Pesticide adjuvants

Abstract

Novel pesticide formulations are described that contain an adjuvant selected from iricicoline and lamellicholic acid anhydride and a pesticidal activity. Concentrates suitable for forming formulations are also provided. Irisicoline and lamellicholic acid anhydride serve as adjuvants in the concentrates and pesticide formulations. Use of the iricicoline and lamellicholic acid anhydride as adjuvants in pesticide formulations is also provided. Methods for making the formulations and treating soil with the formulations are also provided. A method for obtaining iricicoline and lamellicholic acid anhydride from a culture of Cosmospora sp. strain RKDO1747 is also described.

Description

[Technical Field]

[0001] CROSS-REFERENCE TO RELATED APPLICATIONS This application claims priority to U.S. Provisional Application No. 63 / 004,875, entitled "AGROCHEMICAL ADJUVANTS," filed April 3, 2020, the contents of which are incorporated herein by reference in their entirety for all purposes.

[0002] The present invention generally relates to microbially derived adjuvants for pesticidal active formulations and methods of providing adjuvants in pesticide formulations comprising one or more pesticidal adjuvants. The invention also includes treating crops with said formulations.

[0003] Adjuvants are generally defined as chemicals or mixtures of chemicals that can improve the biological activity or efficacy of pesticides. Adjuvants themselves do not control or kill pests. Instead, these additives may interact with molecular targets (e.g., cell walls, ion channels, structural proteins, enzymes) within the target organism or alter the properties of the pesticide formulation (e.g., diffusion, retention, penetration, droplet size), thereby improving the biological activity of the pesticide acting on the organism. Typical types of compounds used as adjuvants include small molecules, surfactants, emulsifiers, oils, and salts. Adjuvants typically do not inhibit the translocation of activity in treated plants. Additionally, adjuvants must not cause undesirable phytotoxic effects on plants.

[0004] Fungi are widespread in terrestrial environments and pose a major challenge to agricultural productivity. Undetected fungal infections can account for over 80% of pre- and post-harvest crop losses. To reduce these losses and meet growing food demands, the use of fungicides to control fungal agricultural pests is, and will continue to be, an important component of agricultural pest management systems.

[0005] New strategies for combating agricultural pests, especially fungal pests, need to be developed. One strategy is to develop adjuvants—safe, nontoxic chemicals that improve the effectiveness of existing fungicides already approved for use in field and greenhouse crops—to prevent or reduce the impact of fungal pests on agricultural productivity. These adjuvants could improve field or postharvest pest control, thereby increasing productivity. They may also reduce the amount of fungicide needed to achieve a desired level of pest control, contributing to the goal of achieving sustainable productivity gains.

[0006] The present invention provides for the use of compounds in pesticide formulations in combination with pesticide actives where the compounds can provide a desired adjuvant, including improved efficacy of the activity. The present invention also provides for the use of pesticide concentrate and dilution formulations that include adjuvants.

[0007] The present invention also aims to provide compounds in agrochemical formulations where the compounds may provide adjuvant properties equivalent to or improved upon existing adjuvants.

[0008] The present invention also aims to provide the use of the compounds as adjuvants and formulations containing the compounds for use in providing adjuvants in pesticide formulations.

[0009] According to a first aspect of the present invention there is provided an agrochemical formulation comprising: i) an adjuvant selected from iricicoline H, hydroxyiricicoline H, iricicoline I, or lamellicholic acid anhydride according to formula (I):

[0010] [ka]

[0011] During the ceremony: R 1 are independently hydrogen or C1-C4 alkyl; R2 independently represent hydrogen, C1-C4 alkyl, hydroxyl, or C1-C4 alkoxyl; R 3 and R 4 independently represent hydrogen or C1-C4 alkyl; R 5 and R 6 independently represent hydrogen, C1-C4 alkyl, hydroxyl, or C1-C4 alkoxyl; and ii) at least one pesticide active substance;

[0012] According to a second aspect of the present invention there is provided a concentrate suitable for producing an agrochemical formulation of the first aspect, said concentrate comprising: i) an adjuvant selected from iricicoline H, hydroxyiricicoline H, iricicoline I, or lamellicholic acid anhydride according to formula (I):

[0013] [ka]

[0014] During the ceremony: R 1 are independently hydrogen or C1-C4 alkyl; R 2 independently represent hydrogen, C1-C4 alkyl, hydroxyl, or C1-C4 alkoxyl; R 3 and R 4 independently represent hydrogen or C1-C4 alkyl; R 5 and R 6 independently represent hydrogen, C1-C4 alkyl, hydroxyl, or C1-C4 alkoxyl; and ii) at least one pesticide active substance;

[0015] According to a third aspect of the present invention, there is provided the use of a compound selected from iricicoline H, hydroxylsilin H, iricicoline I, or lamellicol anhydride according to formula (I) as an adjuvant in an agrochemical formulation comprising at least one agrochemical active:

[0016] [ka]

[0017] During the ceremony: R 1 are independently hydrogen or C1-C4 alkyl; R 2 independently represent hydrogen, C1-C4 alkyl, hydroxyl, or C1 to C4 alkoxyl; R 3 and R 4 independently represent hydrogen or C1-C4 alkyl; R 5 and R 6 independently represent hydrogen, C1 to C4 alkyl, hydroxyl, or C1 to C4 alkoxyl.

[0018] According to a fourth aspect of the present invention there is provided a method of treating vegetation to control pests, the method comprising applying a formulation of the first aspect or a diluted concentrate of the second aspect either to said vegetation or to the immediate environment of said vegetation.

[0019] According to a fifth aspect of the present invention there is provided a method of obtaining an adjuvant according to the first aspect, comprising the steps of: Cultivating Cosmospora sp. RKDO1747 in a medium under conditions that promote the metabolic synthesis of a fungicide adjuvant according to the first aspect from said Cosmospora sp.; and Purify synthetic adjuvants from culture media.

[0020] According to a sixth aspect of the present invention there is provided an organism comprising Cosmospora sp. strain RKDO1747, Agricultural Research Service Culture Collection (NRRL) accession number NRRL-67910.

[0021] According to a seventh aspect of the present invention, there is provided an extract obtained from a microorganism comprising the Cosmospora sp. strain RKDO1747, Agricultural Research Service Culture Collection (NRRL) Accession Number NRL-67910, according to the first aspect, wherein the extract contains iricicoline H, hydroxylsiline H, iricicoline I, or Lamellicholic Anhydride An extract is provided comprising at least one of:

[0022] According to an eighth aspect of the present invention there is provided a method of treating vegetation to control pests comprising applying an organism of the sixth aspect either to said vegetation or to the immediate environment of said vegetation.

[0023] According to a ninth aspect of the present invention there is provided a seed coating composition comprising an adjuvant according to the first aspect or a biological adjuvant according to the sixth aspect.

[0024] According to a tenth aspect of the present invention, there is provided lamellicholic anhydride according to formula (I):

[0025] [ka]

[0026] During the ceremony: R 1 , R 3 , R 4 , and R 6 are all hydrogen, and R 5 is hydroxyl and R 2 is methyl; or R 1 , R 4 , R 5 , and R 6 are all hydrogen, and R 2 and R 3 is methyl; or R 4 , R 5 , and R 6 are all hydrogen, and R 2 , R 3 and R 1 is methyl; or R 3 , R 5 , and R 6 are all hydrogen, and R 2 , R 4 and R 1 is methyl; or R 4 and R 5 is hydrogen and R 2 , R 3 , and R 1 is methyl and R 6 is hydroxyl; or R 3 and R 5 is hydrogen and R 2 , R 4 , and R 1 is methyl and R 6 is hydroxyl.

[0027] The compounds defined herein have been found to provide desirable adjuvant properties when used in pesticide formulations with at least one pesticide activity.Importantly, the identified classes of compounds, lamellicholic anhydride and illicorine, do not exhibit intrinsic insecticidal activity.

[0028] As used herein, the terms "for example," "for instance," "such," or "including" are meant to introduce examples that further clarify more general subject matter. Unless otherwise specified, these examples are provided solely as an aid in understanding the applications illustrated in this disclosure and are not intended to be limiting in any way.

[0029] When describing the number of carbon atoms in a substituent (e.g., "C1-C4 alkyl"), it is understood that the number refers to the total number of carbon atoms present in the substituent, including any carbon atoms present in any branching groups. Furthermore, when describing the number of carbon atoms in, for example, a fatty acid, this refers to the carbon atoms in the carboxylic acid and the total number of carbon atoms present in any branching groups.

[0030] The irisiccoline compound is selected from irisiccoline H, hydroxylsilylcholine H, or irisiccoline I. Preferably, the irisiccoline compound is irisiccoline H.

[0031] The above ilisicoline compounds are understood to be adjuvants and are referred to as such throughout.

[0032] The structure of Iricicolin H is 5-(4-hydroxyphenyl)-2-pyridone, which has a bicyclic decalin system. Iricicolin H compounds are understood to refer to compounds having the structure of formula (A).

