Pesticide adjuvants from Stachybotrys chartarum

Abstract

A novel pesticide formulation comprising an adjuvant selected from phenylspirodrimanes and derivatives thereof, and a pesticide active substance. A concentrate suitable for forming the formulation is also provided. The phenylspirodrimanes provide adjuvancy in the concentrates and pesticide formulations. A method for producing the formulation is also provided, as well as the use of the phenylspirodrimanes as adjuvants in pesticide formulations. A method for obtaining diketopiperazines from a culture of Stachybotrys chartarum, strain RKDO1264, is also described. The extracted phenylspirodrimanes may be used in a formulation for vegetation treatment for pest control or in a seed coating. Alternatively, the strain RKDO1264 itself may be used in a seed coating or in a formulation for vegetation treatment for pest control.

Description

[Technical field]

[0001] The present invention relates generally to microbially derived adjuvants for pesticide active formulations and methods of providing adjuvancy in pesticide formulations that include said adjuvants in combination with one or more pesticide actives. The invention also includes treating crops with said formulations. [Background technology]

[0002] Adjuvants are generally defined as chemicals or mixtures of chemicals that can improve the biological activity or efficacy of pesticide actives. Adjuvants themselves do not control or kill pests. Instead, these additives may interact with molecular targets (cell walls, ion channels, structural proteins, enzymes, etc.) within the target organism or modify the properties of the pesticide formulation (diffusion, retention, penetration, droplet size, etc.), thereby improving the biological activity of the pesticide acting on the organism. Typical types of compounds used as adjuvants can include small molecules, surfactants, emulsifiers, oils, and salts. Adjuvants typically do not inhibit the translocation of activity in the treated plant. In addition, adjuvants must not produce undesirable phytotoxic effects on the plant.

[0003] 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 such losses and meet growing food demands, the use of fungicides to control fungal agricultural pests is and will remain an important component of agricultural pest management systems.

[0004] There is a need to develop new strategies to combat agricultural pests, especially fungal pests. One strategy is to develop adjuvants, safe, non-toxic 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 can improve the control of field or post-harvest pests, 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.

[0005] The present invention aims to provide the use of compounds in pesticide formulations in combination with pesticide active substances, where the compounds may provide desired adjuvants, including improved efficacy of their activity. The present invention also aims to provide the use of pesticide concentrates and dilution formulations comprising said adjuvants.

[0006] The present invention also aims to provide compounds in agrochemical formulations, where the compounds may provide comparable or improved adjuvancy to existing adjuvants.

[0007] The present invention also aims to provide the use of the compounds as adjuvants and formulations containing said compounds in pesticide formulations for use in providing adjuvantcy.

[0008] According to a first aspect of the present invention, there is provided a method for producing a cellular membrane comprising the steps of: i) Formula (I): [ka] {In the formula, R1 independently represents hydrogen, C1-C4 alkyl, hydroxy, methoxy, ethoxy, or amine; R2 independently represents hydrogen, C1-C6 alkyl, phenyl, substituted phenyl, alkylphenyl, or alkyl-substituted phenyl; R3 and R4 independently represent hydrogen or C1-C4 alkyl; or Formula (II): [ka] {In the formula, R 10 each independently represents hydrogen, C1-C4 alkyl, hydroxy, methoxy, ethoxy, or carboxyl; R 11 , R 12 , R 13 , and R 14 each independently represents hydrogen or C1-C4 alkyl; and ii) at least one pesticide active substance; The present invention provides an agrochemical formulation comprising:

[0009] According to a second aspect of the present invention there is provided a concentrate suitable for producing an agrochemical formulation according to the first aspect, said concentrate comprising: i) Formula (I): [ka] {In the formula, R1 independently represents hydrogen, C1-C4 alkyl, hydroxy, methoxy, ethoxy, or amine; R2 independently represents hydrogen, C1-C6 alkyl, phenyl, substituted phenyl, alkylphenyl, or alkyl-substituted phenyl; R3 and R4 independently represent hydrogen or C1-C4 alkyl; or Formula (II): [ka] {In the formula, R 10 each independently represents hydrogen, C1-C4 alkyl, hydroxy, methoxy, ethoxy, or carboxyl; R 11 , R 12 , R 13 , and R 14each independently represents hydrogen or C1-C4 alkyl; and ii) at least one pesticide active substance; Includes.

[0010] According to a third aspect of the present invention, there is provided, as adjuvant in an agrochemical formulation comprising at least one agrochemically active substance, the following: i) Formula (I): [ka] {In the formula, R1 independently represents hydrogen, C1-C4 alkyl, hydroxy, methoxy, ethoxy, or amine; R2 independently represents hydrogen, C1-C6 alkyl, phenyl, substituted phenyl, alkylphenyl, or alkyl-substituted phenyl; R3 and R4 independently represent hydrogen or C1-C4 alkyl; or Formula (II): [ka] {In the formula, R 10 each independently represents hydrogen, C1-C4 alkyl, hydroxy, methoxy, ethoxy, or carboxyl; R 11 , R 12 , R 13 , and R 14 each independently represents hydrogen or C1-C4 alkyl}.

[0011] According to a fourth aspect of the present invention there is provided a method of treating vegetation to control pests comprising applying a formulation according to the first aspect or a diluted concentrated formulation according to the second aspect either to said vegetation or to the environment surrounding said vegetation.

[0012] According to a fifth aspect of the present invention, there is provided a method for obtaining an adjuvant according to the first aspect, comprising the following steps: Cultivating Stachybotrys chartarum RKDO1264 in a medium under conditions that promote metabolic synthesis of an adjuvant according to the first aspect from said Stachybotrys chartarum; and purifying the synthetic adjuvant from the culture medium; The method further comprises:

[0013] According to a sixth aspect of the present invention there is provided an organism comprising Stachybotrys chartarum, strain RKDO1264, Agricultural Research Service Type Culture Collection (NRRL) Accession No. NRRL-67988.

[0014] According to a seventh aspect of the present invention there is provided an extract obtained from an organism consisting of Stachybotrys chartarum, strain RKDO1264, Agricultural Research Service Type Culture Collection (NRRL) accession number NRRL 67988, said extract comprising a phenylspirodrimane according to the first aspect.

[0015] According to an eighth aspect of the present invention there is provided a method of treating vegetation to control pests comprising applying an organism according to the sixth aspect either to the vegetation or to the environment surrounding the vegetation.

