Beauveria bassiana, Metarhizium anisopliae, Methoprene, Pyriproxfen, Fenoxycarb, Diflubenzuron, Sarin, Tabun, Soman, VX, Sulfur, and Diatomaceous Earth.. [000172] A one hundred and eighth embodiment comprises: the method of embodiment 107, wherein the spinosyn comprises at least two chemically distinct spinosyns. [000173] A one hundred and ninth embodiment comprises: the method of embodiment 108, wherein the at least two chemically distinct spinosyns are spinosyn A and spinosyn D. [000174] A one hundred and tenth embodiment comprises: the method of cany of embodiments 83-105, wherein the crop-protective compound is an herbicidal compound. [000175] A one hundred and eleventh embodiment comprises: the method of embodiment 110, wherein the herbicidal compound comprises one or more of a photosynthesis inhibitor, an auxin mimic, a microtubule assembly inhibitor, a cell membrane disruptor, or an amino acid synthesis inhibitor. [000176] A one hundred and twelfth embodiment comprises: the method of embodiment 111, wherein the amino acid synthesis inhibitor is glyphosate. [000177] A one hundred and thirteenth embodiment comprises: the method of any of embodiments 83-105, wherein the crop protective compound is a fungicide. [000178] A one hundred and fourteenth embodiment comprises: the method of embodiment 113, wherein the fungicidal compound comprises one or more of copper, chlorothalonil, Azoxystrobin, Pyraclostrobin, Trifloxystrobin, Fluoxastobin, Picoxystrobin, Kresoxim methyl, Epoxiconazole, tebuconazole, Propiconazole, Cyproconazole, dienconazole, metconazole, Boscalid,Docket No.10860-10910-US Carboxin, Flutoanil, Sedaxane, Fluopyram, Penflufen, Fluxaapyroxad, Fludioxonil, Cymoxanil, Forety-al, Propamocarb, Mandipropamid, Cyazofamid, Famoxadone, Metafenone, Isoprothiolane, Validymycin, Fenhexamid, Pyroquilon, Carpropamid, Fenamidone, Fluopocolide, Harpin, Oxolinic acid, Aluminum sulfate, Bentonite clay, chitosan, Copper Octanoate, Cuprous Oxide, Peptides, Phosphoric Acids, Potassium Bicarbonate, Potassium iodide, Potassium phosphite, Potassium silicate, Potassium thiocyanate, Sodium bicarbonate, Sulfur, Tribasic copper sulphate, Hydrogen Peroxide, Chenopodium quinoa saponins, citric acid, Clove Oil, Coconut Oil, Esquisetum arvense, Eugenol, Garlic Oil, Gu-lutathione, Lamarin, Melaleuca, Peppermint Oil, Phospholipids, Propolis extract, Propylene glycol, Reynoutria, Rosemary Oil, Saponins, Seaweed Extract, Sodium lautyl, Soybean Oil, Sucrose esters, Terpenese, Thyme Oil, NatamycinPolyoxin D Zinc Salt, Bacteriophage, Metschnikowia, Mancozeb, Thiram, Propineb, Prothioconazole. Probenazole, Tricyclazole, Prochloraz, Copper, Sulphur, Fentin, Oxine Copper, Chlorothalonil, Captan, Folpet, Metaxyl, Benalaxyl, Oxadixyl, Carbendazim, Thiophanate, Thiabendazole, Benomyl, Fuberidazole, Spiroxamine, dimethomorph, Fenpropidin, Fenproprimorph, Cypodinil, Pyrimethanil, Mepanipyrim, Fluazinam, Dithianon, Pencycuron, Iminoctadine, Tolylfuanid, Iprodione, Procymidone, Vincllozolin, Fenarimol, Bupirimate, Triforine, Ferimzone, and Pyrifenox. [000179] A one hundred and fifteenth embodiment comprises: the method of embodiment 114, wherein the copper comprises one or more of copper, copper nitrate, copper sulfate, or copper hydroxide. [000180] A one hundred and sixteenth embodiment comprises: the method of any of embodiments 83-105, wherein the crop-protective compound is a nematicide. [000181] A one hundred and seventeenth embodiment comprises: the method of embodiment 116, wherein the nematocidal compound comprises one or more of methyl bromide, 1,3- Dichloropropene, Dazomet, Aldicarb, Fenamiphos, Oxamyl, Bacillus thuringiensis, Paecilomyces lilacinus, Pochonia chlamydosporia, Azadirachtin, nicotinamides, thiophenes, Terpenoids, Flavenoids, Saponins Burkholderia spp., bacillus fermis, bacillus spp., pasteuria nishizawae, fluopyram, bacillus amyloiquefaciens, pyriflumetofen, and chromobacterium subtsugae. [000182] A one hundred and eighteenth embodiment comprises: the method of any of embodiments 79-117, wherein the composition comprises between about 10 wt% and about 30 wt% nitrogen. [000183] A one hundred and nineteenth embodiment comprises: the method of embodiment 118, wherein the composition comprises about 20 wt% nitrogen. [000184] A one hundred and twentieth embodiment comprises: the method of embodiment 119, wherein at least 50% of the nitrogen is in the chemical form of amine and / or nitrate and / or ammonia.Docket No.10860-10910-US Example 1: Preparation of chitosan using various acids [000185] Structural and compositional engineering of chitosan [000186] Chitosan, a natural biopolymer derived from chitin, possesses remarkable properties but dissolves poorly in water due to its inherent hydrophobic nature. Obtaining a stable chitosan solution with optimum structural properties is crucial for various applications, prompting the need for an efficient dissolution method. For the following sub example chitosan or chitin were derived from or more of the following sources - Chitosan is derived from chitin, a natural polymer that is abundantly found in the exoskeletons of crustaceans, insects, and the cell walls of fungi. Chitin is the second most abundant biopolymer on Earth, following cellulose. The source of chitosan can be Crustacean Shells such as Crab shells, Shrimp shells, Lobster shells, Crawfish shells; Insect Exoskeletons, such as beetles and ants; Fungi; Squid Pens; The pen or gladius of squid contains chitin; Algae; Some types of algae contain chitin. Waste from Seafood Processing; Shrimp and crab shell waste generated from seafood processing can be a valuable source of chitosan. Chitosan can also be produced through the fermentation of certain bacteria. Solution 1.1. [000187] A predetermined quantity of chitosan flakes (e.g., 5 grams) was measured and placed in a clean, dry container. A predetermined volume of peracetic acid (e.g., 50 mL) was added to the container containing chitosan flakes. The mixture was stirred continuously at a moderate temperature (e.g., 40°C) until the chitosan flakes completely dissolved. The pH of the solution was adjusted using sodium hydroxide or acetic acid to achieve the desired range (e.g., pH 4-6). The chitosan solution was diluted with water, if necessary, to ensure the stability of the solution. Solution 1.2. [000188] A desired quantity of chitosan flakes (e.g., 5 grams) was measured and placed in a clean, dry container. A predetermined volume of hydrochloric acid (e.g., 50 mL) was added to the container containing chitosan flakes. The mixture was stirred continuously at a moderate temperature (e.g., 60°C) until the chitosan flakes were completely dissolved. The pH of the solution was adjusted when needed using sodium hydroxide or hydrochloric acid to achieve the desired range (e.g., pH 4-6). The chitosan solution was diluted with water, when necessary, to ensure the stability of the solution. Solution 1.3. [000189] A desired quantity of chitosan flakes (e.g., 5 grams) was measured and placed in a clean, dry container. A predetermined volume of acetic acid (e.g., 100 mL) was added to the container containing chitosan flakes. The mixture was stirred continuously at a moderate temperature (e.g., 50°C) until the chitosan flakes were completely dissolved. The pH of the solution was adjusted whenDocket No.10860-10910-US necessary, using sodium hydroxide or acetic acid to achieve the desired range (e.g., pH 4-6). The chitosan solution was diluted with water, when necessary, to ensure the stability of the solution. Solution 1.4. [000190] A predetermined quantity of chitosan flakes (e.g., 5 grams) was measured and placed in a clean, dry container. A predetermined volume of hydrogen peroxide (e.g., 30% concentration, 50 mL) was added to the container containing chitosan flakes. The mixture was stirred continuously at a moderate temperature (e.g., 35°C) until the chitosan flakes were completely dissolved. The pH of the solution was adjusted using sodium hydroxide or acetic acid to achieve the desired range (e.g., pH 4- 6). The chitosan solution was dissolved with water, if necessary, to ensure the stability of the solution. Solution 1.5. [000191] A predetermined quantity of chitosan flakes (e.g., 5 grams) was measured and placed in a clean, dry container. A predetermined volume of lactic acid (e.g., 50 mL) was added to the container containing chitosan flakes. The mixture was stirred continuously at a moderate temperature (e.g., 40°C) until the chitosan flakes were completely dissolved. The pH of the solution was adjusted using sodium hydroxide or lactic acid to achieve the desired range (e.g., pH 4-6). The chitosan solution was diluted with water when necessary, ensuring the stability of the solution. Solution 1.6. [000192] A predetermined quantity of chitosan flakes (e.g., 5 grams) were measured and placed in a clean, dry container. A predetermined volume of citric acid (e.g., 50 mL) was added to the container containing chitosan flakes. The mixture was stirred continuously at a moderate temperature (e.g., 45°C) until the chitosan flakes were completely dissolved. The pH of the solution was adjusted using sodium hydroxide or citric acid to achieve the desired range (e.g., pH 4-6). The chitosan solution with water if necessary, ensuring the stability of the solution. Solution 1.7. [000193] A predetermined quantity of chitosan flakes (e.g., 5 grams) was measured and placed in a sealed reactor or chamber. CO2 gas was introduced into the reactor under controlled pressure and temperature conditions (e.g., 30°C, 50 bar). The CO was allowed to interact with the chitosan flakes, leading to dissolution without the need for additional solvents. The processing conditions were adjusted to control the dissolution rate and achieve the desired chitosan concentration. The system was vented to release any excess CO2and obtain the stable chitosan solution. Solution 1.8. [000194] A predetermined quantity of chitosan flakes (e.g., 5 grams) was measured and placed in a clean, dry container. A predetermined volume of sulfuric acid (e.g., 50 mL) was added to theDocket No.10860-10910-US container containing chitosan flakes. The mixture was stirred continuously at a controlled temperature (e.g., 25°C) until the chitosan flakes were completely dissolved. The concentration of sulfuric acid was adjusted to modulate the solubility of chitosan based on the desired application. The chitosan solution was adjusted with water if necessary, ensuring the stability of the solution. Solution 1.9 [000195] A predetermined quantity of chitosan flakes (e.g., 5 grams) was measured and placed in a clean, dry container. A predetermined volume of nitric acid (e.g., 50 mL) was added to the container containing chitosan flakes. The mixture was stirred continuously at a controlled temperature (e.g., 30°C) until the chitosan flakes were completely dissolved. The concentration of nitric acid was adjusted to introduce nitrogen into the chitosan matrix. The chitosan solution was diluted with water if necessary, ensuring the stability of the solution. Solution 1.10 [000196] A predetermined quantity of chitosan flakes (e.g., 5 grams) was measured and placed in a clean, dry container. Dissolution with Formic Acid: a predetermined volume of formic acid (e.g., 50 mL) was added to the container containing chitosan flakes. The mixture was stirred continuously at a controlled temperature (e.g., 40°C) until the chitosan flakes were completely dissolved. Dissolution with Malic Acid: After the chitosan flakes were dissolved in formic acid, a predetermined volume of malic acid (e.g., 30 mL) was added to the solution. The solution was stirred at the same temperature until the chitosan was fully dissolved in the malic acid. Dissolution with Tartaric Acid: Following the dissolution in malic acid, add a predetermined volume of tartaric acid (e.g., 20 mL) to the solution. The mixture was stirred until the chitosan flakes were completely dissolved in tartaric acid. Adjustment and Dilution: The pH of the chitosan solution was adjusted using sodium hydroxide or acetic acid to achieve the desired range (e.g., pH 4-6). The chitosan solution was diluted with water, if necessary, to ensure the stability of the solution. Solution 1.11 [000197] A predetermined quantity of chitosan flakes (e.g., 5 grams) was measured and placed in a clean, dry container. A predetermined volume of acid or acid mixture was added. The acid or acid mixture was composed or any one or two or three or four or all organic acids such as (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), Inorganic Acids (Hydrochloric acid, Sulfuric acid, Nitric acid, Phosphoric acid, Hydrofluoric acid, Perchloric acid, Hydrobromic acid, Hydroiodic acid, Chloric acid, Bromic acid, Iodic acid) or Other Acids forms such as: Humic acid, Fulvic acid, Amino acids , Nucleic acids (DNA and RNA), Boric acid, Chromic acid, Cyanuric acid, Hyaluronic acid, Arsenic acid and Carboxylic acids (e.g., 50 mL) to the container containing chitosan flakes. The mixture was stirred continuously at a controlled temperature (e.g., 25°C)Docket No.10860-10910-US until the chitosan flakes were completely dissolved. The concentration of acid or combination of acid was adjusted to modulate the solubility of chitosan-based on the desired application. The chitosan solution was diluted with water, if necessary, to ensure the stability of the solution. Solution 1.12 [000198] The chitin-rich source material or chitosan flake or chitin flake was weighed (e.g., 100 grams) and placed in a clean, dry container. A chitinase enzyme was prepared. The chitinase enzyme was one of [Chitinase A (ChiA), Chitinase B (ChiB), and Chitinase C (ChiC) originate from the bacterium Serratia marcescens. Chitinase D (ChiD) is sourced from Bacillus circulans, while Chitinase E (ChiE) is obtained from Bacillus cereus, both belonging to the bacterial kingdom. Streptomyces coelicolor is the bacterium from which Chitinase F (ChiF) is derived, and Chitinase G (ChiG) is sourced from Streptomyces lividans. Chitinase I (ChiI) comes from Streptomyces griseus, and Arthrobacter sp. serves as the source for Chitinase II (ChiII). Chitinase III (ChiIII) is found in Serratia proteamaculans, and once again, Serratia marcescens is the source for Chitinase IV (ChiIV). Pseudomonas aeruginosa contributes Chitinase V (ChiV), while Bacillus thuringiensis provides Chitinase VI (ChiVI) and Chitinase VIII (ChiVIII). Streptomyces griseus is the source for Chitinase VII (ChiVII). Pseudomonas fluorescens is the bacterium providing Chitinase IX (ChiIX). Fungi also contribute to chitinase diversity, with Chitinase X (ChiX) sourced from Aspergillus fumigatus, Chitinase XI (ChiXI) from Aspergillus oryzae, Chitinase XII (ChiXII) from Trichoderma reesei, and Chitinase XIII (ChiXIII) from Talaromyces flavus.] solution in a buffer (eg phosphate buffer) (having pH between 4-6), maintaining a concentration of 1 mg / ml. The chitin-rich source material (100 g) was mixed with the chitinase enzyme solution. The pH of the mixture was adjusted to 5.0 using buffer (eg. Phosphate buffer). The mixture was incubated at 50°C for 24 hours. The progress of hydrolysis was monitored by sampling aliquots at regular intervals. Termination of Enzymatic Reaction: the enzymatic reaction was stopped by adjusting the pH to 9.0 using sodium hydroxide. Chitosan Separation: the reaction mixture was centrifuged at 10,000 rpm for 15 minutes to separate the chitosan from the solution. The supernatant containing chitosan was collected. Dialysis: the collected supernatant was placed in a dialysis membrane and dialyzed against deionized water to remove small molecules, salts, and residual enzymes. The water was changed periodically until the dialysate became neutral. Chitosan Precipitation: the pH of the dialyzed solution was adjusted to 4-5 using acetic acid. Chitosan precipitated; the precipitate was collected by centrifugation. Neutralization: the chitosan was neutralized, when necessary, with a sodium hydroxide solution to achieve the desired pH. Table 1 shows several chitosan and acid formulations, and several properties of each formulation. Table 1Docket No.10860-10910-USExample 2: Preparation of Coupling Enhancer - Tri sodium Polyphosphate (TPP) solution [000199] Step 1. Take 0.200 gram in 50 mL capacity volumetric flask. Step 2. Add 30 mL water. Step 3. Vortex the solution – resultant solution becomes clear. Step 4. Make up the volume to 50 mL using 20 mL water. The total volume of the solution in mL was 50 mL: 49.821 g. The calculated % TP of the solution was 0.4 % w / v. The calculated P% of the solution was 0.101%. The pH of the solution was 9.76. Table 2 shows several chitosan, linker, and acid formulations, and several properties of each formulation. Table 2Docket No.10860-10910-USExample 3: Preparation of chitosan – crop protection composition [000200] Developing a mixture of chitosan with various fungicides, nematicides, insecticides, and herbicides involves careful consideration of compatibility, efficacy, and environmental impact. Here's a general method along with potential examples: [000201] Chitosan Solution Preparation: Prepare the chitosan solution as described in Example 1. [000202] Individual Pesticide Solutions: Prepare individual solutions of the fungicide, nematicide, insecticide, and herbicide in their respective recommended solvents (see the list in Table 1). [000203] Mixture Preparation: Combine the chitosan solution with individual pesticide solutions in a suitable mixing vessel. Stir gently to achieve a homogeneous mixture. [000204] Adjuvant or coupling enhancer Addition: Adjuvants / linkers may be used for enhanced efficacy, incorporate them into the mixture and ensure thorough mixing. [000205] The potential adjuvants include - Surfactants: Non-ionic 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), Penetrants: Crop oil concentrates, Methylated seed oils. Buffering Agents: Ammonium sulfate, Potassium dihydrogen phosphate, Compatibility Agents: Polyvinyl alcohol, Polyacrylic acid; Antifoaming Agents: Silicone-based antifoams, Polyethylene glycol-based antifoams; pH Adjusters: Ammonium hydroxide, Citric acid; Thickeners: Guar gum, Xanthan gum; Humectants: Glycerol, Propylene glycol. [000206] Chitosan cross-linkers (such as Glutaraldehyde, Genipin, Epichlorohydrin, Tripolyphosphate or sodium tripolyphosphate, Sodium Hexametaphosphate, Polyphosphates. Sufates such as ammonium sulphate, dextran Sulfate, Ethylenediamine, Tartaric Acid, Urea, sodium trimetaphosphate) are chemical agents or compounds that facilitate the formation of cross-links between chitosan molecules, resulting in the creation of a three-dimensional network or structure. ThisDocket No.10860-10910-US cross-linking enhances the mechanical, chemical, and thermal properties of chitosan, making it suitable for various applications. [000207] pH Adjustment: Check and adjust the pH of the mixture to ensure compatibility and stability. Chitosan solutions typically work well in a slightly acidic to neutral pH range. Table 3 shows several pesticide formulations, some with chitosan, linker, and acid formulations, and several properties of each formulation. Table 3Docket No.10860-10910-USDocket No.10860-10910-USDocket No.10860-10910-USDocket No.10860-10910-USExample 4: Formulation of Copper-Chitosan Fungicides [000208] SOP of TGBCNP2 – composition:19.7% Nitrate, 10.1% Copper (II), 1% Chitosan, 0.4% Acetic Acid, 0.01% STPP, 0.0001% Tween 80 (Polysorbate 80). [000209] Method (1000g batch) [000210] The dry mass of a 2L beaker was measured. A water bath was heated to 90°C. Deionized water was preheated to 75-85°C in a separate container. 384.3g of copper nitrate (II) trihydrate was dissolved into 460g of preheated deionized water. The solution was mixed at 300 rpm (50mm prop; overhead) until homogeneous in the 90°C water bath. Homogenization took less than 5 minutes.10g of chitosan LMW was added. The chitosan was soaked in salt solution at temperature for 30 minutes with continuous mixing (600 rpm; overhead 50mm prop mixer). The Chitosan mass addedDocket No.10860-10910-US was not adjusted for moisture content, which is between 10-14% for CSL-2-2156. 5g of 80% acetic acid was added. This was blended for 3 hours (600 rpm). Acetic acid was added by transfer pipette over about 15 seconds. During blending, the solution evaporated. 1000g of deionized water was kept in the water bath to regularly replace evaporated mass to avoid overconcentration. The water was added slowly by pipette and hot to avoid precipitation of copper complexes. The solution was filtered while hot using 80 µm nylon filter (cool metal mesh will cause crystallization unless preheated). The beaker was returned to the water bath. Filtration was done by prewarming a 80µm nylon mesh bag, then positioning over a secondary beaker (dry mass recorded), and hand pouring the sample through the bag. The secondary beaker containing the filtered solution was returned to the water and mixer. This process took less than 3 minutes from the time the sample is removed from the water bath to the time it is returned to the water in the secondary beaker. A 150-200g stock sodium tripolyphosphate solution was made at 714.3 ppm concentration in deionized water. Mix until dissolved. Preheat stock solution in water at (90°C).10g of stock tween 80 (polysorbate 80) solution at 1µL tween per gram of solution was made. Mixed gently to avoid foaming. Dropwise with mixing (600 rpm; overhead 50mm prop mixer), 140g of the preheated STPP solution prepared in step 7 was added to the filtered chitosan-copper solution. 999g of final solution was made, correcting for mass loss with hot water while mixing. The mixing was reduced to 300 rpm and add 1g of tween 80 solution prepared in step 8. The solution is cooled to room temperature. Then filtered by 80µm and 5µm filtration. Example 5: Chitosan Compatibility Test with Agrochemicals [000211] The compatibility test between chitosan and crop protection agrochemicals such as fungicide, insecticide, herbicide and nematicide, encompassing both liquid and dry formulations, is a crucial step to ascertain the efficacy and stability of the resulting mixture. For the liquid formulation assessment, a chitosan solution is prepared and diluted with the liquid agrochemical, followed by observation for potential signs of precipitation, flocculation, or phase separation. pH adjustment is performed if necessary, and the mixture is scrutinized for any changes in appearance after a designated observation period. Similar steps are undertaken for the dry formulation compatibility test, where chitosan powder is evenly mixed with the agrochemical dry formulation, and the resulting mixture is inspected for uniformity and absence of clumps. Both assessments consider visual indicators, pH measurements, and overall stability to determine the compatibility of chitosan with agrochemicals. These tests ensure that the combination maintains the desired properties for effective agricultural applications, providing valuable insights into the formulation's suitability and performance in the field. Example 6: Application and uses of the chitosan and / or chitosan / nitrogen nanoparticles [000212] The resultant chitosan and / or chitosan loaded nitrogen nanoparticle products or mixtures of products obtained as a result of the above Examples were used for the multidimensional effects on plants such as growth, protection, and nutrition. The multidimensional effects on plants wereDocket No.10860-10910-US obtained by applying the resultant product of the above Examples onto the seed surface, mixing in the soil, exposing root and / or exposing leaves. The application rate ranged between 0.01% to 50% v / w or v / v. Furthermore, the resultant products of the above Examples were used to mix with other agrochemicals to act as growth promoter, symbiont, nutrient, protection agent against pathogens or non- desired plants for synergistic effects. Example 7: Efficacy Testing. [000213] Evaluating the efficacy of chitosan combined with fungicides, insecticides, herbicides, and nematicides involves a comprehensive approach encompassing both in vitro and in vivo assessments. In vitro studies focus on antifungal, insecticidal, herbicidal, and nematocidal activities through controlled experiments with target organisms or plants. Parameters such as inhibition zones, mortality rates, and growth patterns serve as indicators of efficacy. Moving to in vivo trials, greenhouse and field experiments provide insights into the real-world impact on crops. Greenhouse trials allow for controlled conditions, monitoring disease, insect, or nematode incidence, while field trials assess crop yield, overall health, and economic damage thresholds. Additional analyses, including soil microbial studies and residue analysis, offer a holistic understanding of the formulations' effects on the agroecosystem. Throughout these studies, careful consideration of replication, control groups, monitoring periods, and statistical analyses ensures robust and reliable assessments, guiding the development and optimization of chitosan-based formulations for sustainable and effective agricultural practices. [000214] Chitosan and Herbicide formulations [000215] Experimental Procedure: Chitosan flake acetate flake (Mw 141 kDa) was dissolved in an acidic solution and heated for targeted time periods to reduce the average Mw to A) 40 kDa or B) 8 kDa. The resulting solutions contained 1.7 wt% chitosan + 1 wt% acetic acid and were used as is for the following examples. [000216] Herbicide example 1: Herbicide control (Chitosan solution without herbicide): [000217] Experimental Procedure: [000218] Greenhouse flats filled with potting soil were sowed with 7 common crops: wheat, corn, ryegrass, mustard, sunflower, melon and beans. Each row (oriented across the flats) contained between 8 - 20 seeds depending on the seed size. For each treatment, three flats were sowed. After two weeks in the greenhouse, the seedlings were of sufficient size to be sprayed. Chitosan solutions (1.7 wt%, 1 wt% acidic acid) were diluted with tap water to 1063 ppm chitosan and applied to the trays (get spray rate). After spraying, the flat continued to be watered and fertilized for 3 weeks at regular intervals. During this period, visible herbicide damage (necrosis and stunting) was assessed. Herbicide Table 1a: Effects of Chitosan Mw on Wheat125Docket No.10860-10910-USHerbicide Table 1b: Effects of Chitosan Mw on CornHerbicide Table 1c: Effects of Chitosan Mw on RyegrassHerbicide Table 1d: Effects of Chitosan Mw on MustardHerbicide Table 1e: Effects of Chitosan Mw on SunflowerHerbicide Table 1f: Effects of Chitosan Mw on MelonHerbicide Table 1g: Effects of Chitosan Mw on BeansDocket No.10860-10910-US [000219] The chitosan solution had a bio-stimulant effect on the crops. Generally, 40 kDa acidic chitosan solution had a larger bio-stimulant effect on all crops than the 8 kDa acidic chitosan solution. [000220] Herbicide example 2: [000221] Experimental Procedure: [000222] Greenhouse flats filled with potting soil were sowed with 7 common crops: wheat, corn, ryegrass, mustard, sunflower, melon and beans. Each row (oriented across the flats) contained between 8 - 20 seeds depending on the seed size. For each treatment, three