[0033] [ka]

[0034] The structure of hydroxyl-iricicoline H is the same as iricicoline H with an additional hydroxyl group. It is understood that hydroxyl-iricicoline H compounds refer to compounds having the structure of formula (B).

[0035] [ka]

[0036] The structure of Iricicolin I is a further variation of the bicyclic decalin system. Iricicolin I compounds are understood to refer to compounds having the structure of formula (C):

[0037] [ka]

[0038] The adjuvant of the present invention can also be selected from lamellicholic anhydrides. It is understood that the adjuvant is preferably selected from lamellicholic anhydrides.

[0039] The lamellicholic anhydride is selected from those having the structure of formula (I):

[0040] [ka]

[0041] During the ceremony: R 1 are independently hydrogen or C1-C4 alkyl; R 2 independently represent hydrogen, C1-C4 alkyl, hydroxyl, or C1-C4 alkoxyl; R 3 and R 4 independently represent hydrogen or C1-C4 alkyl; R 5 and R 6 independently represent hydrogen, C1-C4 alkyl, hydroxyl, or C1-C4 alkoxyl.

[0042] The term "C1-C4 alkyl," as used herein, unless otherwise defined, refers to a saturated hydrocarbon radical that is straight or branched and contains from 1 to 4 carbon atoms. 1 From R 6 represents C1-C4 alkyl, the alkyl may be independently selected from methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, sec-butyl, tert-butyl, etc. Preferably, the C1-C4 alkyl is methyl or ethyl. More preferably, the C1-C4 alkyl is methyl.

[0043] The term "C1-C4 alkoxyl," as used herein, unless otherwise defined, refers to an alkyl group attached to an oxygen atom that is attached to an adjacent radical through the oxygen atom, forming an alkoxy radical having the structure -O-Alk, where Alk represents a C1-C4 alkyl group as defined herein.

[0044] R 2 , R 5 , and R 6 When any of the above represents a C1-C4 alkoxyl, said alkoxyl may be independently selected from methoxy, ethoxy, butoxy, propanoxy, etc. Preferably, the C1-C4 alkoxyl is methoxy or ethoxy. More preferably, the C1-C4 alkoxyl is methoxy.

[0045] Preferably, the compound of formula (I) is selected from: R 1 independently represent hydrogen or methyl; R 2 independently represent hydrogen or methyl, preferably methyl; R 3 and R 4 independently represent hydrogen or methyl, preferably both are hydrogen or one is hydrogen and the other is methyl; R 5 and R 6 independently represent hydrogen or hydroxyl, preferably both are hydrogen or one is hydrogen and the other is hydroxyl.

[0046] More preferably, the compound of formula (I) is selected from: R 1 , R 3 , R 4 , R 5 , and R 6 are all hydrogen, and R 2 is methyl; or R 1 , R 3 , R 4 , and R 6 are all hydrogen, and R5 is hydroxyl and R 2 is methyl; or R 1 , R 4 , R 5 , and R 6 are all hydrogen, and R 2 and R 3 is methyl; or R 4 , R 5 , and R 6 are all hydrogen, and R 2 , R 3 , and R 1 is methyl; or R 3 , R 5 , and R 6 are all hydrogen, and R 2 , R 4 , and R 1 is methyl; or R 4 and R 5 is hydrogen and R 2 , R 3 , and R 1 is methyl and R 6 is a hydroxyl group; or R 3 and R 5 is hydrogen and R 2 , R 4 , and R 1 is methyl and R 6 is a hydroxyl group.

[0047] Particularly preferred are lamellicholic anhydrides selected from:

[0048] [ka]

[0049] [ka]

[0050] The organism used for fermentation is a filamentous fungus belonging to the genus Cosmospora. A particular strain of Cosmospora species, RKDO1747, has been found to be particularly useful for the production of novel adjuvants, and this strain has been the subject of a deposit under the Budapest Treaty at the Agricultural Research Service Culture Collection (NRRL), Peoria, Illinois, USA.

[0051] Species Strain Accession number Date of accession Cosmospora species RKDO1747 67910 January 10, 2020

[0052] NRRL-67910 is particularly preferred in providing the fungicide adjuvant activity of the present invention.

[0053] Ilicicolin or lamellicholic acid anhydride can be formed and extracted from the culture of Cosmospora sp., particularly RKDO1747, respectively. The desired compound can be extracted and purified from the culture medium or fungal biomass by any means typically used to recover microbial metabolites. For example, various ion exchange resins, non-ionic adsorption resins, gel filtration chromatography, chromatography using adsorbents such as activated carbon, alumina, and silica gel, or high-performance liquid chromatography, crystallization, vacuum concentration, or freeze-drying. This means that these means can be used alone, in appropriate combinations, or repeatedly.

[0054] Cultures of Cosmospora sp. RKDO1747 can be obtained from natural sources or from culture collections such as the National Agricultural Research Service Culture Collection (NRRL). Cosmospora sp. isolate RKDO1747 can be cultured by methods known in the art of mycology.

[0055] As a means for producing the compounds of the present invention, the production organisms can be grown on any suitable synthetic or natural medium, so long as it contains an appropriate carbon source, nitrogen source, and inorganic salts. If necessary, the medium may be appropriately supplemented with vitamins and other nutrients.

[0056] Common carbon sources include, but are not limited to, sugars such as glucose, maltose, fructose, sucrose, and starch; alcohols such as glycerol and mannitol; amino acids such as glycine, alanine, and asparagine; and oils and fats such as soybean oil and olive oil. Nitrogen sources include organic nitrogen-containing compounds such as soybean flour, corn steep liquor, beef extract, peptone, yeast extract, amino acid mixtures, and fish meal, as well as inorganic nitrogen compounds such as ammonium salts and nitrates. Similarly, inorganic salts of micronutrients, such as calcium carbonate, sodium chloride, potassium chloride, magnesium sulfate, copper sulfate, manganese chloride, zinc sulfate, cobalt chloride, and various phosphates, can also be used.

[0057] The microorganism can be grown at an appropriate culture temperature within a range that allows for growth of the microorganism and effective production of the compound of the present invention. The culture temperature is preferably 10 to 32°C, more preferably 20 to 25°C, and the pH at the start of culture is preferably about 4 to 6. The culture period is generally about one day to several weeks.

[0058] The culture can be terminated when the amount of the compound of the present invention produced reaches an amount suitable for recovery, preferably when it reaches its maximum amount. As the culture method, any method can be suitably used, including commonly used methods such as solid-state culture and conventional agitation culture.

[0059] For example, Cosmospora sp. isolate RKDO1747 can be plated on nutrient-containing (e.g., YM (yeast malt extract)) agar and incubated at room temperature for several days until observable colonies appear. Individual Cosmospora sp. RKDO1747 colonies on the agar can be assayed for the production of ilisicoline or lamellicholic anhydride.

[0060] These colonies producing the desired molecule can be used to inoculate a liquid medium (e.g., YM broth medium), which can be cultured under appropriate conditions (e.g., at room temperature for several days with shaking) to obtain seed inoculum. The seed inoculum can be used to start a larger liquid culture (e.g., potato dextrose broth), which can be incubated at about room temperature for several days (e.g., 4-28 days) to expand the Cosmospora sp. culture.

[0061] Iricicolin and lamellicholic anhydride have been found to be excreted into liquid media (e.g., potato dextrose broth). Iricicolin and lamellicholic anhydride can be isolated from the fermentation broth using liquid extraction with ethyl acetate and water, and binding the compounds to an absorbent resin (e.g., Diaion™ HP20), washing the resin with water, and then eluting lamellicholic anhydride and irricicolin with an appropriate solvent (e.g., methanol or ethanol). Due to the difference in polarity between irricolin and lamellicholic anhydride, the individual compounds can be easily separated using chromatography, such as flash chromatography and reversed-phase stationary phases (e.g., C-18).

[0062] The resulting extracted irisiccoline and lamellicholic acid anhydride can be purified and used as individual homogenous compounds. In another embodiment, the extracted material can be a combination of irisiccoline and lamellicholic acid anhydride according to the first aspect, and can be used in combination in agrochemical formulations.

[0063] In other embodiments, ilisicoline and lamellicholic anhydride can be obtained from other available sources, typically other fungi.

[0064] Ilicicolin or lamellicholic anhydride can also be prepared by synthetic techniques. Lamellicholic anhydride and its derivatives can be prepared by chemical synthesis by those skilled in the art of organic chemistry using commercially available materials and synthetic methods described in the scientific literature. Commercially available materials structurally related to lamellicholic anhydride that can be used as starting materials include, but are not limited to, 1,8-naphthalenedicarboxylic acid, 1,8-naphthalic anhydride, acenaphthene, and 5-bromoacenaphthene. When using naphthalene-based starting materials, the carboxylic acid functionality can be protected, if necessary, using standard protecting group chemistry before completing the aromatic functionalization reaction.