[0016] 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 an organism according to the sixth aspect.

[0017] The compounds defined herein have been found to provide the desired adjuvancy when used in an agrochemical formulation having at least one agrochemical activity.Importantly, the identified class of compounds, phenylspirodrimanes, do not exhibit inherent insecticidal activity.

[0018] As used herein, the terms "for example," "for instance," "such as," or "including" are meant to introduce examples that further clarify a more general subject matter. Unless otherwise specified, these examples are provided only as an aid in understanding the applications presented in this disclosure and are not intended to be limiting in any way.

[0019] When describing the number of carbon atoms in a substituent (e.g., "C1-C4 alkyl"), this number refers to the total number of carbon atoms present in the substituent, including those present in any branching groups. Further, when describing the number of carbon atoms in, for example, a fatty acid, this refers to the total number of carbon atoms, including those located in the carboxylic acid and those present in any branching groups.

[0020] The adjuvants of the present invention have the structure of formula (I): [ka] {In the formula, R1 independently represents hydrogen, C1-C4 alkyl, hydroxy, methoxy, ethoxy, or amine; R2 independently represents hydrogen, C1-C6 alkyl, phenyl, substituted phenyl, alkylphenyl, or substituted alkylphenyl; R3 and R4 independently represent hydrogen or C1-C4 alkyl; or Structure of formula (II); [ka] {In the formula, R 10 each independently represents hydrogen, C1-C4 alkyl, hydroxy, methoxy, ethoxy, or carboxyl; R 11 , R 12 , R 13 , and R 14 each independently represents hydrogen or C1-C4 alkyl. The phenyl spirodrimanes are selected from the group consisting of

[0021] As used herein, the term "C1-C4 alkyl" refers to a saturated hydrocarbon radical that is straight or branched chain containing 1 to 4 carbon atoms, unless otherwise defined. When any of R represents C1-C4 alkyl, said alkyl may be independently selected from methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, sec-butyl, tert-butyl, and the like. Preferred is methyl, ethyl, n-butyl, or isobutyl. More preferred is methyl or isobutyl.

[0022] As used herein, the term "C1-C6 alkyl" refers to a saturated hydrocarbon radical that is straight or branched chain containing 1-6 carbon atoms, unless otherwise defined. When any of R represents C1-C6 alkyl, said alkyl may be independently selected from methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, sec-butyl, tert-butyl, 2-methyl-butyl, pentyl, hexyl, cyclohexyl, and the like. Preferably, it is methyl or ethyl. More preferably, it is methyl.

[0023] As used herein, unless otherwise defined, the term "hydroxyl" refers to a hydroxyl radical having the structure --OH, which contains one oxygen and one hydrogen atom and is bonded to an adjacent radical through its oxygen.

[0024] As used herein, unless otherwise defined, the term "phenyl" refers to a C6H5 organic radical derived from the benzene aromatic hydrocarbon by removal of one hydrogen.

[0025] As used herein, the term "phenyl," unless otherwise defined, refers to phenyl substituted with methyl, ethyl, methoxy, ethoxy, or halo, said substitutions may be at any position on the aromatic ring, preferably at the ortho or para position.

[0026] As used herein, the term "alkylphenyl" refers to a phenyl bearing an alkyl group, preferably methyl or ethyl, and more preferably methyl, through which it is attached to an adjacent radical.

[0027] As used herein, unless otherwise defined, the term "substituted alkylphenyl" refers to a phenyl that includes the alkylphenyl and substituted phenyl characteristics described herein. When the R2 group is a phenyl-containing group, phenyl or alkylphenyl may be preferred, especially alkylphenyl.

[0028] The terms "methoxy" and "ethoxy," as used herein, unless otherwise defined, refer to methyl and ethyl groups linked to an oxygen forming an alkoxy radical having the structures -O-Me and -O-Et and which are bonded to an adjacent radical through that oxygen.

[0029] As used herein, unless otherwise defined, the term “amine” refers to the structure —NR 15 R 16 R 17 and which is bonded to an adjacent radical through a nitrogen, and is a primary, secondary, or tertiary amine. R 15 , R 16 , and R 17 Each represents hydrogen or C1-C4 alkyl as defined herein.

[0030] As used herein, unless otherwise defined, the term "carboxyl" refers to a carboxyl radical having the structure -C(=O)-OH and which is bonded to an adjacent radical through a carbon.

[0031] When the phenylspirodrimane has the structure of formula (I), preferably, R1 represents hydroxy, methoxy, ethoxy, or amine. More preferably, R1 represents hydroxy, methoxy, or amine. Even more preferably, R1 represents hydroxy.

[0032] Preferably, R2 represents C1-C4 alkyl, phenyl, or alkylphenyl. More preferably, R2 represents methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, sec-butyl, phenyl, methylphenyl, or ethylphenyl. More preferably, R2 represents n-butyl, isobutyl, phenyl, or methylphenyl. More preferably, R2 represents isobutyl or methylphenyl.

[0033] Preferably, R3 and R4 independently represent hydrogen, methyl or ethyl, more preferably hydrogen.

[0034] When the phenylspirodrimane has the structure of formula (II), preferably R 10 represents hydrogen, hydroxy, methoxy, or carboxyl, more preferably represents hydrogen, methoxy, or carboxyl, and even more preferably represents carboxyl.

[0035] Preferably, R 11 , R 12 , R 13 , and R 14 each independently represents hydrogen, methyl or ethyl, more preferably hydrogen.

[0036] The compounds of formula (I) and (II) may have optical isomerism around a chiral carbon, and it is understood that both R and S isomers are included within the definition of the phenylspirodrimane of formula (I).

[0037] In particular, phenyl spirodrimer selected from the following may be preferred: [ka]

[0038] The organisms used for fermentation are filamentous fungi belonging to the genus Stachybotrys. A particular strain of Stachybotrys chartarum, RKDO1264, 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 Type Collection (NRRL) in Peoria, Illinois, USA:

[0039] [Table 1]

[0040] NRRL-67988 is particularly preferred for providing adjuvant activity in the present invention. Preferably, there is provided a process according to the fifth aspect for obtaining a phenylspirodrimane adjuvant of formula (I) or (II).

[0041] Phenylspirodrimanes can be formed and extracted from the culture of Stachybotrys chartarum, particularly RKDO1264, respectively. The desired compounds can be extracted and purified from the culture or fungal biomass by any means typically used to recover microbial metabolites in general. For example, separation methods using 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, which means that the means can be used alone or in suitable combinations thereof, or repeatedly.

