Description:TITLE OF THE INVENTION:
FUNGICIDAL COMPOSITION

FIELD OF THE INVENTION:
The present invention relates to a stable, synergistic fungicidal composition comprising Fluxapyroxad, Azoxystrobin, Kasugamycin and Soy protein, for control of fungal pests in various agricultural crops. The present invention further relates to a process for preparing said fungicidal composition.

BACKGROUND OF THE INVENTION:
Fungal diseases of plants can cause severe pre- and post-harvest losses in agricultural crops. Fungi cause a great majority of infectious plant diseases and over 8,000 fungal species have been shown to cause disease.

Fungal infections cause pre-harvest damage to crop by killing them outright or weakening them thereby decreasing yields and rendering the plants susceptible to other infections. Post-harvest, fungal infection also results in significant loss of agricultural products during storage, processing and handling. Clearly, there is a significant need to control the fungal infection of plants and plant products and numerous chemical agents have been developed for this purpose, but till date no fully satisfactory agents, i.e. agents that completely control the fungus while at the same time being devoid of undesirable side effects, have been found.

Use of chemical fungicides is the primary method to prevent fungal diseases in plants. Excessive use of synthetic fungicides, however, may cause development of fungicide resistance in the pest population, resulting in the need for higher quantities of the pesticide for effective control. Fungicide residues have been found, for example, in groundwater, animal feed and food for human consumption as a result of pesticide use and can be harmful to animals, including humans. Fungicides may also eliminate beneficial microorganisms which again may result in emergence of “new” diseases. Alternative ways to control plant pathogens are therefore needed so that reduced amounts of chemical fungicides are used while maintaining the same protection against pre- and post-harvest loss caused by fungi.

Therefore, it is necessary to have a stable combination of fungicidal active ingredients with decreased phytotoxicity and with increased fungicidal activity. Also, it is necessary to have a composition which is easy to handle by the end user in terms of calculation of dose and which does not cause any handling problems or toxicity to the end user.

Surprisingly, the inventors of the present invention have found that the synergistic combination of Fluxapyroxad, Azoxystrobin, Kasugamycin and Soy protein as described herein can provide a much-needed solution to all the above-mentioned problems.

OBJECT(S) OF THE INVENTION:
It is an object of the present invention to provide a novel, synergistic fungicidal combination.

It is another object of the present invention to provide a fungicidal composition comprising as its components, Fluxapyroxad, Azoxystrobin, Kasugamycin and Soy protein for control of fungal pests in various agricultural crops.

It is yet another object of the present invention to provide a synergistic fungicidal composition with enhanced efficacy against fungal pests and enhanced crop yield without any phytotoxicity.

It is yet another object of the present invention to provide a process for preparing the fungicidal composition.

It is yet another object of the present invention to provide a method for controlling pests in various agriculture crops.

SUMMARY OF THE INVENTION:
In one aspect, the present invention provides a fungicidal composition comprising:
a. Fluxapyroxad having weight concentration in the range of 10-30% w/w;
b. Azoxystrobin having weight concentration in the range of 10-30% w/w;
c. Kasugamycin having weight concentration in the range of 1-10% w/w;
d. Soy protein having weight concentration in the range of 0.05-5% w/w; and
e. at least one agriculturally acceptable excipient.

In another aspect, the present invention provides a process for preparing the fungicidal composition as a suspension concentrates formulation, said process comprising:
(i) mixing surfactant, anti-freezing agent in D.M. water, followed by solubilizing in high-speed mixer at 500-1000rpm at room temperature;
(ii) adding anti foaming agent, Fluxapyroxad technical and Azoxystrobin technical, followed by addition of Kasugamycin and rest of the adjuvants to the mixture of step (i) to obtain a homogenous mass;
(iii) grinding the homogenous mass of step (ii) in sand mill until a mean particle size of <5 micron is obtained at a temperature of 15-25ºC;
(iv) adding 2% solution of xanthan gum and biocide to step (iii) under low stirring and mixing well at the speed of 400-700 rpm;
(v) adding soy protein and vegetable oil to the mixture obtained in step (iv) in continuous stirring till a homogenous mixture is obtained in suspension concentrate (SC) formulation. Rest of the steps are carried out at room temperature.

In yet another aspect, the present invention provides a method for controlling pests in various agriculture crops, said method comprising applying a fungicidally effective amount of a fungicidal composition to the target crop or portion thereof.

TECHNICAL ADVANCEMENT OF THE INVENTION:

The fungicidal composition of the present invention comprising as its components, Fluxapyroxad, Azoxystrobin, Kasugamycin and Soy protein as described herein above has several associated technical advantages including but not limited to:
? enhanced disease control;
? synergistic effects;
? plant health benefits; and
? reduced environmental impact.

DETAILED DESCRIPTION OF THE INVENTION:
Abbreviations
D.M. Demineralised Water
RTU Ready to Use
SC Suspension Concentrate
QS Quantum satis
A.I/ a.i Active Ingredients
PDI Percent Disease Index
DAA Days After Application

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, compositions 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 ‘disease control’ as used herein denotes control and prevention of a disease. Controlling effects include all deviation from natural development, for example: killing, retardation, decrease of the pests. The term ‘plants’ refers to all physical parts of a plant, including seeds, seedlings, saplings, roots, tubers, stems, stalks, foliage and fruits.

