Description:FIELD OF THE INVENTION
The present disclosure relates to the field of agrochemical compositions. The present disclosure in particular relates to fungicidal compositions comprising Kasugamycin, Flusilazole and Azoxystrobin that exhibits antifungal and antibacterial properties. The present disclosure further relates to the process of preparation of said composition and uses thereof.
BACKGROUND OF THE INVENTION
Protection of crops and its produce from insect pest damage is essential for enhancement of agriculture produce. Even with the availability of alternative types of pest control, chemical control remains an important tool for the prevention and control of pests in agriculture, especially insect pests. Fungicides of varied types and structural groups have been reported in literature and a large number are in commercial use. However, judicious use of such chemical pesticides is absolutely needed to protect the environment and natural ecosystems. In this regard, application of fungicidal combinations has emerged as an attractive approach to ensure that the chemical load in the environment is reduced. However, not all chemical fungicidal actives are compatible with each other, and therefore requires a great amount of experimentation and effort to develop an efficacious pesticidal combination, which is stable and effective in controlling the insect pests in economically important Synergistic crops.
In order to achieve the high crop productivity, it is important to control the plant from damages which can be caused by plant fungi, bacteria and other pathogenic (disease-causing) organisms. Although there are available numerous chemical compounds (fungicides), which aid in preventing diseases of plants, each of these have practical deficiencies, which restrict its use or warrants repeated applications.
Further demands on fungicidal compositions and antibacterial compositions include reduced phytotoxicity, reduced dosage, increased uptake of active ingredients for faster control, increased shelf-life and stability of the fungicidal formulation, substantial broadening of spectrum and increased safety, to name a few.
The biological properties of known available agrochemical composition are not entirely satisfactory in the areas of plant pest control, environmental and worker exposure, for example in particular, it has been observed that pathogens become resistant to pesticides which are at times administered in higher dosages to achieve the desired control, thereby leading to soil toxicity and other environmental hazards, besides higher costs.
Moreover, conventional fungicide compositions sometimes do not show a practically sufficient control effect on plant pests depending on an applying situation, since it has an insufficient effect on particular plant pathogenesis either by working slowly against the targeted pathogens or its residual effectiveness lasts for a relatively short period. In view of the above, obtaining a fungicidal combination demonstrates no cross-resistance to the existing fungicidal agents, no toxicity problems and little negative impact on the environment with improved rain fastness and reduced pest resistance, is extremely difficult.
Reference may be made to US patent no. US9538761, wherein it discloses a pesticidal composition comprising sulphur, a fungicide selected from the group consisting of cymoxanil, fenhexamid, fenamidone, cyazofamid, chlorothalonil, kresoxim methyl, azoxystrobin, trifloxystrobin, pyraclostrobin, iprodione, validamycin, kasugamycin, cyprodinil, pencycuron, hexaconazole, prochloraz, epoxiconazole, prothioconaozole, trifloxystrobin, thiophanate methyl, spiroxamine, metrafenone or their salts thereof and at least one agricultural acceptable Additives .
There is therefore a continued need to develop fungicidal composition that provide multiple advantages in an agrochemical composition including - enabling increase in the crop yields, to provide a composition with high synergistic action, to provide a combination where the dosage of the active ingredients is reduced as compared to the individual recommended dosage, to avoid excess loading of the toxicant to the environment and to negligibly impact environmental safety.
Kasugamycin is a compound with CAS registration number 6980-18-3and is a derivative of an amino cyclitol glycoside that is isolated from Streptomyces kasugaensis. Kasugamycinexhibits antibiotic and fungicidal properties. It has a role as a bacterial metabolite, a protein synthesis inhibitor and an antifungal agrochemical. It is an amino cyclitol glycoside, an aminoglycoside antibiotic, a monosaccharide derivative, a carboxamidine and an antibiotic Fungicide. Kasugamycin is quite valued both at a domestic and at the industrial level. Unfortunately, no single bactericide and fungicide is useful in all situations and repeated usage of a single bactericide or fungicide such as kasugamycin frequently leads to the development of resistance in phytopathogens towards kasugamycin and its related bactericide and fungicide compounds. Accordingly, significant efforts have been put forward towards finding new and improved combinations of bactericide, fungicide and other related material of natural or synthetic origin that are safer, have better performance, require lower dosages, easier to use and are cost effective. Combinations have also been studied that produce synergism, i.e., the activity of two, or more, agents exceed the activities of the agents when used alone. However, none of the current approaches or reports seem to satisfy the existing needs.
Kasugamycinis well known with IUPAC name is 2-amino-2-[(2R,3S,5S,6R)-5-amino-2-methyl-6-[(2R,3S,5S,6S)-2,3,4,5,6-pentahydro- xycyclohexyl]oxyoxan-3-yl]iminoaceticacid.and has following structure

KASUGAMYCIN

Flusilazole is an organosilicon compound with CAS registration number 85509-19-9.This compound has dimethyl silane in which the hydrogens attached to the silicon are replaced by p-fluorophenyl groups and a hydrogen attached to one of the methyl groups is replaced by a 1H-1,2,4-triazol-1-yl group.
It is a broad-spectrum fungicide used to protect a variety of crops. It has a role as a xenobiotic, an environmental contaminant, an EC 1.14.13.70 (sterol 14alpha-demethylase) inhibitor and is an antifungal agrochemical. It is a member of monofluorobenzenes, a member of triazoles, an organosilicon compound, a conazole fungicide and a triazole fungicide.
Flusilazole is an organosilicon fungicide, which is used to control fungal infections on a variety of fruit and vegetable crops. It is moderately toxic to animals and has been shown to produce birth defects and embryotoxicity at high doses. It is systemic fungicide with protective and curative action. Its resistance to wash-off, re-distribution by rainfall and vapour phase activity are important components in its biological activity.
Flusilazole has a IUPAC name -1-[[bis(4-fluorophenyl)(methyl)silyl]methyl]-1H-1,2,4-and has following structure:

FLUSILAZOLE

Azoxystrobin is an aryloxy pyrimidine that has a CAS no131860-33-8.Azoxystrobin has a 4,6-diphenoxypyrimidine skeleton in which one of the phenyl rings is cyano-substituted at C-2 and the other carries a 2-methoxy-1-(methoxycarbonyl) vinyl substituent, also at C-2. An inhibitor of mitochondrial respiration by blocking electron transfer between cytochromes b and c1, it is used widely as a fungicide in agriculture.
Azoxystrobin has a role as a mitochondrial cytochrome-bc1 complex inhibitor, a xenobiotic, an environmental contaminant, an antifungal agrochemical and a quinone outside inhibitor. It is a nitrile, an aryloxy pyrimidine, an enol ether, a methyl ester and a methoxy acrylate strobilurin antifungal agent.
Azoxystrobin has a IUPAC name as- methyl (E)-2-[2-[6-(2-cyanophenoxy)pyrimidin-4-yl]oxyphenyl]-3-methoxyprop -2-enoate.and has following structure:

AZOXYSTROBIN

It has been found that, as a solution to the above mentioned problems, a combination comprising Kasugamycin, Flusilazole and Azoxystrobin provides an effective composition in controlling a wide variety of undesired pathogenic microorganisms. The present invention provides a combination of Kasugamycin, Flusilazole and Azoxystrobin with enhanced efficacy and spectrum as compared to the use of Kasugamycin, Flusilazole and Azoxystrobin alone.

OBJECTS OF THE INVENTION
It is an object of the present invention to provide a novel, stable, synergistic, composition of Kasugamycin, Flusilazole and Azoxystrobin demonstrating high efficacy and high selectivity.
It is another object of the present invention to provide a novel and effective fungicidal composition for controlling the harmful pests and fungi in plants.
It is another object of the present invention to provide a novel and effective synergistic fungicidal composition which can be easily formulated.
It is yet another object of the present invention to provide the fungicidal composition with enhanced bioefficacy at reduced dosage as compared to individual insecticides when used in isolation.
It is yet another object of the present invention to provide the fungicidal composition that result in good enhancement in crop yield.
It is yet another object of the present invention to provide the fungicidal composition that is environmentally safe and non-phytotoxic.
It is yet another object of the present invention to provide fungicidal composition that has broad spectrum activity and can be used in a wide variety of Synergistic crops.