[0042] A culture of Stachybotrys chartarum RKDO1264 can be obtained from natural sources or from culture collections such as the National Agriculture Research Service Type Culture Collection (NRRL). Stachybotrys chartarum isolate RKDO1264 can be cultured by methods known in the art of mycology.

[0043] As a means of producing the compounds of the invention, the production organisms can be grown on any suitable synthetic or natural medium, so long as it contains an appropriate source of carbon, nitrogen, and inorganic salts. If necessary, the medium can be appropriately supplemented with vitamins and other nutrients.

[0044] Common carbon sources include, but are not limited to, sugars such as glucose, maltose, fructose, sucrose, starch, alcohols such as glycerol, mannitol, amino acids such as glycine, alanine, asparagine, oils and fats such as soybean oil and olive oil. Examples of 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, micronutrients in the form of inorganic salts can be used, such as calcium carbonate, sodium chloride, potassium chloride, magnesium sulfate, copper sulfate, manganese chloride, zinc sulfate, cobalt chloride, and various phosphates.

[0045] The microorganism can be grown at an appropriate culture temperature within a range that allows the growth of the microorganism and effective production of the compound of the present invention. The preferred culture temperature is 10 to 32° C., more preferably 20 to 25° C., and the pH at the start of the culture is preferably about 6 to 8. The culture period is generally about one day to several weeks.

[0046] 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 the maximum amount. As the culture method, any method can be appropriately used, including a commonly used method such as solid-phase culture or ordinary agitation culture.

[0047] For example, Stachybotrys chartarum isolate RKDO1264 can be plated onto nutrient-containing (e.g., YM (yeast malt extract) agar) and incubated at room temperature for several days until observable colonies appear. Individual Stachybotrys chartarum RKDO1264 colonies on the agar can be assayed for production of phenylspirodrimanes.

[0048] Those colonies producing the desired molecule can be used to inoculate liquid media (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 larger liquid cultures (e.g., YES broth supplemented with 10 g / L L-leucine), which can be incubated at about room temperature for several days (e.g., 4-28 days) to expand the Stachybotrys chartarum culture.

[0049] Phenylspirodrimane is found to be excreted into liquid media (e.g., YES broth). Phenylspirodrimane can be isolated from the fermentation broth using liquid:liquid extraction with ethyl acetate and water, and binding the compound to an absorbent resin (such as Diaion™ HP20), washing the resin with water, and then eluting the phenylspirodrimane with a suitable solvent (such as methanol or ethanol). Due to the difference in polarity of the phenylspirodrimane, the individual compounds can be separated using chromatography, such as flash chromatography and reversed-phase stationary phase (such as C-18).

[0050] The resulting extracted phenylspirodrimanes can be purified and used as individual homogenous compounds. In another embodiment, the extracted materials may be in combination with phenylspirodrimanes according to the first aspect and may be used in combination in agrochemical formulations.

[0051] In other embodiments, the phenylspirodrimanes can be obtained from other available sources, typically other fungi.

[0052] Phenylspirodrimanes can also be prepared by synthetic techniques: Phenylspirodrimanes and their derivatives can be prepared by chemical synthesis by one skilled in the art of organic chemistry using commercially available materials and synthetic methods described in the scientific literature.

[0053] It is understood that the properties of the adjuvant itself provide the same advantageous benefits to the agrochemical formulation that contains the adjuvant. Thus, there is provided an agrochemical formulation, when it contains the adjuvant of the present invention, that has the advantage of the properties of the adjuvant itself.

[0054] Agrochemically active compounds, including insecticides and fungicides, require a formulation that allows the active compound to be taken up by the plant / target organism.

[0055] The term "pesticidal formulation" as used herein refers to a composition containing an active pesticide and is intended to include any form of composition, including concentrates and spray formulations. Unless otherwise specified, the pesticide formulation of the present invention can be in the form of a concentrate, a diluted concentrate, or a sprayable formulation.

[0056] The adjuvants of the present invention can be combined with other ingredients to form pesticide formulations containing at least one pesticide active.

[0057] Thus, the pesticidally active compound may be formulated as an emulsifiable concentrate (EC), emulsion concentrate (EW), suspension concentrate (SC), soluble liquid (SL), oil-based suspension concentrate (OD), and / or suspension emulsion (SE).

[0058] 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.

[0059] The adjuvants of the invention are particularly contemplated for use in EC, EW, SC, SL, OD, or SE formulations.

[0060] Agrochemical concentrates are agrochemical compositions that can be aqueous or non-aqueous and are designed to be diluted with water (or a water-based liquid) to form a corresponding spray formulation. Such compositions include liquid forms (such as solutions, emulsions or dispersions) and solid forms (particularly water-dispersible solid forms), such as granules or dusts.

[0061] 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 the individual ingredients, or by a combination of diluting a concentrate and then adding the individual ingredients or mixtures of ingredients. Typically, such end-use mixing is done in the tank where the formulation is sprayed, or alternatively in a holding tank for filling the spray tank. Such mixing and mixing are typically called tank mixes and tank mixes.

[0062] Thus, adjuvants may be incorporated into the formulation of the pesticidal active compound (in-can / built-in formulation) or may be added after dilution of a concentrated formulation of the spray liquid (tank mix). In order to avoid dosing errors and to improve user safety during application of the pesticide product, it is advantageous to incorporate the adjuvant into the formulation. This also avoids unnecessary use of packaging material for tank-mix products.

[0063] Depending on the customer's needs, the concentrate thus formed can typically contain up to 95% by weight of pesticide active. The concentrate can be diluted for use, resulting in a diluted composition having a pesticide active concentration of about 0.5% to about 1% by weight. The diluted composition (e.g., at a spray application rate of 10 to 500 l.ha) can be used in a wide range of applications. -1In the spray formulation, which may be a spray formulation, the pesticide active concentration may be in the range of about 0.001% to about 1% by weight of the total formulation as sprayed.

[0064] The adjuvants of the present invention are typically used in an amount proportional to the amount of active pesticide in the formulation. In pesticide formulation concentrates, 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, it is 1% to 70% by weight. More preferably, it is 3% to 50% by weight.

[0065] For example, upon dilution to form a spray formulation, the adjuvant is typically present at a concentration of 0.01% to 2% by weight of the spray formulation, more typically 0.03% to 0.5% by weight of the formulation, and more preferably 0.12% to 0.4% by weight of the spray formulation.