It is to be understood that the singular forms “a,” “an,” and “the” include plural referents unless the context clearly dictates otherwise.

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.

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 "fungicidal" also called antimycotic, any toxic substance used to kill or inhibit the growth of fungi.

The term “fungicidally effective amount” refers to the quantity of a fungicide needed to inhibit the growth of fungi in a given environment. This amount is sufficient to achieve the desired level of control over the fungal population without causing significant harm to the host organism, be it a plant, animal, or an ecosystem.

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 composition 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.

As used herein, the term “agriculturally acceptable excipient” refers to an ingredient, additive, component or supplement, liquid or solid, suitable for incorporation in agricultural compositions.

Fluxapyroxad is a broad-spectrum pyrazole-carboxamide fungicide. Fluxapyroxad is chemically known as 3-(difluoromethyl)-1-methyl-N-(3',4',5'-trifluorobiphenyl-2-yl)pyrazole-4-carboxamide having chemical structure as below:

Fluxapyroxad is a succinate dehydrogenase inhibitor (SDHI). It interferes with a number of key fungal life functions, including spore germination, germ tube growth, and mycelium growth. Specifically, it interferes with the production of succinate dehydrogenase, the complex II in the mitochondrial respiration chain, which in turn interferes with the tricarboxylic cycle and mitochondrial electron transport.
Azoxystrobin is a broad-spectrum systemic fungicide. Azoxystrobin is chemically known as Methyl (2E)-2-(2-{[6-(2-cyanophenoxy) pyrimidin-4-yl]oxy}phenyl)-3-methoxyprop-2-enoate having chemical structure as below:

Azoxystrobin inhibits mitochondrial respiration by blocking electron transport. They bind at the quinol outer binding site of the cytochrome b-c1 complex, where ubiquinone (coenzyme Q10) would normally bind when carrying electrons to that protein. Thus, production of ATP is prevented. The generic name for this mode of action is "Quinone Outside Inhibitors" QoI.
Kasugamycin (Ksg) is an aminoglycoside antibiotic that was originally isolated in 1965. Kasugamycin is chemically known as 2-amino-2-[(2R,3S,5S,6R)-5-amino-2-methyl-6-[(2R,3S,5S,6S)-2,3,4,5,6-pentahydroxycyclohexyl]oxyoxan-3-yl]iminoacetic acid having chemical structure as below:

Kasugamycin inhibits protein synthesis at the step of translation initiation. It indirectly induces dissociation of P-site-bound fMet-tRNAfMet from 30S subunits through perturbation of the mRNA, thereby interfering with translation initiation.

The details of biological material used in the present invention:

Soy Protein was procured from Sales Office 1900 Crown Colony 10 Drive, 3rd Floor, Quincy, Massachusetts, USA.

Soy protein basically serves as plant-derived bio-stimulant which is readily available commercially. Traditionally, it is made from soybean meal that has been dehulled and defatted. Soy protein extract contains various amino acids – Aspartic, Threonine, Serine, Glutamic acid, Proline, Glycine, Valine, Cystine, Isoleucine, Methionine, Tyrosine, Phenylalanine, Lysine, Histidine, Arginine, and the percentage of each amino acid may vary in the mixture.

Increasing frequency of adverse environmental conditions for crop growth, have driven the development of bio stimulants. These are substances whose function is to stimulate nutrient uptake, nutrient use efficiency, and tolerance to abiotic stresses and crop quality. Soy protein has a bio stimulating effect on the health of the crops.

Soy protein functions as micronutrients and act synergistically along with the active ingredients in the composition in improving plant immune system. Soy protein improves the formulation, stability, and adhesion of the fungicide to plant surfaces and helps the fungicides spread more effectively and adhere to the plant, which can enhance its overall effectiveness. Further, the addition of Soy protein enhances protein synthesis in the meristematic tissues. The enhanced protein synthesis increases photosynthetic pigments in plants and that in turn results in accelerated photosynthetic reaction. Hence, the addition of Soy protein to the active ingredients additionally restores plant health.

The best results are however obtained when Soy protein is incorporated in the composition in an optimum range and such optimum range needs to be determined. Excess amount of Soy protein in the biological system will affect plant cells leading to adverse effects on the plant physiology, finally leading to phytotoxicity. Hence, the optimum range of Soy protein in the composition is very significant.

Many types of formulations for various compositions are known and commercially available. Suspension Concentrates have grown in popularity due to benefits such as absence of dust, ease of use and effectiveness when compared to other formulation types such as emulsifiable concentrate (EC) and wettable powder (WP). These formulations are water based which provide good safety and user convenience. Additionally, these formulations are suitable for active ingredients with low water solubility and smaller particle size.

