WO2023137245A1

WO2023137245A1 - Compositions and methods for controlling plant parasitic nematodes

Info

Publication number: WO2023137245A1
Application number: PCT/US2023/060202
Authority: WO
Inventors: Samuel L. CLOETE; Ahsan Habib
Original assignee: Kannar Earth Science, Ltd.
Priority date: 2022-01-11
Filing date: 2023-01-06
Publication date: 2023-07-20
Also published as: AR128252A1

Abstract

A nematicide composition is provided that includes: (i) swertiamarin; (ii) berberine; (iii) chitosan, (iv) glutathione; and (v) cellulose-based polymer. The composition can kill root knot nematodes within a minute of contact. This result is surprising, given that each of swertiamarin, berberine, chitosan, and glutathione when applied individually, repels, but does not kill nematodes. In addition, coleoptile length resulting from corn seed treated with the composition was significantly increased. Further, the composition applied in furrow in cotton and soybean field trials reduced reniform and root knot nematodes, respectively, and increased yield. The nematicide composition can also exhibit antifungal and insecticidal activities.

Description

COMPOSITIONS AND METHODS FOR CONTROLLING PLANT PARASITIC

NEMATODES

CROSS-REFERENCE TO RELATED APPLICATION

This application claims the benefit of priority of U.S. provisional patent application no. 63/298,257 filed on January 11, 2022, which is incorporated herein in its entirety by this reference.

TECHNICAL FIELD

The present disclosure relates to the field of agricultural pest control to reduce crop damage and loss of yield. Specifically, the present disclosure relates to compositions and methods for control of plant pathogens, including parasitic nematodes, in soil or any other growing media where plants are grown.

BACKGROUND

Nematodes are miniscule soil inhabitants that are parasitic to almost all plants on earth. These parasites, commonly known as Plant Parasitic nematodes (PPNs), cause an estimated loss of yields over $ 100 billion annually (Thoden et al., 2011). Most notorious plant nematodes include root knot, cyst, ring, and lesion nematodes. Since these PPNs cannot be eradicated completely, different controlling strategies are practiced reducing their infestation. These strategies include chemical nematicides, agronomic practices (crop rotation, cover crops or biofumigation), biopesticides (fungi, bacteria, or their derivatives) and plant derived formulations (Nicola et al., 2021). Despite there being so many strategies, current management practices are not enough to significantly reduce nematode infestation. Use of chemical nematicides is also being limited in recent years due to their adverse effect on the environment (Fereidoun and Abolfazl, 2020). In recent years, biopesticides are getting a lot of attention owing to their safety profiles and ease of application, but formulation stability, compatibility and on-seed survival of the active ingredients have limited such product development (Abd-Elgawad, 2016). Use of phytochemicals or botanicals antagonistic to PPN include alkaloids, glycosides, terpene, and phenolics. These phytochemicals are safe and promising tools as fungicides, bactericides, insecticide, and nematicides (Mohamed Khalil, 2014).

There remains a need for agronomically durable, sustainable, and economically feasible alternatives to toxic chemical control of plant pathogens, including parasitic nematodes. The present disclosure provides such alternatives.

SUMMARY

In some respects, the present disclosure is directed to a nematicide composition including: (i) swerdamarin: (ii) berberine; (iii) chitosan; (iv) glutathione: and (v) cellulose- based polymer.

In other respects, a liquid composition is provided including: (i) swertiamarin; (ii) berberine; (iii) chitosan; (iv) glutathione; and (v) cellulose-based polymer, wherein the composition has activity against one or more plant pathogens.

The one or more plant pathogens can be one or a combination of a nematode, an insect, or a fungus.

The composition of the present disclosure can be formulated as a flowable liquid.

The composition of the invention can be in the form of an emulsion or a suspension.

In the liquid composition of the present disclosure, the cellulose-based polymer can include hydroxyethy cellulose or any cellulose-based polymer that keeps the components of the composition in an emulsion or a suspension. The cellulose-based polymer can include hydroxyethylcellulose, hydroxyethyl propyl methyl cellulose (HPMC), quillaja extract, xanthan gum, corn starch, or yucca extract, and combinations thereof.

The emulsion or suspension can be an oil-based or an aqueous-based emulsion or suspension.

The composition of the invention can have a total amount of component (i) in the range from 0.1 to 1.0 g/L, the total amount of component (ii) in the range from 0.1 to 1.0 g/L, the total amount of component (iii) in the range from 0.1 to 1.0 g/L, and the total amount of component (iv) in the range from 0.1 to 0.5 g/L.

The total amount of component (i) in the composition can be equal to or less than 1.0 g/L, the total amount of component (ii) can be equal to or less than 0.5 g/L, the total amount of component (iii) can be equal to or less than 0.5 g/L, and the total amount of component (iv) can be less than or equal to 0.2 g/L.

