A composition comprising chitooligosaccharides

Abstract

The present disclosure relates to a composition comprising a chitooligosaccharide. More particularly, the present disclosure relates to a fungicidal composition comprising a chitooligosaccharide and a wetting agent. Also provided herein are the methods of making the composition and uses thereof.

Description

A COMPOSITION COMPRISING CHITOOLIGOSACCHARIDESFIELD OF INVENTION

[0001] The present invention relates to a composition comprising a chitooligosaccharide. Particularly, the present invention relates to a fungicidal composition comprising a chitooligosaccharide and a wetting agent. The present invention also relates to method of making the composition and methods of uses thereof.BACKGROUND

[0002] Fungal diseases are a threat to pre-harvest and post-harvest stages of agriculture and cause massive damage to the produce and the health of a plant. Numerous solutions are sought after by agriculturists to counter the rampant fungal damage to agricultural produce. Biological solutions are gaining more attention over conventional chemical crop protection agents due to the numerous advantages offered over the latter, in terms of safety to the health of growers, consumers and environment.

[0003] Chitooligosaccharides (CHOS) which encompass chitopoly- or chitooligosaccharides are oligomers prepared from chitosan, and sometimes from chitin, either mechanically, chemically or enzymatically. Low molecular weight chitosan or chitooligosaccharides are known for their fungicidal properties. It is essential that chitooligosaccharides are delivered to the target in a suitable formulation, wherein the chitooligosaccharides are formulated appropriately which increases the stability and efficacy of the active ingredient. However, there is a lack of evidence and research on formulating and delivering chitooligosaccharides in form of an effective composition.

[0004] Thus, there is a need for developing stable and effective compositions of chitooligosaccharides. The present invention aims to solve this aforementioned need and provide a stable composition of chitooligosaccharides. An objective of the present invention is to increase the efficacy of the active ingredient by formulating the chitooligosaccharides with suitable excipients. Another objective is to provide a composition of chitooligosaccharides which can be safely and effectively delivered or applied to the target locus. Thus, an object of the present invention is to provide a composition comprising a chitooligosaccharide and a wetting agent to fulfill the need reverberated herein.BRIEF SUMMARY

[0005] In an aspect, there is provided a composition comprising a chitooligosaccharide and a wetting agent. In an embodiment, said wetting agent is a surfactant selected from a nonionic surfactant, a cationic surfactant, an anionic surfactant, or combinations thereof.

[0006] In an aspect, there is provided a composition comprising a chitooligosaccharide and an anionic surfactant as a wetting agent.

[0007] In an aspect, there is provided a composition comprising a chitooligosaccharide and a wetting agent comprising a combination of a nonionic surfactant, a cationic surfactant, and an anionic surfactant.

[0008] In another aspect, there is provided a method of fungal pathogen and disease control in a plant or a part thereof, said method comprising administering to the plant or a part thereof a composition comprising a chitooligosaccharide and a wetting agent. In an embodiment, said composition may be combined or applied with an additional active ingredient.

[0009] An aspect also provides a use of a composition comprising a chitooligosaccharide and a wetting agent as a fungicide for controlling fungal pathogen and disease in a plant or a part thereof.DETAILED DESCRIPTION

[0010] For the purposes of the following detailed description, it is to be understood that the invention may assume various alternative variations and step sequences, except where expressly specified to the contrary. Moreover, other than in any operating examples, or where otherwise indicated, all numbers expressing, for example, quantities of materials / ingredients used in the specification are to be understood as being modified in all instances by the term "about".

[0011] Unless otherwise stated, all exact values provided herein are representative of corresponding approximate values (e.g., all exact exemplary values provided with respect to a particular factor or measurement can be considered to also provide a corresponding approximate measurement, modified by "about," where appropriate). As used herein, the term "about" refers to a measurable value such as a parameter, an amount, a temporal duration, and the like and is meant to include variations of + / -15% or less, specifically variations of + / -10% or less, more specifically variations of + / -5% or less, even more specifically variations of + / -1% or less, and still more specifically variations of + / -0.1% or less of and from the particularly recited value, in so far as such variations are appropriate to perform in the disclosure described herein.

[0012] Recitation of ranges of values are merely intended to serve as a shorthand method of referring individually to each separate value falling within the range, unless otherwise indicated herein, and each separate value is incorporated into the specification as if it were individually recited herein. All provided ranges of values are intended to include the end points of the ranges, as well as values between the end points, and are independently combinable. As used herein, all numerical values or numerical ranges include integers within such ranges and fractions of the values or the integers within ranges unless the context clearly indicates otherwise. Thus, for example, reference to a range of 90-100%, includes 91%, 92%, 93%, 94%, 95%, 95%, 97%, etc., as well as 91.1%, 91.2%, 91.3%, 91.4%, 91.5%, etc., 92.1%, 92.2%, 92.3%, 92.4%, 92.5%, etc., and so forth. All methods described herein can be performed in a suitable order unless otherwise indicated herein or otherwise clearly contradicted by context.