SUMMARY OF THE INVENTION
In an aspect, Synergistic composition, comprising 0.1% to about 30.0% Kasugamycin, by weight of the composition about 0.1% to about 30.0% flusilazole by weight of the composition about 0.1% to about 30.0% Azoxystrobin by weight of the composition; and an agricultural accepted additive(s)
In another aspect, the present invention provides a combination of Kasugamycin, Flusilazole and Azoxystrobin to control a wide variety of undesired pathogenic microorganisms.
In yet another aspect, the present invention provides a synergistic composition comprising Kasugamycin, Flusilazole and Azoxystrobin, the composition possesses fungicidal activity.
In a further aspect, the present invention provides a method for effective control of various fungi in plants.
In another aspect, the agricultural accepted additive(s) are selected from the group comprising an emulsifier; dispersing agent; w; antifoaming agent; rheology modifier; solvent; biocide; and an antifreezing agent; wherein the composition has about 0.1% to about 20.0% emulsifier; about 0.1% to about 20.0% dispersing agent; about 0.1% to about 15.0% ;antifoaming agent; about 0.01% to about 12.0% rheology modifier; about 1.0% to about 95.0% solvent; about 0% to 3 % biocide; and about 0% to 15 % antifreezing agent;
In another aspect,the emulsifier is selected from the group comprising ethoxylated propoxylated alcohols, alkylphenolethoxylates, alkoxylated tristyrylphenols, ethoxylated propoxylatedpolyaryl phenol, ethoxylated fatty acids, Fatty alcohol ethoxylates, ethoxylatedricinoleic acid triglycerides, sorbitan trioleate, tridecyl alcohol ethoxylate, castor oil ethoxylate, alkoxylated phosphate ester or mixtures thereof.
In another aspect,the dispersing agent is selected from the group comprising naphthalene formaldehyde condensates, acrylic copolymer, nonionic proprietary surfactant blend, polycarboxylates, calcium dodecylbenzene sulfonate, polystyrenated acrylated co-polymer, salts of phenol sulfonic acids, random co-polymer of alcoxylated polyethylene glycol or mixtures thereof.
In another aspect, the antifoaming agent is selected from the group comprising perfluroalkylphosphonic acids, polydimethylsiloxane, polydimethyl siloxane emulsion or mixtures thereof.
In another aspect, the rheology modifier is selected from the group comprising precipitated silica, modified fumed silica, bentonite, hydroxymethyl cellulose, carboxymethyl cellulose, xanthan gum, thickening silica, hydrated clay minerals, magnesium aluminium silicates, organic derivative of hectorite clay, hydrophobic fumed silica, polyvinylpyrrolidone (PVP) or mixture thereof.
In another aspect,the solvent is selected from the group comprising Demineralized (DM) water, N-alkyl-pyrrolidone, oil medium selected from the group comprising, esterified fatty acids selected from methyl ester of triglycerides containing C12–C22 saturated and unsaturated fatty acids, ethyl ester of triglycerides containing C12 –C22 saturated and unsaturated fatty acids such as methyl soyate, ethyl soyate, rapeseed methyl ester, rapeseed ethyl ester or mixtures thereof.
In another aspect, the biocide is selected from the group comprising 1,2-benzisothiazolin-3-one, formaldehyde, dipropyl glycol solution of 1,2-benzisothiazolin-3-one or mixtures thereof.
In another aspect, the antifreezing agent is selected from the group comprising ethylene glycol, 1,2-propanediol, propane-1,2,3-triol, urea or mixtures thereof.
In another aspect, the coloring agent is selected from dye and pigment.
In yet another embodiment of the present invention, the invention further provides the process for preparation of said compositionthat can be one or more of capsule suspension (CS), Dispersible concentrate (DC), Dustable powder (DP), Powder for dry seed treatment (DS), Emulsifiable concentrate (EC), Emulsifiable granule (EG) Emulsifiable water-in-oil (EO), Emulsifiable powder (EP), Emulsifiable for seed treatment (ES), Emulsifiable oil-in-water (EW), flowable concentrate for seed treatment (FS), Suspension Concentrate (SC), Suspo-emulsion (SE), Water dispersible powder for slurry seed treatment (WS), Water dispersible granules (WDG) oil Dispersible (OD) and wettable powders (WP), a mixed formulation of CS and SC (ZC), soluble liquid (SL).
Various objects, features, aspects and advantages of the inventive subject matter will become more apparent from the following detailed description of preferred embodiments