The total amount of component (i) in the composition can be in the range from 0.01 to 1.0 g/L, the total amount of component (ii) can be in the range from 0.005 to 0.5 g/L, the total amount of component (iii) can be in the range from 0.0025 to 0.25 g/L, and the total amount of component (iv) can be in the range from 0.002 to 0.2 g/L.

In one embodiment, an aqueous composition is provided including a ratio by weight of: (i) about 0.05% to about 0.2% swertiamarin; (ii) about 0.025% to about 0.1% berberine; (iii) about 0.01% to about 0.05% chitosan; (iv) about 0.01% to about 0.04% L-glutathione; and (v) about 80-99% cellulose-based polymer, wherein the composition has activity against one or more plant pathogens.

In another instance, a composition is provided including a ratio by weight of: (i) about 0.1% swertiamarin; (ii) about 0.05% berberine; (iii) about 0.025% chitosan; (iv) about 0.02% L-glutathione; (v) about 2% DMSO; (vi) about 90% hydroxy ethylcellulose carrier; and (vii) about 7.805% water, wherein the composition has activity against one or more plant pathogens.

The composition of the present disclosure can include one or more plant bio stimulants. The biostimulants can include amino acids, humic acid, fulvic acid, seaweed extract, laminarin, alginates, or polysaccharides, and combinations thereof.

The composition of the present disclosure can include one or more antioxidants. The antioxidants can include vitamin C, vitamin E, coenzyme Q10, lipoic acid, flavonoids, phenols, polyphenols, or phytoestrogens, and combinations thereof.

In other embodiments, the present disclosure is directed to a plant seed coated with a composition including: (i) swertiamarin; (ii) berberine; (iii) chitosan; (iv) glutathione; and (v) cellulose-based polymer, wherein the composition is present in the coating in an amount suitable to improve plant growth and/or health in the presence of a plant pathogen, or both.

In some aspects, the present disclosure is direct to a method of plant seed treatment including: applying to seeds of a plant a coating of a liquid composition including: (i) swertiamarin; (ii) berberine; (iii) chitosan; (iv) glutathione; and (v) cellulose-based polymer, wherein the liquid composition is applied to the seeds in the form of a suspension or an emulsion and is present in the coating in an amount suitable to improve plant growth and/or health in the presence of a plant pathogen, or both.

In other instances, the present disclosure is directed to a method for improving plant growth and/or health including: planting a plant or a seed of a plant in a suitable growth medium, the plant or the seed having a coating or partial coating of a composition including: (i) swertiamarin; (ii) berberine: (iii) chitosan; (iv) glutathione; and (v) cellulose- based polymer, wherein the composition is present in an amount suitable to improve plant growth and/or health in the presence of a plant pathogen, or both.

In one embodiment of the present disclosure, a method is provided for improving plant growth and/or health including: delivering to seed of a plant, foliage of a plant, roots of a plant, or soil or growth medium surrounding a plant, a liquid composition in the form of an emulsion or a suspension including: (i) swertiamarin; (ii) berberine; (iii) L- glutathione; (iv) chitosan; and (v) cellulose-based polymer, wherein the composition improves plant growth and/or health in the presence of a plant pathogen, or both.

In some embodiments of the presently disclosed compositions and methods, the composition can exhibit control of one or more of plant parasitic nematodes, fungi, or insects.

In various aspects of the present disclosure, the composition in which the seeds or plant parts are coated includes: component (i) in the range from 0.1 to 1.0 g/L, component (ii) in the range from 0.5 to 1.0 g/L, component (iii) in the range from 0.1 to 1.0 g/L, and component (iv) in the range from 0.1 to 0.5 g/L.

In other embodiments of the present disclosure, the composition in which the seeds or plant parts are coated includes: component (i) in an amount equal to or less than 1.0 g/L, component (ii) in an amount equal to or less than 0.5 g/L, component (iii) in an amount equal to or less than 0.5 g/L, and component (iv) in an amount equal to or less than 0.2 g/L.

In the compositions and methods of the present disclosure, the plants or plant seeds can include, but are not limited to, cotton, com, peanut, wheat, soybean, sorghum, or canola, and combinations thereof. In some embodiments, the plants or plant seeds include legumes. In other embodiments, the plants or plant seeds include non-legumes. BRIEF DESCRIPTION OF THE DRAWINGS

Figure 1 is graft showing the effect over time of dilutions of a nematicide composition on mortality of dagger nematodes (Xiphinema sp.).

Figure 2 is a graph showing the positive impact on corn seedling growth after seed treatment with a nematicide composition.

Figure 3 is a graph showing the effect of a nematicide composition as an in- furrow application in a cotton field trial on reniform nematodes and its impact on improving seed cotton yield.

Figure 4A is a graph showing the effect of in-furrow application of a nematicide composition in a soybean field trial on reducing root-knot nematode induced galling.

Figure 4B is a graph showing the increased soybean yield in the trial of Figure 4A.