[0013] It must be noted that, as used in this specification, the singular forms “a,” “an” and “the” include singular and plural referents unless the content clearly dictates otherwise.

[0014] The terms “preferred” and “preferably” refer to embodiments of the invention that may afford certain benefits, under certain circumstances.

[0015] As used herein, the terms “comprising” “including,” “having,” “containing,” “involving,” and the like are to be understood to be open-ended, i.e., to mean including but not limited to.

[0016] The term “chitooligosaccharide” refers to any chitopoly- or chitooligosaccharides and encompasses hydrolysis or cleaved products obtained from chitosan or chitin by chemical, physical, enzymatic means or combinations of any of these processes. It encompasses chitopoly - or chitooligosaccharides of degree of polymerization less than 250, degree of acetylation less than or equal to 40% and / or molecular weight less than 42000 Da. Furthermore, it will be appreciated that structurally and functionally equivalent poly- or oligosaccharides i.e. comprising deacetylated and acetylated units in the preparations described herein, may be derived from sources other than chitosan (or chitin), or they may be produced synthetically. Said term also includes plural variants. Said chitooligosaccharide can be obtained by a process as described in entirety in W02014064119 and may have the properties as described therein.

[0017] "Degree of acetylation" (DA) (i.e. N-acetylation, expressed as a percentage) and "fraction of acetylated residues" (FA, expressed as a value between 0 and 1) are used interchangeably herein and describe the molar fraction of residual N-acetyl-D- glucosamine groups in chitosan or the chitooligosaccharides derived from chitosan. Degree of N -acetylationrefers to the fraction of N-acetyl- D-glucosamine sugars in the molecule (i.e. 10% N-acetylation is reflected as 0.10); the other sugars are D-glucosamine units. As referred to herein "monomers", "units" and "residues" are used interchangeably to refer to individual saccharide molecules that are linked to form the polymer.

[0018] The "average degree of polymerization" (DPn) of a chitosan or chitooligosaccharide derived therefrom is defined herein as the average number of D- glucosamine and N-acetyl-D- glucosamine monomeric units in a chitosan or polysaccharide or oligosaccharide molecule derived from chitosan. The DPn values recited herein represent the DPn of a chitosan / chitooligosaccharide preparation and thus encompass both variability in the range of molecules present in the preparation and furthermore in the detection accuracy. In relation to the latter, variation in determining the DPn may be ±10, e.g. ±5%. Thus for example, a preparation with a DPn of 30 may have a DPn ranging from 28.5 to 31.5 depending on the measurement system used. Preferably the measurement is in accordance with the methods used conventionally (i.e. by 1H- NMR spectroscopy). The variation in the range of molecules present in the preparation exists and the extent of variation may vary. Thus, depending on the preparation method, the molecules in the preparation may be widely divergent in size or the molecules may be of a similar size (e.g. when a size exclusion separation step is used). Preferably the preparation includes molecules of a similar size, e.g. more than 50%, e.g. more than 70, 80, 90, 95 or 99% of the chitosan or chitooligosaccharide molecules in the preparation have a size that varies no more than 20% (e.g. less than 15, 10, 5 or 1 %) from the average DPn. Thus, for example, in a preparation with a DPn of 30 more than 50% of the molecules preferably have a size (DP) between 24 and 36 (20% deviation).

[0019] The term “plant” or “target plant” as used herein refers to any vegetation to which the compositions of the present disclosure can be applied for any purposes or any treatment. The term covers a whole plant or a part thereof, such as stems, branches, bark, pistils, flowers, petals, sepals, roots, rhizomes, buds, bulbs, tubers, petioles, nodes, internodes, leaves, leaflets, meristems, root tips, shoot tips, fruits, reproductive organs such as anther, stamen, carpel, ovary, style, stigma, and so forth. The term also covers seeds, seedlings, or any plant propagation material.

[0020] The term "plant propagation material" is to be understood to denote all the generative parts of the plant such as seeds and vegetative plant material such as cuttings and tubers (e. g. potatoes), which can be used for the multiplication of the plant. This includes seeds, roots, fruits,tubers, bulbs, rhizomes, shoots, sprouts and other parts of plants. Seedlings and young plants, which are to be transplanted after germination or after emergence from soil, may also be included. These plant propagation materials may be treated prophylactically with a plant protection compound either at or before planting or transplanting. In an embodiment, the term includes agronomically useful plants, for example vegetable, fruit and cereal crops, and ornamental plants.

[0021] The term "locus" as used herein, refers to an area, a crop or a part thereof which is infested by or prone to pest infestation and to which a method or composition or combination according to the present invention is applied. It includes any habitat, breeding ground, plant or a part thereof, plant propagation material, soil, area, material or environment in which a plant is growing or may grow, or where the pest breeds, attacks, may breed or may attack. The term covers the vicinity of a desired plant / plants in which control is desired.