[0065] The arylmethyl group of lamellicol anhydride can be introduced by electrophilic aromatic substitution, such as Friedel-Crafts alkylation. Phenol groups can be subsequently introduced by electrophilic aromatic halogenation, for example, using chlorine or bromine, followed by nucleophilic aromatic substitution using sodium hydroxide. To introduce arylmethoxy groups, such as those found in lamellicol anhydrides Ib and Ic, nucleophilic aromatic substitution using sodium methoxide can be performed. Protection and deprotection of hydroxyl and carboxylic acid functionalities can be completed as needed to allow for the generation of anhydride moieties, which can occur, for example, by dehydration of the 1,8-dicarboxylic acid reaction intermediate using acetic anhydride. The synthesis of lamellicol anhydride can also be contemplated using acenaphthene or its derivatives as the starting material. In this approach, the anhydride moiety can be introduced by oxidation of the ethylene bridge. Prior to this oxidation, methyl, methoxy, and / or hydroxyl functionalities can be introduced by a variety of available synthetic means, including, but not limited to, electrophilic and nucleophilic aromatic substitution, as described above. However, these reactions can also be completed after oxidation of the ethylene bridge to adjust the aromatic directing effect, depending on the desired outcome of the aromatic functionalization reaction.

[0066] Biocatalytic or chemoenzymatic methodologies can also be utilized to functionalize the aromatic position of acenaphthene prior to synthesizing the anhydride moiety via oxidation. These reaction steps can lead to lamellicholic anhydrides Ia–Ic or other derivatives in which the positions of the methyl, methoxy, and / or hydroxyl functional groups are varied.

[0067] It is understood that the properties of the adjuvant itself provide the same advantages to the pesticide formulation that contains the adjuvant. Thus, there is provided a pesticide formulation when it contains the adjuvant of the present invention that takes advantage of the properties of the adjuvant itself.

[0068] Agrochemically active compounds, including insecticides and fungicides, require formulations that allow the active compound to be taken up by the plant / target organism.

[0069] As used herein, the term "pesticidal formulation" refers to a composition containing an active pesticide and is intended to include all forms of the composition, including concentrates and spray formulations. Unless specifically stated, the pesticide formulations of the present invention may be in the form of a concentrate, a diluted concentrate, or a sprayable formulation.

[0070] The adjuvants of the present invention can be combined with other ingredients to form pesticide formulations containing at least one pesticidal active.

[0071] Thus, the pesticidally active compound can be formulated as an emulsifiable concentrate (EC), emulsifiable concentrate (EW), suspension concentrate (SC), soluble liquid (SL), oil suspension concentrate (OD), and / or suspension emulsion (SE).

[0072] In EC and SL formulations, the active compound may be present in dissolved form, whereas in OD, SC, EW, or SE formulations, the active compound may be present as a solid or an emulsified liquid.

[0073] It is envisaged that the adjuvants of the invention will find use in particular in EC, EW, SC, SL, OD or SE formulations.

[0074] Agrochemical concentrates are agrochemical compositions, which may be aqueous or non-aqueous, designed to be diluted with water (or an aqueous liquid) to form a corresponding spray formulation. Such compositions include those in liquid form (e.g., solutions, emulsions, or dispersions) and solid form (e.g., granules or powders), particularly water-dispersible solid forms.

[0075] A spray formulation is an aqueous pesticide formulation that contains all the ingredients desired to be applied to plants or their environment. Spray formulations can be prepared by simple dilution of a concentrate containing the desired ingredients (except water), or by mixing individual ingredients, or by a combination of diluting a concentrate and then adding individual ingredients or mixtures of ingredients. Typically, such end-use mixing occurs in the tank where the formulation will be sprayed, or alternatively, in a holding tank for filling the spray tank. Such mixing and blending are typically referred to as tank mixes and tank mixes.

[0076] Thus, adjuvants may be incorporated into the formulation of the pesticide active compound (in-can / incorporated formulation) or added after dilution of a concentrated spray formulation (tank mix). Incorporating adjuvants into the formulation is advantageous to avoid dosing errors and improve user safety during application of the pesticide product. This also avoids unnecessary use of packaging material for tank-mix products.

[0077] Depending on customer needs, the concentrates thus formed typically contain up to 95% by weight of the pesticidal active material. The concentrates are diluted for use to produce diluted compositions having an agrochemically active concentration of about 0.5% to about 1% by weight. The diluted compositions (e.g., spray application rates of 10 to 500 l.ha) are typically used for application in a variety of applications. -1 In spray formulations of the present invention, the pesticidal activity concentration can range from about 0.001% to about 1% by weight of the total formulation sprayed.

[0078] The adjuvants of the present invention are typically used in an amount proportional to the amount of active pesticide in the formulation. In concentrates of pesticide formulations, the proportion of adjuvant depends on the solubility of the components in the liquid carrier. Typically, the concentration of adjuvant in such concentrates is 1% to 99% by weight. Preferably, the concentration of adjuvant in such concentrates is 1% to 70% by weight. More preferably, the concentration of adjuvant in such concentrates is 3% to 50% by weight.

[0079] Upon dilution to form, for example, a spray formulation, the adjuvant is typically present at a concentration of 0.01% to 2% by weight of the spray formulation, more usually 0.03% to 0.5% by weight, and more preferably, the adjuvant is present at a concentration of 0.12% to 0.4% by weight of the spray formulation.

[0080] The ratio of adjuvant to active pesticide in the pesticide composition is preferably about 1:40 to about 1:1. More preferably, the ratio of adjuvant to active pesticide in the pesticide formulation is about 1:20 to about 1:1. Even more preferably, the ratio of adjuvant to active pesticide in the pesticide formulation is about 1:5 to 1:1. This ratio range is generally maintained in concentrated formulations (e.g., when the adjuvant is contained in a dispersible liquid concentrate or dispersible solid granule formulation) and spray formulations.

[0081] When a concentrate (solid or liquid) is used as the source of the active pesticide and / or adjuvant, the concentrate is typically diluted to form a spray formulation. Dilutions can be 1 to 10,000 times, and especially 10 to 1,000 times, the total weight of the concentrate in water to form a spray formulation.

[0082] When the pesticidal active is present as a solid particle in the aqueous end-use formulation, it will most often be present primarily as particles of the active pesticide. However, if desired, the active pesticide can be supported on a solid carrier, such as silica or diatomaceous earth, which can be a solid carrier, filler, or diluent material as described above.

[0083] Spray formulations typically have a pH in the range of moderately acidic (e.g., about 3) to moderately alkaline (e.g., about 10), especially neutral (e.g., about 5 to 8). More concentrated formulations have a similar degree of acidity / alkalinity, but because they are largely non-aqueous, pH is not necessarily an appropriate measure of this.

[0084] The pesticide formulation may contain a solvent (other than water), such as monopropylene glycol, a vegetable oil, or a mineral oil, such as spray oil (the oil included in the spray formulation as a non-surfactant adjuvant), in association with the primary adjuvant and co-adjuvant. Such a solvent may be included as a solvent for the adjuvant and / or as a humectant, e.g., propylene glycol. When used, such a solvent is typically included in the adjuvant in an amount of 5% to 500% by weight, preferably 10% to 100% by weight. Such combinations may also include salts, such as ammonium chloride and / or sodium benzoate, and / or urea, particularly gel inhibitors.

[0085] In another embodiment, the adjuvant of the present invention, or any of the organisms of the sixth aspect, can be included in a seed coating composition suitable for application to seeds. Preferably, the adjuvant of the present invention can be included in a seed coating composition.

[0086] The adjuvant is suitably present in the seed coating composition at a concentration in the range 0.5 to 25% by weight, preferably 2 to 18% by weight, more preferably 5 to 15% by weight, especially 8 to 12% by weight, based on the total weight of the composition.

[0087] Coating may include film coating, pelletizing, and encapsulation, or combinations of these techniques known in the art. It is envisioned that the present invention applies to all such coating types, preferably film coating.

[0088] The seed coating compositions of the present invention can be applied to seeds by conventional methods.

[0089] The seeds may be primed or unprimed (they may have been subjected to a treatment to improve germination rate, such as osmopriming, hydropriming, matrix priming).

[0090] In one embodiment, seeds are not provided with artificial layers before applying the seed coating composition of the present invention, for example, a primer layer containing a binder such as polymer.Therefore, the seed coating composition is preferably applied directly to the natural outer surface of seeds.Nevertheless, before applying the seed coating composition, the seed surface may be subjected to surface treatment.

[0091] Preferably, the seed coating composition is applied as a liquid composition and / or emulsion and / or dispersion and / or latex composition, which is then solidified (including cured and / or dried) to form the seed coating. As used in this application, the term "liquid coating composition" is meant to include coating compositions in the form of suspensions, emulsions, and / or dispersions, preferably dispersions.