Figure 5A is a grayscale image showing the anti-fungal activity in an in vitro petri plate assay of cellulose-based carrier hydroxyethyl cellulose alone against peanut white mold (Sclerotium roljsii).

Figure 5B is a grayscale image showing the anti-fungal activity in an in vitro petri plate assay of a IX concentration of nematicide composition against peanut white mold (Sclerotium roljsii).

Figure 5C is a grayscale image showing the anti-fungal activity in an in vitro petri plate assay of a 2X concentration of nematicide composition against peanut white mold (Sclerotium rol sii).

Figure 5D is a grayscale image showing the anti-fungal activity in an in vitro petri plate assay of commercially available fungicide RANCONA VPD against peanut white mold (Sclerotium roljsii).

Figure 6A is a grayscale image showing the anti-fungal activity in an in vitro petri plate assay of a IX concentration of the nematicide composition against peanut black leaf spot (Nothopassalora personata).

Figure 6B is a grayscale image showing the anti-fungal activity in an in vitro petri plate assay of a 2X concentration of the nematicide composition against peanut black leaf spot (Nothopassalora personata). Figure 6C is a grayscale image showing the anti-fungal activity in an in vitro petri plate assay of commercially available fungicide RANCONA VPD against peanut black leaf spot (Nothopassalora per sonata) in comparison to (c).

DETAILED DESCRIPTION

The present disclosure, in some embodiments, provides a nematicide composition that includes extremely bitter plant based saponins, swertiamarin and berberine, chitosan (an EPA approved adjuvant) and an antioxidant for application to plant seeds to kill or deter plant parasitic nematodes. Swertiamarin and berberine are two bitter phytochemicals from Swertia and Berberis plants, respectively. The nematicide composition comprises swertiamarin from Swertia, berberine from Berberis, and chitosan from crustaceans. In one embodiment, these components are provided in a liquid solution of a cellulose-based polymer containing an antioxidant such as, for example, glutathione, to protect the active ingredients from possible oxidative degradation.

In some embodiments, the nematicide composition can reduce the locomotion and kill root-knot nematodes within a few seconds upon contact. This is unexpected given that when swertiamarin, berberine, glutathione, or chitosan are applied individually, nematodes are repelled, but the application is not fatal at the time of application. An example of the killing activity of a water-soluble nematicide composition of the present disclosure is described in Example 1. This composition kills needle nematodes (Longidorus sp.) within a few seconds of contact (see Table 1 for a summary of the results). In this example, the nematicide composition includes a ratio by weight of: (i) about 0.1% swertiamarin; (ii) about 0.05% berberine; (iii) about 0.025% water soluble chitosan; (iv) about 0.02% by weight L-glutathione; (v) about 2% by weight of DMSO; and (vi) about 97.805% water.

Table 1. Two-minute contact of live needle nematodes (Longidorus sp. with water-based nematicide composition and with each individual component.

Additional experiments are described in Example 1 in which the water earner was replaced with a cellulose-based carrier to keep the components in suspension because of precipitation of some components. Specifically, when a cellulose-based polymer was used as a carrier in the composition described in Example 1, water was replaced by 90% cellulose-based liquid polymer and 7.805% water. The liquid cellulose-based composition killed dagger (Xiphinema sp.) nematodes with a 100% killing rate (see Table 2). As can be seen in Table 2, various sub-combinations of the components repelled the nematodes, but only a combination of swertiamarin, berberine, chitosan, and glutathione in the cellulose- based polymer resulted in death of the dagger nematodes.

Table 2. Two-minute contact of live dagger nematodes (Xiphinema sp.) with liquid nematicide composition in cellulose-based polymer and with sub-combinations of components.

SWTM = swertiamarin, BRBN = berberine, GLTN = glutathione and CHT = chitosan

The composition including swertiamarin, berberine, chitosan, and glutathione in the cellulose-based polymer described above was stored and kept at room temperature (23 °C) for a month and then tested again on live dagger nematodes. In the test, which included four replicates of live swimming dagger nematodes (Xiphinema sp.), addition of the composition instantly (30-60 seconds) killed all the nematodes. The rate of nematode fatality was 100% upon contact with the composition. A summary of the results of the four replicates of this experiment at various dilutions is shown in Figure 1. In one embodiment of the present disclosure, the total amount of component (i) in the nematicide composition is in the range from 0.01 to 1.00 g/L, the total amount of component (ii) is in the range from 0.005 to 0.5 g/L, the total amount of component (iii) is in the range from 0.0025 to 0.25 g/L, and the total amount of component (iv) is in the range from 0.002 to 0.2

In other embodiments, the total amount of component (i) in the nematicide composition is in the range from 0.001 to 1.00 g/L, the total amount of component (ii) is in the range from 0.0005 to 0.5 g/L, the total amount of component (iii) is in the range from 0.00025 to 0.25 g/L, and the total amount of component (iv) is in the range from 0.0002 to 0.2

In one aspect of the present disclosure, the total amount of component (i) in the nematicide composition is equal to or less than 1.0 g/L, the total amount of component (ii) is equal to or less than 0.5 g/L, the total amount of component (iii) is equal to or less than 0.5 g/L, and the total amount of component (iv) is equal to or less than 0.2 g/L.