In a preferred embodiment, the fungicidal composition of the present invention is in the form of Ready to Use Formulation (RTU). The ready to use formulation has various advantages over tank mixture of the active ingredients. These advantages include very less or no inhalation hazards, easy handling, lower chances of sedimentation, environmentally safer, good efficacy without phytotoxicity, lower transportation costs and is a scientific practice as against the usual farmer practice of tank mixing as the formulation is prepared after stability, compatibility studies.

The present invention provides a novel, synergistic fungicidal composition comprising a) Fluxapyroxad as the first component b) Azoxystrobin as the second component c) Kasugamycin as the third component and d) Soy protein as the fourth component wherein Fluxapyroxad is present in an amount ranging from 10-30% w/w, Azoxystrobin is present in an amount ranging from 10-30% w/w, Kasugamycin is present in an amount ranging from 1-10% w/w and Soy protein is present in an amount ranging from 0.05-5% w/w of the fungicidal composition.

In a preferred embodiment of the present invention, Fluxapyroxad is present in an amount of 20% w/w, Azoxystrobin is present in an amount of 22% w/w, Kasugamycin is present in an amount of 6% w/w and Soy protein is present in an amount of 0.2% w/w of the fungicidal composition.

In an embodiment of the present invention, said fungicidal composition further comprises at least one agriculturally acceptable excipient. In an embodiment, at least one excipient is selected from the group consisting of at least one anti-freezing agent, at least one dispersing agent, at least one wetting agent, at least one anti foaming agent at least one biocide, at least one vegetable oil, at least on wetting cum dispersing agent, at least one stabilizer, at least one thickener, and a combination thereof.

In a preferred embodiment of the present invention, said excipient is selected from at least one anti-freezing agent, at least one dispersing agent, at least one wetting agent, at least one vegetable oil, at least one anti foaming agent, at least one biocide, at least one thickener and combination thereof.

In an embodiment of the present invention, said anti-freezing agent is selected from the group consisting of mono ethylene glycol, diethylene glycol, polypropylene glycol, ethylene alcohols, polyethylene glycols, glycerine, epichlorohydrin, urea, glycerol, sorbitol and combination thereof.

In a preferred embodiment of the present invention, said anti-freezing agent is propylene glycol.

In an embodiment of the present invention, said anti-freezing agent is present in an amount ranging from 5-10% w/w.

In a preferred embodiment of the present invention, said anti-freezing agent is present in an amount of 7% w/w.

In an embodiment of the present invention, said dispersing agent is selected from the group consisting of sodium lignosulphonates, sodium naphthalene sulphonate- formaldehyde condensates, aliphatic alcohol ethoxylates, tristyrylphenol ethoxylates potassium salt and esters, ethylene oxide/propylene oxide block copolymers and combination thereof.

In a preferred embodiment of the present invention, said dispersing agent is alkyl phenol ethylene oxide ammonium sulphonates.

In an embodiment of the present invention, said dispersing agent is present in an amount ranging from 2-10%w/w.

In a preferred embodiment of the present invention, said dispersing agent is present in an amount of 3.2% w/w.

In an embodiment of the present invention, said wetting agent is selected from the group consisting of blend of alkyl naphthalene sulfonate, sodium salt, amine salt of poly aryl phenyl ether phosphate, Sodium laurel sulphate, poly alkyl aryl ethylene oxide phosphate ester and combination thereof.

In a preferred embodiment of the present invention, said wetting agent is poly alkyl aryl ethylene oxide phosphate ester.

In an embodiment of the present invention, said wetting agent is present in an amount ranging from 1-7%w/w.
In a preferred embodiment of the present invention, said wetting agent is present in an amount of 2% w/w.

In an embodiment of the present invention, said vegetable oil is selected from the group consisting of castor oil, mustard oil, cotton seed oil, soya bean oil, ground nut oil, sunflower oil, neem seed oil and combination thereof.

In an embodiment of the present invention, said vegetable oil is present in an amount ranging from 0.1-6% w/w.

In a preferred embodiment of the present invention, said vegetable oil is present in amount of 0.2% w/w.

The vegetable oil was procured from 4825 Jalan Permatang Pauh, 13400 Butterworth, P.W. Malaysia.

In an embodiment of the present invention, said antifoaming agent is selected from the group consisting of silicone emulsions, long-chain alcohols, fatty acids, organic fluorine compounds silicon based and combination thereof.

In a preferred embodiment of the present invention, said anti foaming agent is silicon based.
In an embodiment of the present invention, said anti foaming agent is present in an amount ranging from 0.20-3% w/w.

In a preferred embodiment of the present invention, said anti foaming agent is present in an amount of 0.50% w/w.

In an embodiment of the present invention, said biocide is selected from the group consisting of Clavulanic acid, Avibactam, stigmasterol, formaldehyde and combination thereof.
In a preferred embodiment of the present invention, said biocide is formaldehyde.

In an embodiment of the present invention, said biocide is present in an amount ranging from 0.05-2.0% w/w.
In a preferred embodiment of the present, said biocide is present in an amount of 0.20% w/w.

In an embodiment of the present invention, said thickener is selected from the group consisting of polyacrylamide, polyethylene polymers, polysaccharides (long-chain sugars), or vegetable oils, xanthan gum and combination thereof.