[0092] Conventional coating means can be used to coat seeds. A variety of coating machines are available to those skilled in the art. Some well-known techniques include the use of drum coaters, fluidized bed techniques, rotary coaters (with or without integral drying), and spouted beds. Suitably, the seed coating composition is applied to seeds by rotary coaters, rotary dry coaters, pan coaters, or continuous processing equipment.

[0093] The seed coating composition can be applied, for example, by film coating, spray application, dipping or brush application of the seed coating composition.Preferably, this method comprises applying the seed coating composition to form a film or seed coating layer.Seed coating typically comprises forming a firmly adhered, moisture-permeable coating on the surface of the seed.This method typically comprises applying a liquid seed coating composition to the seed before sowing.

[0094] Additional film coating layers may optionally be applied on top of the coating layer of the present invention to provide additional benefits including, but not limited to, cosmetics, coverage, actives, nutrients, and processing improvements such as faster drying, seed flow, durability, etc.

[0095] The pesticide formulation or seed coating composition may also optionally contain other ingredients. These other components may be selected from those including: • Binders that are readily water soluble to give low viscosity solutions at high binder concentrations, such as polyvinylpyrrolidone; polyvinyl alcohol; carboxymethyl cellulose; gum arabic; sugars, e.g., sucrose or sorbitol; starch; ethylene-vinyl acetate copolymers, sucrose, and alginates. • Diluents, absorbents or carriers such as carbon black; talc; diatomaceous earth; kaolin; aluminum, calcium or magnesium stearate; sodium triphosphate; sodium tetraborate; sodium sulfate; sodium, aluminum and mixed sodium silicates; and sodium benzoate; Disintegrants, such as surfactants, materials that swell in water, such as carboxymethylcellulose, collodion, polyvinylpyrrolidone and microcrystalline cellulose swelling agents; salts, such as sodium or potassium acetate, sodium carbonate, bicarbonate or sesquicarbonate, ammonium sulfate and dipotassium hydrogen phosphate; • Wetting agents such as alcohol ethoxylate and alcohol ethoxylate / propoxylate wetting agents; • dispersants such as sulfonated naphthalene formaldehyde condensates and acrylic copolymers such as comb copolymers with polyethylene glycol side chains capped onto a polyacrylic backbone; • Emulsifiers such as alcohol ethoxylates, ABA block copolymers, or castor oil ethoxylates; anti-foaming agents, such as polysiloxane anti-foaming agents, typically in amounts of 0.005% to 10% by weight of the formulation; viscosity modifiers such as commercially available water-soluble or miscible gums, e.g., xanthan gum and / or celluloses, e.g., carboxymethyl, ethyl, or propyl cellulose; and / or Preservatives and / or antimicrobial agents such as organic acids, or their esters or salts, for example ascorbyl palmitate, potassium sorbate such as potassium sorbate, benzoic acid and methyl and propyl 4-hydroxybenzoate, sodium propionate, for example phenols, for example 2-phenylphenol; 1,2-benzisothiazolin-3-one; or formaldehyde, for example paraformaldehyde; or inorganic substances, for example sulfuric acid and its salts, typically in an amount of 0.01% to 1% by weight of the formulation.

[0096] The pesticide formulation or seed coating composition according to the present invention may also contain components such as surfactant materials that form part of the emulsifier system. The surfactants may include surfactant dispersants. Other adjuvants outside the scope of the present invention, such as surfactant adjuvants, can be included in the compositions and formulations of the present invention and used in the present invention. Examples include alkyl polysaccharides (more appropriately called alkyl oligosaccharides); fatty amine ethoxylates, such as coconut alkylamine 2EO; and derivatives of alkyl (or alkenyl) succinic anhydrides, particularly those described in PCT applications WO 94 / 00508 and WO 96 / 16930.

[0097] The formulation / composition may contain one or more biologically active ingredients, including plant enhancers, particularly plant protection products (also called PPPs). Suitable examples of active ingredients, particularly plant enhancers, are antimicrobials, fungicides, insecticides, nematicides, molluscicides, biologicals, miticides or acaricides, pesticides, and biocides. Further possible active ingredients include disinfectants, microorganisms, rodenticides, weed killers (herbicides), attractants, (bird) repellents, plant growth regulators (such as gibberellic acid, auxins or cytokinins), nutrients (such as potassium nitrate, magnesium sulfate, iron chelates), plant hormones, minerals, plant extracts, germination stimulants, pheromones, biologicals, etc.

[0098] The pesticidal active substances suitable for use in the formulations or seed coating compositions according to the invention are all pesticidal active compounds that may be solid or liquid at room temperature. The adjuvants of the invention are believed to be broadly applicable to all types of pesticidal activity.

[0099] In the context of the present invention, pesticide active agents refer to plant protection agents, more specifically biocides, which are chemical substances capable of killing different forms of living organisms and are used in fields such as medicine, agriculture, forestry, and mosquito control. Also included in the group of biocides are so-called plant growth regulators.

[0100] Biocides for use in the pesticide formulations or seed coating compositions of the present invention are typically divided into two subgroups. pesticides such as fungicides, herbicides, insecticides, analgesics, molluscicides, poison mitigators and rodenticides; Antimicrobial agents such as germicides, antibiotics, antibacterial agents, antivirals, antifungals, antiprotozoals, and antiparasitics.

[0101] In particular, biocides selected from insecticides, fungicides, or herbicides are particularly preferred.

[0102] The term "pesticide" is understood to mean a substance or mixture thereof intended to prevent, destroy, repel or mitigate pests. Pesticides can be chemical or biological agents (such as viruses or bacteria) used against pests, including insects, plant pathogens, weeds, mollusks, birds, mammals, fish, nematodes (roundworms) and microorganisms that compete with humans for food, destroy property, spread disease or are harmful to humans. The following examples illustrate pesticides suitable for the pesticide composition of the present invention.

[0103] Fungicides are the chemical control of fungi. Fungicides are chemicals used to prevent fungi from spreading in gardens and crops. Fungicides are also used to fight fungal infections. Fungicides can be contact or systemic. Contact fungicides kill fungi when they come into contact with the fungicide, which is held on the leaf surface. Systemic fungicides are absorbed into plant tissue and kill fungi as they attempt to invade the host.