[0022] While the invention has been illustrated and described in detail in the foregoing description, the same is to be considered as illustrative and not restrictive in character, it being understood that only the preferred embodiments have been shown and described and that all changes and modifications that come within the spirit of the invention are desired to be protected.

[0023] In an embodiment, there is provided a composition comprising a chitooligosaccharide and a wetting agent.

[0024] In an embodiment, there is provided a composition comprising a chitooligosaccharide, a wetting agent and / or agriculturally acceptable excipient.

[0025] In an embodiment, the chitooligosaccharides are characterised by any one or more of DPn (degree of polymerization), DA (degree of acetylation), and MW (molecular weight). In an embodiment, the chitooligosaccharides are characterized by the specification of W02014064119.

[0026] In an embodiment, the chitooligosaccharides are characterised by a DPn of about 250 or lesser. In another embodiment, the chitooligosaccharides are characterised by a DPn of about 200 or lesser. In another embodiment, the chitooligosaccharides are characterised by a DPn of about 150 or lesser. In another embodiment, the chitooligosaccharides are characterised by a DPn of about 100 or lesser. In another embodiment, the chitooligosaccharides are characterised by a DPn of about 75 or lesser. In another embodiment, the chitooligosaccharides are characterised by a DPn of about 60 or lesser. In a preferred embodiment, the chitooligosaccharides are characterised by a DPn in a range of about 10 to about 60. In a preferred embodiment, the chitooligosaccharides are characterised by a DPn in a range of about 15 to about 50. In a preferredembodiment, the chitooligosaccharides are characterised by a DPn in a range of about 20 to about 40.

[0027] In an embodiment, the chitooligosaccharides are characterised by a DA in a range of about 0.01 and about 0.40 or in a range of about 0.05 and about 0.40. Preferably the DA ranges from about 0.05 to 0.20, preferably from 0.1 to 0.20.

[0028] In an embodiment, the chitooligosaccharides are characterized by MW of about 42,000 Da or lower. In an embodiment, the chitooligosaccharides are characterized by MW in the range of about 1,680 - 42,000 Da. In another embodiment, the chitooligosaccharides are characterized by MW in the range of about 1,680 - 25,000 Da. In another embodiment, the chitooligosaccharides are characterized by MW in the range of about 1,680 - 15,000 Da. In another embodiment, the chitooligosaccharides are characterized by MW in the range of about 1,680 - 10,000 Da. In a preferred embodiment, the chitooligosaccharides are characterized by MW in the range of about 1,680 - 8,000 Da. In a preferred embodiment, the chitooligosaccharides are characterized by MW in the range of about 3,300 - 8,000 Da. In a preferred embodiment, the chitooligosaccharides are characterized by MW in the range of about 3,300 - 7,000 Da. In a preferred embodiment, the chitooligosaccharides are characterized by MW in the range of about 5,000 - 7,000 Da. In a most preferred embodiment, the chitooligosaccharides are characterized by MW in the range of about 5,900 - 6,700 Da.

[0029] In an embodiment, chitooligosaccharides are characterised by an average degree of polymerization (DPn) of about 250 or lesser, preferably in a range of about 10 to about 60, more preferably in a range of about 20 to 40; degree of acetylation (DA) in a range of about 0.01 to about 0.40, preferably in a range of about 0.05 to 0.20, more preferably in a range of about 0.1 to 0.20; and molecular weight (MW) of about 42,000 Da or lower, in the range of about 1,680 - 10,000 Da, more preferably in the range of about 5,900 - 6,700 Da.

[0030] In order to characterize chitooligosaccharides in terms of DPn, DA and MW, several techniques known in the art may be used, primarily nuclear magnetic resonance (NMR) and mass spectrometry (MS).

[0031] Thus, in an embodiment, the chitooligosaccharides are characterised by one or more of: i. a DPn of about 250 or lesser; ii. a DA in a range of about 0.01 and about 0.40 or in range of about 0.05 and about 0.40; and / oriii. MW of about 42,000 Da or lower.

[0032] In an alternate embodiment, the chitooligosaccharides are characterised by one or more of: i. a DPn in a range of about 10 to about 60; ii. a DA in a range of about 0.01 and about 0.40 or in a range of about 0.05 and about 0.40; and / or iii. MW in the range of about 1,680 - 15,000 Da.

[0033] In an embodiment, the chitooligosaccharides may be produced from chitin or chitosan using chemical, mechanical or enzymatic processes. In another embodiment, the chitooligosaccharides may be produced by a process as embodied in the application W02014064119.

[0034] Examples of wetting agents that can be selected for use in connection with the invention include, without limitation, anionic, cationic and / or nonionic (including silicone based) surfactants, oil, methylated oil or combinations thereof.