[0066] The ratio of adjuvant to active pesticide in the pesticide formulation is preferably about 1:40 to about 1:1. More preferably, it is about 1:20 to about 1:1. Even more preferably, it is about 1:5 to 1:1. This ratio range is generally maintained in the case of concentrated formulations (e.g., when the adjuvant is contained in a dispersible liquid concentrate or dispersible solid granule) and in spray formulations.

[0067] When a concentrate (solid or liquid) is used as a source of active pesticide and / or adjuvant, the concentrate is typically diluted to form a spray formulation. The dilution can be with 1 to 10,000 times, particularly 10 to 1,000 times, the total mass of the concentrate with water to form a spray formulation.

[0068] When the pesticide active is present as a solid particle in the aqueous end-use formulation, in most cases it will 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 support, filler or diluent material as described above.

[0069] Spray formulations typically have a pH ranging from moderately acidic (e.g., about 3) to moderately alkaline (e.g., about 10), especially closer to neutral (e.g., about 5 to 8). More concentrated formulations have a similar degree of acidity / alkalinity, but pH is not necessarily an appropriate measure of this, as they may be largely non-aqueous.

[0070] The agrochemical formulation may include, in association with the adjuvant, a solvent (other than water), such as monopropylene glycol, a vegetable oil or a mineral oil, such as an oil that may be a spray oil (an oil included in the spray formulation as a non-surfactant adjuvant). Such a solvent may be included as a solvent for the adjuvant and / or as a humectant, such as propylene glycol in particular. When used, such a solvent is typically included in an amount of 5% to 500% by weight, desirably 10% to 100% by weight, based on the weight of the adjuvant. Such a combination may also include salts, such as ammonium chloride and / or sodium benzoate, and / or urea, in particular as a gel suppression aid.

[0071] In another embodiment, the adjuvant of the present invention, or any of the organisms according to the sixth aspect, can be included in a seed application composition suitable for application to a seed. Preferably, the adjuvant of the present invention can be included in a seed coating composition.

[0072] The adjuvant is suitably present in the seed coating composition in 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.

[0073] Coating may include film coating, pelleting, 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 to film coating.

[0074] The seed coating composition of the present invention can be applied to the seeds in a conventional manner.

[0075] The seeds may be primed or unprimed (subjected to a treatment to improve germination rate, e.g. osmopriming, hydropriming, matrix priming).

[0076] In one embodiment, before applying the seed coating composition of the present invention, for example a primer layer containing a binder such as a polymer, the seed is not provided with an artificial layer.Therefore, the seed coating composition is preferably applied directly to the natural outer surface of the seed.Nevertheless, before applying the seed coating composition, the seed surface may be subjected to a surface treatment.

[0077] 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. The term "liquid coating composition" as used in this application is meant to include coating compositions in the form of suspensions, emulsions, and / or dispersions, preferably dispersions.

[0078] Conventional coating means can be used to coat the 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 the seeds by a rotary coater, a rotary dry coater, a pan coater, or a continuous processing device.

[0079] The seed coating composition can be applied, for example, by film coating, spray application, dipping or brush application of the seed coating composition.Preferably, the method includes applying the seed coating composition to form a film or seed coating layer.Seed coating typically includes forming a tightly adherent moisture-permeable coating on the surface of the seed.The method typically includes applying a liquid seed coating composition to the seed before sowing.

[0080] Additional film coating layers may optionally be applied on top of the coating layers 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.

[0081] The pesticide formulation or seed coating composition may optionally contain other ingredients. These other ingredients may be selected from the following: Binders, especially those which dissolve readily in water and give solutions with low viscosity at high binder concentrations, such as polyvinylpyrrolidone; polyvinyl alcohol; carboxymethylcellulose; gum arabic; sugars, such as sucrose or sorbitol; starch; ethylene-vinyl acetate copolymers, sucrose, and alginates; Diluents, absorbents or carriers, such as carbon black; talc; diatomaceous earth; kaolin; aluminium stearate, calcium stearate or magnesium stearate; sodium tripolyphosphate; sodium tetraborate; sodium sulfate; sodium silicate, aluminium silicate and sodium aluminium silicate mixtures; sodium benzoate and the like; Disintegrants, such as surfactants, materials which swell in water, such as carboxymethylcellulose, collodion, polyvinylpyrrolidone, and microcrystalline cellulose swelling agents; salts, such as sodium or potassium acetate, sodium carbonate, sodium bicarbonate or sodium sesquicarbonate, ammonium sulfate, and dipotassium hydrogen phosphate; Wetting agents, such as alcohol ethoxylate and alcohol ethoxylate / propoxylate wetting agents; Adjuvants, such as sulfonated naphthalene formaldehyde condensates and acrylic acid copolymers, such as comb copolymers with capped polyethylene glycol side chains on a polyacrylic acid backbone; Emulsifiers, such as alcohol ethoxylates, ABA block copolymers, castor oil ethoxylates, etc.; · Antifoaming agents, such as polysiloxane antifoaming agents, typically in an amount 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 cellulosic materials, such as, for example, carboxy-, methyl-, ethyl- or propyl-cellulose; and / or preservatives and / or antimicrobial agents, for example organic acids or their esters or salts, for example ascorbic acid based, for example ascorbyl palmitate, sorbic acid based, for example potassium sorbate, benzoic acid based, for example benzoic acid, methyl 4-hydroxybenzoate and propyl 4-hydroxybenzoate, propionic acid based, for example sodium propionate, phenolic based, for example sodium 2-phenylphenate; 1,2-benzisothiazolin-3-one; or formaldehyde itself or paraformaldehyde; or inorganic materials, for example sulfurous acid and its salts, typically in an amount of 0.01% to 1% by weight of the formulation.

[0082] The pesticide formulation or seed coating composition according to the invention may comprise ingredients such as, for example, surfactants which form part of an emulsifier system. Said surfactants include surface active adjuvants.

[0083] Other adjuvants outside the scope of the present invention, such as surface active adjuvants, may be included in the compositions and formulations of the present invention and those 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(en)yl succinic anhydrides, particularly those described in PCT applications WO94 / 00508 and WO96 / 16930.