[0104] According to the present invention, examples of suitable fungicides include (3-ethoxypropyl)mercuric bromide, 2-methoxyethylmercuric chloride, 2-phenylphenol, 8-hydroxyquinoline sulfate, 8-phenylmercuryoxyquinoline, acibenzolar, acyl amino acid fungicides, acipetac, aldimorph, aliphatic nitrogen fungicides, allyl alcohol, amide fungicides, ampropylphos, anilazine, anilide fungicides, antibiotic fungicides, aromatic fungicides, aureofungin, azaconazole, adithiram, azoxystrobin, barium polysulfide, benalaxyl-M, benodanil, benomyl, benzyl benzoate, benzophenone, benzophenone-1, benzophenone-2, benzophenone-3, benzophenone-4, benzophenone-5, benzophenone-6, benzophenone-7, benzophenone-8, benzophenone-9, benzophenone-10, benzophenone-11, benzophenone-12, benzophenone-13, benzophenone-14, benzophenone-15, benzophenone-16, benzophenone-17, benzophenone-18, benzophenone-19, benzophenone-20, benzophenone-21, benzophenone-22, benzophenone-23, benzophenone-24, benzophenone-25, benzophenone-26, benzophenone-27, benzophenone-28, benzophenone-29, benzophenone-30, benzophenone-31, benzophenone-32, benzophenone-33, benzophenone-34, benzophenone-35, benzophenone-36, benzophenone-37, benzophenone-38, benzophenone-39, benzophenone-40, benzophen Inquinox, bentalon, benthiavalicarb, benzalkonium chloride, benzamacryl, benzamide fungicides, benzamorph, benzanilide fungicides, benzimidazole fungicides, benzimidazole precursor fungicides, benzimidazolylcarbamate fungicides, benzohydroxamic acid, benzothiazole fungicides, bethoxadin, binapacryl, biphenyl, bitertanol, bithionol, blasticidin-S, Bordeaux mixture, boscalid, bridged diphenyl fungicides, bromuconazole, bupirimate, Burgundy mixture, butiobe Butylamine, calcium polysulfide, captafol, captan, carbamate fungicides, carbamorph, carbanilic acid fungicides, carbendazim, carboxin, carpropamid, carvone, cheshunt mixture, chinomethionate, clobenziazone, chloraniformethane, chloranil, chlorphenazole, chlordinitronaphthalene, chloroneb, chloropicrin, chlorthalonil, chlorquinox, chlozolinate, ciclopirox, clivazol, clotrimazole, conazole fungicide, conazole fungicide (imidazole), Nazole fungicide (triazole), copper (II) acetate, copper (II) carbonate, basic, copper fungicide, copper hydroxide, copper naphthenate, copper oleate, copper oxychloride, copper (II) sulfate, copper sulfate, basic, zinc copper chromate, cresol, khufuraneb, cuprobam, cuprous oxide, cyazofamid, cyclafuramid, cyclic dithiocarbamate fungicide, cycloheximide, cyflufenamid, cymoxanil, cypendazole, cyproconazole, cyprodinil, dazomet, DBCP, debacarb, decafentin, dehydroacetic acid, dicarboximide fungicide, dichlofluanidDichloron, dichlorophen, dichlorophenyl, dicarboximide fungicides, dichlozolin, diclobutrazol, diclocymet, diclomedine, dicloran, diethofencarb, diethyl pyrocarbonate, difenoconazole, diflumetrim, dimethirimol, dimethomorph, dimoxystrobin, diniconazole, dinitrophenol fungicides, dinobuton, dinocap, dinocton, dinopenton, dinosulfone, dinotervon, diphenylamine, dipyrithione, disulfiram, ditalinphos, dithianon, dithiocarbamate fungicides, DNOC, Dodemorph, Dodicine, Dodine, Donatodine, Drazoxolone, Edifenphos, Epoxiconazole, Etaconazole, Ethem, Ethaboxam, Ethirimol, Ethoxyquin, Ethylmercury 2,3-dihydroxypropyl mercaptide, Ethylmercury acetate, Ethylmercury bromide, Ethylmercury chloride, Ethylmercury phosphate, Etridiazole, Famoxadone, Fennamidone, Fenapanil, Fenarimol, Fenbuconazole, Fenfuram, Fenhexamid, Fenitropan, Fenoxanil, Fenpiclonil, Fenpropidin, Fenpropimol F, fentin, ferbam, ferimzone, fluazinam, fludioxonil, flumetover, fluopicolide, fluoroimide, flutrimazole, fluoxastrobin, fluquinconazole, flusilazole, flusulfamide, flutolanil, flutriafol, folpet, formaldehyde, fosetyl, fuberidazole, furalaxyl, furametpyr, furamide fungicide, furanilide fungicide, flucarbanil, fluconazole, fluconazolecis, furfural, flumecyclox, furophanate, gliodin, griseofur Bin, guazatine, halacrinate, hexachlorobenzene, hexachlorobutadiene, hexachlorophene, hexaconazole, hexylthiophos, hydralgafen, hymexazole, imazalil, imibenconazole, imidazole fungicides, iminoctadine, inorganic fungicides, inorganic mercury fungicides, iodomethane, ipconazole, iprobenfos, iprodione, iprovalicarb, isoprothiolane, isovaledione, kasugamycin, kresoxim methyl, lime sulfur, mancopper, mancozeb, maneb, mebenil, mecarbinzide, mepanipyrim,Mepronil, mercuric chloride, mercuric oxide, mercurous chloride, mercury fungicides, metalaxyl, metalaxyl-M, metam, metazoxolone, metconazole, metasulfocarb, metofloxam, methyl bromide, methyl isothiocyanate, methylmercuric benzoate, methylmercuric dicyandiamide, methylmercuric pentachlorophenoxide, metiram, metominostrobin, metrafenone, metsulfobax, milneb, morpholine fungicides, myclobutanil, myclozolin, N-(ethylmercury)-p-toluenesulfonanilide, nabam, natamycin, nitros Stylene, nitrotar isopropyl, nuarimol, OCH, octhilinone, ofrace, organomercurial fungicides, organophosphate fungicides, organotin fungicides, orysastrobin, oxadixyl, oxathiin fungicides, oxazole fungicides, oxine copper, oxpoconazole, oxycarboxin, pefurazoate, penconazole, pencycuron, pentachlorophenol, penthiopyrad, phenylmercuric urea, phenylmercuric acetate, phenylmercuric chloride, phenylmercuric derivatives of pyrocatechol, phenylmercuric nitrate, phenylmercuric salicylate, phenylsulfa Mido fungicides, phosdifen, phthalide, phthalimide fungicides, picoxystrobin, piperalin, polycarbamate, polymeric dithiocarbamate fungicides, polyoxin, polyoxorim, polysulfide fungicides, potassium azide, potassium polysulfide, potassium thiocyanate, probenazole, prochloraz, procymidone, propamocarb, propiconazole, propineb, proquinazide, prothiocarb, prothioconazole, pyracarbollide, pyraclostrobin, pyrazole fungicides, pyrazophos, pyridine fungicides, pyridinnitrile, pyrife Nox, pyrimethanil, pyrimidine fungicides, pyroquilon, pyroxchlor, piroxifil, pyrrole fungicides, quinacetol, quinazamide, quinconazole, quinoline fungicides, quinone fungicides, quinoxaline fungicides, quinoxyfen, quintozene, labenzazole, salicylanilide, silthiofam, simeconazole, sodium azide, sodium orthophenylphenoxide, sodium pentachlorophenoxide, sodium polysulfide, spiroxamine, streptomycin, strobilurin fungicides, sulfonanilide fungicides, sulfur,Sultropene, TCMTB, tebuconazole, tecloftalam, tecnazene, tecoram, tetraconazole, thiabendazole, thiadifluor, thiazole fungicides, tichofen, thifluzamide, thiocarbamate fungicides, thiochlorfenfim, thiomersal, thiophanate, thiophanate methyl, thiophene fungicides, thioquinox, thiram, tiadinil, thi, midoxytibutrclofos methyl, tolnaftate, tolylfluanid, tolylmercuric acetate, triadimefon , triadimenol, triamiphos, triarimol, triazbutyl, triazine fungicides, triazole fungicides, triazoxide, tributyltin oxide, triclamide, tricyclazole, trifloxystrobin, triflumizole, triforin, triticonazole, unclassified fungicides, undecylenic acid, uniconazole, urea fungicides, validamycin, valiamide fungicides, vinclozolin, zaliramide, zinc naphthenate, zineb, ziram, zoxamide, and mixtures thereof.

[0105] Herbicides are pesticides used to kill unwanted plants. Selective herbicides kill specific targets while leaving desired crops relatively unharmed. Some of these act by preventing weed growth and are often based on plant hormones. Herbicides used to remove waste soil are non-selective, killing all plant material they come into contact with. Herbicides are widely used in agricultural and lawn landscape management. They are applied in integrated vegetation management (TVC) programs for highway and railroad maintenance. Smaller amounts are used in the management of forests, pasture systems, and areas reserved for wildlife habitat.

[0106] Suitable herbicides are, for example, triazine herbicides such as aryloxycarboxylic acids, for example MCPA, aryloxyphenoxypropionates, for example clodinafop, cyclohexanedione oximes, for example sethoxydim, hydroxybenzonitriles, for example bromoxynil, sulfonylureas, for example nicosulfuron, triazolopyrimidines, for example penoxsulam, trikethiones, for example mesotrione, metribuzin, hexaxinone, or atrazine; sulfonylurea herbicides, for example chlorsulfuron; uracils, for example lenacil, bromacil, terbacil; urea herbicides, for example linuron, diuron, siduron, or nebron; acetanilide herbicides, for example alachlor, or metolachlor; benthio thiocarbamate herbicides such as carb, triallate; oxadiazolone herbicides such as oxadiazon; isoxazolidone herbicides, phenoxyacetic acid; diphenyl ether herbicides such as fluazifop, acifluorfen, bifenox, or oxyfluorfen; dinitroaniline herbicides such as trifluralin; organophosphonate herbicides such as glufosinate salts and esters and glyphosate salts and esters; and / or dihalobenzonitrile herbicides such as bromoxynil, or dipyridylium herbicides such as ioxynil, benzoic acid herbicides, paraquat; other herbicides such as clomazone, carfentrazone, saflufenacil, pyroxasulfone.

[0107] Particularly preferred herbicides may be selected from 2,4-dichlorophenoxyacetic acid (2,4-D), atrazine, dicamba as a benzoic acid, glyphosate, glufosinate, imazapic as an imidazolinone, metolachlor as a chloroacetamide, picloram as a pyridine carboxylic acid, clopyralid, and triclopyr or synthetic auxins, their respective water-soluble salts and esters, and mixtures thereof.

[0108] Insecticides are insecticides used to control insects in all their developmental stages, including insecticides and larvicides used to control insect eggs and larvae. Insecticides are used in agriculture, medicine, industry, and the home.