In a preferred embodiment of the present invention, said thickener is xanthan gum.

In an embodiment of the present invention, said thickener is present in an amount ranging from 0.05-10% w/w.

In a preferred embodiment of the present invention, said thickener is present in an amount of 4% w/w.

The present composition may work well as several formulations 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), macro granules (GG), dry flowable (DF), aqueous Suspo-emulsions (SE), capsule suspensions (CS) and microgranules (MG), while the preferred formulation according to the present invention is a suspension concentrate (SC).

SC offers certain advantages not provided by other formulations, particularly the ones listed below:

1) It is safe and ecofriendly pesticide formulation,
2) It is easy to mix as it is water-based formulation.
3) Enhanced efficacy.
4) Better adhesion and penetration on target.
5) Organic solvents are not used. Solvent may increase toxicity to operator/environment and degrades spray equipment parts, freezing may initiate crystal formation.

In another embodiment, the present invention further provides a process of preparing a fungicidal composition comprising a) Fluxapyroxad b) Azoxystrobin c) Kasugamycin and d) Soy protein, wherein Fluxapyroxad is present in an amount ranging from 10-30% w/w, Azoxystrobin is present in an amount ranging from 10-30% w/w, Kasugamycin is present in an amount ranging from 1-10% w/w and Soy protein is present in an amount ranging from 0.05-5% w/w of the fungicidal composition; as a suspension concentrates formulation, said process comprising the following steps:

(i) mixing surfactant, anti-freezing agent in D.M. water, followed by solubilizing in high speed mixer at 500-1000rpm at room temperature,
(ii) adding anti foaming agent, Fluxapyroxad technical and Azoxystrobin technical, followed by the addition of Kasugamycin and rest of the adjuvants to the mixture of step (i) to obtain a homogenous mass;
(iii) grinding the homogenous mass of step (ii) in sand mill until a mean particle size of <5 micron is obtained at a temperature of 15-25ºC;
(iv) adding 2% solution of xanthan gum and biocide to step (iii) under low stirring and mix well at the speed of 400-700 rpm;
(v) adding soy protein and vegetable oil to the mixture obtained in step (iv) in continuous stirring till homogenous mixture is obtained in suspension formulation.

There was no effluent generation during the formulation of this product.

The present invention also provides a method for controlling pests in various agriculture crops, comprises applying fungicidally effective amount of a fungicidal composition to the target crop or portion thereof.

In a preferred embodiment of the present invention, said effective amount of the fungicidal composition comprises:
a. Fluxapyroxad in an amount ranging from 10-30% w/w;
b. Azoxystrobin in an amount ranging from 10-30% w/w;
c. Kasugamycin in an amount ranging from 1-10% w/w;
d. Soy protein in an amount ranging from 0.05-5% w/w; and
e. at least one agriculturally acceptable excipient.

In a preferred embodiment of the present invention, said agriculturally acceptable excipient is selected from the group comprising at least one anti-freezing agent, at least one dispersing agent, at least one wetting agent, at least one vegetable oil, at least one anti foaming agent, at least one biocide, at least one thickener and combination thereof.

In a preferred embodiment, the present invention provides a method of treating the blight disease in rice crop, said method comprising applying an effective amount of the fungicidal composition to rice crop in spraying mode.

In an embodiment of the present invention, said fungicidal composition is effective against a range of plant diseases caused by fungi including but not limited to rice sheath blight disease, rice flax spot, rice bakanae disease, rice seedling rot and others.

In a preferred embodiment of the present invention, the fungicidal composition is particularly effective against blight disease in rice crop.

The present invention provides protection against diseases caused by species including but not limited to Fusarium graminearum; Gaeumannomyces graminis; Helminthosporium; Microdochium nivale; Mycosphaerella species, Plasmopara viticola; Podosphaera leucotricha; Pseudocercosporella herpotrichoides; Pseudoperonospora, Pyricularia oryzae, Pyricularia grisea, Ramularia areola; Rhizoctonia species.

Surprisingly, inventors of the present invention have found that the presently claimed fungicidal composition when applied at significantly lesser dosages than the recommended dosages of the individual active ingredients when applied separately, exhibit superior efficacy as measured in terms of percent control of target pest expressed in terms of PDI.

In other words, it was a surprise finding in the present invention that these conventional fungicides when combined at desired weight percentages, which is reduced as compared to their individual AI (Active Ingredient) recommended dosages, the fungicidal effect of the composition is unexpectedly enhanced.
Further, it has been found that the active compound combination of the presently claimed composition exhibits a synergistic effect and not merely an additive effect. Thus, clearly, the customary dosages of the individual actives are substantially reduced, leading to lesser amount of soil residues and environmental pollution when the composition of the present invention is used.

The foregoing description of the embodiments has been provided for purposes of illustration and not intended to limit the scope of the present invention. Individual components of a particular embodiment are generally not limited to that embodiment but are interchangeable. Such variations are not to be regarded as a departure from the present invention, and all such modifications are considered to be within the scope of the present invention.