[0084] The formulation / composition may contain one or more biologically active ingredients, including plant enhancers, especially plant protection products (also called PPPs). Suitable examples of active ingredients, especially plant enhancers, are antimicrobial agents, fungicides, insecticidal agents, nematicides, molluscicides, biological agents, acaricides or miticides, 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, biological agents, etc.

[0085] The pesticide active substances suitable for use in the formulation or seed coating composition according to the invention are all pesticide active compounds that can be solid or liquid at room temperature. It is envisaged that the adjuvants of the invention have a broad applicability to all types of pesticide active substances.

[0086] Agrochemical active ingredients in the context of the present invention are plant protection agents, more specifically biocides, which are chemicals capable of killing various forms of living organisms, for example, in the fields of medicine, agriculture, forestry, mosquito control, etc. The group of biocides also includes so-called plant growth regulators.

[0087] The biocides used in the pesticide formulations or seed coating compositions of the invention are typically divided into two subgroups: pesticides, including, for example, fungicides, herbicides, insecticides, algaecides, molluscicides, acaricides, and rodenticides; and Antimicrobial agents, including for example germicides, antibiotics, antibacterial agents, antiviral agents, antifungal agents, antiprotozoal agents and antiparasitic agents.

[0088] In particular, biocides selected from insecticides, fungicides or herbicides may be particularly preferred.

[0089] The term "pesticides" is understood to refer to substances or mixtures of substances intended to prevent, destroy, repel or mitigate pests. Pesticides are used to control insects, plant pathogens, weeds, mollusks, and other pests. A pesticide is a chemical or biological agent (such as a virus or bacteria) used against pests, including animals, birds, mammals, fish, nematodes (roundworms), and microorganisms that compete with humans for food, destroy property, spread disease, or cause nuisance. The following examples illustrate pesticides suitable for use in the pesticide composition according to the present invention.

[0090] Fungicides are chemical control agents for fungi. Fungicides are compounds used to prevent the spread of fungi in gardens and crops. Fungicides are also used to fight fungal infections. Fungicides are either contact or systemic. Contact fungicides kill fungi when they come into contact with the fungicide that is held on the leaf surface. Systemic fungicides are absorbed into the plant tissue and kill the fungus as it attempts to invade the host.

[0091] According to the present invention, examples of suitable fungicides are (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, benozom ... Benquinox, bentaluron, benthiavalicarb, benzalkonium chloride, benzamacryl, benzamide fungicides, benzamorph, benzanilide fungicides, benzimidazole fungicides, benzimidazole precursor fungicides, benzimidazolylcarbamate fungicides, benzohydroxamic acid, benzothiazole fungicides, bethoxazin, binapacryl, biphenyl, bitertanol, bithionol, blasticidin-S, Bordeaux mixture, boscalid, bridged diphenyl fungicides, bromuconazole, bupirimate, Burgundy mixture, buthiobacter , butylamine, calcium polysulfide, captafol, captan, carbamate fungicides, carbamorph, carbanilate fungicides, carbendazim, carboxin, carpropamid, carvone, chechant mixture, chinomethionate, clobenziazone, chloraniformethane, chloranil, chlorphenazole, chlordinitronaphthalene, chloroneb, chloropicrin, chlorthalonil, chlorquinox, clozolinate, ciclopirox, clivazol, clotrimazole, conazole fungicide, conazole fungicide (imidazole), co Nazole fungicides (triazoles), copper (II) acetate, copper (II) carbonate, basic, copper fungicides, 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 fungicides, cycloheximide, cyflufenamid, cymoxanil, cypendazole, cyproconazole, cyprodinil, dazomet, DBCP, debacarb, decafentin, dehydroacetic acid, dicarboximide fungicides, dichlofluanid,Dicloron, dichlorophen, dichlorophenyl, dicarboximide fungicides, diclozolin, diclobutrazol, diclocymet, diclomedine, dicloran, diethofencarb, diethyl pyrocarbonate, difenoconazole, diflumetrim, dimethirimol, dimethomorph, dimoxystrobin, diniconazole, dinitrophenol fungicides, dinobuton, dinocap, dinokton, dinopenton, dinosulfone, dinotervone, diphenylamine, dipyrithione, disulfiram, ditalinphos, dithianon, dithiocarbamate fungicides, DNOC , dodemorph, dodicine, dodine, donatodine, drazoxolone, edifenphos, epoxiconazole, etaconazole, ethem, ethaboxam, ethirimol, ethoxyquin, ethylmercuric 2,3-dihydroxypropyl mercaptide, ethylmercuric acetate, ethylmercuric bromide, ethylmercuric chloride, ethylmercuric phosphate, etridiazole, famoxadone, fennamidone, fenapanil, fenarimol, fenbuconazole, fenfuram, fenhexamid, fenitropan, fenoxanil, fenpiclonil, fenpropidin, fenpropimol F, Fentin, Ferbam, Ferimzone, Fluazinam, Fludioxonil, Flumetover, Fluopicolide, Fluorimide, Flutrimazole, Fluoxastrobin, Fluquinconazole, Flusilazole, Flusulfamide, Flutolanil, Flutriafol, Folpet, Formaldehyde, Fosetyl, Fuberidazole, Furalaxyl, Furametopyr, Furamide Fungicide, Furanilide Fungicide, Flucarbanil, Fluconazole, Fluconazolecis, Furfural, Flumesilox, Furofunate, Gliodin, Griseoful Bin, guazatine, halacrinate, hexachlorobenzene, hexachlorobutadiene, hexachlorophene, hexaconazole, hexylthiophos, hydrargafen, hymexazole, imazalil, imibenconazole, imidazole fungicide, iminoctadine, inorganic fungicide, inorganic mercury fungicide, iodomethane, ipconazole, iprobenfos, iprodione, iprovalicarb, isoprothiolane, isovaledion, 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, methasulfocarb, metofloxam, methyl bromide, methyl isothiocyanate, methylmercuric benzoate, methylmercuric dicyandiamide, methylmercuric pentachlorophenoxide, metiram, metominostrobin, metrafenone, methsulfovax, milneb, morpholine fungicides, myclobutanil, myclozolin, N-(ethylmercury)-p-toluenesulfonanilide, nabam, natamycin, nitro Styrene, nitrotar isopropyl, nuarimol, OCH, octhilinone, ofrace, organomercurial fungicides, organophosphate fungicides, organotin fungicides, orysastrobin, oxadixyl, oxathiin fungicides, oxazole fungicides, auxin copper, oxpoconazole, oxycarboxin, pefurazoate, penconazole, pencycuron, pentachlorophenol, penthiopyrad, phenylmercuric urea, phenylmercuric acetate, phenylmercuric chloride, phenylmercuric derivatives of pyrocatechol, phenylmercuric nitrate, phenylmercuric salicylate, phenylsulfonyl Phamid 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, pyridinitrile, pyri Fenox, 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, tichophen, thifluzamide, thiocarbamate fungicides, thiochlorfenfim, thiomersal, thiophanate, thiophanate methyl, thiophene fungicides, thioquinox, thiram, thiadinil, thi, midoxytibutrclofos methyl, tolnaftate, tolylfluanid, tolylmercuric acetate, triazimef , 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, valinamide fungicides, vinclozolin, zaliramide, zinc naphthenate, zineb, ziram, zoxamide, and mixtures thereof.