[0109] Suitable insecticides include, for example, chlorinated insecticides such as camphechlor, DDT, hexachlorocyclohexane, gamma-hexachlorocyclohexane, methoxychlor, pentachlorophenol, TDE, aldrin, chlordane, chlordecone, dieldrin, endosulfan, endrin, heptachlor, mirex and mixtures thereof; organophosphorus compounds such as acephate, azinphos methyl, bensulide, chlorethoxyphos, chlorpyrifos, chlorpyrifos methyl, diazinon, dichlorvos (DDVP), dicrotophos, dimethoate, disulfon, ethoprop, fenamiphos, fenitrothion, fenthion, fosthiazate, malathion, methamidophos, methidathion, methyl parathion, mevinphos, naled, omethoate, oxydemeton methyl, parathion, folate, phosalone, phosmet, phostebe carbamates such as aldicarb, carbofuran, carbaryl, methomyl, 2-(1-methylpropyl)phenylmethylcarbamate, and mixtures thereof; pyrethroids such as allethrin, bifenthrin, deltamethrin, permethrin, resmethrin, sumithrin, tetramethrin, tralomethrin, transfluthrin, and mixtures thereof; plant toxin-derived compounds such as derris (rotenone), pyrethrum, neem (azadirachtin), nicotine, caffeine, and mixtures thereof; neonicotinoids such as imidacloprid; abamectin, emamacutin; oxadiazines such as indoxacarb; and / or anthranildiamides such as rynaxypyr.

[0110] Insecticides are insecticides that kill mites. Antibiotic insecticides, carbamate insecticides, formamidine insecticides, mite growth regulators, organochlorines, permethrin, and organophosphate insecticides all belong to this category. Molluscicides are insecticides used to control mollusks such as moths, slugs, and snails. These substances include metaldehyde, methiocarb, and aluminum sulfate. Nematicides are a type of chemical pesticide used to kill parasitic nematodes (Nematoda).

[0111] Most preferably, the active substance present in the pesticide formulation or seed coating composition of the present invention is selected from triazole fungicides, strobilurin fungicides, or combinations thereof, in particular tebuconazole, flutriafol, carbendazim, azoxystrobin, crezoxim-methyl, cyproconazole, or pyraclostrobin.

[0112] Nutrients may be present in addition to or as an alternative to pesticides. In such formulations / compositions, the nutrients are typically in dry form.

[0113] The nutrient may preferably be a solid-phase nutrient. In the present invention, solid nutrients should be understood to mean substances with a melting point above 20°C (standard pressure). Solid nutrients also include insoluble nutrient components, i.e., nutrient components whose solubility in water is such that significant solids are present in the concentrate after addition.

[0114] Nutrients refer to chemical elements and compounds that are desirable or necessary to promote or improve plant growth. Suitable nutrients are generally described as macronutrients or micronutrients. The nutrients suitable for use in the concentrate according to the present invention are all nutritional compounds.

[0115] Micronutrients usually refer to trace metals or trace elements, and are often applied in low doses.Suitable micronutrients include trace elements selected from zinc, boron, chlorine, copper, iron, molybdenum and manganese.Micronutrients can be in soluble form or can be included as insoluble solid, and can be salt or chelated.

[0116] Macronutrients typically refer to those containing nitrogen, phosphorus, and potassium, and include fertilizers such as ammonium sulfate, and water conditioners. Suitable macronutrients include fertilizers and other nitrogen-, phosphorus-, potassium-, calcium-, magnesium-, sulfur-containing compounds, and water conditioners.

[0117] Suitable fertilizers include inorganic fertilizers that provide nutrients such as nitrogen, phosphorus, potassium, or sulfur. The fertilizers may be included in relatively low concentration, diluted formulations, or as more concentrated solutions, which may be very concentrated and include solid fertilizers and solutions.

[0118] It is expected that the nutrient content will depend on the particular nutrient, with micronutrients typically being present in lower concentrations, while macronutrients are typically present in higher concentrations.

[0119] Biostimulatory ingredients can be added to formulations or seed coating compositions to promote the growth of crop plants. Biostimulatory ingredients can include or consist of one or more biostimulants.

[0120] Examples of useful biostimulants include, but are not limited to, plant growth hormones and plant growth regulators, such as cytokinins, auxins, gibberellins, ethylene, and abscisic acid. Other biostimulants include protein hydrolysate derivatives, seaweed extracts, amino acids, plant extracts, chitosan derivatives, biopolymers, inorganic compounds, humic substances, microbial inoculants, and microbial products, or mixtures thereof.

[0121] The adjuvants of the present invention provide adjuvants for pesticide formulations in which they are included, and may find particular use in providing fungicide adjuvancy.

[0122] As used herein, the term "adjuvant" or "adjuvant" refers to a compound that, when added to a pesticide formulation, improves the desired effect of the pesticide. Adjuvants can affect the diluent, mixture, active substance, or target by improving the performance of the active substance. Adjuvants can be used to attach pesticides to the area where they function, alter the epidermal layer of the leaf surface that allows the pesticide to penetrate, and / or sensitize the target pesticide to the active pesticide in the pesticide formulation.

[0123] Specific adjuvant effects may include surfactants, emulsifiers (dispersants and suspending agents), oils, emulsifiable oils, compatibilizers, buffers and conditioning agents, defoamers, deposition agents, drift control agents, thickeners, spreading agents (wetting agents), stickers (builders and extenders), plant penetrants, translocators, soil penetrants, stabilizers (UV filters), and / or pest sensitization to the active pesticide.

[0124] Preferably, the adjuvants of the present invention find use as the sole component or primary functionalizing agent in adjuvants formulated for tank addition or directly in agrochemical concentrates.

[0125] As a measure of adjuvant activity relative to the activity of a fungicide alone (e.g., pyraclostrobin) against Botrytis cinerea, percent inhibition (adjuvant and fungicide) divided by percent inhibition (fungicide) can be defined, with the higher value being more desirable. Thus, a value of 1 would represent equivalent activity of the adjuvant / fungicide combination to the fungicide alone, while a value greater than 1 would represent greater activity by the adjuvant / fungicide combination than the fungicide alone. Actives of the present invention can have values ​​greater than 1. Preferably, actives of the present invention have values ​​greater than 1.5, and most preferably greater than 2.

[0126] All features described herein can be combined with any of the above aspects in any combination. [Example]

[0127] In order that the present invention may be more readily understood, reference is made by way of example to the following description.

[0128] It is understood that all tests and physical properties described are measured at atmospheric pressure and room temperature (i.e., 20-25°C) unless otherwise stated herein or in the referenced test methods and procedures.

[0129] Formation and Extraction Bioassay-guided fractionation of culture extracts of the fungus Cosmospora sp. RKDO1747 led to the isolation of lamellicholic anhydride and silycorine. Isolate RKDO1747 was cultured on YM (yeast extract malt extract) agar medium and incubated at 22°C for 14 days. Eight colony explants (approximately 3 mm) were collected. 3 The seed inoculum was prepared by inoculating 15 mL of YM broth into a sterile 50 mL test tube with the culture medium and shaking at 200 RPM at 22 °C for 5 days. The seed culture was then used to inoculate sterile rice fermentation medium (1% brown rice and 2.5% YNB broth (0.67% YNB powder, 0.5% sucrose)) prepared in an Erlenmeyer flask.

[0130] After 21 days at 22°C, the fermentation was extracted with 1 volume of ethyl acetate and shaken at 175 RPM for 60 minutes. The extract was clarified by filtration through Whatman #3 filter paper, and the solvent was removed under reduced pressure prior to chemical purification.

[0131] The RKDO1747 fermentation extract was purified on a Silasep C18 flash cartridge (43 g C-18) using a gradient of 10% MeOH:90% HO to 100% MeOH on a Teledyne Nextgen 300 + Fractionation was performed on a Combiflash column for 20 minutes. Fractions were analyzed on a Thermo Scientific Accela UHPLC coupled to a Thermo Exactive Electrospray Mass Spectrometer (ESI-MS) equipped with a SEDEX 80LT ELSD and a thermophotodiode array (PDA) detector. Fractions containing lamellicholic anhydride or iliticolin were purified using reverse-phase C-18 HPLC (Kinetex 5 μm C18 column, 10 × 250 mm) on a Waters HPLC system equipped with an evaporative light scattering detector (Waters 2424) and a mass spectrometer (Waters 3100). Initial purification of lamellicholic anhydride was performed with an isocratic elution of 60% MeCN in water at a flow rate of 3 mL / min, and iliticolin H was eluted with an isocratic elution of 80% MeCN in water.

[0132] A combination of mass spectrometry and NMR analysis was used to elucidate the structures of lamellicol anhydride and iricicoline H. The NMR spectra were 1 H and 13 C were recorded on a Bruker Avance III 400 MHz NMR spectrometer operating at 400 MHz and 150 MHz, respectively. Spectra were referenced to residual unaltered solvent peaks.

[0133] m / z 261.0396[M+H] + and m / z 434.2327[M+H] +NMR analysis of the corresponding metabolites was consistent with literature data and confirmed their structures as lamellicol anhydride (Ia) and irsicolin H (A), respectively. The remaining four metabolites (m / z 275.0553 (Ic), 277.0342 (Ib), 289.0707 (Id and Ie), and 305.0657 (If and Ig) [M+H] + ) corresponded to further substituted lamellicholic anhydrides containing additional hydroxyl and / or methoxy substitutions.