The present invention is further described in light of the following experiments which are set forth for illustration purposes only and not to be construed for limiting the scope of the invention. The following experiments can be scaled up to industrial/commercial scale and the results obtained can be extrapolated to industrial scale.

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 Suspension concentrates (SC)
Table no. 1 below provides a fungicidal composition as described in the present invention, formulated as suspension concentrates (SC).

Table 1: Chemical Composition of CF-2262 SC
Ingredients Tentative percentage w/w
Fluxapyroxad (100% basis) 20.0
Azoxystrobin (100% basis) 22.0
Kasugamycin (100% basis) 6.0
Soy protein 0.2
Anti- freezing (poly-propylene glycol) 7.0
Dispersing agent (alkyl phenol ethylene oxide ammonium sulphonates) 3.2
Wetting agent (poly alkyl aryl ethylene oxide phosphate ester) 2.0
Vegetable oil 0.2
Anti foaming agent (silicon based) 0.5
Biocide (formaldehyde) 0.2
Thickener (Xanthan gum) 4.0
D.M water 35
Total 100

Example 2: Field evaluation of the bio efficacy of the present fungicidal composition
The presently disclosed fungicidal mixture (coded as CF-2262 SC) was tested for its bio efficacy against bacterial blight disease and sheath blight disease in rice crop in a local variety during Kharif season 2023 at Raj Mundri, Andhra Pradesh. The plants were aligned in the plot size of 50 vines per treatments. The trial was laid out in a random block design consisting of a total of 10 treatments in three replications. The trial was conducted at a temperature of 34oC under 85% relative humidity, under no winds and optimum soil moisture conditions.

Measured quantity of the chemical was added to required volume of water @ 375 lit. /ha for spraying. 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. The solution was stirred well before application. Knapsack sprayer fitted with boom along with flood jet nozzle was used to apply the fungicidal composition.

Table 2: below provides the treatment details of the rice crop in the trial experiment conducted. For purpose of convenience, the present composition has been represented by the code “CF-2262 SC”.

Treatments Product a.i./Ha Dose/Ha
1. Untreated
2. Azoxystrobin 23 115 500
3. Kasugamycin 3 L 30 1000
4. CF-2226 SC 75 + 82.5 + 22.5 375
5. CF-2262 SC 75 + 82.5 + 22.5 + 0.75 375
6. CF-1514 SC 56.25 + 67.5 + 15 375
7. CF-1512 SC 56.25 + 67.5 + 15 + 0.75 375
8. CF-2248 SC 93.75 + 90 + 30 375
9. CF-2282 SC 93.75 + 90 + 30 + 0.75 375
10. Fluxapyroxad 62.5% + Epoxiconazole 62.5% EC 46.88 + 46.88 750

Note:

CF-2226 SC: Fluxapyroxad 20% + Azoxystrobin 22 % + Kasugamycin 6% SC

CF-2262 SC: Fluxapyroxad 20% + Azoxystrobin 22 % + Kasugamycin 6% + Soy Protein 0.2% SC

CF-1514 SC: Fluxapyroxad 15% + Azoxystrobin 18 % + Kasugamycin 4% SC

CF-1512 SC: Fluxapyroxad 15% + Azoxystrobin 18 % + Kasugamycin 4% + Soy Protein 0.2% SC

CF-2248 SC: Fluxapyroxad 25% + Azoxystrobin 24 % + Kasugamycin 8 % SC

CF-2282 SC: Fluxapyroxad 25% + Azoxystrobin 24 % + Kasugamycin 8 % + Soy Protein 0.2% SC

EVALUATION OF BIO-EFFICACY AGAINST DISEASE MANAGEMENT
Evaluation method for Bacterial blight disease

Randomly selected 75 leaves/replication and 3 replication/plot were observed and were visually rated as per the rating below. The percent disease index (PDI) was calculated as per given formula:

Grade Spots on Leaves
0 No visible symptoms
1 < 1% leaf area affected
3 1-10% leaf area affected
5 11-25% leaf area affected
7 26-50% leaf area affected
9 >50 leaf area affected

Evaluation method for Sheath blight disease
Randomly selected 25 sheath/replication and 3 replication/plot were observed and visually rated each disease as per the rating below. The percent disease index (PDI) was calculated as per given formula:

Grade Spots on Leaves
0 No visible symptoms
1 < 1% Sheath area affected
3 1-10% Sheath area affected
5 11-25% Sheath area affected
7 26-50% Sheath area affected
9 >50 Sheath area affected

Sum of numerical rating
PDI = ---------------------------------------------------------------------------- x 100
Total no. of plants/Leaves observed X Maximum grade

Table 3: Effect of fungicide treatment on Sheath blight disease of rice at 7 and 14 days after application for CF-2262 SC comprising Fluxapyroxad 20% + Azoxystrobin 22% + Kasugamycin 6% + Soy protein 0.2%