[0035] In an embodiment, said wetting agent may be selected from a nonionic surfactant, a cationic surfactant, an anionic surfactant, or combinations thereof.

[0036] The nonionic surfactant(s) may be either water soluble nonionic surfactants, water insoluble nonionic surfactants, or a combination of water soluble nonionic surfactants and water insoluble nonionic surfactants.

[0037] Water insoluble nonionic surfactants: Non-limiting examples of water insoluble nonionic surfactants include alkyl and aryl: poly oxy ethylenated fatty acids, alkanolamine condensates, alkanolamides, tertiary acetylenic glycols, polyoxyethylenated mercaptans, carboxylic acid esters, polyoxyethylenated polyoxyproylene glycols, sorbitan fatty esters, or combinations thereof. Also included are EO / PO block copolymers (EO is ethylene oxide, PO is propylene oxide), EO polymers and copolymers, polyamines, and polyvinylpynolidones.

[0038] Water soluble nonionic surfactants: Non-limiting examples of water soluble nonionic surfactants include - glycerol ethers, glycol ethers, ethanolamides, sulfoanylamides, alcohols, amides, alcohol ethoxylates (ethoxylated alcohol), glycerol esters, glycol esters, ethoxylates of glycerol ester and glycol esters, sugar-based alkyl polyglycosides, sorbitan fatty acid alcohol ethoxylates (ethoxylated alcohol) and sorbitan fatty acid ester ethoxylates.

[0039] Combination of nonionic surfactants: In one embodiment formulations of the compositions described herein comprise at least one nonionic surfactant. In one embodiment, the compositions comprise at least one water insoluble nonionic surfactant and at least one water soluble nonionic surfactant.

[0040] Preferred examples of nonionic surfactants include ethoxylated alcohol, polyarylphenol polyethoxyethers, polyalkylphenol polyethoxy ethers, polyglycol ether derivatives of saturated fatty acids, polyglycol ether derivatives of unsaturated fatty acids, polyglycol ether derivatives of aliphatic alcohols, polyglycol ether derivatives of cycloaliphatic alcohols, fatty acid esters of polyoxyethylene sorbitan, alkoxylated vegetable oils, alkoxylated acetylenic diols, polyalkoxylated alkylphenols, fatty acid alkoxylates, ethoxylated sorbitan esters, sorbitan alkoxylates, sorbitol esters, C8-C22 alkyl or alkenyl polyglycosides, polyalkoxy styrylaryl ethers, alkylamine oxides, block copolymer ethers, polyalkoxylated fatty glyceride, polyalkylene glycol ethers, linear aliphatic or aromatic polyesters, organo silicones, polyaryl phenols, sorbitol ester alkoxylates, polyalkylene oxide block copolymers, acrylic copolymers and mono- and diesters of ethylene glycol and mixtures thereof.

[0041] Preferred nonionic surfactants, according to an embodiment, are ethoxylated sorbitan esters such as ethoxylated sorbitan monolaurate, ethoxylated alcohol such as 2 -ethylhexanol ethoxylate, or combinations thereof.

[0042] The anionic surfactant(s) may be either water soluble anionic surfactants, water insoluble anionic surfactants, or a combination of water soluble anionic surfactants and water insoluble anionic surfactants. Non-limiting examples of anionic surfactants include sulfonic acids, sulfuric acid esters, carboxylic acids, and salts thereof. Non -limiting examples of water soluble anionic surfactants include alkyl sulfates, alkyl ether sulfates, alkyl amido ether sulfates, alkyl aryl polyether sulfates, alkyl aryl sulfates, alkyl aryl sulfonates, monoglyceride sulfates, alkyl sulfonates, alkyl amide sulfonates, alkyl aryl sulfonates, benzene sulfonates, toluene sulfonates, xylene sulfonates, cumene sulfonates, alkyl benzene sulfonates, alkyl diphenyloxide sulfonate, alpha-olefin sulfonates, alkyl naphthalene sulfonates, paraffin sulfonates, lignin sulfonates, alkyl sulfosuccinates, ethoxylated sulfosuccinates, alkyl ether sulfosuccinates, alkylamide sulfosuccinates, alkyl sulfosuccinamate, alkyl sulfoacetates, alkyl phosphates, phosphate ester, alkyl ether phosphates, acyl sarconsinates, acyl isethionates, N-acyl taurates, N- acyl-N-alkyltaurates, alkyl carboxylates, or a combination thereof.

