DESC:FUNGICIDAL COMPOSITION COMPRISING PICOXYSTROBIN AND VALIDAMYCIN
FIELD OF THE INVENTION
The present invention relates to synergistic fungicidal compositions, methods of preparing and using such compositions for control of undesired plant diseases. More particularly, the present invention relates to synergistic fungicidal compositions comprising picoxystrobin, validamycin, and excipients.
BACKGROUND OF THE INVENTION
Fungicides are natural or synthetic compounds that act to protect plants from damage caused by fungi. Current agricultural methods are highly dependent on the use of fungicides. In fact, some crops cannot grow usefully without the use of fungicides. The use of fungicides allows growers to improve the yield and quality of the crop and consequently increase the value of the crop. In most situations, increasing the value of the crop is at least three times the cost of using the fungicide.
Various agricultural and horticultural fungicides have been known, but there are a great variety of diseases to be controlled and it is difficult to specify the kind of the disease in practice and control the disease by choosing a fungicide suitable for the disease. However, not all fungicides are useful in all situations, and repeated use of a single fungicide frequently leads to the development of resistance to the fungicide and related fungicides. As a result, research to produce fungicides and combinations of multiple fungicides that are safer, have better performance, require less dose, are easier to use, and cost less has been implemented. Combinations that produce synergy, i.e., where the activity of two or more compounds exceeds the activity of these compounds when used alone, have also been investigated.
In addition, it is necessary to cope with the appearance of fungi which have acquired tolerance to specific agents and the occurrence of novel diseases due to the change of the mode of agriculture. For these reasons and the like, there is desired a fungicidal composition having a high activity and a wide anti-microbial spectrum.
The present invention provides a fungicidal composition which has a high fungicidal activity and a wide antimicrobial spectrum and is also markedly effective against diseases caused by plant pathogenic fungi which have acquired tolerance to certain fungicides.

OBJECT OF THE INVENTION
It is an objective of the present invention to provide synergistic fungicidal compositions comprising combination of picoxystrobin and validamycin.
It is another objective of the present invention to provide synergistic fungicidal compositions to control broad spectrum of phytopathogenic microorganisms.
It is yet another objective of the present invention to provide a synergistic fungicidal composition with low toxicity to humans or other mammals.
It is yet another objective of the present invention to provide a synergistic fungicidal composition resulting in increased yield of the crop and enhanced fungicidal activity.
It is yet another objective of the present invention to provide a method for preparing fungicidal compositions of the present invention.
SUMMARY OF THE INVENTION
The present invention provides a fungicidal composition comprising as its active ingredient validamycin present in a range from 4% to 20% (w/w), picoxystrobin present in a range from 8% to 20% (w/w) and excipients wherein the excipients are selected from the group comprising a wetting agent, a dispersing agent, an anti-freezing agent, a biocide, a viscosity modifier, a defoamer and a base.
In an embodiment of the present invention, the wetting agent is selected from the group comprising Ethoxylated TSP-phosphate ester, Amine salt of poly aryl phenyl ether phosphate, polyoxyethylenated alkylphenol, dialkyl naphthalene sulphonate sodium salt, alkyl naphthalene sulfonate (ANS) condensate blend, sodium ligno sulphonate, linear alcohol derivative and polyethylene glycol nonyl phenyl ether ammonium sulfate present in an amount of 0.2 to 1.5% (w/w).
In an embodiment of the present invention, the dispersing agent is selected from the group comprising EO/PO polymeric dispersant, Ethoxylated TSP-phosphate ester, Polyoxy alkylene glycol butyl ether, Polyoxyethylene-Polyoxypropylene Block Polymer, Polyethylene glycol 2,4,6-tris(1-phenylethyl)phenyl ether sulfate ammonium salt, Methyl methacrylate graft copolymer, Sodium polyalkyl naphthalene sulfonate, Sodium salt of methyl naphthalene sulfonate, sodium ligno sulphonate, acrylate copolymer, phenol sulfonic acid-formaldehyde-polycondensation as sodium salt, sodium polycarboxylate, sodium methyl oleoyl taurate and sodium lauryl sulphate or combination thereof present in an amount of 0.2 to 5% (w/w).
In an embodiment of the present invention, the anti-freezing agent is selected from propylene glycol, glycerin, and mono ethylene glycol, present in an amount of 1.0% (w/w).
In an embodiment of the present invention, the biocide is selected from 1,2-Benzisothiazol-3(2H)-one and formaldehyde, present in an amount of 0.1% (w/w).
In an embodiment of the present invention, the viscosity modifier is selected from the group comprising of xanthan gum, carboxymethyl cellulose, and polyvinyl pyrrolidine (PVP), present in an amount of 1.0% (w/w).
In an embodiment of the present invention, the defoamer is Polydimethylsiloxane antifoam emulsion, present in an amount of 0.5% (w/w).
In an embodiment of the present invention, the base is water present in an amount of 54% to 85% (w/w).
In an embodiment of the present invention, the composition is formulated as a suspension concentrate.
DESCRIPTION OF THE INVENTION
The following detailed description is presented to enable any person skilled in the art to make and use the invention. For purposes of explanation, specific nomenclature is set forth to provide a thorough understanding of the present application. However, it will be apparent to one skilled in the art that these specific details are not required to practice the invention. Descriptions of specific applications are provided only as representative examples. The present application is not intended to be limited to the embodiments shown but is to be accorded the widest possible scope consistent with the principles and features disclosed herein.
It is to be noted that, as used in the specification, the singular forms "a", "an" and "the" include plural referents unless the context clearly dictates otherwise. Thus, for example, reference to a composition containing “a compound” includes a mixture of two or more compounds. It should also be noted that the term “or” is generally employed in its sense including “and/or” unless the content clearly dictates otherwise.
Similarly, the words "comprise", "comprises", and "comprising" are to be interpreted inclusively rather than exclusively. Likewise, the terms "include", "including", and "or" should all be construed to be inclusive, unless such a construction is clearly prohibited from the context. However, the embodiments provided by the present disclosure may lack any element that is not specifically disclosed herein. Thus, a disclosure of an embodiment defined using the term "comprising" is also a disclosure of embodiments "consisting essentially of” and "consisting of” the disclosed components. Where used herein, the term "example" particularly when followed by a listing of terms, is merely exemplary and illustrative, and should not be deemed to be exclusive or comprehensive. Any embodiment disclosed herein can be combined with any other embodiment disclosed herein unless explicitly indicated otherwise.
The expression of various quantities in terms of “% w/w” or “%” means the percentage by weight, relative to the weight of the total solution or composition unless otherwise specified.
As used herein, the terms "crops" and "vegetation" can include, for instance, dormant seeds, germinant seeds, emerging seedlings, plants emerging from vegetative propagules, immature vegetation, and established vegetation.
As used herein, a "fungicidally effective amount" may be understood to include an amount of an active ingredient that causes a "fungicidal effect," i.e., an adversely modifying effect including, for instance, a deviation from natural growth or development, killing, regulation, desiccation, growth inhibition, growth reduction, and retardation of fungi.
As used herein, immature vegetation may be understood to include small vegetative plants prior to reproductive stage, and mature vegetation may be understood to include vegetative plants during and after the reproductive stage.
In an embodiment of the present invention, the fungicidal composition of the present invention comprises of picoxystrobin and validamycin.
In an embodiment of the present invention, the fungicidal composition of the present invention comprises of picoxystrobin, validamycin and excipients.
In another embodiment of the invention, the present invention provides a method for preparing the compositions of picoxystrobin and validamycin.
Picoxystrobin is a cereal fungicide used to control a wide range of diseases including brown rust, tan spot, powdery mildew and net blotch. It has a role as a mitochondrial cytochrome-bc1 complex inhibitor and an antifungal agrochemical. Picoxystrobin inhibits mitochondrial respiration. Picoxystrobin binds at the Qo centre on cytochrome b and block electron transfer between cytochrome b and cytochrome c1. This disrupts the energy cycle within the fungus by halting production of ATP.
It differs from the existing strobilurins due to its complete biokinetic properties, such as a rapid uptake, systemic and translaminar movements, vapour phase activity, strong affinity with the wax layer, a protection of newly formed organs and a curative activity.
Picoxystrobin, chemically known as (E)-Methyl 3-methoxy-2-(2-(((6-(trifluoromethyl) pyridin-2-yl)oxy)methyl)phenyl)acrylate is the new strobilurin fungicide developed by Syngenta to control cereal diseases in early season. The structure of picoxystrobin is represented as