[0092] Herbicides are pesticides used to kill unwanted plants. Selective herbicides kill specific targets, leaving desirable crops relatively unharmed. Some of these work by preventing the growth of weeds and are often based on plant hormones. Herbicides used to clear wasteland are non-selective, killing all plant material they come into contact with. Herbicides are widely used in agriculture and landscape management. They are applied in total vegetation control (TVC) programs for highway and railroad maintenance. Smaller amounts are used to manage forests, pasture systems, and areas set aside as wildlife habitat.

[0093] Suitable herbicides are, for example, triazine herbicides such as aryloxycarboxylic acids, e.g. MCPA, aryloxyphenoxypropionates, e.g. clodinafop, cyclohexanedione oximes, e.g. setoxydim, hydroxybenzonitriles, e.g. bromoxynil, sulfonylureas, e.g. nicosulfuron, triazolopyrimidines, e.g. penoxsulam, trikethiones, e.g. mesotrione, metribuzin, hexaxinone or atrazine; sulfonylurea herbicides, e.g. chlorsulfuron; uracils, e.g. lenacil, bromacil, terbacil; urea herbicides, e.g. linuron, diuron, siduron or nebulon; acetanilide herbicides, e.g. alachlor or metolachlor; benoxyl herbicides, e.g. phenylalanine ... herbicides such as thiocarbamate herbicides, such as thiocarb, 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.

[0094] 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.

[0095] Insecticides are agricultural chemicals used against insects in all their forms of development, including ovicides and larvicides used against insect eggs and larvae. Insecticides are used in agriculture, medicine, industry and the home.

[0096] 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, phosalone ... 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 delis (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.

[0097] Acaricides are pesticides that kill mites. Antibiotic acaricides, carbamate acaricides, formamidine acaricides, mite growth regulators, organochlorines, permethrin, and organophosphate acaricides all belong to this category. Molluscicides are pesticides 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 (phylum Nematoda).

[0098] Most preferably, the active substances present in the pesticide formulations or seed coating compositions of the invention are selected from triazole fungicides, strobilurin fungicides, or combinations thereof, in particular tebuconazole, flutriafol, carbendazim, azoxystrobin, crezoxim-methyl, cyproconazole, or pyraclostrobin.

[0099] Nutrients may be present in addition to, or as an alternative to, pesticides. In such formulations / compositions, the nutrients are typically in dry form.

[0100] The nutrient may preferably be a solid-phase nutrient. Solid nutrient should be understood in the present invention as meaning a substance with a melting point above 20°C (standard pressure). Solid nutrient also includes insoluble nutrient components, i.e. nutrient components whose solubility in water is such that there is a significant solid content in the concentrate after addition.

[0101] 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. Nutrients suitable for use in the concentrate according to the invention are all nutritional compounds.

[0102] Micronutrients typically 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 may be included in soluble form or as insoluble solids, and may be salts or chelated.

[0103] Macronutrients typically refer to those containing nitrogen, phosphorus and potassium, including, for example, fertilizers such as ammonium sulfate, and water conditioners. Suitable macronutrients include fertilizers and other nitrogen-, phosphorus-, potassium-, calcium-, magnesium-, or sulfur-containing compounds, and water conditioners.

[0104] 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 fertilizer solids and solutions.

[0105] It is expected that nutrient content will depend on the particular nutrient, with micronutrients typically being included in lower concentrations, while macronutrients are typically included in higher concentrations.

[0106] Biostimulant components can be added to formulations or seed coating compositions to promote the growth of crop plants. The biostimulant components can include or consist of one or more biostimulants.

[0107] Examples of useful biostimulants include, but are not limited to, plant growth hormones and plant growth regulators, such as cytokinins, auxins, gibberellins, ethylene, abscisic acid, etc. Other biostimulants include protein hydrolysate derivatives, seaweed extracts, amino acids, plant extracts, chitosan derivatives, biopolymers, inorganic compounds, humic substances, microbial inocula and microbial products, or mixtures thereof.

[0108] The adjuvants of the invention provide adjuvancy to the pesticide formulation in which they are included and may find particular use in providing fungicide adjuvancy.

[0109] As used herein, the term "adjuvant" or "adjuvancy" refers to a compound that improves the desired effect of a pesticide when added to an agrochemical formulation. Adjuvants can affect the diluent, mixture, active, or target by improving the performance of the active. Adjuvants can be used to attach a pesticide to the area where it functions, to change the epidermal layer of the leaf surface that allows the pesticide to penetrate, and / or to sensitize the target pesticide to the active pesticide in the agrochemical formulation.

[0110] Specific adjuvant effects may include surfactants, emulsifiers (dispersing and suspending agents), oils, emulsifiable oils, compatibilizers, buffering 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 active pesticides.

[0111] 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.

[0112] As a measure of adjuvant activity in relation to the activity of fungicide alone (e.g. pyraclostrobin) against B. cinerea, the percentage inhibition (adjuvant and fungicide) divided by the percentage inhibition (fungicide) can be defined with a higher desired value. Thus, a value of 1 will represent the equivalent activity of the adjuvant / fungicide combination to the fungicide alone, while a value greater than 1 will represent a higher activity of the adjuvant / fungicide combination than the fungicide alone. The active substance of the present invention can have a value greater than 1. Preferably, it has a value greater than 1.5, most preferably greater than 2.

[0113] All features described in this specification can be combined with any of the above aspects in any combination. EXAMPLES

[0114] In order that the invention may be more readily understood, reference is now made, by way of example, to the following descriptions.