flats were sowed. After two weeks in the greenhouse, the seedlings were of sufficient size to be sprayed. Chitosan solutions (1.7 wt%, 1 wt% acidic acid) were diluted with tap water to various ppm chitosan concentrations, spiked with the herbicide 1120 ppm Glyphosate, K salt (Roundup Weathermax) and applied to the trays (get spray rate). After spraying, the flat continued to be watered and fertilized for 3 weeks. During this period, visible herbicide damage (necrosis and stunting) was assessed. Herbicide Table 2ai: Effects of Chitosan Mw with Herbicide on WheatHerbicide Table 2aii: Effects of Chitosan Mw with Herbicide on WheatHerbicide Table 2bi: Effects of Chitosan Mw with Herbicide on CornHerbicide Table 2bii: Effects of Chitosan Mw with Herbicide on CornDocket No.10860-10910-USHerbicide Table 2ci: Effects of Chitosan Mw with Herbicide on RyegrassHerbicide Table 2cii: Effects of Chitosan Mw with Herbicide on RyegrassHerbicide Table 2di: Effects of Chitosan Mw with Herbicide on MustardHerbicide Table 2dii: Effects of Chitosan Mw with Herbicide on MustardHerbicide Table 2ei: Effects of Chitosan Mw with Herbicide on SunflowerDocket No.10860-10910-USHerbicide Table 2eii: Effects of Chitosan Mw with Herbicide on SunflowerHerbicide Table 2fi: Effects of Chitosan Mw with Herbicide on MelonHerbicide Table 2fii: Effects of Chitosan Mw with Herbicide on MelonHerbicide Table 2gi: Effects of Chitosan Mw with Herbicide on BeansHerbicide Table 2gii: Effects of Chitosan Mw with Herbicide on BeansDocket No.10860-10910-US [000223] Generally, 40 kDa acidic chitosan solution had reduced stunting rates observed at 14 days vs. the 8 kDA acidic chitosan solution. Stunting tended to continue increasing at 21 days with 8 kDa chitosan, yet, not with 40 kDa chitosan. Interestingly, necrosis ratings for 4 crops tended to decrease with the 8kDa chitosan but not with the 40 kDa chitosan between 14 and 21 days. [000224] Herbicide example 3: [000225] Experimental Procedure: [000226] Greenhouse flats filled with potting soil were sowed with 7 common crops: wheat, corn, ryegrass, mustard, sunflower, melon and beans. Each row (oriented across the flats) contained between 8 - 20 seeds depending on the seed size. For each treatment, three flats were sowed. After two weeks in the greenhouse, the seedlings were of sufficient size to be sprayed. Chitosan solutions (1.7 wt%, 1 wt% acidic acid) were diluted with tap water, followed by A) an herbicide solution, mixed till uniform, addition of the TPP solution or B) TPP solution mixed till uniform followed by an herbicide solution. In both examples, the final solutions contained 531 ppm chitosan, 1120 ppm Glyphosate, K salt and 37 when applied to the trays (get spray rate). After spraying, the flat continued to be watered and fertilized for 3 weeks. During this period, visible herbicide damage (necrosis and stunting) was assessed. Herbicide Table 3ai: Effects of Order of Addition of Coupling Enhancer to Chitosan with Herbicide on WheatOrder of addition: A) Chitosan solution, glyphosate solution, TPP solution; B) Chitosan solution, TPP solution, glyphosate solution. Herbicide Table 3aii: Effects of Order of Addition of Coupling enhancer to Chitosan with Herbicide on WheatOrder of addition: A) Chitosan solution, glyphosate solution, TPP solution; B) Chitosan solution, TPP solution, glyphosate solution. Herbicide Table 3bi: Effects of Order of Addition of Crosslinker to Chitosan with Herbicide on CornDocket No.10860-10910-USOrder of addition: A) Chitosan solution, glyphosate solution, TPP solution; B) Chitosan solution, TPP solution, glyphosate solution Herbicide Table 3bii: Effects of Order of Addition of Coupling enhancer to Chitosan with Herbicide on CornOrder of addition: A) Chitosan solution, glyphosate solution, TPP solution; B) Chitosan solution, TPP solution, glyphosate solution Herbicide Table 3ci: Effects of Order of Addition of Coupling enhancer to Chitosan with Herbicide on RyegrassOrder of addition: A) Chitosan solution, glyphosate solution, TPP solution; B) Chitosan solution, TPP solution, glyphosate solution Herbicide Table 3cii: Effects of Order of Addition of Coupling enhancer to Chitosan with Herbicide on RyegrassOrder of addition: A) Chitosan solution, glyphosate solution, TPP solution; B) Chitosan solution, TPP solution, glyphosate solution Herbicide Table 3di: Effects of Order of Addition of Coupling enhancer to Chitosan with Herbicide on MustardDocket No.10860-10910-US Order of addition: A) Chitosan solution, glyphosate solution, TPP solution; B) Chitosan solution, TPP solution, glyphosate solution Herbicide Table 3dii: Effects of Order of Addition of Coupling enhancer to Chitosan with Herbicide on MustardOrder of addition: A) Chitosan solution, glyphosate solution, TPP solution; B) Chitosan solution, TPP solution, glyphosate solution Herbicide Table 3ei: Effects of Order of Addition of Coupling enhancer to Chitosan with Herbicide on SunflowerOrder of addition: A) Chitosan solution, glyphosate solution, TPP solution; B) Chitosan solution, TPP solution, glyphosate solution Herbicide Table 3eii: Effects of Order of Addition of Coupling enhancer to Chitosan with Herbicide on SunflowerOrder of addition: A) Chitosan solution, glyphosate solution, TPP solution; B) Chitosan solution, TPP solution, glyphosate solution Herbicide Table 3fi: Effects of Order of Addition of Coupling enhancer to Chitosan with Herbicide on MelonOrder of addition: A) Chitosan solution, glyphosate solution, TPP solution; B) Chitosan solution, TPP solution, glyphosate solutionDocket No.10860-10910-US Herbicide Table 3fii: Effects of Order of Addition of Coupling enhancer to Chitosan with Herbicide on MelonOrder of addition: A) Chitosan solution, glyphosate solution, TPP solution; B) Chitosan solution, TPP solution, glyphosate solution Herbicide Table 3gi: Effects of Order of Addition of Coupling enhancer to Chitosan with Herbicide on Beanssolution, glyphosate solution Herbicide Table 3gii: Effects of Order of Addition of Coupling enhancer to Chitosan with Herbicide on BeansOrder of addition: A) Chitosan solution, glyphosate solution, TPP solution; B) Chitosan solution, TPP solution, glyphosate solution [000227] In 8 of 14 experiments resulted in order of addition B chitosan + TPP + herbicide) having more stunting (and to a lesser extend necrosis) than the chitosan + herbicide or chitosan + herbicide + TPP samples. [000228] Herbicide Example 4 [000229] Experimental Procedure [000230] Palmer amaranth (Amaranthus palmeri), common lambsquarters (Chenopodium album), green foxtail (Seraria viridis) and velvetleaf (Abutilon theophrasti) were transplanted into plastic cones with dimensions of 2.5” by 10” (6.4 cm x 25.4 cm) and a volume is 40 in³ or 656 ml. Seeds were initially germinated in aluminum trays (Figure 2). The soil used was ProMix General Purpose growing medium (Premier Tech, Quakertown PA, USA). Plants were watered manually as needed with 5-1-4 fertilizer (Wilbur Ellis, San Francisco, CA, USA) injected into irrigation water at aDocket No.10860-10910-US rate of 0.2%. All pots were treated with Bacillus thuringiensis subsp israelensis (Gnatrol, Valent, Libertyville, IL, USA) at a rate of 0.49 g l-1for control of fungal gnats. Supplemental light (Phillips lighting) was provided as needed to maintain a 16 hour photo period. Planting and treatment dates are given in Table 4.1. Herbicide Table 4.1 Planting, treatment dates, and height of weed species used in this study. Species Run Planted Treated Height (in) Palmer amaranth 1 11 / 19 / 24 12 / 17 / 24 3-5 Palmer amaranth 2 11 / 19 / 24 12 / 17 / 24 3-5 Common lambsquarters 1 11 / 8 / 24 12 / 8 / 24 3-5 Common lambsquarters 2 11 / 8 / 24 12 / 8 / 24 3-5 Green foxtail 1 11 / 19 / 24 12 / 13 / 24 3-4 Green foxtail 2 11 / 19 / 24 12 / 13 / 24 3-4 Velvetleaf 1 11 / 11 / 24 12 / 8 / 24 3-5 Velvetleaf 2 11 / 11 / 24 12 / 8 / 24 3-5 [000231] Treatments (Table 2) were applied with a three nozzle enclosed track sprayer (Devries Manufacturing, Hollandale, MN, USA) (Figure 2) equipped with a AIXR11004 (TeeJet, Wheaton IL, USA) nozzles. Sprayer traveling velocity was 10.56 miles hr-1with a pressure of 40 PSI to deliver a carrier volume application of 15 gallons acre-1. Nozzle height was 15” above the plants. Herbicide Table 4.2. Treatments used in the study. Trt Treatment Rate11. Untreated Check 2. RUPM3 + AMS 20 + 17 3. RUPM3 + AMS + Chitosan 20 + 17 + 0.25 4. RUPM3 + AMS + Chitosan 20 + 17 + 0.5 5. RUPM3 + AMS + Chitosan 20 + 17 + 1.0 6. AMS + Chitosan 17 + 1.0 1Roundup PowerMax 3 in fl oz A-1, AMS in lbs 100 gal-1, Chitosan in v v-1. [000232] The plants were removed from the spray chamber 10 seconds after spraying occurred and relocated to a greenhouse under climate-controlled conditions. Visual evaluations were made and pictures of rep 1 plants taken at 7, 14, and 21 days after treatment (DAT). At 21 DAT, above-ground biomass was harvested and plants were dried to constant biomass. Dry biomass weights were recorded and converted into percentage of biomass reduction compared to non-treated control using equation 1 (in which NT represents the mean biomass of non-treated plants and T represents the biomass of the treated plants):100 [1]Docket No.10860-10910-US [000233] All data were subjected to ANOVA using a mixed model (PROC GLIMMIX) in SAS (Statistical Analysis Software, version 9.4, Cary, NC, USA) with replication set to the random and means separation at α=0.05 using Tukey’s HSD test. [000234] The plants were evaluated visually, with graphs 4.3, 4.4, 4.5, and 4.6 depicting percentage reductions in growth. Photos of the plants, which help illustrate the visual evaluation of plant injury taken at 7, 14, and 21 days after treatment (DAT). Injury used as reference to rate plants include necrosis and stunting (Figures 5A, 5B, 5C) (Figures 6A, 6B.6C) (Figures 7A, 7B, 7C) (Figures 8A, 8B, 8C) (Figures 9A, 9B, 9C) (Figures 10A, 10B, 10C) (Figures 11A, 11B, 11C) (Figures 12A, 12B, 12C). These tests indicate that chitosan has a synergistic effect in at least some situations when used with a glyphosate herbicide. The palmer amaranth study in particular illustrates this synergy. In at the seven day after treatment review of the plants, the formulation which uses 0.5 % v / v chitosan increased the plant injury by 10% over the glyphosate alone. The 1.0 % v / v chitosan formulation increased the plant injury by 7% over the glyphosate alone. Additionally, the dry biomass reduction showed a 5% reduction in biomass with the 0.5 % v / v over the glyphosate alone. Herbicide Table 4.3. Visual estimation of injury and dry biomass reduction of Palmer amaranth. 7 Tr DA 14 Dry Biomass t Product Rate1T2,3DAT 21 DAT Reduction _______________________________________________%_____________________________________ 2RUPM3 + AMS20 + 1786 B 94 B 95 A 90.5 A3 RUPM3 + AMS + 20 + 17 + 0.25 Chitosan 95 A 98 AB 99 A 94.7 A 4 RUPM3 + AMS + 20 + 17 + 0.5 Chitosan 96 A 100 A 100 A 95.4 A 5 RUPM3 + AMS + 20 + 17 + 1.0 Chitosan 93 AB 99 A 99 A 95.0 A 6AMS + Chitosan 17 + 1.01 C 0 C 0 B 0.0 B1Roundup PowerMax 3 in fl oz A-1, AMS in lbs 100 gal-1, Chitosan in v v-1. 2Means followed by the same letter within a column are not different using Tukey’s HSD at alpha=0.05. 3 Visual evaluations at 7 DAT, 14 DAT, and 21 DAT were on a 0 to 100 scale with 0 being no observed injury and 100 being complete plant death. Herbicide Table 4.4. Visual estimation of injury and dry biomass reduction of common lambsquarters. 7 Dry DA 14 Biomass Trt T2,3DAT 21 DAT____________ 20 + 17 2 RUPM3 + AMS 85 B 94 A 95 A 95.2 ADocket No.10860-10910-US RUPM3 20 + 17 + 0.25 3 + AMS +Chitosan 96 A 94.0 A RUPM3 + AMS + 20 + 17 + 0.5 4 Chitosan98 A 95.0 A RUPM3 + AMS + 20 + 17 + 1.0 5 Chitosan94 A 93.5 A AMS + Chitosan 17 + 1.0 60 B 0.0 B 1Roundup PowerMax 3 in fl oz A-1, AMS in lbs 100 gal-1, Chitosan in v v-1. 2Means followed by the same letter within a column are not different using Tukey’s HSD at alpha=0.05. 3 Visual evaluations at 7 DAT, 14 DAT, and 21 DAT were on a 0 to 100 scale with 0 being no observed injury and 100 being complete plant death. Herbicide Table 4.5. Visual estimation of injury and dry biomass reduction of green foxtail. 7 Dry DA 14 Biomass Trt T2,3DAT 21 DAT____________ 2 RUPM3 + AMS 20 + 17 83 A 95 A 100 A 96.9 A 3 RUPM3 + AMS 20 + 17 + 0.25 + Chitosan 79 A 91 A 98 A 94.2 A 4 RUPM3 + AMS 20 + 17 + 0.5 + Chitosan 79 A 92 A 99 A 96.1 A 5 RUPM3 + AMS 20 + 17 + 1.0 + Chitosan 82 A 93 A 100 A 96.2 A 6 AMS + Chitosan 17 + 1.0 0 B 0 B 0 B 0.0 B 1Roundup PowerMax 3 in fl oz A-1, AMS in lbs 100 gal-1, Chitosan in v v-1. 2Means followed by the same letter within a column are not different using Tukey’s HSD at alpha=0.05. 3 Visual evaluations at 7 DAT, 14 DAT, and 21 DAT were on a 0 to 100 scale with 0 being no observed injury and 100 being complete plant death. Herbicide Table 4.6. Visual estimation of injury and dry biomass reduction of velvetleaf. 7 Dry DA 14 Biomass Trt Product Rate1T2,3DAT 21 DAT Reduction______________ 20 + 17 2 RUPM3 + AMS 70 A 86 A 93 A 91.5 A RUP 20 + 17 + 0.25 3 M3 + AMS 64+ Chitosan B 90 A 88.6 A RUPM3 + AMS 20 + 17 + 0.5 4 + Chitosan 68 AB94 A 92.5 ADocket No.10860-10910-US RUPM3 + AMS 20 + 17 + 1.0 5 + Chitosan 67 AB 83 A 91 A 90.5 A AMS + Chitosan 17 + 1.0 6 0 C 0 B 0 B 0.0 B 1Roundup PowerMax 3 in fl oz A-1, AMS in lbs 100 gal-1, Chitosan in v v-1. 2Means followed by the same letter within a column are not different using Tukey’s HSD at alpha=0.05. 