Treatments Product a.i./Ha Dose/Ha 7 DA1A % Control 14 DA1A % Control 7 DA2A % Control 14 DA2A % Control
1 Untreated 17.5 0.00 19.2 0.00 23.7 0.00 29.7 0.00
2 Azoxystrobin 23 115 500 9.4 46.29 10.5 45.31 7.5 68.35 8.3 72.05
3 Kasugamycin 3 L 30 1000 4.3 75.43 4.8 75.00 4.1 82.70 4.3 85.52
4 CF-2226 SC 75 + 82.5 + 22.5 375 2.1 88.00 2.4 87.50 2.2 90.72 2.5 91.58
5 CF-2262 SC 75 + 82.5 + 22.5 + 0.75 375 1.5 91.43 1.7 91.15 1.3 94.51 1.7 94.28
6 CF-1514 SC 56.25 + 67.5 + 15 375 3.5 80.00 3.8 80.21 4.2 82.28 4.8 83.84
7 CF-1512 SC 56.25 + 67.5 + 15 + 0.75 375 3.2 81.71 3.5 81.77 3.9 83.54 4.2 85.86
8 CF-2248 SC 93.75 + 90 + 30 375 2.7 84.57 3.1 83.85 2.9 87.76 3.4 88.55
9 CF-2282 SC 93.75 + 90 + 30 + 0.75 375 1.9 89.14 2.3 88.02 2.7 88.61 2.8 90.57
10 Fluxapyroxad 62.5% + Epoxiconazole 62.5% EC 46.88 + 46.88 750 2.8 84.00 3.4 82.29 3.2 86.50 3.9 86.87

As seen in Table 3, treatment with Azoxystrobin alone at the concentration of 115g a.i/ha results in about 68% control of disease 7 days after second application and treatment with Kasugamycin alone at the concentration of 30g a.i/ha results in about 830% control of disease 7 days after second application . However, unexpectedly and surprisingly, advantageously, the combination of Fluxapyroxad, Azoxystrobin, Kasugamycin and Soy protein at concentration of 75 g a.i/ha + 82.5 g a.i/ha + 22.5 g a.i/ha + 0.75 g a.i/ha respectively results in about 94.51% control of the disease7 days after second application.

Further, treatment with the combination of Fluxapyroxad, Azoxystrobin and Kasugamycin at the concentration of 75 g a.i/ha + 82.5 g a.i/ha + 22.5 g a.i/ha respectively results in about 91% control of the disease 7 days after second application. However, unexpectedly and surprisingly, advantageously, the combination of Fluxapyroxad, Azoxystrobin, Kasugamycin and Soy protein at a concentration of 75 g a.i/ha + 82.5 g a.i/ha + 22.5 g a.i/ha + 0.75 g a.i/ha respectively results in about 94.51% control of the disease 7 days after second application. It can be appreciated that when soy protein is used with the combination of Fluxapyroxad, Azoxystrobin and Kasugamycin, synergistically enhances the activity of fungicidal composition. It can be clearly seen that the result obtained with the formulation code CF-2226 SC were superior to other possible combinations.
Table 4: Bio-efficacy of fungicidal composition (Fluxapyroxad 20% + Azoxystrobin 22% + Kasugamycin 6% + Soy protein 0.2%) on Bacterial leaf blight disease of Rice

Treatments Product a.i./Ha Dose/Ha 7 DA1A % Control 14 DA1A % Control 7 DA2A % Control 14 DA2A % Control
1 Untreated 13.2 0.00 15.3 0.00 17.2 0.00 23.2 0.00
2 Azoxystrobin 23 115 500 9.6 27.27 10.2 33.33 8.1 52.91 8.5 63.36
3 Kasugamycin 3 L 30 1000 4.1 68.94 4.7 69.28 4.1 76.16 4.4 81.03
4 CF-2226 SC 75 + 82.5 + 22.5 375 2 84.85 2.7 82.35 1.6 90.70 1.8 92.24
5 CF-2262 SC 75 + 82.5 + 22.5 + 0.75 375 1.3 90.15 2.3 84.97 1.3 92.44 1.7 92.67
6 CF-1514 SC 56.25 + 67.5 + 15 375 3.2 75.76 4 73.86 4 76.74 4.5 80.60
7 CF-1512 SC 56.25 + 67.5 + 15 + 0.75 375 3.1 76.52 3.7 75.82 3.9 77.33 4.3 81.47
8 CF-2248 SC 93.75 + 90 + 30 375 2.6 80.30 2.9 81.05 2.3 86.63 2.4 89.66
9 CF-2282 SC 93.75 + 90 + 30 + 0.75 375 2.1 84.09 2 86.93 3.1 81.98 3.5 84.91
10 Fluxapyroxad 62.5% + Epoxiconazole 62.5% EC 46.88 + 46.88 750 8.4 36.36 9 41.18 9 47.67 14 39.66

As seen in Table 4 above, treatment with Azoxystrobin alone at the concentration of 115g a.i/ha results in about 53% control of disease 7 days after second application and treatment with Kasugamycin alone at the concentration of 30g a.i/ha results in about 76 % control of the disease 7 days after second application. However, unexpectedly and surprisingly, advantageously, the combination of Fluxapyroxad, Azoxystrobin, Kasugamycin and Soy protein at concentration of 75 g a.i/ha + 82.5 g a.i/ha + 22.5 g a.i/ha + 0.75 g a.i/ha respectively results in 92% control of the disease 7 days after second application.