Example 2 of the present disclosure describes an experiment in which treatment of com seeds with the nematicide composition results in enhanced seedling growth. In the experiment, seedling roots and shoots (coleoptiles) were measured after treated corn seeds were placed on wet germination paper and kept in the dark for seven days. There was no significant difference in seedling main root length but the nematicide composition significantly increased (24%) coleoptile length (see Figure 2).

Thus, in one embodiment, the present disclosure is directed to a seed coated with a composition including: (i) swertiamarin; (ii) berberine; (iii) chitosan; (iv) glutathione; and (v) cellulose-based polymer, wherein the composition is present in the coating in an amount suitable to improve plant growth and/or health in the presence of a plant pathogenic nematode, or both.

In other aspects, the present disclosure is direct to a method of plant seed treatment including applying to seeds of a plant a coating of a composition including: (i) swertiamarin; (ii) berberine; (iii) chitosan; (iv) glutathione and, (v) cellulose-based polymer, wherein the composition is present in the coating in an amount suitable to improve plant growth and/or health in the presence of a plant pathogenic nematode, or both. Examples 3 and 4 of the present disclosure describe field performance of the nematicide composition. Specifically, the composition applied in furrow in cotton and soybean field trials reduced reniform nematodes in cotton soil and root knot nematode induced root galling in soybean and improved cotton and soybean yields. See Figure 3 for the cotton field trial results and Figures 4 A and 4B for the soybean field trial results.

Example 5 of the present disclosure describes an experiment in which the current composition exhibited anti-fungal activity through in vitro assays. The composition was able to show some degree of control of peanut white mold (Sclerotium rolfsii) and black leaf spot fungi (Nothopassarola per sonata). Specifically, for the white mold assay, plates treated with cellulose-based carrier hydroxy ethyl cellulose alone did not exhibit any control of fungus (see Fig. 5a). Fungicide RANCONA VPD showed some control of fungus by zone of inhibition (or zone of clearance) but soon got occupied by fungus (see Fig. 5d). In contrast, the nematicide composition, both at lx (see Fig. 5b) and 2x (see Fig. 5c) concentration restricted fungal growth around the application areas at the center of the culture plates. For Black Eeaf spot fungus, fungicide RANCONA VPD exhibited good control of fungus by producing larger clearing zone around the point of application (see Fig. 6c). The nematicide composition at lx and 2x concentrations also exhibited control of fungal growth but the effect was less than fungicide RANCONA VPD (Fig. 6a & b). In both assays, the nematicide composition showed a similar pattern of control efficacy at 1 x and 2x concentrations. These zone of inhibition assays demonstrate the anti-fungal activity of the composition and its broad-spectrum nature.

Example 6 of the present disclosure describes an experiment in which the composition also exhibited insecticidal activity in laboratory assays. Plant feeding insects, particularly those under the coleoptera order (most beetles and weevils) are very voracious insect for crops. As surrogates of coleoptera insects’ larvae rootworm and wireworm, Superworm (Zophobas morio) and mealworm (Tenebrio molitor) were used in this experiment. The composition when applied directly to the body of the coleoptera insect larvae demonstrated a high degree (100%) of paralysis followed by death to the larvae (data not shown).

These results demonstrate that the composition of the present disclosure can function as a broad spectrum biopesticide. Thus, in some embodiments, the present disclosure is directed to a method for improving plant growth and/or health including: planting a plant or a seed of a plant in a suitable growth medium, the plant or the seed having a coating or partial coating of a composition including: (i) swertiamarin; (ii) berberine; (iii) chitosan; (iv) glutathione; and (v) cellulose-based polymer, wherein the composition is present in an amount suitable to improve plant growth and/or health in the presence of a plant pathogen, or both.

In other embodiments of the present disclosure, a method is provided for improving plant growth and/or health including: delivering to seed of a plant, foliage of a plant, roots of a plant, or soil or growth medium surrounding a plant, a liquid composition in the form of an emulsion or a suspension including: (i) swertiamarin; (ii) berberine; (iii) L- glutathione; (iv) chitosan; and (v) cellulose-based polymer, wherein the composition improves plant growth and/or health in the presence of a plant pathogen, or both.

In the compositions and methods of the present disclosure, the plants or plant seeds can include, but are not limited to, cotton, com, peanut, wheat, soybean, sorghum, or canola, and combinations thereof. In some embodiments, the plants or plant seeds include legumes. In other embodiments, the plants or plant seeds include non-legumes.