3 Visual evaluations at 7 DAT, 14 DAT, and 21 DAT were on a 0 to 100 scale with 0 being no observed injury and 100 being complete plant death. [000235] Chitosan and Fungicide Preparations [000236] Chitosan stock solution preparation example 1: [000237] Experimental Procedure: [000238] Chitosan flake acetate flake (Mw 141 kDa) was dissolved in an acidic solution and heated for targeted time periods to reduce the average Mw to 7.6 kDa. The resulting solution contained 1.7 wt% chitosan + 1 wt% acetic acid and were used as is for the following examples. [000239] Chitosan stock solution preparation example 2: [000240] Experimental Procedure: [000241] Chitosan flake acetate flake (Mw 250 kDa) was dissolved in an acidic solution and heated for targeted time periods to reduce the average Mw to 6.8 kDa. The resulting solution contained 1.7 wt% chitosan + 1 wt% acetic acid and were used as is for the following examples. [000242] Fungicide example 1: [000243] Experimental Procedure: [000244] Potato dextrose broth stock solutions were spiked with technical grade active ingredient mixed with dimethyl sulfoxide as a solubilizing agent. Additionally, potato dextrose broth stock solutions were spiked with an acidic chitosan solution that has been diluted with distilled water. These two types of solutions will each contain 4X the amount of active ingredient or chitosan reported in the final array.50 ul aliquots from both types of solutions were dispensed in triplicate into microtiter plates and stored at -80C until use. On the day of the test, the prepared arrays were thawed and mixed with 100 ul of 5x104CFU / ml of Fusarium oxysporum inoculum prepared in potato dextrose broth. The plates are incubated (get condition) for two days, then the contents of each well were spread onto petri dishes (get specifics) (each condition in triplicate) and allow to incubate two additional days. After 4 days of total experimentation time, the number of colonies were counted per plate. Wells with < 25 colonies were identified has the MFC (the equivalent to 99.9% death of the microorganism in the test).Docket No.10860-10910-US [000245] Chitosan stock solution preparation example 1: [000246] Experimental Procedure: [000247] Chitosan flake acetate flake (Mw 141 kDa) was dissolved in an acidic solution and heated for targeted time periods to reduce the average Mw to 7.6 kDa. The resulting solution contained 1.7 wt% chitosan + 1 wt% acetic acid and were used as is for the following examples. [000248] Chitosan stock solution preparation example 2: [000249] Experimental Procedure: [000250] Chitosan flake acetate flake (Mw 250 kDa) was dissolved in an acidic solution and heated for targeted time periods to reduce the average Mw to 6.8 kDa. The resulting solution contained 1.7 wt% chitosan + 1 wt% acetic acid and were used as is for the following examples. [000251] Fungicide example 1: [000252] Experimental Procedure: Potato dextrose broth stock solutions were spiked with technical grade active ingredient mixed with dimethyl sulfoxide as a solubilizing agent. Additionally, potato dextrose broth stock solutions were spiked with an acidic chitosan solution that has been diluted with distilled water. These two types of solutions will each contain 4X the amount of active ingredient or chitosan reported in the final array. 50 ul aliquots from both types of solutions were dispensed in triplicate into microtiter plates and stored at -80C until use. On the day of the test, the prepared arrays were thawed and mixed with 100 ul of 5x104CFU / ml of Fusarium oxysporum inoculum prepared in potato dextrose broth. The plates are incubated for two days, then the contents of each well were spread onto petri dishes (each condition in triplicate) and allow to incubate two additional days. After 4 days of total experimentation time, the number of colonies were counted per plate. Wells with < 25 colonies were identified has the MFC (the equivalent to 99.9% death of the microorganism in the test). Fungicide Table 1a: Select examples of Azoxystrobin + various chitosan gradesDocket No.10860-10910-US5 Fungicide Table 1b: Select examples of Chlorothalonil + various chitosan gradesDocket No.10860-10910-US Fungicide Table 1c: Select examples of Fludioxonil + various chitosan grades[000253] Fungicide example 2: [000254] Experimental Procedure: [000255] Potato dextrose broth stock solutions were spiked with technical grade active ingredient mixed with dimethyl sulfoxide as a solubilizing agent. Additionally, potato dextrose broth stock solutions were spiked with an acidic chitosan solution that has been diluted with distilled water. Lastly, potato dextrose broth stock solutions were spiked with sodium tripolyphosphate (TPP)Docket No.10860-10910-US solution that has been diluted with distilled water. These three types of solutions will each contain 4X the amount of reagent reported in the final array.50 ul aliquots from the three types of solutions were dispensed in triplicate into microtiter plates and stored at -80C until use. On the day of the test, the prepared arrays were thawed and mixed with 50 ul of 1x105CFU / ml of Fusarium oxysporum inoculum prepared in potato dextrose broth. The plates are incubated (get conditions) for two days, then the contents of each well were spread onto petri dishes (get specifics) (each condition in triplicate) and allow to incubate two additional days. After 4 days of total experimentation time, the number of colonies were counted per plate. Wells with < 25 colonies were identified has the MFC (the equivalent to 99.9% death of the microorganism in the test). Fungicide Table 2a: Select examples of Azoxystrobin + various chitosan grades + TPPDocket No.10860-10910-USFungicide Table 2b: Select examples of Chlorothalonil + various chitosan grades + TPP[000256] Chitosan stock solution preparation example 3: [000257] Experimental Procedure: [000258] Chitosan flake acetate flake (Mw 141 kDa) was dissolved in an acidic solution and heated for targeted time periods to reduce the average Mw to A) 118 kDa; B) 39 kDa and C) 6.8 kDa. The resulting solution contained 1.7 wt% chitosan + 1 wt% acetic acid and were used as is for the following examples. [000259] Chitosan acetate preparation example 4: [000260] Experimental Procedure: [000261] A suitable amount of acidic chitosan solution (Mw 6.8 kDa) was in a freeze dryer and run for 24 hours. The resulting chitosan acetate powder was used as is for the following examples. [000262] Fungicide Example 3 [000263] Potato dextrose broth stock solutions were spiked with technical grade active ingredient mixed with dimethyl sulfoxide as a solubilizing agent. Additionally, potato dextrose broth stock solutions were spiked with an acidic chitosan solution that has been diluted with distilled water. Two separate chitosan types were used. PT-1685 an Ultralow Molecular Weight (ULMW) Papaya Process Chitosan. And TgA a Low Molecular Weight (LMW) Chitosan. Both types of chitosan were prepared in solutions which were diluted to 0.75% chitosan. On the day of the test, the prepared arraysDocket No.10860-10910-US were thawed and mixed with 100 ul of 5x104CFU / ml of Fusarium oxysporum inoculum prepared in potato dextrose broth. The plates are incubated for two days, then the contents of each well were spread onto petri dishes and allowed to incubate two additional days. After 4 days of total experimentation time, the number of colonies were counted per plate. Wells with < 25 colonies were identified has the MFC (the equivalent to 99.9% death of the microorganism in the test). Fungicide Table 3a: Examples of Chitosan and Azoxystrobin fungicide solutionsFungicide Table 3b: Examples of Chitosan and Chlorothalonil fungicide solutionsDocket No.10860-10910-US Fungicide Table 3c: Examples of Chitosan and Pyraclostrobin fungicide solutionsFungicide Table 3d: Examples of Chitosan and Fludioxonil fungicide solutions[000264] Fungicide Example 4Docket No.10860-10910-US [000265] The following examples were used to determine the synergism between chitosan products and fungicides to determine minimum inhibitory concentrations (MIC) of various formulations of chitosan and fungicides. These examples were further used to assess the effect chitosan has on the effectiveness of the fungicides. The MIC measurement studies were performed using a microtiter plate bioassay. The test fungus was Fusarium oxysporum I3-4 seeded at 2.5x104cfu / mL. From the MIC tests, compositions showing effectiveness or antifungal synergism were selected to evaluate minimum fungicidal concentration (MFC). The MFC tests were performed on potato dextrose agar. The test products were combinations of chitosan and a fungicide. The chitosan used in these analysis were EXP- 12960 and EXP-12982. The EXP-12960 chitosan was formulated with glacial acetic acid. The EXP- 12982 chitosan was formulated with hydrochloric acid. The fungicides used were azoxystrobin, pyraclostrobin, propiconazole, prothioconazole, and fluopyram. [000266] In a first MIC test EXP-12960 which is a chitosan preparation with 2 wt% LMW Chitosan and 6 wt% glacial acetic acid was combined with various amounts of azoxystrobin. Additionally, acetic acid and various amounts of azoxystrobin were tested to identify what effect acetic acid had on fungal growth and to show that all effects on fungal growth were because of the LMW chitosan in the composition. The analysis was run in three replicates. The analysis indicates that acetic acid has no effect on reducing fungal growth. Tables 4a, 4b, and 4c show the fungal growth where 0 is no growth, 1 is 25% growth, 2 is 50% growth, 3 is 75% growth, and 4 is 100% growth. Figure 14 is a photo of the first replicate showing that the chitosan composition EXP-12960 inhibits fungal growth, while acetic acid does not inhibit fungal growth. Fungicide Table 4aDocket No.10860-10910-USFungicide Table 4cDocket No.10860-10910-US[000267] The chitosan formulation EXP-12982 which is a chitosan preparation with 2 wt% LMW Chitosan and 0.67 wt% hydrochloric acid was analyzed in combination with azoxystrobin, pyraclostrobin, propiconazole, prothioconazole, and fluopyram. [000268] Table 4d shows the effect of the chitosan formulation EXP-12982 with azoxystrobin, to determine the MIC. The chitosan formulation EXP-12982 in combination with azoxystrobin as shown in Figure 14B is effective beginning with a 16ppm fungicidal concentration. This indicates that the EXP-12982 chitosan formulation in combination with azoxystrobin is more effective than the EXP- 12982 chitosan formulation and at least as effective as azoxystrobin alone. Fungicide Table 4d[000269] Table 4e shows the effect of the chitosan formulation EXP-12982 with pyraclostrobin to determine the MIC. The chitosan formulation EXP-12982 in combination with pyraclostrobin as shown in Figure 14BC is effective beginning with a 16ppm fungicidal concentration. This indicates that the EXP-12982 chitosan formulation in combination with pyraclostrobin is more effective than the EXP-12982 chitosan formulation and at least as effective as pyraclostrobin alone.Docket No.10860-10910-US Fungicide Table 4e[000270] Table 4f shows the effect of the chitosan formulation EXP-12982 with propiconazole to determine the MIC. The chitosan formulation EXP-12982 in combination with propiconazole as shown in Figure 14D is effective beginning with a 0.5-16ppm fungicidal concentration. This indicates that the EXP-12982 chitosan formulation in combination with propiconazole is more effective than the EXP-12982 chitosan formulation and more effective than propiconazole alone. The EXP-12982 chitosan formulation in combination with propiconazole shows synergistic effects over EXP-12982 and propiconazole. Fungicide Table 4fDocket No.10860-10910-US[000271] Table 4g shows the effect of the chitosan formulation EXP-12982 with prothioconazole to determine the MIC. The chitosan formulation EXP-12982 in combination with prothioconazole as shown in Figure 14E is effective beginning with a 0.5-16ppm fungicidal concentration. This indicates that the EXP-12982 chitosan formulation in combination with pyraclostrobin is more effective than the EXP-12982 chitosan formulation and at least as effective as pyraclostrobin alone. Fungicide Table 4g[000272] Table 4h shows the effect of the chitosan formulation EXP-12982 with fluopyram to determine the MIC. The chitosan formulation EXP-12982 in combination with fluopyram as shown in Figure 14F is effective beginning with a 0.5-16ppm fungicidal concentration. This indicates that theDocket No.10860-10910-US EXP-12982 chitosan formulation in combination with fluopyram is more effective than the EXP-12982 chitosan formulation and at least as effective as fluopyram alone. Fungicide Table 4h[000273] Fungicide Example 5 [000274] A composition comprising multiple molecular weights of chitosan may be effective against multiple pathogens. A study was conducted to determine the effectiveness of different molecular weights of chitosan against different pathogens. A fungicide was prepared with three types of chitosan an ULMW, LMW, and HMW chitosan. Tables 5.1 and 5.2. Fungicide Table 5.1 Fungicide Table 5.2Docket No.10860-10910-US [000275] Further compositions were prepared; a composition with ULMW chitosan, a composition with LMW chitosan, and a composition with HMW chitosan. The ULMW chitosan has a molecular weight of between about 3.2 to about 3.9 kDa. The LMW chitosan has a molecular weight of between about 191 to about 256 kDa. The HMW chitosan has a molecular weight of between about 360 and about 543 kDa. Table 5.3. Fungicide Table 5.3[000276] The five compositions were tested against several types of pathogens at several different concentrations. The pathogens include; pythium, phytophthora, fusarium oxysporum, tricoderma. [000277] The compositions were tested against pythium. Concentrations of 100 ppm and 300 ppm were utilized. In