DETAILED DESCRIPTION OF THE INVENTION
Those skilled in the art will be aware that the invention described herein is subject to variations and modifications other than those specifically described. It is to be understood that the invention described herein includes all such variations and modifications. The invention also includes all such steps, features, composition, formulation and compounds referred to or indicated in this specification, individually or collectively, and all combinations of any two or more of said steps or features. Unless otherwise specified, all terms used in disclosing the invention, including technical and scientific terms, have the meaning as commonly understood by one of the ordinary skills in the art to which the invention belongs. For further guidance, term definitions may be included to better appreciate the teaching of the present invention.
As used herein, the term “plant” refers to any plant or part thereof including serial and subterranean parts of the plant. It is contemplated that the parts of the plant may be for example, flowers, fruits or vegetables, shoots, leaves, needles, stalks, stems, fruiting bodies, seeds also roots and that parts of the plants may or may not be attached to the remainder of the plant.
The term “locus” of a plant as used herein is intended to embrace the place on which the plants are growing, where the plant propagation materials of the plants are sown or where the plant propagation materials of the plants will be placed into the soil. The term “plant propagation material” is understood to denote generative parts of a plant, such as seeds, vegetative material such as cuttings or tubers, roots, fruits, tubers, bulbs, rhizomes and parts of plants, germinated plants and young plants which are to be transplanted after germination or after emergence from the soil.
It is to be understood that the singular forms “a,” “an,” and “the” include plural referents unless the context clearly dictates otherwise. It should be emphasized that the term “comprises/comprising” when used in this specification is taken to specify the presence of stated features, steps or components but does not preclude the presence or addition of one or more other features, steps, components or groups thereof. The term "insects" as used herein, includes all organisms in the class "Insecta." The term "Fungicidal" as used herein, refers to the ability of a pesticide to increase mortality or inhibit growth rate of insects.
The terms “weight percent”, “wt%”, “percent by weight”, “% by weight” and variations thereof, as used herein, refer to the concentration of a substance as the weight of that substance divided by the total weight of the formulation and multiplied by 100. It is understood that, as used here, “percent “, “%” and the like are intended to be synonymous with “weight percent”, “wt. %”, etc.
The term “synergist” as used herein refers to a material which enhances the activity of other materials such as insecticides so that the overall activity of the formulation is greater than the sum of the individual ingredients. Features that are described and/or illustrated with respect to one embodiment may be used in the same way or in a similar way in one or more other embodiments and/or in combination with or instead of the features of the other embodiments.
Unless the context requires otherwise, throughout the specification which follow, the word “comprise” and variations thereof, such as, “comprises” and “comprising” are to be construed in an open, inclusive sense that is as “including, but not limited to.”
Reference throughout this specification to “one embodiment” or “an embodiment” means that a particular feature, structure or characteristic described in connection with the embodiment is included in at least one embodiment. Thus, the appearances of the phrases “in one embodiment” or “in an embodiment” in various places throughout this specification are not necessarily all referring to the same embodiment. Furthermore, the particular features, structures, or characteristics may be combined in any suitable manner in one or more embodiments.
In some embodiments, the numbers expressing quantities of ingredients, properties such as concentration, and so forth, used to describe and claim certain embodiments of the invention are to be understood as being modified in some instances by the term “about.” Accordingly, in some embodiments, the numerical parameters set forth in the written description are approximations that can vary depending upon the desired properties sought to be obtained by a particular embodiment. In some embodiments, the numerical parameters should be construed in light of the number of reported significant digits and by applying ordinary rounding techniques. Notwithstanding that the numerical ranges and parameters setting forth the broad scope of some embodiments of the invention are approximations, the numerical values set forth in the specific examples are reported as precisely as practicable.
The recitation of ranges of values herein is merely intended to serve as a shorthand method of referring individually to each separate value falling within the range. Unless otherwise indicated herein, each individual value is incorporated into the specification as if it were individually recited herein.
Various terms as used herein are shown below. To the extent a term used in a claim is not defined below, it should be given the broadest definition persons in the pertinent art have given that term as reflected in printed publications and issued patents at the time of filing.
As used herein, the term “additive(s)” or "auxiliary agent(s)" or “agricultural acceptable carrier(s)” can be used interchangeably and refers to inert substances which are commonly used asdiluent, to provide stability or to increase the activity profile of the composition or formulation with or without having agrochemical activity or direct effect on the undesired phytopathogenic fungi and/or microorganisms.
As used herein, the term "agrochemically acceptable salts" are typically acid addition salts of inorganic or organic acids, preferably of hydrochloric acid, hydrobromic acid, sulfuric acid, nitric acid, perchloric acid, phosphoric acid, formic acid, acetic acid, trifluoroacetic acid, oxalic acid, malonic acid, toluenesulfonic acid or benzoic acid.
As used herein, the term “additive(s)” or "auxiliary agent(s)" or “Synergistically acceptable carrier(s)”can be used interchangeably and refers to inert substances which are commonly used as diluent, to provide stability or to increase the activity profile of the composition or formulation with or without having agrochemical activity.
As used herein, the term “lower” or “minimal” or “minimum” can be used interchangeably and refers to the optimum concentration of active or inactive ingredients of formulation to achieve the expected efficacy.
As used herein, the term "effective amount" means the amount of the active substances in the compositions to achieve an observable effect on growth, including the effects of necrosis, death retardation, prevention, and removal, destruction, or otherwise diminishing the occurrence and activity of the target organism. The effective amount can vary for the various compositions used in the present invention. An effective amount of the compositions will also vary according to the prevailing conditions such as desired pesticidal effect and duration, weather, target species, locus, mode of application, and the like.
As used herein, the term “composition” or "formulation" can be used interchangeably, unless stated otherwise, is meant to encompass, and are not limited to, compositions or formulations containing the combination of Kasugamycin, Flusilazole and Azoxystrobin.
The present disclosure relates to synergistic composition. Particularly, the present disclosure provides the fungicidal composition including Kasugamycin, Flusilazole; Azoxystrobin and Synergistically acceptable excipient(s) that exhibits antifungal and antibacterial properties.
Technical effect:
The present disclosure is based on the premise of an unexpected finding by the applicant that composition including Kasugamycin, Flusilazole and Azoxystrobin and agricultural acceptable additives (s) that aids in reducing dosage of the overall compositions and formulation and consequently aids in reducing or mitigating phytotoxicity.
Bio efficiency and phytotoxicity results are well elaborated and provided in examples later.
The present invention thus provides synergistic composition comprising Kasugamycin, as the first active ingredient; Flusilazole as the second active ingredient; and Azoxystrobin as the third active ingredients and agricultural acceptable additive(s). In the composition-Kasugamycin, is present in an amount ranging from 0.1-30% w/w, Flusilazole is present in an amount ranging from 0.1-30% % w/w and Azoxystrobin is present in an amount ranging from 0.1-30% w/w and agricultural acceptable additives(s)
In another aspect, the agricultural accepted additives are selected from the group comprising an comprising an emulsifier; dispersing agent; w; antifoaming agent; rheology modifier; solvent; biocide; and an antifreezing agent; wherein the composition has about 0.1% to about 20.0% emulsifier; about 0.1% to about 20.0% dispersing agent; about 0.1% to about 15.0% ;antifoaming agent; about 0.01% to about 12.0% rheology modifier; about 1.0% to about 95.0% solvent; about 0% to 3 % biocide; and about 0% to 15 % antifreezing agent;
In another aspect, the emulsifier is selected from the group comprising ethoxylated propoxylated alcohols, alkylphenolethoxylates, alkoxylatedtristyrylphenols, ethoxylated propoxylatedpolyaryl phenol, ethoxylated fatty acids, Ethoxylated Tristyrylphenol , Soprophor 3D33,Fatty alcohol ethoxylates, ethoxylatedricinoleic acid triglycerides, sorbitan trioleate, tridecyl alcohol ethoxylate, castor oil ethoxylate, alkoxylated phosphate ester or mixtures thereof.
In another aspect, the dispersing agent is selected from the group comprising naphthalene formaldehyde condensates, acrylic copolymer, nonionic proprietary surfactant blend, polycarboxylates, calcium dodecylbenzene sulfonate, polystyrenatedacrylated co-polymer, Atlox 4917, salts of phenol sulfonic acids, Soprophor 3D33, Butyl polyalkylene oxide block co-polymer ,random co-polymer of alcoxylated polyethylene glycol or mixtures thereof.
In another aspect, the antifoaming agent is selected from the group comprising perfluroalkylphosphonic acids, polydimethylsiloxane, polydimethylsiloxane emulsion or mixtures thereof.
In another aspect, the rheology modifier is selected from the group comprising precipitated silica, modified fumed silica, bentonite, hydroxymethyl cellulose, carboxymethyl cellulose, xanthan gum, thickening silica, hydrated clay minerals, magnesium aluminium silicates, organic derivative of hectorite clay, hydrophobic fumed silica, polyvinylpyrrolidone (PVP) or mixture thereof.
In another aspect, the solvent is selected from the group comprising Demineralized (DM) water, N-alkyl-pyrrolidone, oil medium selected from the group comprising, esterified fatty acids selected from methyl ester of triglycerides containing C12–C22 saturated and unsaturated fatty acids, ethyl ester of triglycerides containing C12 –C22 saturated and unsaturated fatty acids such as methyl soyate, ethyl soyate, rapeseed methyl ester, rapeseed ethyl ester or mixtures thereof.
In another aspect, the biocide is selected from the group comprising 1,2-benzisothiazolin-3-one, formaldehyde, dipropyl glycol solution of 1,2-benzisothiazolin-3-one or mixtures thereof.
In another aspect, the antifreezing agent is selected from the group comprising ethylene glycol, 1,2-propanediol, propane-1,2,3-triol, urea or mixtures thereof.
In another aspect,the stabilizing agents is selected from the group polysaccharides, carboxymethyl cellulose, Precipitated Silica bentonite clay, aluminum magnesium silicate, citric acid either alone or mixtures thereof.
In an embodiment, the fungicidal composition is formulated in a form selected from the group comprising water-soluble concentrates (SL), emulsifiable concentrates (EC), emulsions (EW), micro-emulsions (ME), Suspension concentrates (SC), oil-based suspension concentrates (OD), flowable suspensions (FS), water-dispersible granules (WG), water-soluble granules (SG), wettable powders (WP), water soluble powders (SP), granules (GR), encapsulated granules (CG), fine granules (FG), macrogranules (GG), dry flowables (DF), aqueous Suspo-emulsions (SE), capsule suspensions (CS) and micro granules (MG). In a preferred embodiment, the fungicidal formulation is in the form of suspension concentrates (SC), water-dispersible granule (WDG) and wettable powder (WP).
In yet another embodiment, the combination or the composition of the present invention is effective for management of fungi or pests selected from one or more of. wheat, rye, barley, triticale, oats or rice; beet, e.g. sugar beet or fodder beet; fruits, such as pomes, stone fruits or soft fruits, e.g. apples, pears, plums, peaches, almonds, cherries, strawberries, raspberries, blackberries or gooseberries; leguminous plants, such as lentils, peas, alfalfa or soybeans; oil plants, such as rape, mustard, olives, sunflowers, coconut, cocoa beans, castor oil plants, oil palms, ground nuts or soybeans; cucurbits, such as squashes, cucumber or melons; fiber plants, such as cotton, flax, hemp or jute; citrus fruit, such as oranges, lemons, grapefruits or mandarins; vegetables, such as spinach, lettuce, asparagus, cabbages, carrots, onions, tomatoes, potatoes, cucurbits or paprika; lauraceous plants, such as avocados, cinnamon or camphor; energy and raw material plants, such as corn, soybean, rape, sugar cane or oil palm; corn; tobacco; nuts; coffee; tea; bananas; vines (table grapes and grape juice grape vines); hop; turf; sweet leaf (also called Stevia); natural rubber plants or ornamental and forestry plants, such as flowers, shrubs, broadleaved trees or evergreens, e.g. conifers; and on the plant propagation material, such as seeds, and the crop material of these plants.
In yet another preferred embodiment, the present invention provides a fungicidal combination or composition comprising Kasugamycin, Flusilazole, Azoxystrobin to control the pathogenic microorganism on economically important crops such as rice, chilli, apple, peppers, soybean, cotton, chick pea, pigeon pea, Grapes, Apple and pomegranate, tea, potato, and tomato.
In another embodiment of the present invention, the invention further provides the process for preparation of the said composition wherein, the said composition can be one or more of as suspension concentrate (SC), wettable granules (WG), wettable powder (WP), a water dispersible granule (WDG), a water dispersible tablet (WT), an ultra-low volu (ULV) liquid (UL), an ultra-low volume (ULV) suspension (SU), a water soluble powder (SP), a suspo-emulsion (SE), granule (GR), an emulsifiable granule (EG), an oil-in-water or water in oil emulsion (EW), an emulsifiable concentrate (EC), a micro-emulsion (ME), an oil dispersion (OD), a capsule suspension (CS), a dustable powder (DP) or an aerosol (AE).
In preferred embodiment of the present invention, the formulation is Suspension Concentrate (SC), Suspo-emulsion (SE), oil dispersion (OD), Water dispersible granules (WDG), Granules (GR) and Wettable powders (WP).
In another preferred embodiment of the invention, the yield of the plants treated according to the method of the invention, is increased synergistically. According to the present invention, "increased yield" of a plant, in particular of anSynergistic, silvicultural and/or horticultural plant means that the yield of a product of the respective plant is increased by a measurable amount over the yield of the same product of the plant produced under the same conditions, but without the application of the mixture according to the invention.
Increase in yield of treated plants means increased plant weight, increased plant height, increased biomass such as higher overall fresh weight (FW), increased number of flowers per plant, higher grain yield, more tillers or side shoots (branches), larger leaves, increased shoot growth, increased protein content, increased oil content, increased starch content, increased pigment content, Increase in yield due to protection against Insect-pest damage and fungal diseases, Increase in yield due to plant growth regulation, check vegetative growth and increase in reproductive parts of plant, Increase in yield due to more number of tillers, more branches and sub branches, more number of fruits, flowers, and grains size, Increase plant vigor, Increase tolerance to insect-pests and fungal damage, Increase tolerance to the weather stress, moisture stress and heat stress, Prevents lodging in susceptible plants (lodging due to biotic and abiotic factors, like heavy rains, winds, insects and diseases damage.
While the foregoing description discloses various embodiments of the disclosure, other and further embodiments of the invention may be devised without departing from the basic scope of the disclosure. The invention is not limited to the described embodiments, versions or examples, which are included to enable a person having ordinary skill in the art to make and use the invention when combined with information and knowledge available to the person having ordinary skill in the art.
Examples-:
The examples below are given solely for the purpose of illustration and are not to be construed as limitations of the present invention, as many variations thereof are possible without departing from the spirit and scope of the invention.
Example 1: Preparation of Fungicidal composition as Suspo-emulsion (SE)formulation:
The chemical composition of the present Fungicidal composition is depicted below in Table 1 below:
Table-1: Fungicidal Composition of the Suspo-emulsion (SE)formulation
Component FORMULATION (%w/w) Remark
Kasugamycin 2.8 Active Ingredient
Flusilazole 8.4 Active Ingredient
Azoxystrobin 8.8 Active Ingredient
Polyalkoxylated butyl ether block co-polymer 2.00 Emulsifier
Ethoxylated Tristyrylphenol 2.00 Emulsifier
Soprophor 3D33 4.00 Dispersing agent
Atlox 4917 2.00 Dispersing agent
Butyl polyalkylene oxide block co-polymer 1.00 Dispersing agent
1,2-propanediol 6.00 Antifreezing Agent
Dipropylene glycol solution of 1,2-benzisothiazolin-3-one 0.10 Biocide
Polydimethylsiloxane emulsion 0.30 Antifoaming agent
Precipitated Silica 2.00 Stabilizing agent
Carboxy methyl cellulose 0.7 Stabilizing agent
Xanthum gum (2 % w/w aqueous of Rhodopol 23) 3.00 Rheology modifier
Solvent C-9 10.0 Solvent
Demineralized Water Balance to 100 Solvent
Process for preparing Suspo-emulsion (SE)formulation:
Preparation of Flusilazole oil phase
The active ingredient Flusilazole was dissolved inasolvent-C9 at room temperature. To this solution an emulsifier Polyalkoxylated butyl ether block co-polymer and Ethoxylated Tristyrylphenol were added with continuousstirringto form Flusilazole oil phase. This was first part of the process.
Preparation of aqueous phase of Kasugamycin and Azoxystrobin:
In demineralizedwater constituents namely dipropylene glycol solution of 1,2-benzisothiazolin-3-one, polydimethylsiloxane emulsion, 1,2-propanediol, Atlox 4917, Soprophor 3D33 and Butyl polyalkylene oxide block co-polymer were added into the clean pre-mixing vessel fitted with the homogenizer and all the ingredients were mixed for sufficient time with the homogenizer to obtain a mixture.
Following the above, the active ingredients, kasugamycin, azoxystrobin along with precipitated silica were added to the obtained mixture and continued to mix for sufficient time with the homogenizer to obtain a slurry.
The obtained slurry was passed through jacketed bead mill with chilled water circulation for particle size reduction to obtain a aqueous phase of Kasugamycin and Azoxystrobin of desirable particle size of d(90)<20 micron. The aqueous phase of Kasugamycin and Azoxystrobin was collected into mixing vessel fitted with the stirrer.
Preparation of Suspo-emulsion (SE) formulation:
The obtained Flusilazole oil phase was added to the obtained aqueous phase of Kasugamycin and Azoxystrobin under a continuous agitation for 2 hrs. at 10-30 deg C temperature.
Then a suitable quantity of rheology modifiers Carboxy Methyl Cellulose and 2 % Xanthun gum was added to the mixture. The obtained mixture was continuously for 1 hr. to form Suspo-emulsion (SE) formulation.
Example 2: Preparation of Fungicidal composition as Suspo-emulsion (SE)formulation:
The chemical composition of the present Fungicidal composition is depicted below in Table 2 below:
Table-2: Fungicidal Composition of the Suspo-emulsion (SE) composition
Component FORMULATION (%w/w) Remark
Kasugamycin 2.8 Active Ingredient
Flusilazole 8.4 Active Ingredient
Azoxystrobin 8.8 Active Ingredient
Ethoxylated Tristyrylphenol 1.00 Emulsifier
Soprophor 3D33 1.00 Emulsifier
Fatty Alcohol Ethoxylate 0.50 Emulsifier
Sorbitan trioleate 0.50 Emulsifier
Nonionic proprietary surfactant blend 1.00 Dispersing agent
Acrylic copolymer 2.00 Dispersing agent
Propane-1,2,3-triol 5.00 Antifreezing Agent
Dipropylene glycol solution of 1,2-benzisothiazolin-3-one 0.30 Biocide
Polydimethylsiloxane emulsion 0.20 Antifoaming agent
Xanthum gum (2 % w/w aqueous of Rhodopol 23) 10.0 Rheology modifier
Solvent C-9 10.0 Solvent
Demineralized Water Balance to 100 Solvent