Further, treatment with the combination of Fluxapyroxad, Azoxystrobin and Kasugamycin at the concentration of 75 g a.i/ha + 82.5 g a.i/ha + 22.5 g a.i/ha respectively results in about 91% control of the disease 7 days after second application. However, unexpectedly and surprisingly, advantageously, the combination of Fluxapyroxad, Azoxystrobin, Kasugamycin and Soy protein at concentration of 75 g a.i/ha + 82.5 g a.i/ha + 22.5 g a.i/ha + 0.75 g a.i/ha respectively results in about 92 % control of the disease 7 days after second application. It can be appreciated that when soy protein is used with the combination of Fluxapyroxad, Azoxystrobin and Kasugamycin, it synergistically enhances the activity of fungicidal composition. It can be clearly seen that the result obtained for the formulation code CF-2262 SC was superior to other possible combinations.

Example 3: Evaluation of Phytotoxicity
Visual observations were recorded 3, 7, and 10 days after the application (DAA) of testing products. The parameters observed were leaf injury on tip/surface, necrosis, vein clearing, epinasty, hyponasty, and wilting. The score scale (1-10) followed for leaf injury on tips/surface is given below in Table.

Table 5: Phytotoxicity symptoms scoring and rating for leaf injury on tip/surface
Leaf injury on tips/surface Rating
0-10% 1
11-20% 2
21-30% 3
31-40% 4
41-50% 5
51-60% 6
61-70% 7
71-80% 8
81-90% 9
91-100% 10

Table 6 Results of phytotoxicity studies conducted for various treatments on rice blight disease

Table 6a: Phytotoxic studies (Necrosis) of various treatments on rice blight disease
Treatments Product a.i./Ha Dose/Ha 1 DA1A 3 DA1A 5 DA1A 10 DA1A 1 DA2A 3 DA2A 5 DA2A 10 DA2A
1 Untreated 0 0 0 0 0 0 0 0
2 Azoxystrobin 23 115 500 0 0 0 0 0 0 0 0
3 Kasugamycin 3 L 30 1000 0 0 0 0 0 0 0 0
4 CF-2226 SC 75 + 82.5 + 22.5 375 0 0 0 0 0 0 0 0
5 CF-2262 SC 75 + 82.5 + 22.5 + 0.75 375 0 0 0 0 0 0 0 0
6 CF-1514 SC 56.25 + 67.5 + 15 375 0 0 0 0 0 0 0 0
7 CF-1512 SC 56.25 + 67.5 + 15 + 0.75 375 0 0 0 0 0 0 0 0
8 CF-2248 SC 93.75 + 90 + 30 375 0 0 0 0 0 0 0 0
9 CF-2282 SC 93.75 + 90 + 30 + 0.75 375 0 0 0 0 0 0 0 0
10 Fluxapyroxad 62.5% + Epoxiconazole 62.5% EC 46.88 + 46.88 750 0 0 0 0 0 0 0 0

Table 6b: Phytotoxic studies (Yellowing) of various treatments on rice blight disease
Treatments Product a.i./Ha Dose/Ha 1 DA1A 3 DA1A 5 DA1A 10 DA1A 1 DA2A 3 DA2A 5 DA2A 10 DA2A
1 Untreated 0 0 0 0 0 0 0 0
2 Azoxystrobin 23 115 500 0 0 0 0 0 0 0 0
3 Kasugamycin 3 L 30 1000 0 0 0 0 0 0 0 0
4 CF-2226 SC 75 + 82.5 + 22.5 375 0 0 0 0 0 0 0 0
5 CF-2262 SC 75 + 82.5 + 22.5 + 0.75 375 0 0 0 0 0 0 0 0
6 CF-1514 SC 56.25 + 67.5 + 15 375 0 0 0 0 0 0 0 0
7 CF-1512 SC 56.25 + 67.5 + 15 + 0.75 375 0 0 0 0 0 0 0 0
8 CF-2248 SC 93.75 + 90 + 30 375 0 0 0 0 0 0 0 0
9 CF-2282 SC 93.75 + 90 + 30 + 0.75 375 0 0 0 0 0 0 0 0
10 Fluxapyroxad 62.5% + Epoxiconazole 62.5% EC 46.88 + 46.88 750 0 0 0 0 0 0 0 0

Table 6c: Phytotoxic studies (Scorching) of various treatments on rice blight disease
Treatments Product a.i./Ha Dose/Ha 1 DA1A 3 DA1A 5 DA1A 10 DA1A 1 DA2A 3 DA2A 5 DA2A 10 DA2A
1 Untreated 0 0 0 0 0 0 0 0
2 Azoxystrobin 23 115 500 0 0 0 0 0 0 0 0
3 Kasugamycin 3 L 30 1000 0 0 0 0 0 0 0 0
4 CF-2226 SC 75 + 82.5 + 22.5 375 0 0 0 0 0 0 0 0
5 CF-2262 SC 75 + 82.5 + 22.5 + 0.75 375 0 0 0 0 0 0 0 0
6 CF-1514 SC 56.25 + 67.5 + 15 375 0 0 0 0 0 0 0 0
7 CF-1512 SC 56.25 + 67.5 + 15 + 0.75 375 0 0 0 0 0 0 0 0
8 CF-2248 SC 93.75 + 90 + 30 375 0 0 0 0 0 0 0 0
9 CF-2282 SC 93.75 + 90 + 30 + 0.75 375 0 0 0 0 0 0 0 0
10 Fluxapyroxad 62.5% + Epoxiconazole 62.5% EC 46.88 + 46.88 750 0 0 0 0 0 0 0 0