the 100ppm test, the ULMW chitosan formulation showed the most growth inhibition of the pythium. With a 25% + growth inhibition of pythium. The LMW chitosan formulation shows no growth inhibition of the pythium. At concentrations of 300 ppm all chitosan formulations show 100% growth inhibition of pythium. [000278] The compositions were tested against phyytophthora. Concentrations of 100 ppm, 300 ppm, and 500 ppm were utilized. In all three tests, the ULMW chitosan formulation showed the most growth inhibition of the phytophthora. The ULMW formulation drives the pathogen control of phytophthora. [000279] The compositions were tested against fusarium oxysporum. Concentrations of 100 ppm, 300 ppm, and 500 ppm were utilized. Chitosan may not be effective at concentrations below 100 ppm against fusarium oxysporum. At concentration of 300 ppm and 500 ppm all compositions performed similarly. There was slight improvement in growth inhibition at the higher 500 ppm concentration. [000280] The compositions were tested against tricoderma viride. Concentrations of 100 ppm, 300 ppm, and 500 ppm were utilized. Chitosan may not be effective at concentrations below 100 ppm against tricoderma viride. At concentration of 300 ppm and 500 ppm all compositions performed similarly. There was slight improvement in growth inhibition at the higher 500 ppm concentration.Docket No.10860-10910-US [000281] The compositions were tested against xanthomonas citri. Concentrations of 100 ppm, 300 ppm, and 500 ppm were utilized. Chitosan may not be effective at any concentrations against xanthomonas citri. There may be slight growth inhibition at 500 ppm concentration. [000282] The compositions were tested against rhizobium sp. Concentrations of 100 ppm, 300 ppm, and 500 ppm were utilized. All concentrations have good pathogen control. All concentrations completely prevent growth of rhizobium sp. [000283] Compositions which include multiple chitosan formulations at multiple different molecular weights may be effective as a crop protective compound against multiple different pathogens. Table 5.4 illustrates the effectiveness of compositions with multiple formulations against different pathogens. The combination composition Spectra 1.2 performs best at 300 ppm and 500 ppm. Fungicide Table 5.4[000284] Bactericide [000285] Suitable amounts of water transferred into 50 ml falcon tubes. For examples the 1.7 wt% acidic chitosan solutions were diluted with a suitable amount of water. [000286] Anti-bacterial assay was used to determine the Minimum Inhibitory Concentration against Xanthomonas citri. The bioassay contained three replicates per treatment. A selected optimal agar medium for the bacterial strain was prepared and sterilized. The diluted bactericidal solutions were added to the growth medium in flasks, mixed gently, and 20 ml media was poured into three plates for each dilution. An untreated medium is taken as a control to validate the colony growth. Streptomycin treatment with concentration of 0.025 mg / ml will be tested as positive control. A single colony was grown in a selected broth medium, and inoculum suspension containing 108CFU / ml ofDocket No.10860-10910-US bacteria was measured and immediately inoculated on the media plate as four spots (1μl per spot) using a multichannel pipettor. The plates were covered and incubated at 30 °C for 72 hours. MIC endpoint will be recorded based on the absence of colony growth. Incubation time was extended for further growth observation of weak colonies. Bactericide Table 1a: Select examples of acidic chitosan solutions controlling X. citri[000287] For examples 4-6, 10-12 below, in 50 ml falcon tubes, 1.7 wt% acidic chitosan solutions were diluted with a suitable amount of water. This was followed by acetic acidic (equivalent to 0.5 wt% of the final mass) to acidify the water. After the copper sulfate was added, the resulting mixture was heated at 50C for X hours to ensure full dissolution of the copper salt. When necessary, the solution was vortexed to assist dissolution. The solutions were allowed to cold and then their pH’s measured. The resulting solutions contained 6 wt% elemental copper and were then suitably diluted further when tested in the anti-bacterial assay below. [000288] For examples 7-9 below, suitable amounts of water followed by acetic acidic (equivalent to 0.5 wt% of the final mass) was added to the 50 ml falcon tubes to acidify the water. Next a dry chitosan acetate salt (6.8 kDa Mw) was added. After the chitosan salt dissolved, the copper sulfate was added. The resulting mixture was heated at 50C for X hours to ensure full dissolution of the copper salt. When necessary, the solution was vortexed to assist dissolution. The solutions were allowed to cold and then their pH’s measured. The resulting solutions contained 6 wt% elemental copper and were then suitably diluted further when tested in the anti-bacterial assay below. Bactericide Table 1b: Select examples of acidic chitosan copper sulfate solutions controlling X. citriDocket No.10860-10910-US [000289] For examples 13-17 below, in 50 ml falcon tubes, a 1.7 wt% acidic chitosan solution (6.8 kDa Mw) was diluted with a suitable amount of water. This was followed by acetic acidic (equivalent to 0.5 wt% of the final mass) to acidify the water. After the copper sulfate was added, the resulting mixture was heated at 50C for X hours to ensure full dissolution of the copper salt. When necessary, the solution was vortexed to assist dissolution. The solutions were allowed to cold and then their pH’s measured. The resulting solutions contained 1 wt% to 0.1 wt% elemental copper and were then suitably diluted further when tested in the anti-bacterial assay below. [000290] For examples 18 and 19 below, suitable amounts of water followed by acetic acidic (equivalent to 0.5 wt% of the final mass) was added to the 50 ml falcon tubes to acidify the water. Next a dry chitosan acetate salt (6.8 kDa Mw) was added. After the chitosan salt dissolved, the copper sulfate was added. The resulting mixture was heated at 50C for X hours to ensure full dissolution of the copper salt. When necessary, the solution was vortexed to assist dissolution. The solutions were allowed to cold and then their pH’s measured. The resulting solutions contained 0.1 wt% elemental copper and were then suitably diluted further when tested in the anti-bacterial assay below. [000291] Anti-bacterial assay was used to determine the Minimum Inhibitory Concentration against Xanthomonas citri. The bioassay contained three replicates per treatment. A selected optimal agar medium for the bacterial strain was prepared and sterilized. The diluted bactericidal solutions were added to the growth medium in flasks, mixed gently, and 20 ml media was poured into three plates for each dilution. An untreated medium is taken as a control to validate the colony growth. Streptomycin treatment with a concentration of 0.025 mg / ml will be tested as positive control. A single colony was grown in a selected broth medium, and inoculum suspension containing 108CFU / ml of bacteria was measured and immediately inoculated on the media plate as four spots (1μl per spot) using a multichannel pipettor. The plates were covered and incubated at 30 °C for 72 hours. MIC endpoint will be recorded based on the absence of colony growth. Incubation time was extended for further growth observation of weak colonies. Bactericide Table 1c: Select examples of acidic chitosan copper sulfate solutions controlling X. citriDocket No.10860-10910-US [000292] Increasing the chitosan > copper improved the efficacy. Next step would be increasing the loading of the solids to get the Cu loading to 1 wt% or 1.7 wt% as a solution and or slurry. Bactericide example 2: [000293] Suitable amounts of copper hydroxide powders (Kocide 3000 or Chem-Impex assay grade) were transferred into 50 ml falcon tubes. In the majority of experiments, a 6.8 kDa Mw chitosan acetate salt was additionally added to the tube. In one instant, sodium tripolyphosphate (TPP) was all so added. All tubes were inverted multiple times to ensure mixing of the solid powders. The total amount of solids in the tube were scaled to ensure that upon addition of ca.40 g of water, that the resulting slurries would contain 1 wt% elemental Copper. The 1 wt% Cu slurries were then suitably diluted further when tested in the anti-bacterial assay below. [000294] Anti-bacterial assay was used to determine the Minimum Inhibitory Concentration against Xanthomonas citri. The bioassay contained three replicates per treatment. A selected optimal agar medium for the bacterial strain was prepared and sterilized. The diluted bactericidal solutions were added to the growth medium in flasks, mixed gently, and 20 ml media was poured into three plates for each dilution. An untreated medium is taken as a control to validate the colony growth. Streptomycin treatment with concentration of 0.025 mg / ml will be tested as positive control. A single colony was grown in a selected broth medium, and inoculum suspension containing 108CFU / ml of bacteria was measured and immediately inoculated on the media plate as four spots (1μl per spot) using a multichannel pipettor. The plates were covered and incubated at 30 °C for 72 hours. MIC endpoint will be recorded based on the absence of colony growth. Incubation time was extended for further growth observation of weak colonies. Bactericide Table 2a: Select examples of chitosan acetate copper hydroxide blends controlling X. citri[000295] Anti-bacterial Field testDocket No.10860-10910-US [000296] An antibacterial field test was conducted to evaluate the relative efficacy of copper products on Xanthomonas citri, Phytophthora spp Septoria citri, Colletotrichum gloeosporioides additional considerations include determining whether product use offers benefits to ecosystem health indicators. TGBCNP3-007 chitosan formulation includes chitosan and copper (II) sulfate pentahydrate. A ULMW chitosan solution (1.56% chitosan PT1685 at approximately 7.1kDa, and 1% acetic acid) was combined with copper (II) sulfate pentahydrate. The solution includes 2564.13 g of chitosan PT- 1685 and 943.77 g copper (II) sulfate pentahydrate. [000297] Citrus orchards, in California, were established into plots in areas with prior citrus canker infections. Each plot contained 5 Hamlin orange trees, ensuring uniformity in tree size and health. Randomize The assignment of different in-season treatments was randomized to each plot, including treatments recommended by the client and standard industry practices. A control plot with no treatment was included to serve as a baseline for comparison. The trial of foliar treatments was initiated in late May and applied monthly. The study was run for 7-8 months to harvest (~Dec 2024). [000298] Citrus orchards (Navel or Valencia oranges) were established, in Florida, into plots in areas with prior citrus canker infections. Each plot contained 5 trees, ensuring uniformity in tree size and health. The assignment of different in-season treatments was randomized to each plot, including treatments recommended by the client and standard industry practices. A control plot with no treatment was included to serve as a baseline for comparison. The trial of foliar treatments was initiated in late November and applied monthly. The study was run for 8-9 months to harvest. [000299] Pathogen severity and incidences were evaluated pre and post application timing of foliage and fruit at regular intervals. The canopy health was qualitatively measured. Fruit abort data was collected preharvest. Total plots and a subsample of 100 fruits per plot were harvested and disease symptomology was measured. Soil samples (n = 5) at 0-12inches per plot (pre-app, midseason and harvest) were taken to assess for copper levels and to quantify taxonomic richness of bacterial, fungi and invertebrate communities. Soils samples were taken three times at pre-application, mid-season and end season. The samples were sent to an ISO certified lab that follows EPA digestion method 3050B. Soil bacteria, fungi, and invertebrate taxonomic richness were sent to CaleDNA. [000300] In-Season Treatments: The assigned treatments were administered according to the study specifications, ensuring precise application timing and dosage accuracy. 1. UTC (Untreated Control): This served as a baseline to compare the efficacy of other treatments against no treatment.2. Standard Practice - Kocide: Applied Kocide according to standard industry practices, to serve as a reference point for comparison with modified treatments. Rate: 2.5lbs / ac; Frequency (app dates) 14-30 day interval.3.Kocide Reduced Rate: Applied Kocide at a 50% lower dosage than the standard practice to assess if a reduced rate still provided effective control against Xanthomonas citri. Rate: 1.25lbs / ac; Frequency (app dates): 7 –14day interval. 4. Kocide Reduced Frequency: Applied Kocide lessDocket No.10860-10910-US frequently than the standard practice to evaluate if fewer applications still maintained adequate control over Xanthomonas citri. Rate: 2.5lbs / ac; Frequency (app dates): 28 day interval. This is on an even odd schedule. 5. Kocide Reduced Frequency + Alternated with Spectra at Low Rate for Plant Health: Alternated applications of Kocide 50% grower standard with Spectra (at 12.0 fl oz / ac) at a reduced rate to assess if this combination enhanced plant health while managing Xanthomonas citri. Rate: 12fl oz / ac of Spectra then 2.5lb / ac Kocide 3000 Frequency (app dates); 14 day spray intervals assuming heavy pressure, even / odd (Kocide / Spectra) applications; further note this is on a 14day off interval between treatments. 