Process for preparing Suspo-emulsion (SE) composition:
Preparation of Kasugamycin aqueous phase
The active ingredient Kasugamycin was dissolved in appropriate amount of demineralized waterat room temperature. To this obtained solution glycerol was added undercontinuous stirring to form Kasugamycin aqueous phase.
Preparation of Flusilazole oil phase
The active ingredient Flusilazole was dissolved in solvent-C9 at room temperature. To this solution an emulsifier Sorbitan trioleate, Fatty Alcohol Ethoxylate, Ethoxylated Tristyrylphenoland Soprophor 3D33 were added under stirring to form Flusilazole oil phase.
Preparation of Kasugamycin and Flusilazole emulsion in water phase
Kasugamycin aqueous phase was added in Flusilazole oil phase with appropriate rate under stirring to form Kasugamycin and Flusilazole emulsion in water phase.
Preparation of aqueous phase of Azoxystrobin:
In demineralized water constituents namely dipropylene glycol solution of 1,2-benzisothiazolin-3-one, polydimethylsiloxane emulsion, Propane-1,2,3-triol, Nonionic proprietary surfactant blend and Acryliccopolymer as mentioned in Table 2 were added into the clean pre-mixing vessel fitted with the homogenizer and all the ingredients were mixed for sufficient time with the homogenizer to obtain a mixture.
The active ingredient azoxystrobin was added to the obtained mixture and continued to mix for sufficient time with the homogenizer to obtain slurry. The slurry was passed through jacketed bead mill with chilled water circulation for particle size reduction to obtain a aqueous phase of Azoxystrobin of desirable particle size of d(90)<20 micron. The aqueous phase of Azoxystrobin was collected into a mixing vessel fitted with the stirrer.
Preparation of Suspo-emulsion (SE)formulation:
The obtained Kasugamycin and Flusilazole emulsion in water phase was added to the aqueous phase of Azoxystrobin in under a continuous agitation for an optimum amount of time at room temperature. Then a suitable quantity of rheology modifier 2 % Xanthum gum was added to the mixture. The obtained mixture was continuously agitated until the appropriate time to form Suspo-emulsion (SE) formulation.
Stability Data
Stability Study (Accelerated Storage test and low-temperature storage test)
According to the FAO/WHO manual, the “accelerated storage test” is considered as an indicative of product stability. That is, accelerated storage test data provides an indication that the product is stable for at least two years at ambient temperature. Further, the FAO/WHO manual indicates storage at 54 ± 2°C for 14 days as the default test conditions. Further, the FAO/WHO manual indicates low-temperature storage at 0 ± 2°C for 7 days.
The “ambient” is the sample at the room temperature which is before subjecting to the accelerated storage test.
The “accelerated storage” is the sample after subjecting the sample to accelerated storage tests at 54±2°C for 14 days.
Table 3: Stability data for Suspo-emulsion (SE) formulation
Sr. no. Test Unit of measurement Results
14 days Ambient 14 days Accelerated storage at 54 °C
1 Appearance -- Off white to creamliquid free from extraneous matter Off white to cream liquid free from extraneous matter
2 Active content % w/w
Kasugamycin 3.02 2.75
Flusilazole 9.22 9.14
Azoxystrobin 9.16 8.91
3 Dispersion stabilityinCIPAC standard water A and D at 23 ± 2°C. % w/w 95.2 95.1
4 pH -- 5.20 5.10
5 Viscosity cPs 653 620
6 Wet Sieve retained on 75µ test sieve % w/w 0.08 0.10
7 Persistent Foam ml 10 12
8 Pourability (as residue) % w/w 4.6 4.4
9 Stability at 0°Cfor 7 days -- After storage at 0 ± 2°C for 7 days, the formulation complies the general criteria of dispersion stability and wet sieve test of formulation

The fungicidal composition of Examples 1 - 2 were found stable after accelerated stability study at 54 ± 2° C for 14 days and low temperature at 0 ± 2°C for 7 days.