For the purposes of this specification and claims, the term “about” when used in connection with one or more numbers or numerical ranges, should be understood to refer to all such numbers, including all numbers in a range and modifies that range by extending the boundaries above and below the numerical values set forth. The recitation of numerical ranges by endpoints includes all numbers, e.g., whole integers, including fractions thereof, subsumed within that range (for example, the recitation of 1 to 5 includes 1, 2, 3, 4, and 5, as well as fractions thereof, e.g., 1.5, 2.25, 3.75, 4.1, and the like) and any range within that range. In addition, as used herein, the term "about", when referring to a value can encompass variations of, in some embodiments +/-100%, in some embodiments +/-50%, in some embodiments +/-20%, in some embodiments +/-10%, in some embodiments +/- 5%, in some embodiments +/-1%, in some embodiments +/-0.5%, and in some embodiments +/-0.1 % , from the specified amount, as such variations are appropriate in the disclosed compositions and methods. Where particular values are described in the application and claims, unless otherwise stated, the term “about” meaning within an acceptable error range for the particular value should be assumed.

Following long-standing patent law convention, the terms “a,” “an,” and “the” refer to “one or more” when used in this application, including the claims. Thus, for example, reference to “a seed” includes a plurality of seeds, unless the context clearly is to the contrary, and so forth.

Throughout this specification and the claims, the terms “comprise,” “comprises,” and “comprising” are used in a non-exclusive sense, except where the context requires otherwise. Likewise, the terms “having” and “including” and their grammatical variants are intended to be non-limiting, such that recitation of items in a list is not to the exclusion of other like items that can be substituted or added to the listed items.

This disclosure describes inventive concepts with reference to specific examples. However, the intent is to cover all modifications, equivalents, and alternatives of the inventive concepts that are consistent with this disclosure.

For the purposes of promoting an understanding of the principles of the present disclosure, reference will now be made to preferred embodiments and specific language will be used to describe the same. It will nevertheless be understood that no limitation of the scope of the disclosure is thereby intended, such alteration and further modifications of the disclosure as illustrated herein, being contemplated as would normally occur to one skilled in the art to which the disclosure relates.

EXAMPLES

Example 1

Composition Kills Nematodes within Seconds of Contact

Live root knot nematodes were collected from soil and kept in shallow water in small glass beakers at room temperature. The individual components and combined composition were prepared according to the following procedures.

Stock solutions of individual components were prepared in water at the concentrations as follows: 1000 ppm swertiamarin (95% purity; Xi’an Best Biotech Co, China), 500 ppm berberine HC1 (98% purity; Shaanxi Jintai Biological Engineering Co Ltd, China), 250 ppm water soluble chitosan (85% purity; Shaanxi Rainwood Biotech Co Ltd, China) and 200 ppm L-glutathione (99% purity; Hunan Insen Biotech Co Ltd, China). Swertiamarin and berberine were dissolved in 0.5 and 1.0 ml of DMSO separately (total 1.5 ml DMSO) prior to dilution in water. The pH of the composition was adjusted to 5.7. The combined composition included: (i) about 0.1% swertiamarin; (ii) about 0.05% berberine; (iii) about 0.025% water soluble chitosan; (iv) about 0.02% by weight L- glutathione; (v) about 2% by weight of DMSO. This is the composition used in the experiment described below.

The effect of each individual component and the combined composition on live nematodes was tested as follows. An individual Needle nematode (Longidorus .s .) was placed in a watch glass (55 mm) containing 2 ml water and kept under a digital inspection microscope (TAGARNO). One drop of the stock solution of one of the individual components described above was added to the watch glass with the swimming nematode and the reaction of the nematode was recorded for a period of 1-2 minutes. This procedure was repeated 4 to 7 times for each of the individual components and for the combination of all components (i.e., the composition). The results are summarized in Table 1. Each of the four agents swertiamarin, berberine, glutathione, and chitosan when applied alone using the water-based composition exhibited reactions by the needle nematodes via faster locomotion and swimming away from the point of application in the measured 1-2 minutes period but did not kill the nematodes. This reaction is a repellent effect. In contrast, the composition including all the components together instantly (10-30 seconds) killed the needle nematode upon contact.