6. Kocide Reduced Rate with Spectra (Tank Mix): Kocide and Spectra were mixed and applied at a reduced rate to evaluate the combined effect of both products on disease control and plant health. Rate: 1.25lb / ac | 6fl oz / ac; Frequency (app dates) 14 – 30 day intervals. 7. Kocide Reduced Frequency with Spectra (Tank Mix) for Slow Release: Tank-mix Kocide and Spectra were applied together at a 28 days frequency, to evaluate for slow release of the active ingredients to sustain disease control and promote plant health over an extended period. Rate: 12fl oz / ac of Spectra then 2.5lb / ac Kocide 3000 Frequency (app dates); 28 day spray intervals. 8. TGBCNP3-007: applied every 14 –30 days, allowed for a direct comparison of efficacy versus Kocide and CS 2005 controls. 9. CS 2005 Control: Applied at a frequency that matches TGBCNP3-007, which allowed for a direct comparison of efficacy. [000301] Figures 14A, 14B, and 14C are graphs depicting the results of the tests at three different locations. The TGBCNP3-007 chitosan formulation was the most effective. Therefore, chitosan and copper have a synergistic effect. The Kocide reduced with, Spectra 50 / 50 was the next most effective. [000302] Chitosan and Insecticide Preparations [000303] Chitosan stock solution preparation example 1: [000304] Experimental Procedure 1: [000305] Chitosan acetate flake (Mw 141 kDa) was dissolved in an acetic acid solution containing 0.17 wt% peroxyacetic acid and heated for targeted time periods to reduce the average Mw to A) 78 kDa, B) 32 kDa or C) 8 kDa as measured by GPC. The resulting solutions contained 1.7 wt% chitosan + 1 wt% acetic acid + and were used as is for the following examples. [000306] Insecticide example 1: [000307] Experimental Procedure: [000308] 2.5 ml of an acidic chitosan solution containing 50 millimolar sucrose was transferred into 40 ml scintillation vial. A plug (made from Whatman filter paper) was pressed into the bottom of the vial to ensure all the sugar solution was absorbed (no free liquid). Ten sexed Drosophila Melanogaster were added to the vial followed by a cellulose plug. The flies reside in anDocket No.10860-10910-US approximately 2-3 cm headspace between the filter paper plug soaked with solution and the cellulose plug. [000309] Once the plug is in place, the experimental clock starts. The vial was observed for D. Melanogaster mortality hourly for the first day, then lest frequently over the course of two weeks. Once all D. Melanogaster have died or 2 weeks, the experiment was stopped. Efficacy differences between solutions was ranked by calculating the median survival (or LC50). Insecticide Table 1: Effects of chitosan Mw on Median Survival of D. Melanogaster[000310] Insecticide example 2: [000311] Experimental Procedure: [000312] 2.5 ml of an acidic chitosan solution containing 50 millimolar sucrose with or without 1.1 wt% 2-phenylethanol was transferred into a 40 ml scintillation vial. A plug (made from Whatman filter paper) was pressed into the bottom of the vial to ensure all the sugar solution was absorbed (no free liquid). Ten male Drosophila Melanogaster were added to the vial followed by a cellulose plug. The flies reside in an approximately 2-3 cm headspace between the filter paper plug soaked with solution and the cellulose plug. [000313] Once the plug is in place, the experimental clock starts. The vial was observed for D. Melanogaster mortality hourly for the first day, then lest frequently over the course of two weeks. Once all D. Melanogaster have died or 2 weeks, the experiment was stopped. Efficacy differences between solutions was ranked by calculating the median survival (or LC50). Insecticide Table 2: Effects of Chitosan and 1-Phenylethanol on Median Survival of Male D.MelanogasterDocket No.10860-10910-US[000314] Insecticide example 3: [000315] Experimental Procedure: [000316] 2.5 ml of an acidic chitosan solution containing 50 millimolar sucrose with or without 1.1 wt% 1-phenylethanol was transferred into a 40 ml scintillation vial. A plug (made from Whatman filter paper) was pressed into the bottom of the vial to ensure all the sugar solution was absorbed (no free liquid). Ten female Drosophila Melanogaster were added to the vial followed by a cellulose plug. The flies reside in an approximately 2-3 cm headspace between the filter paper plug soaked with solution and the cellulose plug. [000317] Once the plug is in place, the experimental clock starts. The vial was observed for D. Melanogaster mortality hourly for the first day, then lest frequently over the course of two weeks. Once all D. Melanogaster have died or 2 weeks, the experiment was stopped. Efficacy differences between solutions was ranked by calculating the median survival (or LC50). Insecticide Table 3: Effects of Chitosan and 1-Phenylethanol on Median Survival of Female D. Melanogaster[000318] Insecticide example 4:Docket No.10860-10910-US [000319] Experimental Procedure: [000320] 2.5 ml of an acidic chitosan solution containing 50 millimolar sucrose was transferred into a 40 ml scintillation vial. The control was the sucrose solution without any additives. Additional controls of 225 ppm Chitosan composition 12100 and 3 ppm TPP individually and in combination were also used, as was a control of Spinosad on its own. Further solutions include a Chitosan 12100 + Spinosad solution and a Chitosan 12100 + TPP + Spinosad solution. A plug (made from Whatman filter paper) was pressed into the bottom of the vial to ensure all the sugar solution was absorbed (no free liquid). twenty male Drosophila Melanogaster were added to the vial followed by a cellulose plug. The flies reside in an approximately 2-3 cm headspace between the filter paper plug soaked with solution and the cellulose plug. Figures 15A and 15B. [000321] 2.5 ml of an acidic chitosan solution containing 50 millimolar sucrose was transferred into a 40 ml scintillation vial. The control was the sucrose solution without any additives. Additional controls of 225 ppm Chitosan composition 12100 and 3 ppm TPP individually and in combination were also used, as was a control of 2-phenylethanol (2PE) on its own. Further solutions include a Chitosan 12100 + (2PE) solution and a Chitosan 12100 + TPP + (2PE) solution. A plug (made from Whatman filter paper) was pressed into the bottom of the vial to ensure all the sugar solution was absorbed (no free liquid). twenty male Drosophila Melanogaster were added to the vial followed by a cellulose plug. The flies reside in an approximately 2-3 cm headspace between the filter paper plug soaked with solution and the cellulose plug. Figures 15C and 15D. [000322] 2.5 ml of an acidic chitosan solution containing 50 millimolar sucrose was transferred into a 40 ml scintillation vial. The control was the sucrose solution without any additives. Additional controls of 225 ppm Chitosan composition 12126 and 3 ppm TPP individually and in combination were also used, as was a control of (2PE) on its own. Further solutions include a Chitosan + (2PE) solution and a Chitosan + TPP + (2PE) solution. A plug (made from Whatman filter paper) was pressed into the bottom of the vial to ensure all the sugar solution was absorbed (no free liquid). twenty male Drosophila Melanogaster were added to the vial followed by a cellulose plug. The flies reside in an approximately 2-3 cm headspace between the filter paper plug soaked with solution and the cellulose plug. Figures 15E and 15F. [000323] 2.5 ml of an acidic chitosan solution containing 50 millimolar sucrose was transferred into a 40 ml scintillation vial. The control was the sucrose solution without any additives. Additional controls of 225 ppm Chitosan and 3 ppm TPP individually and in combination were also used, as was a control of (2PE) on its own. Further solutions include a Chitosan + (2PE) solution and a Chitosan + TPP + Spinosad solution. A plug (made from Whatman filter paper) was pressed into the bottom of the vial to ensure all the sugar solution was absorbed (no free liquid). twenty male Drosophila Melanogaster were added to the vial followed by a cellulose plug. The flies reside in anDocket No.10860-10910-US approximately 2-3 cm headspace between the filter paper plug soaked with solution and the cellulose plug. Figures 15G and 15H. [000324] 2.5 ml of an acidic chitosan solution containing 50 millimolar sucrose was transferred into a 40 ml scintillation vial. The control was the sucrose solution without any additives. Additional controls of 225 ppm (2PE) and 3 ppm TPP individually and in combination were also used, as was a control of (2PE) on its own. Further solutions include a Chitosan + Spinosad solution and a Chitosan + TPP + (2PE) solution. A plug (made from Whatman filter paper) was pressed into the bottom of the vial to ensure all the sugar solution was absorbed (no free liquid). twenty male Drosophila Melanogaster were added to the vial followed by a cellulose plug. The flies reside in an approximately 2-3 cm headspace between the filter paper plug soaked with solution and the cellulose plug. Figures 151 and 15J [000325] Once the plug is in place, the experimental clock starts. The vial was observed for D. Melanogaster mortality hourly for the first day, then lest frequently over the course of two weeks. Once all D. Melanogaster have died or 2 weeks, the experiment was stopped. Efficacy differences between solutions was ranked by calculating the median survival (or LC50). [000326] Referring to Figures 15A and 15B which are graphs showing the effect of Spinosad chitosan combinations on male and female D. Melanogaster. The controls of sucrose, Chitosan, and TPP show no effect on either the male or female D. Melanogaster. Spinosad on its own results in total death of female D. Melanogaster at 45 hours and in total death of male D. Melanogaster at 20 hours. Spinosad combined with chitosan results in total death of female D. Melanogaster by about 2 hours and in total death of male D. Melanogaster by about 2 hours. Spinosad combined with chitosan and TPP results in total death of female D. Melanogaster by about 2 hours and in total death of male D. Melanogaster by about 2 hours. [000327] Insecticide example 5: [000328] Experimental Procedure: [000329] To evaluate effective dosages of chitosan 12100 in combination with Spinosad at 2.5 ppm various concentrations of 12100 chitosan were tested. Figures 16A-16F. 2.5 ml of an acidic chitosan solution containing 50 millimolar sucrose was transferred into a 40 ml scintillation vial. The control was the spinosad without any additives. Solutions of Spinosad at 2.5 ppm were prepared with concentrations of chitosan 12100 at 28.12 ppm, 56.25 ppm, 112.5 ppm 225 ppm 450 ppm, and 1800 ppm. A plug (made from Whatman filter paper) was pressed into the bottom of the vial to ensure all the sugar solution was absorbed (no free liquid). twenty male Drosophila Melanogaster were added to the vial followed by a cellulose plug. The flies reside in an approximately 2-3 cm headspace between the filter paper plug soaked with solution and the cellulose plug. Figures 16A-16F.All compositions with chitosan 12100 at concentrations over 225 ppm showed improvement over Spinosad on its own.Docket No.10860-10910-US [000330] Insecticide example 6 [000331] To determine the potential pesticidal synergism of the chitosan, a linear polysaccharide composed of randomly distributed β-(1→4)-linked D-glucosamine (deacetylated unit) and N-acetyl-D- glucosamine (acetylated unit), on natural spinosad, natural pyrethrins and synthetic pyrethroids (permethrin) against the fruit fly (Drosophila melanogaster). The fruit fly (Drosophila melanogaster) was supplied by Department of Entomology, University of California at Riverside (Riverside, CA), or commercial suppliers from Amazon.com. [000332] A full range bioassay was conducted after initial range finder test to generate dose – response curves, 24-hour LC50 (LD50) and LC90 (LD90) and their 95% confidential intervals (CIs) were computerized by probit analysis. Target stages, doses, way of application and replications [000333] A: Target stages: The last larval stages of the fruit fly (3rdinstar) will be used in bioassay. [000334] B: Larvae will be treated by systemic / diet (ingestion and cuticle absorption ad libitum) to determine the main route of synergism by chitosan. [000335] C: Doses: Five doses plus untreated control (UTC) to induce the 24-hour mortality range of approximately 5-95%. [000336] D: Replications: Each dose and UTC will have three replications. Table 6.1 illustrates the effect of pesticides alone on drosophila larvae dose response curves are shown in Figures 19A-19C. Insecticide Table 6.1Docket No.10860-10910-US[000338] Table 6.2 shows the results of each pesticide with chitosan. The chitosan formulation 12965 was used with each pesticide. Dose response curves are shown in Figures 20A-20C. Insecticide Table 6.2Docket No.10860-10910-US[000339] The results are then turned into dose response curves as seen in table 6.3. Insecticide Table 6.3[000340] Insecticide example 7 [000341] Further testing of the effectiveness of Spinosad in combination with chitosan and TPP indicates that the combination is more effective than Spinosad on its own. The combination is more effective than Spinosad on its own against male and female drosophila as can be seen in Figures 21A and 21B.Docket No.10860-10910-US [000342] The invention has been described with reference to various specific and preferred embodiments and techniques. Nevertheless, it is understood that many variations and modifications may be made while remaining within the spirit and scope of the invention.