[0134] Lamellicol Anhydrous (Ia): 1 H NMR (MeOD, 400MHz): δ2.78(s, 3H), 6.35(s, H), 6.78(s, H)ppm; HRMS(ESI) m / z:C 13 Calculated value for H8O6[M+H]+: 261.0394, found value: 261.0396. Lamellicol Anhydrous 277(Ib): 1 H NMR (MeOD, 400MHz): δ2.84(s, 3H), 6.78(s, H)ppm; HRMS (ESI) m / z:C 13 Calculated value for H8O7[M+H]+: 277.0343, found value: 277.0342. Lamellicol Anhydrous 275 (Ic): 1 H NMR (MeOD, 400MHz): δ2.67(s, 3H), 3.96(s, H), 6.35(s, H), 6.62(s, H)ppm; HRMS(ESI) m / z:C 14 H 10 Calculated value for O6[M+H]+: 275.0550, found value: 275.0553. Lamellicol Anhydrous 289 (Id and Ie): 1 H NMR (MeOD, 400MHz): δ2.96(s, 3H), 3.82(s, 3H), 3.90(s, 3H), 6.79(s, H), 6.34(s, H)ppm; HRMS(ESI) m / z:C 15 H 12 Calculated value for O6[M+H]+: 289.0707, found value: 289.0708. Lamellicol Anhydrous 305 (If and Ig): 1H NMR (MeOD, 400MHz): δ2.66(s, 3H), 3.77(s, 3H), 3.96(s, 3H), 6.37(s, H)ppm;z:C 15 H 12 Calculated value for O7[M+H]+: 305.0656, found value: 305.0657.

[0135] Supporting examples The parameters "percent inhibition" and "fold change" are understood to represent ad and are calculated as follows: Percent Inhibition - Percent inhibition is understood to represent the amount by which a disinfectant and / or adjuvant inhibits the visible growth of a microorganism after 48 hours of incubation at 22°C compared to a vehicle-treated control. It is calculated using the following formula: [(Θ C -Θ T ) / Θ C ]×100% where: Θ C = diameter of colonies grown on agar supplemented with solvent (solvent-treated control), and τ = diameter of colonies grown on agar supplemented with fungicides and / or adjuvants formulated in an appropriate vehicle (i.e., solvents such as water, EtOH, MeOH, MeCN, DMSO, etc.).

[0136] Fold Change - The fold change is a measure of the adjuvant / bactericide combination in inhibiting the microorganism compared to the bactericide alone. This indicates how the adjuvant performs compared to the bactericide alone. It is calculated using the following formula: INH AF / INH F where: Inhibition of fungal growth when treated with fungicides and adjuvants AF = percent inhibition, and INH F = Percent fungal growth inhibition when treated with fungicide alone.

[0137] Example - Adjunctive therapy with pyraclostrobin Botrytis cinerea (ATCC 90479) was cultured on potato dextrose agar (PDA) for 7 days under a diurnal UV cycle (12 hours UV light and 12 hours darkness). Spores were harvested in buffered sterile saline (0.9% saline, 1% Tween 80) and counted using a hemocytometer. The spore suspension contained 8.5 × 10 6 A standardized inoculum was prepared by adjusting the final concentration of spores / mL.

[0138] To prepare mycelial fragments for adjuvant testing, 8.5 x 10 4 Spores were used to inoculate 10 mL of potato dextrose broth in a 150 × 25 mm test tube. The tube was incubated at 220 RPM and 22°C for 48 hours. To generate mycelial fragments from the culture, the culture was transferred to a 50 mL plastic conical tube containing approximately 20 sterile 5 mm diameter glass beads and vortexed for 5 minutes. The tube was then allowed to stand for 5 minutes to allow large mycelial clumps to settle. The upper layer containing the mycelial fragments was then removed and used as inoculum for growth inhibition assays.

[0139] The disinfectant and adjuvant were dissolved in methanol and added to molten PDA (approximately 50°C), and the agar was then distributed into the wells of a 12-well multiwell plate (1 mL / well). The plates were cooled to room temperature, and 10 μL of mycelial inoculum was added to the center of each well. The plates were incubated at 22°C for 48 hours, and colony diameters were measured using a digital caliper. The biological growth control consisted of mycelia and medium (0.07% methanol), and the negative control was medium and medium (0.07% methanol).

[0140] The results of the adjuvant activity of the present invention are shown in Table 1. Adjuvant activity of pyraclostrobin against B. cinerea was observed for each compound tested, most notably for lamellichol anhydrides 289 (Id and Ie) and 305 (If and Ig), which showed a 7.59- and 6.10-fold increase in bactericidal activity compared to 0.1 μg / mL pyraclostrobin alone. Lamellicholic anhydride 289 (Id and Ie), lamellicholic anhydride 305 (If and Ig), and irisicolin H (A) did not exhibit intrinsic bactericidal activity at any of the concentrations tested.

[0141] [Table 1]

[0142] The effect of adjuvants on fungicidal activity is expressed as fold increase in growth inhibition, with a value of 1 indicating no increase in fungicidal activity, a value less than 1 indicating a decrease in fungicidal activity, and a value greater than 1 indicating an increase in fungicidal activity.

[0143] Example - Cytotoxicity The cytotoxicity of lamellicholic anhydride and iricicoline H was evaluated in vitro against African green monkey Vero kidney cells (ATCC CCL-81). The cells were cultured in 10% fetal bovine serum (VWR#CA95043-976), 100 μU penicillin, and 75 cm 2 Cells were grown and maintained in 15 mL of Eagle's minimum essential medium (Sigma M5650) supplemented with 0.1 mg / mL streptomycin in cell culture flasks. Cells were incubated at 37 °C in a humidified atmosphere of 5% CO for 24 h. Culture medium was refreshed every 2–3 days, and cells were not allowed to exceed 80% confluence.

[0144] At 80% confluence, cells were counted, diluted, and plated into 96-well cell culture plates (VWR#29442-054) at a cell density of 10,000 cells per well in 90 μL of growth medium. The plates were incubated at 37°C in a humidified atmosphere of 5% CO2 for 24 hours to allow cells to attach to the plates before treatment. After 24 hours, adjuvants were dissolved in DMSO, serially diluted, and added to the wells at final concentrations ranging from 1 μg / mL to 128 μg / mL. DMSO was used as a vehicle at a final concentration of 1% in the wells.

[0145] Plates were incubated at 37°C in a humidified atmosphere of 5% CO2 for 24 hours, after which AlamarBlue (Invitrogen) was added to each well at 10% of the culture volume. Fluorescence was monitored at 0 and 4 hours after AlamarBlue addition using a Thermo Scientific Varioskan Flash plate reader at excitation 560 / 12 and emission 590 nm. The estimated percentage of cell viability relative to vehicle control wells was calculated after subtracting the time-zero emission 590 nm reading from the final reading.

[0146] Lamellicol Anhydrous 289 (Id and Ie), Lamellicol Anhydrous 305 (If and Ig), and Irsicolin H (A) showed no cytotoxicity to Vero cells at the highest concentration tested (128 mg / mL), indicating that the adjuvants were not toxic to mammalian cells at the concentrations tested.

[0147] Example - Adjuvancy with other fungicides against B. cinerea To extend the functional characterization of lamellicholic anhydride adjuvant (Ia), it was tested in combination with seven fungicides belonging to six different fungicide groups classified by the Fungicide Resistance Action Committee (FRAC Code List 2018). The effect of adjuvant-fungicide combinations on fungicidal activity against B. cinerea is shown in Table 2.

[0148] [Table 2]

[0149] Numbers indicate fold change in fungal growth inhibition compared to fungal test organisms treated with fungicide and adjuvant vehicle alone; nt indicates not tested.

[0150] Adjuvant (Ia), along with six of the seven fungicides, increased fungicidal activity against B. cinerea by 16-45 fold. The lowest effect was observed with mancozeb, but even in this case, a 1.9-3.1 fold increase in fungicidal activity was observed.

[0151] These results clearly demonstrate that Lamellicol anhydrous adjuvant (Ia) provides an adjuvant effect applicable to a number of fungicides belonging to different chemical classes with different modes of action.

[0152] This improved control was not due to an additive fungicidal effect, as the adjuvant alone did not inhibit the growth of B. cinerea, indicating that it did not affect fungal growth.

[0153] Example - Adjuvant with other fungicides against other fungi To assess the biological spectrum of activity, Lamericol Anhydrous Adjuvant (Ia) was tested in combination with seven fungicides against six different fungal test organisms belonging to five different genera.

[0154] Prior to assessing the adjuvant effect of (Ia), the minimum inhibitory concentration (MIC) of each bactericide against each test organism was determined by measuring growth on agar plates containing sterile concentrations.

[0155] Once the MICs were determined, adjuvant tests were performed using a fixed (Ia) concentration of 4 μg / mL and each fungicide at a concentration that inhibited fungal growth by 10–20%.

[0156] The results are shown in Table 3.