Example-3: Evaluation of Bio-efficacy & Phytotoxicity of Fungicidal composition and thereof (Kasugamycin, Flusilazole and Azoxystrobin) against Powdery mildew (Leveillulataurica) and Anthracnose (Colletotrichumcapsici) disease on Chilli crop.
Chilli (Capsicum annum L.) is an important spice as well as a vegetable crop grown globally. Pungent forms are used as green chilli, dry chilli, chilli powder, chilli paste, chilli sauce, chilli oleoresin, or in mixed forms. Dried fruits are extensively used as a spice. Disease infestation is one of the major factors for yield loss under irrigated conditions. Chilli is usually planted at wider spacing and grows very slowly during the fruiting stage. Diseases that emerge and become established during the fruiting stages of chilli growth can be very competitive and significantly reduce the chilli yield potential. Powdery mildew (Leveillulataurica) and anthracnose (Colletotrichumcapsici)are economically important disease affecting chilli crop.
Field evaluation of the bio - efficacy of present fungicidal composition:
The presently disclosed fungicidal (Kasugamycin and Flusilazole and Azoxystrobin) composition were tested for its bio-efficacy against Powdery mildew(Leveillulataurica)and anthracnose (Colletotrichumcapsici) diseases on chilli crop, trial was laid out in randomized block design consisting of eight treatments including control in three replications.
Measured quantity of the chemical was added to required volume of water @ 500 lit. /ha was added for foliar spray. The spray tank was filled with ½ the quantity of clean required volume of water and then the measured chemical (according to the dose) was added followed by the rest half quantity of water was added. Knapsack sprayer fitted with boom along with flood jet nozzle was used to apply the spray of fungicidal composition.

Synergistic effect of the different Fungicidal Composition:
Synergistic effect was checked using Colby’s method for ternary mixes. In the Colby’s method, for a given combination of three active components, E (expected efficacy) can be expressed as:
E = A+B+C – (AB+AC+BC) + (ABC)
100 1000

Where, E = expected efficacy,
A, B and C = the efficacy of three active ingredients A, B and C at a given dose.
Synergy ratio (R) = Experimentally observed efficacy (O)
Expected efficacy (E)

If the synergism ratio (R) between observed and expected is > 1 then synergy is exhibited, if R = 1 then the effect is additive and if R<1 then the mix is antagonistic.
The experimental data was statistically analysed by Randomized Block Design (RBD) (One factor analysis) using OPSTAT HAU statistical software. The results are expressed as Mean +SE (standard error) and data was statistically analysed by one-way analysis of variance (ANOVA), with the level of significance set at p < 0.01.
Evaluation of Bio-efficacy against Powdery mildew and Anthracnose disease in Chilli-
Disease rating scale:
Table.4 - Disease rating scale for Powdery mildew disease in Chilli:
S. No Rating Scale Per cent infected leaf area (%)
1 0 0
2 1 1 %
3 3 1 -10 %
4 5 11-25 %
5 7 26-50 %
6 9 51-75 %

Table.5 - Disease rating scale for Anthracnose disease in Chilli:
S. No Rating Scale Per cent fruit area covered (%)
1 0 0
2 1 Slightly -10
3 3 11-25 %
4 5 26-50 %
5 7 51-75 %
6 9 >75 %

The per cent disease index (PDI) for above mentioned diseases was calculated according to the following formula:
Sum of all numerical rating
Per cent disease index = ×100
Total No. of Leaves/Fruits × Maximum Rating Grade

Phytotoxicity observations:
Phytotoxicity observations were recorded from all the treatments on 0-10 scale for Epinasty, Hyponasty, Chlorosis, Necrosis, vein clearing and stunting etc. after 3, 5, 7 and 10 days of treatments.
Table. 6 - Description of Phytotoxicity scale (0-10):
Rating Crop Injury (%) Description
0 - No symptoms
1 1-10 Slight discoloration
2 11-20 More severe, but not lasting
3 21-30 Moderate and more lasting
4 31-40 Medium and lasting
5 41-50 Moderately heavy
6 51-60 Heavy
7 61-70 Very heavy
8 71-80 Nearly destroyed
9 81-90 Destroyed
10 91-100 Completely destroyed

Table 7. Evaluation of bio-efficacy of Kasugamycin 2.4 % and Flusilazole 7.5 % and Azoxystrobin 8% against Powdery mildew (Leveillulataurica) disease of Chilli
T. No. Treatments Dose/ha
a.i.(g/ml)
Formulation
(g/ml)/ha
5 DAT Colby,sValue 10 DAT Colby,sValue
PDI %Control - PDI %Control
-

1 Kasugamycin 3 % SL 37.5 1250 26.67
(31.07) 34.96 32.67
(34.83) 37.18
2 Flusilazole 40 % EC 120 300 25.33
(30.19) 38.21 31.00
(33.81) 40.38
3 Azoxystrobin 23%SC 125 500 24.00
(29.31) 41.46 29.33
(32.77) 43.59
4 Kasugamycin 3% SL + Flusilazole 40% EC + Azoxystrobin 23% SC 37.5+120+125 1250+300+500 7.67
(16.04) 81.30 1.06 8.33
(16.76) 83.97 1.06
5 Kasugamycin 2.4% + Flusilazole 7.5% + Azoxystrobin 8% 18+56.25+60 750 14.00
(21.95) 65.85 0.86 16.33
(23.81) 68.59 0.87
6 Kasugamycin 2.4% + Flusilazole 7.5% + Azoxystrobin 8% 24+75+80 1000 12.67
(20.82) 69.11 0.90 14.00
(21.95) 73.08 0.93
7 Kasugamycin 2.4% + Flusilazole 7.5% + Azoxystrobin 8% 30+93.75+100 1250 10.00
(18.41) 75.61 0.99 11.67
(19.96) 77.56 0.98
8 Untreated Control - - 41.00
(39.79) - - 52.00
(46.12) - -
CD (5%) - - 1.14 - - 1.33 - -
SE(m) - - 0.37 - - 0.43 - -
Figures in parenthesis are angular transformed values.

Results of Table 7:
• Results presented in Table 7 showed that incidence of powdery mildew disease reduced in all the treatments. At 5th and 10th DAT, Tank mix formulation of Kasugamycin 3% SL + Flusilazole 40% EC + Azoxystrobin 23% SC @ 1250+300+500 g/ml/ha was found as the best treatment in reducing the incidence to 81.30 and 83.97 %, respectively.

• This was followed by applying-Kasugamycin 2.4% and Flusilazole 7.5% and Azoxystrobin 8%@1250 ml/ha with 75.61 and 77.56% control, respectively.

• Kasugamycin 2.4% and Flusilazole 7.5% and Azoxystrobin 8%@1000 ml/ha resulted in 69.11 and 73.08% control whereas, Kasugamycin 2.4% and Flusilazole 7.5% and Azoxystrobin 8%@750 ml/ha provided 65.85 and 68.59% control, respectively after 5th and 10th DAT, which were significantly superior in comparison to the solo application of Kasugamycin 3% SL @ 1250 ml/ha, Flusilazole 40% EC @ 300 ml/ha and Azoxystrobin 23% SC @ 500 gm/ha against powdery mildew disease of Chilli.

• Among the treatments mentioned in Table 7, only tank mix formulation of Kasugamycin 3% SL + Flusilazole 40% EC + Azoxystrobin 23% SC @ 1250+300+500 gm/ml/ha showed synergistic effect with Colby’s synergistic value of 1.06 on 5th and 10th DAT.