[0043] Preferred examples of anionic surfactants include ethoxylated tristyrylphenol phosphate, tristyrylphenol ethoxylate phosphate ester, tristyryl phenol ethoxylate sulphate ester, Polyoxyethylene alkyl ether phosphate, alcohol sulfates, alcohol ether sulfates, alkyl ether sulfates, alkylaryl ether sulfates, alkylaryl sulfonates such as alkylbenzene sulfonates and alkylnaphthalene sulfonates and salts thereof, alkyl sulfonates, mono- or di-phosphate esters of polyalkoxylated alkyl alcohols or alkylphenols , mono- or di-sulfosuccinate esters of C12-C15 alkanols or polyalkoxylated C12-C15 alkanols, alcohol ether carboxylates, phenolic ether carboxylates, polybasic acid esters of ethoxylated poly oxyalkylene glycols consisting of oxybutylene or the residue of tetrahydrofuran, sulfoalkylamides and salts thereof such as N- methyl-N-oleoyltaurate Na salt, polyoxyalkylene alkylphenol carboxylates, polyoxyalkylene alcohol carboxylates alkyl polyglycoside / alkenyl succinic anhydride condensation products, alkyl ester sulfates, napthalene sulfonates, naphthalene formaldehyde condensates, alkyl sulfonamides, sulfonated aliphatic polyesters, sulfate esters of styrylphenyl alkoxy lates, and sulfonate esters of styrylphenyl alkoxylates and their corresponding sodium, potassium, calcium, magnesium, zinc, ammonium, alkylammonium, diethanolammonium, or triethanolammonium salts, salts of ligninsulfonic acid such as the sodium, potassium, magnesium, calcium or ammonium salt, polyarylphenol polyalkoxyether sulfates and polyarylphenol polyalkoxyether phosphates, and sulfated alkyl phenol ethoxylates and phosphated alkyl phenol ethoxylates, sodium stearate, potassium palmitate, sodium cetyl sulfate, sodium lauryl phosphate, sodium polyoxyethylene lauryl sulfate, triethanolamine palmitate, polyoxyethylene sodium lauryl phosphate, sodium N- acyl glutamate, and combinations thereof.

[0044] Preferred anionic surfactants, according to an embodiment, are ethoxylated tristyrylphenol phosphate, tristyrylphenol ethoxylate phosphate ester, alkyl ether sulfates, or combinations thereof.

[0045] Cationic surfactants include alkanol amides of Cs-Cis fatty acids and Cs-Cis fatty amine polyalkoxylates, Cio-Cis alkyldimethylbenzylammonium chlorides, coconut alkyldimethylaminoacetic acids, alkyl trimethyl ammonium chloride and phosphate esters of Cs- Cis fatty amine polyalkoxylates.

[0046] Preferred cationic surfactant, according to an embodiment, includes alkyl trimethyl ammonium chloride such as hexadecyltrimethylammonium chloride.

[0047] In an embodiment, there is provided a composition comprising a chitooligosaccharide and an anionic surfactant as a wetting agent. Said anionic surfactant may be selected fromethoxylated tristyrylphenol phosphate, tristyrylphenol ethoxylate phosphate ester, alkyl ether sulfates, or combinations thereof.

[0048] In an embodiment, there is provided a composition comprising a chitooligosaccharide and a combination of wetting agents selected from a nonionic surfactant, a cationic surfactant, and an anionic surfactant. In an embodiment, the nonionic surfactant may be selected from ethoxylated sorbitan esters such as ethoxylated sorbitan monolaurate, ethoxylated alcohol such as 2-ethylhexanol ethoxylate, or combinations thereof. In an embodiment, the cationic surfactant is alkyl trimethyl ammonium chloride such as hexadecyltrimethyl ammonium chloride. In an embodiment, the anionic surfactant may be selected from ethoxylated tristyrylphenol phosphate, tristyrylphenol ethoxylate phosphate ester, alkyl ether sulfates, or combinations thereof.

[0049] In an embodiment, the amount of the chitooligosaccharide in the composition ranges from about 1% w / w to 99.9 % w / w of the total weight of the composition. In an embodiment, the amount of the chitooligosaccharide in the composition ranges from about 1% w / w to 80 % w / w of the total weight of the composition. In an embodiment, the amount of the chitooligosaccharide in the composition ranges from about 1% w / w to 50 % w / w of the total weight of the composition. Preferably, the amount of the chitooligosaccharide in the composition ranges from about 1% w / w to 20 % w / w of the total weight of the composition. More preferably, the amount of the chitooligosaccharide in the composition ranges from about 1% w / w to 10 % w / w of the total weight of the composition.

[0050] In an embodiment, the amount of the wetting agent in the composition ranges from about 0.1 % w / w to 99 % w / w of the total weight of the composition. In an embodiment, the amount of the wetting agent in the composition ranges from about 0.1 % w / w to 50 % w / w of the total weight of the composition. In an embodiment, the amount of the wetting agent in the composition ranges from about 0.5 % w / w to 25 % w / w of the total weight of the composition. In a preferred embodiment, the amount of the wetting agent in the composition ranges from about 0.5 % w / w to 15 % w / w of the total weight of the composition.