Claims

Docket No.10860-10910-US WHAT IS CLAIMED IS:

1. A composition comprising: chitosan, and a fungicidal compound.

2. The composition of claim 1, wherein the chitosan is present in the composition in an amount sufficient to enhance the antifungal activity of the fungicidal compound.

3. The composition of claim 1, wherein the chitosan and the fungicidal compound, together, have synergistic antifungal activity.

4. The composition of any of claims 1-3, wherein the chitosan and the antifungal compound are the same.

5. The composition of any of claims 1-4, wherein the chitosan comprises ultra-low molecular weight, low molecular weight, or high molecular weight chitosan.

6. The composition of any of claims 1-5, wherein the majority of the chitosan in the composition by weight is ultra-low molecular weight, low molecular weight, or high molecular weight chitosan.

7. The composition of any of claims 1-6, wherein the composition comprises chitosan from at least two of ultra-low molecular weight, low molecular weight, and high molecular weight chitosan.

8. The composition of claim 5, wherein the ultra-low molecular weight chitosan has a molecular weight between about .05 kDa and about 10 kDa.

9. The composition of claim 5, wherein the ultra-low molecular weight chitosan has a molecular weight between about 3 kDa and about 6 kDa.

10. The composition of any of claims 1-9, wherein the fungicidal compound comprises one or more of copper, Azoxystrobin, Pyraclostrobin, Trifloxystrobin, Fluoxastobin, Picoxystrobin, Kresoxim methyl, Epoxiconazole, tebuconazole, Propiconazole, Cyproconazole, dienconazole, metconazole, Boscalid, Carboxin, Flutoanil, Sedaxane, Fluopyram, Penflufen, Fluxaapyroxad, Fludioxonil, Cymoxanil, Forety-al, Propamocarb, Mandipropamid, Cyazofamid, Famoxadone, Metafenone, Isoprothiolane, Validymycin, Fenhexamid, Pyroquilon, Carpropamid, Fenamidone, Fluopocolide, Harpin, Oxolinic acid, Aluminum sulfate, Bentonite clay, chitosan, Copper Octanoate, Cuprous Oxide, Peptides, Phosphoric Acids, Potassium Bicarbonate, Potassium iodide, Potassium phosphite, Potassium silicate, Potassium thiocyanate, Sodium bicarbonate, Sulfur, Tribasic copper sulphate, Hydrogen Peroxide, Chenopodium quinoa saponins, citric acid, Clove Oil, Coconut Oil, Esquisetum arvense, Eugenol, Garlic Oil, Gu-lutathione, Lamarin, Melaleuca, Peppermint Oil, Phospholipids, Propolis extract, Propylene glycol, Reynoutria , Rosemary Oil, Saponins, Seaweed Extract, Sodium lautyl, Soybean Oil, Sucrose esters, Terpenese, Thyme Oil, Natamycin, Agrobacterium Gliocladium, Polyoxin D Zinc Salt, Bacteriophage, Metschnikowia, Mancozeb, Thiram, Propineb, Prothioconazole. Probenazole, Tricyclazole, Prochloraz, Copper, Sulphur, Fentin, Oxine Copper, Chlorothalonil, Captan, Folpet, Metaxyl, Benalaxyl, Oxadixyl, Carbendazim, Thiophanate, Thiabendazole, Benomyl, Fuberidazole, Spiroxamine, dimethomorph, Fenpropidin, Fenproprimorph, Cypodinil, Pyrimethanil, Mepanipyrim, Fluazinam, Dithianon, Pencycuron,Docket No.10860-10910-US Iminoctadine, Tolylfuanid, Iprodione, Procymidone, Vincllozolin, Fenarimol, Bupirimate, Triforine, Ferimzone, and Pyrifenox.