As swertiamarin and berberine were observed to precipitate in water within a few hours, an alternative composition was generated in which a cellulose-based carrier was used to keep the components suspended. For this composition, stock solutions of individual components were prepared in a cellulose-based carrier (88.76% Natrosol 250-LR from Ashland) at the concentrations as follows: 1000 ppm swertiamarin (95% purity; Xi’an Best Biotech Co, China), 500 ppm berberine HC1 (98% purity; Shaanxi Jintai Biological Engineering Co Ltd, China), 250 ppm water soluble chitosan (85% purity; Shaanxi Rainwood Biotech Co Ltd, China) and 200 ppm L-glutathione (99% purity; Hunan Insen Biotech Co Ltd, China). Swertiamarin and berberine were dissolved in 0.5 and 1.0 ml of DMSO separately (total 1.5 ml DMSO) prior to addition to the carrier. Chitosan and glutathione were dissolved in 7.8 ml water prior to addition to the 90.0 ml carrier. The pH of the composition was adjusted to 5.7. The combined composition included: (i) about 0.1% swertiamarin; (ii) about 0.05% berberine; (iii) about 0.025% water soluble chitosan; (iv) about 0.02% by weight L-glutathione; (v) about 2% by weight of DMSO; (vi) about 90% cellulose-based liquid polymer; and (vii) about 7.805% by weight water. The effect of this composition, and various sub-combinations of the individual components, on live dagger nematodes Xiphinema .sp.) was determined using the same procedures as described above for the water-based composition and the results are shown in Table 2. As can be seen in Table 2, various sub-combinations of the components repelled the nematodes, but only a combination of swertiamarin, berberine, chitosan, and glutathione in the cellulose-based polymer resulted in death of the dagger nematodes.

The nematicide composition in cellulose-based polymer described above was stored and kept at room temperature (23 °C) for a month and then tested again on live dagger nematodes (Xiphinema .sp.). In the test, there were four replicates each containing a live dagger nematode swimming in water in circular glass plates. A drop or two of the composition was added to the swimming nematodes, which instantly (30-60 seconds) killed all the nematodes. The rate of nematode fatality was 100% upon contact that occurred from 0.5 min to 1 hour depending on various dilutions of the nematicide concentrate (ready-to-use) composition. A summary of the results of the experiment is shown in Figure 1. The composition repeatedly killed dagger nematodes at time of contact (within 1 minute).

Example 2 Composition Applied as a Seed Treatment Improves Plant Growth

An experiment was conducted to investigate the response of the nematicide composition when applied on seed. Hybrid com seeds (UPL variety, 2020) were treated with nematicide composition or water as a control at the rate of 25 fl oz/100 lbs of seeds. For a small amount, about 100-gram corn seeds were treated with 1.63 ml of composition in a ZIPLOC bag and then were shaken vigorously for 1 minutes to have a good seed coating. Treated seeds were placed on wet germination paper in plastic trays and kept in the dark for seven days. Seedling roots and shoots (coleoptiles) were measured using WINRHIZO root scanner (Regent Instrument, Canada). There was no significant difference in seedling main root length but the nematicide composition significantly increased (24%) coleoptile length (Figure 2). Specifically, Figure 2 shows the positive impact on com seedling growth after seed treatment with nematicide composition. There was no negative impact on root growth while a significant enhancement (@ 0.05 level) of com seedling shoot (coleoptile) growth was observed.

Example 3

Composition Applied in-furrow Controls Nematodes and Improves Cotton Yield

A filed trial was conducted by the University of Arkansas, Department of Agriculture (UADA) with cotton in 2022. Nematicide composition was applied as an infurrow application at the rate of 1.0 quart/A mixed with 10 gallons of water after seed sowing. Before planting, all of the cotton seed in the trial was treated with an industry standard fungicide seed treatment (GAUCHO + EVERGOL XTEND) In the trial, the control consisted of seed treated with GAUCHO + EVERGOL XTEND alone. It was a replicated field plot trial with 4 replicates arranged in a completely randomized design. Cotton was harvested at the end of the season and data was analyzed by ARM software. Reniform nematode counts in field plots were made 30 days after planting. The nematicide composition reduced the number of reniform nematodes (Rotylenchulus .sp.) by 26.37% and increased seed cotton production by 348 Ibs/A, equal to 14% increase as compared to control (Figure 3).

Example 4

Composition Applied in-furrow Controls Nematodes and Improves Soybean Yield

A filed trial was conducted by the University of Arkansas, Department of Agriculture (UADA) with soybean in 2022. Nematicide composition was applied as an infurrow application at the rate of 1.0 quart/A mixed with 10 gallons of water after seed sowing. Before planting, all of the soybean seed in the trial was treated with an industry standard fungicide seed treatment (CRUISERMAXX VIBRANT + UNICOAT NEUTRAL + red color). In the trial, the control consisted of seed treated with CRUISERMAXX VIBRANT, UNICOAT NEUTRAL, and red color alone. It was a replicated field plot trial with 4 replicates arranged in a completely randomized design. Soybean was harvested at the end of the season and data were analyzed by ARM software. The composition reduced the number of galls caused by root knot nematodes (Meloidogyne .sp.) by 62.5% and yielded soybean production equal to control (Figure 4 A & B).