[0051] In an embodiment, there is provided a composition comprising a chitooligosaccharide, a wetting agent and / or agriculturally acceptable excipient, wherein, the agriculturally acceptable excipient includes but not limited to antimicrobial agents, preservatives, deodorizers, coloring agents, fragrances, additional emulsifiers, additional solubilizers, corrosion inhibitors, biocides and additional solvents.

[0052] In an embodiment, said agriculturally acceptable excipient comprises antimicrobial agents, preservatives, deodorizers, coloring agents, fragrances, additional emulsifiers, additional solubilizers, corrosion inhibitors, biocides and additional solvents.

[0053] In an embodiment, said agriculturally acceptable excipient comprises a biocide. According to an embodiment, biocides may be selected from benzothiazoles, 1,2- benzisothiazolin-3-one, sodium dichloro-s-triazinetrione, sodium benzoate, potassium sorbate, l,2-phenyl-isothiazolin-3-one, chloroxylenol, paraoxybenzoate butyl.

[0054] In an embodiment, the amount of biocide in the composition ranges from about 0.01 % w / w to about 20 % w / w of the total weight of the composition.

[0055] In an embodiment, said agriculturally acceptable excipient comprises a biocide ranges from about 0.01 % w / w to about 20 % w / w of the total weight of the composition; wherein said biocide is selected from the group consisting of benzothiazoles, l,2-benzisothiazolin-3-one, sodium dichloro-s-triazinetrione, sodium benzoate, potassium sorbate, 1,2-phenyl-isothiazolin- 3 -one, chloroxylenol, paraoxybenzoate butyl.

[0056] Thus, an embodiment provides a composition comprising a chitooligosaccharide, a wetting agent and a biocide.

[0057] In an embodiment, said agriculturally acceptable excipient comprises a solvent. Said solvent is an optional component in the composition of the present invention. In a preferred embodiment, the solvent is water.

[0058] In an embodiment, said agriculturally acceptable excipient comprises a solvent, wherein the solvent is water.

[0059] Thus, an embodiment provides a composition comprising a chitooligosaccharide, a wetting agent, a biocide, and a solvent.

[0060] According to an embodiment, it is possible to add various kinds of oils, adjuvants, fertilizers or micronutrients and further pesticides (e.g. herbicides, insecticides, fungicides, growth regulators, safeners) to the composition of the present invention in the form of a premix or, if appropriate, not until shortly before use (tank-mix).

[0061] In an embodiment, said composition may be in any agriculturally suitable form for storage and application to a locus, a surface, a plant propagation material, foliage or the ground.

[0062] The compositions of the present invention may be formulated into solid, or liquid formulations, including but not limited to wettable powders, granules, dusts, soluble liquidconcentrates, suspension concentrates, oil in water emulsion, water in oil emulsion, emulsifiable concentrates, capsule suspensions, ZC formulations, oil dispersions or other known formulation types.

[0063] In an embodiment, the composition is formulated into solid or liquid formulations selected from wettable powders, granules, dusts, soluble liquid concentrates, suspension concentrates, oil in water emulsion, water in oil emulsion, emulsifiable concentrates, capsule suspensions, ZC formulations and oil dispersions.

[0064] In a preferred embodiment, said composition is an SL formulation.

[0065] An embodiment of the present invention provides a soluble liquid concentrate composition comprising a chitooligosaccharide and a wetting agent.

[0066] In an embodiment, the composition has a pH in the range of about 2 to about 6. Preferably, the composition has a pH in the range of about 2.5 to about 5.5.

[0067] In an embodiment, the composition is stored at a temperature of about 15° C to about 50° C to avoid color change. Preferably the composition is stored at a temperature of about 35° C to 40° C to avoid color change.

[0068] In another embodiment, there is provided a method of fungal pathogen and disease control at a locus or in a plant or a part thereof, said method comprising administering to the locus, or the plant or a part thereof a composition comprising a chitooligosaccharide and a wetting agent.

[0069] According to an embodiment, the composition can be administered in undiluted form or diluted with a solvent. According to an embodiment, the composition can be administered, for example, in the formulation forms customary for liquid preparations, either as such or after prior dilution with water or any solvent. Application is effected by customary methods, i.e., for example by spraying, pouring or injecting.

[0070] Said compositions of the present invention can advantageously be mixed with water, and optionally one or more additional ingredients, at or near the location where the end user intends to apply the composition to a locus.

[0071] The composition may also be used for application to or treatment or coating of a plant propagation material such as seeds etc.

[0072] An embodiment of the present invention provides the use of the composition comprising a chitooligosaccharide and a wetting agent as a fungicide for controlling fungal pathogen and disease in a plant or a part thereof.