Table 8. Evaluation of bio-efficacy of Kasugamycin 2.8% and Flusilazole 8.4% and Azoxystrobin 8.8% against Powdery mildew (Leveillulataurica) disease of Chilli.
T. No. Treatments Dose/ha
a.i.(g/ml)
Formulation
(g/ml)/ha
5 DAT Colby,sValue 10 DAT Colby,sValue
PDI %Control
-

PDI %Control -

1 Kasugamycin 3 % SL 37.5 1250 27.00
(31.29) 34.15 32.67
(34.84) 37.18
2 Flusilazole 40 % EC 120 300 25.33
(30.19) 38.21 30.33
(33.40) 41.67
3 Azoxystrobin 23%SC 125 500 23.33
(28.87) 43.09 28.67
(32.35) 44.87
4 Kasugamycin 3% SL + Flusilazole 40% EC + Azoxystrobin 23% SC 37.5+120+125 1250+300+500 8.00
(16.40) 80.49 1.04 8.00
(16.40) 84.62 1.06
5 Kasugamycin 2.8% + Flusilazole 8.4 % + Azoxystrobin 8.8% 21+63+66 750 12.00
(20.24) 70.73 0.92 14.33
(22.20) 72.44 0.90
6 Kasugamycin 2.8% + Flusilazole 8.4 % + Azoxystrobin 8.8% 28+84+88 1000 7.33
(15.59) 82.11 1.06 7.00
(15.23) 86.54 1.08
7 Kasugamycin 2.8% + Flusilazole 8.4 % + Azoxystrobin 8.8% 35+105+110 1250 6.67
(14.92) 83.74 1.09 6.67
(14.94) 87.18 1.09
8 Untreated Control - - 41.00
(39.79) - - 52.00
(46.12) - -
CD (5%) - - 1.54 - - 2.09 - -
SE(m) - - 0.50 - - 0.68 - -
Figures in parenthesis are angular transformed values.
Results of Table 8:
• Results presented in Table 8 showed that Kasugamycin 2.8% and Flusilazole 8.4 % and Azoxystrobin 8.8% @1250 g/ml/ha was found most effective with 87.18% control followed by Kasugamycin 2.8% and Flusilazole 8.4 % and Azoxystrobin 8.8% @1000 ml/ha with 86.54% control, both of the treatments were found at par with tank mix formulation of Kasugamycin 3% SL and Flusilazole 40% EC and Azoxystrobin 23% SC @ 1250+300+500 g/ml/ha with 84.62% control.
• Kasugamycin 2.8% and Flusilazole 8.4 % and Azoxystrobin 8.8% @750 g/ml/ha resulted in 72.44% control after 10th DAT, which were significantly superior in comparison to the solo application of Kasugamycin 3% SL @ 1250 g/ml/ha, Flusilazole 40% EC @ 300 g/ml/ha and Azoxystrobin 23%SC@ 500 g/ml/ha against powdery mildew disease of Chilli.
• Among the treatments mentioned in Table 8, Kasugamycin 2.8% and Flusilazole 8.4 % andAzoxystrobin 8.8% @1250 ml/ha.
• Kasugamycin 2.8% and Flusilazole 8.4 % and Azoxystrobin 8.8% @1000 ml/ha and Kasugamycin 3% SL and Flusilazole 40% EC and Azoxystrobin 23% SC @ 1250+300+500 g/ml/ha showed synergistic effect.
• On 5th and 10thDAT, Colby’s synergistic value of 1.06 and 1.08 was calculated for Kasugamycin 2.8% and Flusilazole 8.4 % and Azoxystrobin 8.8% @1000 ml/ha.

Table 9. Evaluation of bio-efficacy of Kasugamycin 3.0% and Flusilazole 9.5% and Azoxystrobin 9.9% against Powdery mildew (Leveillulataurica) disease of Chilli.
T. No. Treatments Dose/ha
a.i.(g/ml)
Formulation
(g/ml)/ha
5 DAT Colby,sValue 10 DAT Colby,sValue
PDI %Control -
PDI %Control
-


1 Kasugamycin 3 % SL 37.5 1250 27.00
(31.27) 34.15 32.67
(34.83) 37.18
2 Flusilazole 40 % EC 120 300 25.33
(30.20) 38.21 31.00
(33.81) 40.38
3 Azoxystrobin 23%SC 125 500 24.00
(29.31) 41.46 28.33
(32.13) 45.51
4 Kasugamycin 3% SL + Flusilazole 40% EC + Azoxystrobin 23% SC 37.5+120+125 1250+300+500 7.50
(15.88) 81.71 1.07 8.17
(16.51) 84.29 1.06
5 Kasugamycin 3.0% + Flusilazole 9.5% + Azoxystrobin 9.9% 22.5+71.25+74.25 750 11.67
(19.96) 71.54 0.93 13.00
(21.11) 75.00 0.94
6 Kasugamycin 3.0% + Flusilazole 9.5% + Azoxystrobin 9.9% 30+95+99 1000 6.67
(14.94) 83.74 1.09 6.33
(14.43) 87.82 1.10
7 Kasugamycin 3.0% + Flusilazole 9.5% + Azoxystrobin 9.9% 37.5+118.75+123.75 1250 6.33
(14.56) 84.55 1.11 6.00
(14.04) 88.46 1.11
8 Untreated Control - - 41.00
(39.79) - - 52.00
(46.12) - -
CD (5%) - - 1.546 - - 3.085 - -
SE(m) - - 0.505 - - 1.007 - -
Figures in parenthesis are angular transformed values.

Results of Table 9:
• Results presented in Table 9 showed that Kasugamycin 3.0% and Flusilazole 9.5% and Azoxystrobin 9.9%@1250 g/ml/ha was found most effective with 88.46% control.
• Kasugamycin 3.0% and Flusilazole 9.5% and Azoxystrobin 9.9%@1000 g/ml/ha found effective with 87.82% control,both of the treatments were found at par with tank mix formulation of Kasugamycin 3% SL and Flusilazole 40% EC and Azoxystrobin 23% SC @ 1250+300+500 g/ml/ha with 84.29% control.
• Kasugamycin 3.0% and Flusilazole 9.5% and Azoxystrobin 9.9%@750 ml/ha resulted in 75.00% control after 10th DAT, which were significantly superior in comparison to the solo application of Kasugamycin 3% SL @ 1250 ml/ha, Flusilazole 40% EC @ 300 ml/ha and Azoxystrobin 23%SC @ 500 g/ml/ha against powdery mildew disease of Chilli.
• Kasugamycin 3.0% and Flusilazole 9.5% and Azoxystrobin 9.9%@1000 g/ml/ha also showed synergistic effect with Colby’s synergistic value of 1.09 and 1.10 at 5th and 10th DAT, respectively.

Table 10. Evaluation of bio-efficacy of Kasugamycin 2.4 % and Flusilazole 7.5 % andAzoxystrobin 8% against Anthracnose (Colletotrichumcapsici) disease of Chilli.
T. No. Treatments Dose/ha
a.i.(gm/ml)
Formulation
(g/ml)/ha
5 DAT Colby,sValue 10 DAT Colby,sValue
PDI %Control - PDI %Control -
1 Kasugamycin 3 % SL 37.5 1250 26.33
(30.86) 34.17 31.67
(32.35) 37.50
2 Flusilazole 40 % EC 120 300 25.00
(29.97) 37.50 30.67
(31.65) 39.47
3 Azoxystrobin 23%SC 125 500 24.00
(29.31) 40.00 29.00
(31.00) 42.76
4 Kasugamycin 3% SL + Flusilazole 40% EC + Azoxystrobin 23% SC 37.5+120+125 1250+300+500 8.00
(16.36) 80.00 1.06 8.67
(16.31) 82.89 1.06
5 Kasugamycin 2.4% + Flusilazole 7.5% + Azoxystrobin 8% 18+56.25+60 750 14.33
(22.23) 64.17 0.85 15.67
(23.28) 69.08 0.88
6 Kasugamycin 2.4% + Flusilazole 7.5% + Azoxystrobin 8% 24+75+80 1000 12.67
(20.82) 68.33 0.90 14.00
(21.23) 72.37 0.92
7 Kasugamycin 2.4% + Flusilazole 7.5% + Azoxystrobin 8% 30+93.75+100 1250 10.33
(18.71) 74.17 0.98 12.00
(19.00) 76.32 0.97
8 Untreated Control - - 40.00
(39.21) - - 50.67
(46.34) - -
CD (5%) - - 1.390 - - 2.346 - -
SE(m) - - 0.454 - - 0.654 - -
Figures in parenthesis are angular transformed values

Results of Table 10:
• Results presented in Table 10 showed that incidence of anthracnose disease reduced in all the treatments.
• At 5th and 10th DAT, Tank mix formulation of Kasugamycin 3% SL and Flusilazole 40% EC and Azoxystrobin 23% SC @ 1250+300+500 gm/ml/ha was found as the best treatment in reducing the incidence to 80.00 and 82.89 %, respectively.
• This followed by Kasugamycin 2.4% and Flusilazole 7.5% and Azoxystrobin 8%@1250 ml/ha with 74.17 and 76.32% control respectively.
• Kasugamycin 2.4% and Flusilazole 7.5% and Azoxystrobin 8%@1000 g/ml/ha resulted in 68.33 and 72.37% control whereas, Kasugamycin 2.4% and Flusilazole 7.5% and Azoxystrobin 8%@750 ml/ha provided 64.17 and 69.08% control, respectively after 5th and 10th DAT, which were significantly superior in comparison to the solo application of Kasugamycin 3% SL @ 1250 ml/ha, Flusilazole 40% EC @ 300 ml/ha and Azoxystrobin 23% SC @ 500 gm/ha against anthracnose disease of Chilli.
• Among the treatments mentioned in Table 10, only tank mix formulation of Kasugamycin 3% SL and Flusilazole 40% EC and Azoxystrobin 23% SC @ 1250+300+500 gm/ml/ha showed synergistic effect with Colby’s synergistic value of 1.06 on 5th and 10th DAT.