[0157] [Table 3]

[0158] The fungal targets are FT1-Fusarium solani, FT2-Fusarium oxysporum, FT3-Alternaria infectorium, FT4-Penicillium roqueforti, FT5-Aspergillus flavus, FT6-Cladosporium sp.

[0159] Numbers indicate the fold change in fungal growth inhibition compared to fungal test organisms treated with fungicide and adjuvant vehicle only. A value of 1 indicates no effect on fungicidal activity, while a value >1 indicates an improvement in fungicidal activity. Values ​​indicate no effect on fungicidal activity.

[0160] Adjuvant activity was observed with dimethomorph against Fusarium solani (1.63-fold increase), pyraclostrobin against Fusarium oxysporum (1.63-fold increase), and mancozeb, dimethomorph, prothioconazole, and pyraclostrobin against Alternaria infectorium (2.45-, 1.25-, 1.50-, and 1.18-fold increases, respectively). Pyrimethanil, mancozeb, dimethomorph, prothioconazole, iprodione, pyraclostrobin, and captan increased by 1.31, 1.63, 1.85, 1.52, 1.65, 1.27, and 1.17 times against P. roqueforti, mancozeb, dimethomorph, prothioconazole, and iprodione increased by 2.01, 1.27, 1.28, and 1.72 times against A. flavus, and pyrimethanil increased by 1.67 times against Cladosporium sp.

[0161] These results demonstrate that Lamellicol Anhydrous Adjuvant (Ia) can be combined with multiple fungicides with different chemical structures and modes of action to improve the control of several fungal species, thus further broadening the practical application of the adjuvants described herein.

[0162] Example - Phytotoxicity Plant safety is an important feature of fungicide adjuvants, so that application of the adjuvant does not adversely affect plant health. To determine safe use levels of lamellicholic anhydride, phytotoxicity studies were conducted on soybean (Glycine max, cv Alexa).

[0163] Soybean plants were grown in 9 cm x 9 cm plastic pots from single seeds inoculated with Bradyrhizobium japonicum (Novozymes GlyciMax) in Levington's M3 compost. The greenhouse temperature was set at 22 ± 2°C during the day and 19 ± 2°C at night. The photoperiod was set at 14 hours.

[0164] Supplemental lighting was provided to soybean plants from SON-T bulbs. Biological controls were used to prevent thrips (Bioline-Amblyseius cucumeris) damage.

[0165] All plants were maintained in a well-hydrated state. Adjuvant treatments were applied to soybean plants that had reached the V4 stage (6 weeks). Leaves were placed on a platform made from pot racks to hold the foliar treatments immediately after application. Frames were stacked high enough to allow the leaves to lie flat on top. Stacked card sheets were taped to the top of the frames to form a support for the leaves.

[0166] For soybean plants, one fully expanded trifoliate central leaf was held flat by carefully taping down the edges with micropore tape.

[0167] Two preparations were evaluated for phytotoxicity: a crude fermentation extract containing illicorine and lamellicholic anhydride prepared by extracting the fermentation of solid rice with ethyl acetate, and purified fractions (If and Ig) containing lamellicholic anhydride 305. These materials were prepared as described above.

[0168] The test substance was dissolved in 20% (v / v) DMSO at a concentration of 10 mg / mL, and a dilution series of 0.05 to 1 g / L was prepared while maintaining the DMSO concentration at 2% (v / v).

[0169] The positive phytotoxicity control was Synperonic A11 (1 g / L; available from Croda), while the negative control was Atplus UEP-100 (1 g / L; available from Croda).

[0170] Treatments were randomly assigned to leaf sites and marked with a fine-tip marker pen. For any dilution rate, all treatments could be applied to a single leaf. A total of eight replicate leaves on separate plants were used for each treatment dilution. Treatments were applied to the leaves as 10 μL drops. The overhead plant lights were turned off during treatment and were not turned on until the drops had completely dried.

[0171] Phytotoxicity was scored and plants were assessed for phytotoxic tissue damage at 1 day after treatment (DAT), 7 days and 14 days.

[0172] The results are shown in Table 4.

[0173] [Table 4]

[0174] Phytotoxicity scale 0~3: 0 - no damage / necrosis; 1 - slight spot-like necrosis in areas wetted by the drop; 2 - ring-shaped necrosis; 3 - expanding necrosis.

[0175] The observed phytotoxicity of the crude extract and lamellicholic anhydride 305 was less than 0.4 when tested up to 0.25 mg / L / spot, indicating that the phytotoxic effect was minor and limited to the spot wetted by the drop. No phytotoxic effect was observed in any of the test samples below 0.1 mg / mL. As expected, Atplus UEP-100 exhibited only a slight phytotoxic effect, which manifested only 14 DAT.

[0176] These results demonstrate the applicability of adjuvant preparations prepared from Cosmospora sp. RKDO 1747 did not damage soybean tissue when applied at concentrations below 0.25 g / L, and the crude extract and lamellicholic anhydride 305 samples showed no phytotoxic effects when applied at 0.1 or 0.05 g / L, respectively. Because the adjuvant activity of lamellicholic anhydride is generally achieved at concentrations below 0.01 g / L, the rate required to achieve adjuvant activity is significantly lower than the rate at which phytotoxic effects are observed. It should be understood that the present invention is not limited to the details of the above embodiments, which are described by way of example only. Many variations are possible.

Claims

1. below: i) Formula (I): 【Chemistry 1】 [In the formula, R １ , R ３ , R ４ , R ５ , and R ６ are all hydrogen, and R ２ is methyl; or R １ , R ３ , R ４ , and R ６ are all hydrogen, and R ５ is hydroxyl, and R ２ is methyl; or R １ , R ４ , R ５ , and R ６ are all hydrogen, and R ２ and R ３ is methyl; or R ４ , R ５ , and R ６ are all hydrogen, and R ２ , R ３ , and R １ is methyl; or R ３ , R ５ , and R ６ are all hydrogen, and R ２ , R ４ , and R １ is methyl; or R ４ and R ５ is hydrogen, and R ２ , R ３ , and R １ is methyl, and R ６ is a hydroxyl group; or R ３ and R ５ is hydrogen, and R ２ , R ４ , and R １ is methyl, R ６ is a hydroxyl group] an adjuvant which is lamellicholic anhydride according to ii) at least one pesticidal active substance; Pesticide formulations containing

2. The compound of formula (I) R １ independently represent hydrogen or methyl; R ２ independently represent hydrogen or methyl; R ３ and R ４ independently represent hydrogen or methyl; and R ５ and R ６ independently represent hydrogen or hydroxyl The formulation of claim 1 , wherein the compound is selected from the group consisting of:

3. Lamellicholic anhydride is as follows: 【Chemistry 2】 【Transformation 3】 The formulation according to claim 1 or 2, wherein the formulation is selected from the group consisting of:

4. As an adjuvant in a pesticide formulation containing at least one pesticidal active substance, a compound of formula (I): 【Chemistry 4】 [In the formula, R 1 , R 3 , R 4 , R 5 , and R 6 are all hydrogen and R 2 is methyl; or R 1 , R 3 , R 4 , and R 6 are all hydrogen, R 5 is hydroxyl, and R 2 is methyl; or R 1 , R 4 , R 5 , and R 6 are all hydrogen and R 2 and R 3 are methyl; or R 4 , R 5 , and R 6 are all hydrogen, and R 2 , R 3 , and R 1 are methyl; or R 3 , R 5 , and R 6 are all hydrogen and R 2 , R 4 , and R 1 are methyl; or R 4 and R 5 are hydrogen, R 2 , R 3 , and R 1 are methyl, and R 6 is a hydroxyl group; or R 3 and R 5 are hydrogen, R 2 , R 4 , and R 1 are methyl, and R 6 is a hydroxyl group. Use of Lamellicol Anhydrous by.

5. A method for treating plants to control pests, comprising applying a pesticide formulation according to any one of claims 1 to 3 either to the vegetation or to the immediate environment of the vegetation.

6. A method for obtaining the adjuvant according to any one of claims 1 to 3, comprising the following steps: Cultivating Cosmospora sp. RKDO1747 in a culture medium under conditions that promote the metabolic synthesis of the adjuvant according to any one of claims 1 to 3 from said Cosmospora sp.; and Purifying the synthetic adjuvant from the culture medium A method comprising:

7. An organism consisting of Cosmospora sp. strain RKDO1747, Agricultural Research Service Culture Collection (NRRL) accession number NRRL-67910.

8. 4. An extract obtained from an organism consisting of Cosmospora sp. strain RKDO1747, Agricultural Research Service Culture Collection (NRRL) accession number NRRL-67910, comprising at least one of the lamellicol anhydrates according to any one of claims 1 to 3.

9. A seed coating composition comprising the adjuvant according to any one of claims 1 to 3. 【Request Item 10】 【Chemistry 5】 【Transformation 6】 【Transformation 7】 Lamellicholic anhydride selected from:

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