11. The composition of claim 10, wherein the copper comprises one or more of copper, copper nitrate, copper sulfate, or copper hydroxide.

12. The composition of any of the preceding claims, wherein the chitosan comprises between about 1.2 wt% and about 2.2 wt% of the composition.

13. The composition of any of the preceding claims, wherein the chitosan comprises about 1.7 wt% of the composition.

14. The composition of any of the preceding claims, further comprising an acid.

15. The composition of claim 14, wherein the acid comprises one or more of 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.

16. The composition of any of the preceding claims, further comprising a coupling enhancer.

17. The composition of claim 16, wherein the coupling enhancer is sodium tripolyphosphate.

18. The composition of claim 17, wherein the fungicidal compound is comprised in a nanoparticle.

19. The composition of any of the preceding claims, wherein the fungicidal activity of the composition is increased by between about 5% and about 50% as compared to the same fungicide not in a composition with chitosan.

20. The composition of any of the preceding claims, wherein the composition comprises between about 10 wt% and about 40 wt% nitrogen.

21. The composition of claim 20, wherein the composition comprises about 20 wt% nitrogen.

22. The composition of claim 21, wherein at least 50% of the nitrogen is in the chemical form of amine and / or nitrate and / or ammonia.

23. The composition of any of the preceding claims, wherein the composition has antifungal activity against pythium irregulare.

24. A composition comprising: chitosan, and an insecticidal compound.

25. The composition of claim 24, wherein the chitosan is present in the composition in an amount sufficient to enhance the insecticidal activity of the insecticidal compound.

26. The composition of claim 24, wherein the chitosan and the insecticidal compound, together have synergistic insecticidal activity.

27. The composition of any of claims 24-26, wherein the chitosan comprises ultra-low molecular weight, low molecular weight, or high molecular weight chitosan.Docket No.10860-10910-US 28. The composition of claims 24-27, wherein the majority of the chitosan in the composition by weight is ultra-low molecular weight, low molecular weight, or high molecular weight chitosan.

29. The composition of claim 24, wherein the chitosan comprises between about 1.2 wt% and about 2.2 wt% of the composition.

30. The composition of claim 29, wherein the chitosan is about 1.7 wt% of the composition.

31. The composition of claim 24, wherein the chitosan has a molecular weight between about 4 kDa and about 500 kDa.

32. The composition of claim 31, wherein the chitosan has a molecular weight between about 20 kDa and about 380 kDa.

33. The composition of claim 32, wherein the chitosan has a molecular weight between about 50 kDa and about 300 kDa.

34. The composition of claim 33, wherein the chitosan has a molecular weight about 100 kDa and about 250.

35. The composition of claim 34, wherein the chitosan has a molecular weight between about 150 kDa and about 200 kDa.

36. The composition of claim 24, further comprising an acid.

37. The composition of claim 36, wherein the acid comprises one or more of 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.

38. The composition of claim 24, further comprising a coupling enhancer.

39. The composition of claim 38, wherein the coupling enhancer is sodium tripolyphosphate.

40. The composition of any of claims 24-39, wherein the insecticidal compound comprises one or more of a spinosyn, a phenylethanol, Malathion, Parathion, Diazinon, Chlorpyrifos, Carbaryl, Methomyl, Aldicarb, Propoxur, Permethrin, Cypermethrin, Deltamethrin, Lambda-cyhalothrin, Imidacloprid, Clothianidin, Thiamethoxam, Acetamiprid, Dichlorodiphenyltrichloroethane (DDT), Lindane, Chlorane, Pyrethrins, Rotenone, Azadirachtin, Beauveria bassiana, Metarhizium anisopliae, Methoprene, Pyriproxfen, Fenoxycarb, Diflubenzuron, Sarin, Tabun, Soman, VX, Sulfur, and Diatomaceous Earth..

41. The composition of claim 40, wherein the spinosyn comprises at least two chemically distinct spinosyns.

42. The composition of claim 41, wherein the at least two chemically distinct spinosyns are spinosyn A and spinosyn D.Docket No.10860-10910-US 43. The composition of claim 24, wherein the composition comprises between about 10 wt% and about 30 wt% nitrogen.

44. The composition of claim 24, wherein the composition comprises about 20 wt% nitrogen.

45. The composition of claim 44, wherein at least 50% of the nitrogen is in the chemical form of amine and / or nitrate and / or ammonia.

46. A composition comprising: chitosan and an herbicidal compound.

47. The composition of claim 46, wherein the chitosan comprises between about 1.2 wt% and about 2.2 wt% of the composition.

48. The composition of claim 46, wherein the chitosan is about 1.7 wt% of the composition.

49. The composition of claim 46, wherein the chitosan has a molecular weight between about 4 kDa and about 60 kDa.

50. The composition of claim 49, wherein the chitosan has a molecular weight between about 4 kDa and about 16 kDa.

51. The composition of claim 50, wherein the chitosan has a molecular weight between about 6 kDa and about 12 kDa.

52. The composition of claim 51, wherein the chitosan has a molecular weight of about 8 kDa.

53. The composition of claim 50, wherein the chitosan has a molecular weight between about 20 kDa and about 60 kDa.

54. The composition of claim 53, wherein the chitosan has a molecular weight between about 30 kDa and about 50 kDa.

55. The composition of claim 54, wherein the chitosan has a molecular weight of about 40 kDa.

56. The composition of claim 55, further comprising an acid.

57. The composition of claim 56, wherein the acid comprises one or more of 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.

58. The composition of claim 57, further comprising a coupling enhancer.

59. The composition of claim 58, wherein the coupling enhancer is sodium tripolyphosphate.

60. The composition of any of claims 46-59 wherein the herbicidal compound comprises one or more of a photosynthesis inhibitor, an auxin mimic, a microtubule assembly inhibitor, a cell membrane disruptor, or an amino acid synthesis inhibitor.

61. The composition of claim 60, wherein the amino acid synthesis inhibitor is glyphosate.

62. The composition of any of claims 46-61, wherein the composition comprises between about 10 wt% and about 40 wt% nitrogen.Docket No.10860-10910-US 63. The composition of claim 62, wherein the composition comprises about 20 wt% nitrogen.

64. The composition of claim 63, wherein at least 50% of the nitrogen is in the chemical form of amine and / or nitrate and / or ammonia.

65. A method of producing crop-protective chitosan nanoparticles, the method comprising: dissolving chitosan in acidic water, where the water is made acidic by an acid selected from the group of 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; dissolving a crop-protective compound in the acidic water; and forming nanoparticles from the dissolved chitosan and the crop protective compound.

66. The method of claim 65, wherein the chitosan comprises between about 1.2 wt% and about 2.2 wt% of the composition.

67. The method of claim 66, wherein the chitosan is about 1.7 wt% of the composition.

68. The method of claim 67, wherein the chitosan has a molecular weight between about 4 kDa and about 160 kDa.

69. The method of claim 68, wherein the chitosan has a molecular weight between about 4 kDa and about 16 kDa.

70. The method of claim 69, wherein the chitosan has a molecular weight between about 6 kDa and about 12 kDa.

71. The method of claim 70, wherein the chitosan has a molecular weight of about 8 kDa.

72. The method of claim 71, wherein the chitosan has a molecular weight between about 20 kDa and about 50 kDa.

73. The method of claim 72, wherein the chitosan has a molecular weight between about 25 kDa and about 40 kDa.

74. The method of claim 73, wherein the chitosan has a molecular weight of about 32 kDa.

75. The method of claim 68, wherein the chitosan has a molecular weight between about 60 kDa and about 100 kDa.

76. The method of claim 75, wherein the chitosan has a molecular weight between about 75 kDa and about 85 kDa.

77. The method of claim 76 wherein the chitosan has a molecular weight of about 78 kDa.

78. The method of claim 77, wherein chitosan has a molecular weight between about 120 kDa and about 160 kDa.

79. The method of claim 78, wherein chitosan has a molecular weight between about 130 kDa and about 150 kDa.Docket No.10860-10910-US 80. The method of claim 79, wherein the chitosan has a molecular weight of about 141 kDa.

81. The method of claim 66, further comprising a coupling enhancer.

82. The method of claim 81, wherein the coupling enhancer is sodium tripolyphosphate.

83. The method of any of claims 66-82, wherein the crop-protective compound is an insecticidal compound.

84. The method of claim 83, wherein the insecticidal compound comprises one or more of a spinosyn, a phenylethanol, Malathion, Parathion, Diazinon, Chlorpyrifos, Carbaryl, Methomyl, Aldicarb, Propoxur, Permethrin, Cypermethrin, Deltamethrin, Lambda-cyhalothrin, Imidacloprid, Clothianidin, Thiamethoxam, Acetamiprid, Dichlorodiphenyltrichloroethane (DDT), Lindane, Chlorane, Pyrethrins, Rotenone, Azadirachtin, Beauveria bassiana, Metarhizium anisopliae, Methoprene, Pyriproxfen, Fenoxycarb, Diflubenzuron, Sarin, Tabun, Soman, VX, Sulfur, and Diatomaceous Earth..

85. The method of claim 84, wherein the spinosyn comprises at least two chemically distinct spinosyns.

86. The method of claim 85, wherein the at least two chemically distinct spinosyns are spinosyn A and spinosyn D.

87. The method of any of claims 66-82, wherein the crop-protective compound is an herbicidal compound.

88. The method of claim 87, wherein the herbicidal compound comprises one or more of a photosynthesis inhibitor, an auxin mimic, a microtubule assembly inhibitor, a cell membrane disruptor, or an amino acid synthesis inhibitor.

89. The method of claim 88, wherein the amino acid synthesis inhibitor is glyphosate.

90. The method of any of claims 66-83, wherein the crop protective compound is a fungicide.

91. The method of any of claim 90, wherein the fungicidal compound comprises one or more of copper, chlorothalonil, azoxystrobin, prothioconazole, iprodione, or fludioxonil.

92. The method of claim 91, wherein the copper comprises one or more of copper, copper nitrate, copper sulfate, or copper hydroxide.

93. The method of any of claims 66-83, wherein the crop-protective compound is a nematicide.

94. The method of claim 93, wherein the nematocidal compound comprises one or more of methyl bromide, 1,3-Dichloropropene, Dazomet, Aldicarb, Fenamiphos, Oxamyl, Bacillus thuringiensis, Paecilomyces lilacinus, Pochonia chlamydosporia, Azadirachtin, nicotinamides, thiophenes, Terpenoids, Flavenoids, and Saponins.

95. The method of any of claims 66-94, wherein the composition comprises between about 10 wt% and about 30 wt% nitrogen.

96. The method of claim 95, wherein the composition comprises about 20 wt% nitrogen.

97. The method of claim 96, wherein at least 50% of the nitrogen is in the chemical form of amine and / or nitrate and / or ammonia.

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