[0073] Surface tension is a property of liquids that causes them to resist spreading out and behaving as if they are coated with an elastic film. This is due to the cohesive forces between the molecules of the liquid, which cause them to behave as if they are attracted to one another. The surface tension of a liquid is determined by the strength of these cohesive forces and is typically measured in units of mN per meter (mN / m). In the context of agrochemicals, surface tension is important because it determines how well the chemicals spread and penetrate through the surface of plants and soil. Chemicals with low surface tension can spread more easily and penetrate deeper into the surface, making them more effective at controlling pests and promoting plant growth. On the other hand, chemicals with high surface tension may be less effective because they tend to bead up and roll off of surfaces rather than spreading out and penetrating the surface of a leaf. A liquid with a lower surface tension will wet the surface more easily.

[0074] In an embodiment, the surface tension or the dynamic surface tension of the compositions of the present invention is reduced as compared to a composition of chitooligosaccharides without a wetting agent. The compositions of the present invention improves wetting of chitooligosachcharide by reducing dynamic surface tension when applied on crops at field dilution. The reduction in the dynamic surface tension of the compositions of the present invention compared to a composition of chitooligosaccharides without a wetting agent is atleast 10% or more. Said reduction in dynamic surface tension ranges from about 10% to about 50%, preferably about 10% to about 30%. In an embodiment, the dynamic surface tension of the compositions of the present invention is evaluated by conventional and other suitable methods known to a skilled person in the art.

[0075] In an embodiment, the compositions of the present invention may be combined or mixed with an additional active ingredient or a composition comprising the additional active ingredient. Said combination may be premixed, tank mixed at the time of application or may involve sequential application of the compositions of the present invention. Thus, in an embodiment, the compositions of the present invention and an additional active ingredient or a composition comprising the additional active ingredient may be applied jointly or separately.

[0076] In an embodiment, the additional active ingredient may be a fungicide, a biological agent, a biostimulant, a synergist, an insecticide, or any other agent compatible with the compositions of the present invention.

[0077] In an embodiment, the composition further comprising an additional active ingredient selected from the group consisting of fungicide, a biological agent, a biostimulant, a synergist or an insecticide.

[0078] Specifically, the invention includes embodiments in which particular subject matter is excluded, in full or in part, such as substances or materials, method steps and conditions, protocols, procedures, assays or analysis. Thus, even though the invention is generally not expressed herein in terms of what the invention does not include aspects that are not expressly included in the invention are nevertheless disclosed herein. Although a number of embodiments of the invention have been described, it will be understood that various modifications may be made without departing from the spirit and scope of the invention.

[0079] These and other advantages of the invention may become more apparent from the examples set forth. These examples are provided merely as illustrations of the invention and are not intended to be construed as a limitation thereof.EXAMPLESExample 1 - Compositions and stability

[0080] Chitooligosaccharides prepared by the process as described in W02014064119 were formulated into following compositions described in Table 1 . The formulation process was carried out by charging required quantity of water into a mixing vessel followed by agitation. Chitooligosaccharide was charged into the vessel and agitated for 60 minutes for dissolving the chitooligosaccharides. The wetting agent(s) was / were charged into the vessel, followed by the preservative and further agitated for 60-90 min for obtaining a homogenous and clear solution. The solution was filtered and packed into a container. Stability analysis of the compositions was conducted. The stability of the compositions was tested at (i) 25±2°C and 0°C for a period of 7 days and at (ii) 35 °C for a period of 12 weeks.Table 1 : Compositions of chitooligosaccharides with wetting agents.Example 2: Composition of chitooligosaccharide

[0081] Chitooligosaccharides prepared by the process as described in W02014064119 were formulated into an SL composition described in Table 2. Table 2: Composition of chitooligosaccharide with anionic surfactant as wetting agent.Example 3: Composition of chitooligosaccharide

[0082] Chitooligosaccharides prepared by the process as described in W02014064119 were formulated into an SL composition described in Table 3. Table 3: Composition of chitooligosaccharide with combination of surfactants as wetting agentsExample 4: Efficacy of compositions of Example 2 and Example 3

[0083] The efficacy of compositions of Example 2 in controlling cucumber powdery mildew was compared with a composition of chitooligosaccharide 10 SL formulated with water and without a wetting agent, provided in table 4.Table 4: Composition of chitooligosaccharide without wetting agent

[0084] The two compositions were applied at a dosage of 2 L / ha and 4 L / ha each (each corresponding to 186 g / ha of chitooligosaccharide) on potted cucumber plants infected with powdery mildew. The experiment was conducted in 3 repetitions. Water volume was maintained at 500L / ha. 3 applications of each composition were made at an interval of 8 days. Assessment was done to identify % disease severity and % disease control which is presented in table 5.Table 5: Efficacy of chitooligosaccharide compositions with and without wetting agents against powdery mildew of cucumber