[0115] It is understood that all tests and physical properties described herein are measured at atmospheric pressure and room temperature (i.e., 20-25°C) unless otherwise noted herein or in the referenced test methods and procedures.

[0116] Formation and Extraction Bioassay-guided fractionation of culture extracts of the fungus Stachybotrys chartarum RKDO1264 led to the isolation of phenylspirodrimane and stachybocin A. The isolate RKDO1264 was cultured on YM agar medium (w / v: 1% malt extract, 0.2% yeast extract, 1% glucose, 2% agar) and incubated at 22°C for 14 days. Eight colony explants (approximately 3 mm 3 ) was used to inoculate 15 mL of YM broth (w / v: 1% malt extract, 0.2% yeast extract, 1% glucose) in a sterile 50 mL test tube and shaken at 200 RPM at 22° C. for 5 days to generate a seed inoculum. The seed culture was used to inoculate fermentation medium (w / v: 15% sucrose, 2% yeast extract, 0.05% magnesium sulfate heptahydrate, and 1% L-leucine) prepared in an Erlenmeyer flask.

[0117] After 21 days at 22° C., the fermentation was extracted with one 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.

[0118] RKDO1264 fermentation extract was incubated with Teledyne Nextgen 300 +Fractionation was performed on a Silasep C18 flash cartridge (43 g C-18) using a gradient of 10% MeOH:90% HO to 100% MeOH over 20 min in a Combiflash. Fractions were analyzed by 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 phenylspirodrimane and stachybocin A were purified using reversed-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 mass spectrometer (Waters 3100). Initial purification of phenylspirodrimane and stachybocin A was performed using isocratic elution with 60% MeCN in water at a flow rate of 3 mL / min.

[0119] A combination of mass spectrometry and NMR analysis was used to elucidate the structures of phenylspirodrimane and stachybocin A. 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 non-deuterated solvent peaks.

[0120] NMR analysis of the metabolites was consistent with literature data and confirmed the structures as phenylspirodrimane (Ia), phenylspirodrimane (Ib), and stachybosin A (IIa).

[0121] Adjuvant Examples The parameters "percent inhibition" and "fold change" are understood to be expressed and calculated as follows: Inhibition rate - Inhibition rate 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. This is calculated using the following formula:

number

[0122] Fold Change - The fold change is a measure of the adjuvant / bactericide combination in inhibiting the microorganism compared to the bactericide alone. It indicates how the adjuvant performs compared to the bactericide alone. It is calculated using the following formula: INH AF / INH F {In the formula, INH AF = percent inhibition of fungal growth when treated with fungicide and adjuvant, and INH F = Inhibition rate of fungal growth when treated with fungicide alone} It is calculated using

[0123] Example - Adjuvancy Botrytis cinerea (ATCC 90479) was cultured on Difco potato dextrose agar (PDA) for 7 days with a diurnal UV cycle (12 hours UV light and 12 hours dark). Spores were harvested in buffered sterile saline (w / v: 0.9% NaCl with 1% Tween 80) and counted using a hemocytometer. Spore suspensions were cultured at 8.5 × 10 6 A standardized inoculum was prepared by adjusting the final concentration of spores / mL.

[0124] To prepare mycelial fragments for adjuvant testing, 8.5 x 10 4Spores were used to inoculate 10 mL of Difco potato dextrose broth in a 150 x 25 mm 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. After vortexing, the tube was left for 5 minutes to allow large mycelial clumps to settle, and the top layer containing the mycelial fragments was then removed and used as inoculum for the growth inhibition assay.

[0125] Bactericides and adjuvants were dissolved in methanol and added to molten PDA (approximately 50° C.), then the agar was distributed into the wells of a 12-well multiwell plate (1 mL / well). Plates were cooled to room temperature and 10 μL of mycelial inoculum was added to the center of each well. Plates were incubated at 22° C. for 48 hours and colony diameters were measured using digital calipers. Biological growth controls consisted of mycelia and media (0.07% methanol) and negative controls were culture medium and media (0.07% methanol).

[0126] The results of the adjuvant activity of the present invention are shown in Tables 1 to 3. [Table 2]

[0127] [Table 3]

[0128] [Table 4]

[0129] 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, values ​​less than 1 indicating decreased fungicidal activity and values ​​greater than 1 indicating increased fungicidal activity.

[0130] Adjuvant activity with pyraclostrobin against B. cinerea was observed for each of the compounds tested, most notably phenylspirodrimane (Ia) and phenylspirodrimane (Ib), which showed a 33.83- and 17.87-fold increase in fungicidal activity compared to pyraclostrobin alone at 0.1 μg / mL.

[0131] Phenylspirodrimane (Ia) showed a 39.68-fold increase in fungicidal activity compared to tebuconazole alone at 0.05 μg / mL. Phenylspirodrimanes (Ia) and (Ib), as well as stachybocin A (IIa), showed enhanced fungicidal activity compared to dimethomorph, prothioconazole, mancozeb, metalaxyl and pyrimethanil. Phenylspirodrimanes (Ia) and (Ib), and stachybocin A (IIa), did not demonstrate intrinsic fungicidal activity by themselves at any of the concentrations tested.

[0132] Example - Cytotoxicity The cytotoxicity of phenylspirodrimane and stachybocin A was evaluated in vitro against African green monkey Vero kidney cells (ATCC CCL-81). 2 Cells were cultured in 15 mL of Eagle's minimum essential medium (Sigma) supplemented with 0.1 mg / mL streptomycin in cell culture flasks. Cells were incubated for 24 h at 37 °C in a humidified atmosphere of 5% CO2. Culture medium was refreshed every 2-3 days and cells were never allowed to exceed 80% confluence.

[0133] At 80% confluence, cells were counted, diluted, and plated in 96-well flat-bottom cell culture plates (Corning) at a cell density of 10,000 cells per well in 90 μL of growth medium. 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 with a final concentration of 1% in the wells.

[0134] 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, 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.

[0135] Phenylspirodrimanes (Ia) and (Ib), as well as stachybocin A (IIa), 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.

[0136] Example - Phytotoxicity The phytotoxicity of phenylspirodrimane (Ia) was evaluated in soybean and butterhead lettuce leaves. Soybean seeds were inoculated with Bradyrhizobium japnicum and then cultured at 9 cm. 2Soybean plants sown in Levington's M3 compost in plastic pots were given supplemental lighting using SON-T bulbs. Biological control was used to prevent thrips damage (Biolone-Amblyseius cucumeris). Glasshouse temperature was set to maintain 22°C ± 2 during the day and 19°C ± 1 at night. Lettuce plants were grown in the same manner without inoculation with Rhizobacteria. Supplemental lighting was provided by LED bulbs. All plants were kept well hydrated and grown for 14 days.