Table 11. Evaluation of bio-efficacy of Kasugamycin 2.8% and Flusilazole 8.4 % and Azoxystrobin 8.8% against Anthracnose (Colletotrichumcapsici) disease of Chilli.
T. No. Treatments Dose/ha
a.i.(gm/ml)
Formulation
(g/ml)/ha
5 DAT Colby,s Value 10 DAT Colby,s Value

PDI % Control
-
PDI %C ontrol
-


1 Kasugamycin 3 % SL 37.5 1250 27.00
(31.29) 32.50 33.00
(35.04) 34.00
2 Flusilazole 40 % EC 120 300 25.67
(30.42) 35.83 30.33
(33.39) 39.33
3 Azoxystrobin 23%SC 125 500 23.67
(29.09) 40.83 28.67
(32.35) 42.67
4 Kasugamycin 3% SL + Flusilazole 40% EC + Azoxystrobin 23% SC 37.5+120+ 125 1250+300+ 500 7.50
(15.87) 81.25 1.09 8.33
(16.68) 83.33 1.08
5 Kasugamycin 2.8% + Flusilazole 8.4 % + Azoxystrobin 8.8% 21+63+66 750 13.00
(21.11) 67.50 0.91 13.67
(21.60) 72.67 0.94
6 Kasugamycin 2.8% + Flusilazole 8.4 % + Azoxystrobin 8.8% 28+84+88 1000 7.00
(15.31) 82.50 1.11 8.00
(21.60) 84.00 1.09
7 Kasugamycin 2.8% + Flusilazole 8.4 % + Azoxystrobin 8.8% 35+105+110 1250 6.67
(14.92) 83.33 1.12 7.33
(15.67) 85.33 1.11
8 Untreated Control - - 40.00
(39.21) - - 50.00
(44.98) - -
CD (5%) - - 1.47 - - 2.56 - -
SE(m) - - 0.48 - - 0.83 - -
Figures in parenthesis are angular transformed values.

Results of Table 11:
• Results presented in Table 11 showed that Kasugamycin 2.8% and Flusilazole 8.4 % and Azoxystrobin 8.8% @1250 g/ml/ha was found most effective with 85.33% control followed by Kasugamycin 2.8% and Flusilazole 8.4 % and Azoxystrobin 8.8% @1000 ml/ha with 84.00% control.
• Both of the treatments were found at par with tank mix formulation of Kasugamycin 3% SL and Flusilazole 40% EC and Azoxystrobin 23% SC @ 1250+300+500 g/ml/ha with 83.33% control.
• Kasugamycin 2.8% and Flusilazole 8.4 % and Azoxystrobin 8.8% @750 g/ml/ha resulted in 72.67% control after 10th DAT, which were significantly superior in comparison to the solo application of Kasugamycin 3% SL @ 1250 g/ml/ha, Flusilazole 40% EC @ 300 g/ml/ha and Azoxystrobin 23% SC@ 500 g/ml/ha against anthracnose disease of Chilli.
• Among the treatments mentioned in Table 11-
Kasugamycin 2.8% and Flusilazole 8.4 % and Azoxystrobin 8.8% @1250 ml/ha,
Kasugamycin 2.8% and Flusilazole 8.4 % + Azoxystrobin 8.8% @1000 ml/ha, and
Kasugamycin 3% SL and Flusilazole 40% EC and Azoxystrobin 23% SC @ 1250+300+500 g/ml/ha showed synergistic effect.
• On 5th and 10th DAT, Colby’s synergistic value of 1.11 and 1.09 was calculated for Kasugamycin 2.8% and Flusilazole 8.4 % and Azoxystrobin 8.8% @1000 ml/ha.

Table 12. Evaluation of bio-efficacy of Kasugamycin 3.0% and Flusilazole 9.5% and Azoxystrobin 9.9% against Anthracnose (Colletotrichumcapsici) disease of Chilli.
T. No. Treatments Dose/ha
a.i.(gm/ml)
Formulation
(g/ml)/ha
5 DAT Colby,s Value 10 DAT Colby,s Value

PDI % Control


- PDI % Control


-
1 Kasugamycin 3 % SL 37.5 1250 26.33
(30.85) 34.17 31.67
(34.23) 36.67
2 Flusilazole 40 % EC 120 300 25.50
(30.31) 36.25 30.33
(33.40) 39.33
3 Azoxystrobin 23%SC 125 500 24.00
(29.31) 40.00 28.33
(32.13) 43.33
4 Kasugamycin 3% SL + Flusilazole 40% EC + Azoxystrobin 23% SC 37.5+120+125 1250+300+ 500 7.33
(15.65) 81.67 1.09 8.17
(16.57) 83.67 1.07
5 Kasugamycin 3.0% + Flusilazole 9.5% + Azoxystrobin 9.9% 22.5+71.25+74.25 750 11.33
(19.64) 71.67 0.96 12.67
(20.76) 74.67 0.95
6 Kasugamycin 3.0% + Flusilazole 9.5% + Azoxystrobin 9.9% 30+95+99 1000 6.33
(14.56) 84.17 1.13 7.17
(15.36) 85.67 1.10
7 Kasugamycin 3.0% + Flusilazole 9.5% + Azoxystrobin 9.9% 37.5+118.75+ 123.75 1250 6.00
(14.04) 85.00 1.14 6.83
(15.06) 86.33 1.10
8 Untreated Control - - 40.00
(39.21) - - 50.00
(44.98) - -
CD (5%) - - 2.20 - - 2.71 - -
SE(m) - - 0.72 - - 0.89 - -
Figures in parenthesis are angular transformed values

Results of Table 12:
• Results presented in Table 12 showed that Kasugamycin 3.0% and Flusilazole 9.5% and Azoxystrobin 9.9%@1250 g/ml/ha was found most effective with 86.33% control followed by Kasugamycin 3.0% and Flusilazole 9.5% and Azoxystrobin 9.9%@1000 g/ml/ha found effective with 85.67% control.
• Both of the treatments were found at par with tank mix formulation of Kasugamycin 3% SL and Flusilazole 40% EC and Azoxystrobin 23% SC @ 1250+300+500 g/ml/ha with 83.67% control and Kasugamycin 3.0% and Flusilazole 9.5% and Azoxystrobin 9.9%@750 ml/ha resulted in 74.67% control after 10th DAT, which were significantly superior in comparison to the solo application of Kasugamycin 3% SL @ 1250 ml/ha, Flusilazole 40% EC @ 300 ml/ha and Azoxystrobin 23%SC @ 500 g/ml/ha against anthracnose disease of Chilli.
• Kasugamycin 3.0% and Flusilazole 9.5% and Azoxystrobin 9.9%@1000 g/ml/ha also showed synergistic effect with Colby’s synergistic value of 1.13 and 1.10 at 5th and 10th DAT, respectively.

Phytotoxicity observations
• Various Fungicidal composition of the present invention provided good control of Powdery mildew &Anthracnosedisease of chilli as compared to the reference products.
• Further, the use of these Fungicidal composition resulted in better crop condition i.e. fresh green leaves and didn’t produce any phytotoxic symptoms on the plants.
• Table 10-12 depicts the phytotoxic effects of the various fungicidal Formulation of the present invention on chilli at 3rd, 5th, 7th and 10th DAT.