Example 5 Composition Exhibits Control of Plant Parasitic Fungi

The nematicide composition was also tested against plant pathogenic fungi. White mold (Sclerotium rolfsii) and black leaf spot fungi (Nothopassarola personata) were collected from peanut fields of University of Georgia, Tifton. Through in vitro petri plate assays, nematicide composition was tested at two concentrations lx and 2x, against the growth of fungal spores on culture medium. An industry standard fungicide, RANCONA VPD (from UPL), was used as a positive control. The white mold assay is shown in Figure 5 and the black leaf spot fungi assay in Figure 6. For the white mold assay, the cellulose- based carrier hydroxyethyl cellulose alone did not exhibit any control of fungus (Fig. 5a). Fungicide VPD showed some control of fungus by zone of inhibition (or zone of clearance) but soon got occupied by fungus (Fig. 5d). The nematicide composition, both at lx (Fig. 5b) and 2x (Fig. 5c) concentration restricted fungal growth around the application areas at the center of the culture plates. For Black Leaf spot fungus, fungicide VPD showed a good control of fungus by producing larger clearing zone around the point of application (Fig. 6c). The nematicide composition at lx and 2x concentrations also exhibited control of fungal growth but the effect was less than fungicide VPD (Fig. 6a & b).

In both assays, the nematicide composition showed a similar pattern of control efficacy at 1 x and 2x concentrations. These zone of inhibition assays demonstrate the antifungal activity of the composition and its broad- spectrum nature.

Example 6

Composition Exhibits Control of Plant Parasitic Insects The nematicide composition was also tested against insects. Plant feeding insects, particularly those under the coleoptera order (most beetles and weevils) are very voracious insect for crops. As surrogates of coleoptera insects’ larvae rootworm and wireworm, Superworm (Zophobas mor io) and mealworm (Tenebrio molitor) were used in this experiment. Individual live insect larva was placed in a glass beaker or a plastic weighing tray, and about one milliliter of the nematicide composition was applied onto the body of a larva. At contact, the larva reacted by shaking its body and turning sideway or upside, and then it became paralyzed within 2 minutes and eventually died within 15-20 minutes (data not shown). These results demonstrate the insecticidal effect of the composition as a broad spectrum biopesticide.

Accordingly, while the compositions and methods have been described in reference to specific embodiments, features, and illustrative embodiments, it will be appreciated that the utility of the subject matter is not thus limited, but rather extends to and encompasses numerous other variations, modifications and alternative embodiments, as will suggest themselves to those of ordinary skill in the field of the present subject matter, based on the disclosure herein.

Various combinations and sub-combinations of the compositions, methods, and features described herein are contemplated and will be apparent to a skilled person having knowledge of this disclosure. Any of the various features and elements as disclosed herein may be combined with one or more other disclosed features and elements unless indicated to the contrary herein. Correspondingly, the subject matter as hereinafter claimed is intended to be broadly construed and interpreted, as including all such variations, modifications and alternative embodiments, within its scope and including equivalents of the claims.

Claims

CLAIMS What is claimed is:

1. A liquid composition comprising:

(i) swertiamarin:

(ii) berberine;

(iii) chitosan;

(iv) glutathione; and

(v) cellulose-based polymer, wherein the composition is in the form of an emulsion or a suspension and has activity against one or more plant pathogens.

2. The liquid composition of claim 1, wherein the total amount of component (i) is in the range from 0.01 to 1.0 g/L, the total amount of component (ii) is in the range from 0.005 to 0.5 g/L, the total amount of component (iii) is in the range from 0.0025 to 0.25 g/L, and the total amount of component (iv) is in the range from 0.002 to 0.2 g/L.

3. The liquid composition of claim 1, wherein the plant pathogen is one or a combination of a nematode, an insect, or a fungus.

4. The liquid composition of claim 1, wherein the emulsion or suspension is an oilbased or an aqueous-based emulsion or suspension.

5. The liquid composition ofclaim 1, wherein the cellulose-based polymer comprises hydroxyethylcellulose, hydroxyethyl propyl methyl cellulose (HPMC), quillaja extract, xanthan gum, corn starch, or yucca extract, and combinations thereof.

6. The liquid composition of claim 1, wherein the composition further comprises one or more plant biostimulants.

7. The liquid composition of claim 6, wherein the plant biostimulant comprises amino acids, humic acid, fulvic acid, seaweed extract, laminarin, alginates, or polysaccharides, and combinations thereof.

8. The liquid composition of any of claim 1, wherein the composition further comprises one or more antioxidants.

9. The liquid composition of claim 8, wherein the antioxidant comprises vitamin C, vitamin E, coenzyme Q10, lipoic acid, flavonoids, phenols, polyphenols, or phytoestrogens, and combinations thereof.

10. A plant seed coated with a composition comprising:

(i) swertiamarin;

(ii) berberine;

(iii) chitosan;

(iv) glutathione; and

(v) cellulose-based polymer, wherein the composition is present in the coating in an amount suitable to improve plant growth and/or health in the presence of a plant pathogen, or both.

11. The plant seed of claim 10, wherein the total amount of component (i) is in the range from 0.01 to 1.0 g/L, the total amount of component (ii) is in the range from 0.005 to 0.5 g/L, the total amount of component (iii) is in the range from 0.0025 to 0.25 g/L, and the total amount of component (iv) is in the range from 0.002 to 0.2 g/L.