[0137] Test compounds / extracts were resuspended in 2% DMSO (0.10%, 1.0 g / L). Synperonic A11 LQ was used as a positive control and Atplus UEP-100 was used as a negative control. Treatments were applied as six 10 μL droplets onto the foliage. Phytotoxic tissue damage was assessed by a score of 0 to 3 1 and 7 days after treatment. The results are shown in Table 4.

[0138] [Table 5]

[0139] Phytotoxicity of 1 g / L of compound / extract is expressed as the average phytotoxicity necrosis score, where 0=no necrosis; 1=minor spot-like necrosis at the inoculation site; 2=annular necrosis; 3=expanding necrosis. Phenylspirodrimane (Ia) showed no phytotoxicity to soybean or butterhead lettuce leaves after 7 days. The RKDO1264 crude extract showed very little phytotoxicity to soybean and butterhead lettuce after 7 days (scores of 0.4 and 0.2, respectively).

[0140] It should be understood that the invention should not be limited to the details of the above embodiments, which are given purely by way of example, but that many variations are possible.

Claims

1. below: i) a compound of formula (I): 【Chemistry 1】 {During the ceremony, R 1 are independently hydrogen, C 1 ~C 4 represents alkyl, hydroxy, methoxy, ethoxy, or amine; R 2 are independently hydrogen, C 1 ~C 6 represents alkyl, phenyl, substituted phenyl, alkylphenyl or alkyl-substituted phenyl; R 3 and R 4 are independently hydrogen or C 1 ~C 4 represents alkyl} or The following formula (II): 【Chemistry 2】 {During the ceremony, R 10 are independently hydrogen, C 1 ~C 4 represents alkyl, hydroxy, methoxy, ethoxy, or carboxyl; R 11 , R 12 , R 13 , and R 14 are each independently hydrogen or C 1 ~C 4 an adjuvant selected from phenylspirodrimanes, ii) at least one pesticide active substance; A pesticide formulation comprising:

2. The adjuvant is according to formula (I), R 1 represents hydroxy, methoxy, or amine; R 2 represents n-butyl, isobutyl, phenyl, or methylphenyl; and R 3 and R 4 independently represent hydrogen, methyl, or ethyl; 2. The formulation of claim 1.

3. The adjuvant is according to formula (I), R 1 represents hydroxy; R 2 represents isobutyl or methylphenyl; and R 3 and R 4 each independently represents hydrogen; 2. The formulation of claim 1.

4. The adjuvant is according to formula (II), R 10 represents hydrogen, hydroxy, methoxy, or carboxyl; and R 11 , R 12 , R 13 , and R 14 each independently represents hydrogen, methyl, or ethyl; 2. The formulation of claim 1.

5. The adjuvant is according to formula (II); and R 10 represents carboxyl; and R 11 , R 12 , R 13 , and R 14 each independently represents hydrogen; 2. The formulation of claim 1.

6. The adjuvant has the following formula: 【Chemistry 3】 6. A formulation according to any one of claims 1 to 5, wherein the phenylspirodrimanes are selected from the group consisting of phenylspirodrimanes.

7. 2. A concentrated formulation suitable for producing the pesticide formulation according to claim 1, comprising: i) a compound of formula (I): 【Chemistry 4】 {During the ceremony, R 1 are independently hydrogen, C 1 ~C 4 represents alkyl, hydroxy, methoxy, ethoxy, or amine; R 2 are independently hydrogen, C 1 ~C 6 represents alkyl, phenyl, substituted phenyl, alkylphenyl or alkyl-substituted phenyl; R 3 and R 4 are independently hydrogen or C 1 ~C 4 represents alkyl} or The following formula (II): 【Chemistry 5】 {During the ceremony, R 10 are independently hydrogen, C 1 ~C 4 represents alkyl, hydroxy, methoxy, ethoxy, or carboxyl; R 11 , R 12 , R 13 , and R 14 are each independently hydrogen or C 1 ~C 4 an adjuvant selected from phenylspirodrimanes, ii) at least one pesticide active substance; The concentrated formulation comprising:

8. As adjuvants in pesticide formulations containing at least one pesticide active substance, the following: i) a compound of formula (I): 【Chemistry 6】 {During the ceremony, R 1 are independently hydrogen, C 1 ~C 4 represents alkyl, hydroxy, methoxy, ethoxy, or amine; R 2 are independently hydrogen, C 1 ~C 6 represents alkyl, phenyl, substituted phenyl, alkylphenyl or alkyl-substituted phenyl; R 3 and R 4 are independently hydrogen or C 1 ~C 4 represents alkyl} or The following formula (II): 【Chemistry 7】 {During the ceremony, R 10 are independently hydrogen, C 1 ~C 4 represents alkyl, hydroxy, methoxy, ethoxy, or carboxyl; R 11 , R 12 , R 13 , and R 14 are each independently hydrogen or C 1 ~C 4 The use of a compound selected from phenylspirodrimanes, wherein R represents an alkyl group.

9. 13. A method of treating vegetation to control pests, comprising applying the formulation of claim 1 or the diluted concentrated formulation of claim 7 to either the vegetation or the surrounding environment of the vegetation.

10. A method for obtaining the adjuvant of claim 1, comprising the steps of: Cultivating Stachybotrys chartarum RKDO1264 in a medium under conditions that promote metabolic synthesis of the adjuvant of claim 1 from said Stachybotrys chartarum; and purifying the synthetic adjuvant from the culture medium; The method comprising:

11. An organism consisting of Stachybotrys chartarum, strain RKDO1264, Agricultural Research Service Type Culture Collection (NRRL) accession number NRRL-67988.

12. 2. An extract obtained from an organism consisting of Stachybotrys chartarum, strain RKDO1264, National Agriculture Research Service Type Culture Collection (NRRL) Accession No. NRRL67988, said extract comprising the phenylspirodrimane of claim 1.

13. 13. A method for treating vegetation to control pests, comprising applying either an organism according to claim 11 or an extract according to claim 12 to the vegetation or to the surrounding environment of the vegetation.

14. 12. A seed coating composition comprising the adjuvant of claim 1 or the organism of claim 11.

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