Table 13. Evaluation of Phytotoxic effects of Kasugamycin 2.4 % and Flusilazole 7.5 % and Azoxystrobin 8% against Powdery mildew (Leveillulataurica) and Anthracnose (Colletotrichumcapsici) disease of Chilli

Treatments Phytotoxicity rating
Yellowing Wilting Necrosis Epinasty Hyponasty
Days After Treatment
3 5 7 10 3 5 7 10 3 5 7 10 3 5 7 10 3 5 7 10
T1 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0
T2 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0
T3 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0
T4 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0
T5 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0
T6 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0
T7 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0

Table 14. Evaluation of Phytotoxicity effect of Kasugamycin 2.8% and Flusilazole 8.4% and Azoxystrobin 8.8% against Powdery mildew (Leveillulataurica) and Anthracnose (Colletotrichumcapsici) disease of Chilli
Treatments Phytotoxicity rating
Yellowing Wilting Necrosis Epinasty Hyponasty
Days After Treatment
3 5 7 10 3 5 7 10 3 5 7 10 3 5 7 10 3 5 7 10
T1 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0
T2 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0
T3 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0
T4 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0
T5 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0
T6 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0
T7 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0

Table 15. Evaluation of Phytotoxicity effect of Kasugamycin 3.0% and Flusilazole 9.5% and Azoxystrobin 9.9% against Powdery mildew (Leveillulataurica) and Anthracnose (Colletotrichumcapsici) disease of Chilli
Treatments Phytotoxicity rating
Yellowing Wilting Necrosis Epinasty Hyponasty
Days After Treatment
3 5 7 10 3 5 7 10 3 5 7 10 3 5 7 10 3 5 7 10
T1 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0
T2 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0
T3 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0
T4 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0
T5 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0
T6 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0
T7 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0

Conclusion:
• After evaluating three compositions of Kasugamycin, Flusilazole and Azoxystrobin-
? Kasugamycin 2.4 % and Flusilazole 7.5 % and Azoxystrobin 8%;
? Kasugamycin 2.8% and Flusilazole 8.4 % and Azoxystrobin8.8% and
? Kasugamycin 3.0% and Flusilazole 9.5% and Azoxystrobin 9.9%) at three different doses @ 750 ml/ha, 1000 ml/ha and 1250 ml/ha.
It was found that the combination of Kasugamycin 2.8% and Flusilazole 8.4% and Azoxystrobin 8.8% @ 1000 g/ml/ha was the most effective against both Powdery Mildew and Anthracnose diseases of Chilli.
• Kasugamycin 2.8% and Flusilazole 8.4% and Azoxystrobin 8.8% @ 1000 g/ml/ha was observed at par with higher formulation, higher dose and higher active ingredient and also showed a strong synergistic effect in controlling the Powdery Mildew and Anthracnose diseases of chilli.
• Therefore, the application of Kasugamycin 2.8% and Flusilazole 8.4% and Azoxystrobin 8.8% at 1000 ml/ha (28+84+88 gm a.i/ha) can be recommended for effective control of Powdery Mildew and Anthracnose diseases in Chilli.
ADVANTAGES OF THE PRESENT COMPOSITION
• The present disclosure provides a new and improved Synergistic composition that may overcome the limitations associated with the conventional fungicidal composition.
• The present disclosure provides a Synergistic composition that exhibits broad spectrum antifungal and antibacterial properties.
• The present disclosure provides a Synergistic composition that exhibits desired antifungal and antibacterial effect at lower dosage.
• The present disclosure provides a Synergistic composition that aid in precluding development of resistance of phytopathogens towards Kasugamycin and Flusilazole and Azoxystrobin or salt thereof.
• The present disclosure provides Synergistic composition that is safe to use.
• The present disclosure provides Synergistic composition that is cost-effective.
• The present disclosure provides Synergistic composition that is easy to prepare.
, Claims:
1. A Synergistic composition, comprising:
(a) 0.1% to about 30.0% kasugamycin, by weight of the composition;
(b) about 0.1% to about 30.0% flusilazole by weight of the composition;
(c) about 0.1% to about 30.0% azoxystrobin by weight of the compositionand ;
(d) agriculturally acceptable additives(s) .
2. The Synergistic composition as claimed in claim 1, wherein the Kasugamycin is in 2.8%w/w, Flusilazole is in 8.4 %w/wand Azoxystrobin is in 8.8% w/w of the formulation.

3. The Synergistic composition as claimed in claim 1, wherein the composition is selected from a wettable powder (WP), a water dispersible granule (WDG), a water dispersible tablet (WT), an ultra-low volume (ULV) liquid (UL), an ultra-low volume (ULV) suspension (SU), a water soluble powder (SP), a soluble concentrate (SL), a water soluble granule (SG), a suspo-emulsion (SE), granule (GR), an emulsifiable granule (EG), an oil-in- water emulsion (EW), an emulsifiable concentrate (EC), a micro-emulsion (ME), an oil dispersion (OD), a capsule suspension (CS), an aerosol (AE) or a mixed formulation of CS and SC (ZC).

4. The synergistic composition as claimed in claim 3, wherein said synergistic composition is a suspo-emulsion (SE).

5. The Synergistic composition as claimed in claim 1, wherein agriculturally acceptable additives is selected from the group comprising an emulsifier; dispersing agent; antifoaming agent; rheology modifier; solvent; biocide; Stabilising agent and an antifreezing agent.
Wherein the composition comprises-
(a) about 0.1% to about 20.0% emulsifier;
(b) about 0.1% to about 20.0% dispersing agent;
(d) about 0.01% to about 7.0% antifoaming agent;
(e) about 0.01% to about 12.0% rheology modifier;
(f) about 1.0% to about 95.0% solvent;
(g) about 0.1% to about 5% Stabilising;
(h) about 0% to 3 % biocide; and
(i) about 0% to 15 % antifreezing agent.

6. The Synergistic composition as claimed in claim 5, wherein the emulsifier is selected from the group comprising ethoxylated propoxylated alcohols, alkylphenolethoxylates, alkoxylatedtristyrylphenols, ethoxylated propoxylatedpolyaryl phenol, ethoxylated fatty acids, Ethoxylated Tristyrylphenol , Soprophor 3D33,Fatty alcohol ethoxylates, ethoxylatedricinoleic acid triglycerides, sorbitan trioleate, tridecyl alcohol ethoxylate, castor oil ethoxylate, alkoxylated phosphate ester or mixtures thereof.

7. The Synergistic composition as claimed in claim 5, wherein the dispersing agent is selected from the group comprising naphthalene formaldehyde condensates, acrylic copolymer, nonionic proprietary surfactant blend, polycarboxylates, calcium dodecylbenzene sulfonate, polystyrenatedacrylated co-polymer, Atlox 4917, salts of phenol sulfonic acids, Nonionic proprietary surfactant blend, Soprophor 3D33, Butyl polyalkylene oxide block co-polymer ,random co-polymer of alcoxylated polyethylene glycol or mixtures thereof.

8. The Synergistic composition as claimed in claim 5, wherein the antifoaming agent is selected from the group comprising perfluroalkylphosphonic acids, polydimethylsiloxane, polydimethylsiloxane emulsion or mixtures thereof.
9. The Synergistic composition as claimed in claim 5,wherein the rheology modifier is selected from the group comprising precipitated silica, modified fumed silica, bentonite, hydroxymethyl cellulose, carboxymethyl cellulose, xanthan gum, thickening silica, hydrated clay minerals, magnesium aluminium silicates, organic derivative of hectorite clay, hydrophobic fumed silica, polyvinylpyrrolidone (PVP) or mixture thereof.

10. The Synergistic composition as claimed in claim 5,wherein the solvent is selected from the group comprising of Demineralized (DM) water, N-alkyl-pyrrolidone, oil medium selected from the group comprising, esterified fatty acids selected from methyl ester of triglycerides containing C12–C22 saturated and unsaturated fatty acids, ethyl ester of triglycerides containing C12 –C22 saturated and unsaturated fatty acids such as methyl soyate, ethyl soyate, rapeseed methyl ester, rapeseed ethyl ester or mixtures thereof.

11. The Synergistic composition as claimed in claim 5,wherein the biocide is selected from the group comprising 1,2-benzisothiazolin-3-one, formaldehyde, dipropyl glycol solution of 1,2-benzisothiazolin-3-one or mixtures thereof.

12. The Synergistic composition as claimed in claim 5,wherein the antifreezing agent is selected from the group comprising ethylene glycol, 1,2-propanediol, propane-1,2,3-triol, urea or mixtures thereof.

13. The Synergistic composition as claimed in claim 5,wherein the stabilizing agents is selected from the group polysaccharides, carboxymethyl cellulose,Precipitated Silica bentonite clay, aluminum magnesium silicate, citric acid either alone or mixtures thereof.
14. A process for preparing the synergistic composition of claim 1, comprising mixing the kasugamycin, flusilazole and azoxystrobin with agriculturally acceptable formulation additives.