[0085] From the above table it is evident that the compositions of the present invention provide better disease control compared to a composition comprising chitooligosaccharide and no wetting agent, despite all the compositions comprising equivalent amount of active ingredient.Example 5: Control of fungal pathogens by composition of example 2, example 3 and table 4.Efficacy of the compositions of the example 2, 3 and table 4 was evaluated in petriplates against various fungi which cause fungal diseases in fruits, vegetables and cereals by applying said compositions with concentration of chitooligosaccharides ranging from 100 ppm to 600 ppm.The experiment was conducted in 3 replicates of each treatment and efficacy calculated using Minimum Inhibitory Concentration (MIC) method.Table 6: Efficacy of composition of example 2 against fungal pathogens (MIC)= not conducted Table 7: Efficacy of composition of example 3 against fungal pathogens (MIC)= not conductedIt is observed from the above tables that wetting agent plays an important role in increasing the efficacy of the chitooligosaccharides against fungal pathogens which cause diseases across various crops.Example 6: Dynamic surface tension analysis.Estimation of dynamic surface tension was done by using Bubble pressure tensiometer, model-BPlOO (KRUSS, Germany). Working sample for analysis was prepared by diluting formulation product (chitooligosaacharide without wetting agent for control, composition of example 2 and composition of example 3) in water at a concentration of 150 - 400 g active ingredient per liter of chitooligosaccharides. Dynamic surface tension was measured from bubble age of 10 mili Second to 500 mili second at 25±2°C and rH = 65 - 75%. Dynamic surface tension under lab condition is very much corelated with actual field spray condition on crop at application dose rate.Table 8: % change (decrease) in surface tension

Claims

I / WE CLAIM,1) A composition comprising a chitooligosaccharide, a wetting agent and / or agriculturally acceptable excipient.2) The composition of claim 1, wherein chitooligosaccharides are characterised by an average degree of polymerization (DPn) of about 250 or lesser, preferably in a range of about 10 to about 60, more preferably in a range of about 20 to 40; degree of acetylation (DA) in a range of about 0.01 to about 0.40, preferably in a range of about 0.05 to 0.20, more preferably in a range of about 0.1 to 0.20; and molecular weight (MW) of about 42,000 Da or lower, in the range of about 1,680 - 10,000 Da, more preferably in the range of about 5,900 - 6,700 Da.3) The composition of claim 1, wherein the wetting agent is selected from a nonionic surfactant, a cationic surfactant, an anionic surfactant, or combinations thereof.4) The composition of claim 1, wherein the nonionic surfactant(s) is a water soluble nonionic surfactants, water insoluble nonionic surfactants, or a combination of water soluble nonionic surfactants and water insoluble nonionic surfactants.5) The composition of claim 1, wherein the chitooligosaccharide is present in the composition in a range of about 1 % w / w to about 99.9 % w / w, preferably in the range of the about 1% w / w to about 50 % w / w, more preferably in the range of about 1% w / w to about 10 % w / w of the total weight of the composition.6) The composition of claim 1, wherein the wetting agent is present in the composition in a range of about 0.1 % w / w to about 99 % w / w, preferably in the range of the about 0.1 % w / w to about 50 % w / w, more preferably in the range of about 0.5% w / w to about 15 % w / w of the total weight of the composition.7) The composition of claim 1 , wherein the agriculturally acceptable excipient is selected from antimicrobial agents, preservatives, deodorizers, coloring agents, fragrances, additional emulsifiers, additional solubilizers, corrosion inhibitors, biocides and additional solvents.8) The composition of claim 1, wherein the agriculturally acceptable excipient comprises a biocide ranges from about 0.01 % w / w to about 20 % w / w of the total weight of the composition; wherein said biocide is selected from the group consisting of benzothiazoles, l,2-benzisothiazolin-3-one, sodium dichloro-s-triazinetrione, sodium benzoate, potassium sorbate, l,2-phenyl-isothiazolin-3-one, chloroxylenol, paraoxybenzoate butyl.9) The composition of claim 1, wherein the agriculturally acceptable excipient comprises a solvent, wherein the solvent is water.10) The composition of claim 1, wherein the composition is formulated into solid or liquid formulations selected from wettable powders, granules, dusts, soluble liquid concentrates, suspension concentrates, oil in water emulsion, water in oil emulsion, emulsifiable concentrates, capsule suspensions, ZC formulations and oil dispersions.11) The composition of claim 1, wherein the composition has a pH in the range of about 2 to about 6, preferably, the composition has a pH in the range of about 2.5 to about 5.5.12) A method of fungal pathogen and disease control at a locus or in a plant or a part thereof, said method comprising administering to the locus, or the plant or a part thereof a composition as claimed in claim 1.13) The composition of claim 1, wherein said composition further comprising an additional active ingredient selected from the group consisting of fungicide, a biological agent, a biostimulant, a synergist or an insecticide. 14) Use of the composition as claimed in claim 1 as a fungicide for controlling fungal pathogen and disease in a plant or a part thereof.

Images