As seen in Table 6a-c, no visible phytotoxicity was observed associated with the treatment of any of the ingredients to the plants, suggesting that said composition is completely safe to use.

The present invention thus provides a novel and synergistic fungicidal combination of Fluxapyroxad as the first component, Azoxystrobin as the second component, Kasugamycin as the third component and Soy protein as the fourth component, which significantly enhances the economic value of crops. While the present specification illustrates various concentrations across which enhanced results are seen, in particular, the combination of Fluxapyroxad, Azoxystrobin, Kasugamycin and Soy protein at an active concentration of 75 g a.i/ha + 82.5 g a.i/ha + 22.5 g a.i/ha + 0.75 g a.i/ha respectively shows the best results in controlling sheath blight disease and bacterial leaf blight disease on rice crop. There is a clear and significant enhancement in efficacy as compared to solo application of actives, further enhanced by addition of Soyprotein in desired weight percentage which is unexpected and surprising.
, Claims:We Claim:
1. A fungicidal composition, wherein said fungicidal composition comprises:
a. Fluxapyroxad in an amount ranging from 10-30% w/w;
b. Azoxystrobin in an amount ranging from 10-30% w/w;
c. Kasugamycin in an amount ranging from 1-10% w/w;
d. Soy protein in an amount ranging from 0.05-5% w/w; and
e. at least one agriculturally acceptable excipient.

2. The fungicidal composition as claimed in claim 1, wherein said at least one agriculturally acceptable excipient is selected from the group comprising anti-freezing agent, dispersing agent, wetting agent, anti-foaming agent, a thickener, one biocide, at least one vegetable oil, demineralized water and combinations thereof.

3. The fungicidal composition as claimed in claim 2, wherein said anti-freezing is present in an amount ranging from 5-15% w/w, said dispersing agent is present in an amount ranging from 2-10% w/w, said wetting agent is present in an amount ranging from 1-7% w/w, said vegetable oil is present in an amount ranging from 0.1-6% w/w, said anti- foaming agent is present in an amount ranging from 0.2-3% w/w, said biocide is present in amount ranging from 0.05-2% w/w, said thickener is present in an amount ranging from 0.5-10% w/w and demineralized water is present in amount ranging from 20-70% w/w.

4. The fungicidal composition as claimed in claim 1, wherein said fungicidal composition is formulated in a form selected from the group consisting 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), macro granules (GG), dry flowable (DF), aqueous Suspo-emulsions (SE), capsule suspensions (CS) and microgranules (MG).

5. The fungicidal composition as claimed in claim 4, wherein said fungicidal composition is in the form of suspension concentrates (SC) formulation.

6. The fungicidal composition as claimed in claim 1, wherein Fluxapyroxad is present in an amount of 20% w/w, Azoxystrobin is present in an amount of 22% w/w, Kasugamycin is present in an amount of 6% and Soy protein is present in an amount of 0.2% w/w of the fungicidal composition.

7. A process for preparing the fungicidal composition as claimed in claim 1 in the form of a suspension concentrate formulation, said process comprising:
(i) mixing surfactant, anti-freezing agent in D.M. water, followed by solubilizing in high-speed mixer at 500-1000rpm at room temperature;
(ii) adding anti foaming agent, Fluxapyroxad technical and Azoxystrobin technical, followed by addition of Kasugamycin and rest of the adjuvants to the mixture of step (i) to obtain a homogenous mass;
(iii) grinding the homogenous mass of step (ii) in a sand mill until a mean particle size of <5 micron is obtained at a temperature of 15-25ºC;
(iv) adding 2% solution of xanthan gum and biocide to step (iii) under low stirring and mixing well at the speed of 400-700 rpm;
(v) adding soy protein and vegetable oil to the mixture obtained in step (iv) in continuous stirring till homogenous mixture is obtained in suspension formulation.

8. A method for controlling pests in various agriculture crops, said method comprising applying fungicidally effective amount of a fungicidal composition as claimed in claims 1-6 to the target crop or portion thereof.

9. The method as claimed in claim 8, wherein said fungicidally effective amount of the fungicidal composition comprises:
a. Fluxapyroxad in an amount ranging from of 10-30% w/w;
b. Azoxystrobin in an amount ranging from10-30% w/w;
c. Kasugamycin in an amount ranging from 1-10% w/w;
d. Soy protein in an amount ranging from 0.05-5% w/w; and
e. at least one agriculturally acceptable excipient.