12. The plant seed of claim 10, wherein the plant pathogen is one or a combination of a nematode, an insect, or a fungus.

13. The plant seed of claim 10, wherein the cellulose-based polymer comprises hydroxyethylcellulose, hydroxyethyl propyl methyl cellulose (HPMC), quillaja extract, xanthan gum, corn starch, or yucca extract, and combinations thereof.

14. The plant seed of claim 10, wherein the composition further comprises one or more plant biostimulants.

15. The plant seed of claim 14, wherein the plant biostimulant comprises amino acids, humic acid, fulvic acid, seaweed extract, laminarin, alginates, or polysaccharides, and combinations thereof.

16. The plant seed of claim 10, wherein the composition further comprises one or more antioxidants.

17. The plant seed of claim 16, wherein the antioxidant comprises one or more of vitamin C, vitamin E, coenzyme Q10, lipoic acid, flavonoids, phenols, polyphenols, or phytoestrogens, and combinations thereof.

18. A method for improving plant growth and/or health comprising: planting a plant or a seed of a plant in a suitable growing medium, the plant or the seed having a coating or partial coating of a composition comprising:

(i) swertiamarin;

(ii) berberine;

(iii) chitosan;

(iv) glutathione; and

(v) cellulose-based polymer, wherein the composition is present in an amount suitable to improve plant growth and/or health in the presence of a plant pathogen, or both.

19. The method of claim 18, wherein the total amount of component (i) is in the range from 0.01 to 1.0 g/L, the total amount of component (ii) is in the range from 0.005 to 0.5 g/L, the total amount of component (iii) is in the range from 0.0025 to 0.25 g/L, and the total amount of component (iv) is in the range from 0.002 to 0.2 g/L.

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20. The method of claim 18, wherein the plant pathogen is one or a combination of a nematode, an insect, or a fungus.

21. The method of claim 18, wherein the cellulose-based polymer comprises hydroxyethylcellulose, hydroxyethyl propyl methyl cellulose (HPMC), quillaja extract, xanthan gum, corn starch, or yucca extract, and combinations thereof.

22. The method of claim 18, wherein the composition further comprises one or more plant biostimulants.

23. The method of claim 23, wherein the plant biostimulant comprises amino acids, humic acid, fulvic acid, seaweed extract, laminarin, alginates, or polysaccharides, and combinations thereof.

24. The method of claim 18, wherein the composition further comprises one or more antioxidants.

25. The method of claim 24, wherein the antioxidant comprises vitamin C, vitamin E, coenzyme Q10, lipoic acid, flavonoids, phenols, polyphenols, or phytoestrogens, and combinations thereof.

26. A method for improving plant growth and/or health comprising: delivering to seed of a plant, foliage of a plant, roots of a plant, or soil or growth medium surrounding a plant, a liquid composition in the form of an emulsion or a suspension comprising:

(i) swertiamarin;

(ii) berberine;

(iii) chitosan;

(iv) glutathione; and

(v) cellulose-based polymer,

20 wherein the composition improves plant growth and/or health in the presence of a plant pathogen, or both.

27. The method of claim 26, wherein the total amount of component (i) is in the range from 0.01 to 1.0 g/L, the total amount of component (ii) is in the range from 0.005 to 0.5 g/L, the total amount of component (iii) is in the range from 0.0025 to 0.25 g/L, and the total amount of component (iv) is in the range from 0.002 to 0.2 g/L.

28. The method of claim 26, wherein the plant pathogen is one or a combination of a nematode, an insect, or a fungus.

29. The method of claims 26, wherein the emulsion or suspension is an oil-based or an aqueous-based emulsion or suspension.

30. The method of claim 26, wherein the cellulose-based polymer comprises hydroxyethylcellulose, hydroxyethyl propyl methyl cellulose (HPMC), quillaja extract, xanthan gum, corn starch, or yucca extract, and combinations thereof.

31. The method of claim 26, wherein the composition further comprises one or more plant biostimulants.

32. The method of claim 31, wherein the plant biostimulant comprises amino acids, humic acid, fulvic acid, seaweed extract, laminarin, alginates, or polysaccharides, and combinations thereof.

33. The method of claim 26, wherein the composition further comprises one or more antioxidants.

34. The method of claim 33, wherein the antioxidant comprises vitamin C, vitamin E, coenzyme Q10, lipoic acid, flavonoids, phenols, polyphenols, or phytoestrogens, and combinations thereof.

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35. The liquid composition, plant seeds, or methods of any of claims 1-34, wherein the plants or plant seeds comprise cotton, corn, peanut, wheat, soybean, sorghum, or canola, and combinations thereof.

36. The liquid composition, plant seeds, or methods of any of claims 1-34, wherein the plants or plant seeds comprise legumes.

37. The liquid composition, plant seeds, or methods of any of claims 1-34, wherein the plants or plant seeds comprise non-legumes.

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