Validamycin is an antibiotic and fungicide produced by Streptomyces hygroscopicus. It is used as an inhibitor of trehalase. It is produced from fermentation of Streptomyces hygroscopicus variety limoneus (207, 243). It is most effective against soil borne diseases and is used for the control of Rhizoctonia solani in rice, potatoes, vegetables, and others as well as damping off diseases in vegetable seedlings, cotton, sugar beets, rice and other plants. Validamycin is structurally similar to trehalose and acts as a competitive inhibitor for the enzyme trehalase, which is widely spread among animals, plants, insects, and microorganisms. Trehalase enzymes are responsible for trehalose degradation and serves an important role in regulation of such mechanisms as the active transport of glucose into the intestines, reserve supply of energy, germination of spores, etc. It acts on hyphae and destroys the fungus by its contact action and controls the spread of diseases. The structure of validamycin is represented as

The active ingredients constituting the fungicidal composition of the present invention are, in the same manner as conventional agricultural chemicals, mixed with various adjuvants.
Suitable adjuvants may be a solid or liquid and are generally a substance commonly used in formulation processing process, for example, natural or regenerated minerals, solvents, dispersing agents, wetting agents, adhesives, thickeners or viscosity modifiers, biocide, anti-freezing agent, defoamer, stabilizers, binders or fertilizers.
The compositions of the present invention are formulated into various formulations such as Granular composition (GR), Capsule suspension (CS), Dispersible concentrate (DC), Dustable powder (DP), Powder for dry seed treatment (DS), Emulsifiable concentrate (EC), Emulsifiable granule (EG), Emulsion water-in-oil (EO), Emulsifiable powder (EP), Emulsion for seed treatment (ES), Emulsion oil-in-water (EW), Flowable concentrate for seed treatment (FS), Granules (GR), Micro-emulsion (ME), Oil-dispersion (OD), Oil miscible flowable concentrate (OF), Oil miscible liquid (OL), Oil dispersible powder (OP), Suspension concentrate (SC), Suspension concentrate for direct application (SD), Suspo-emulsion (SE), Water soluble granule (SG), Soluble concentrate (SL), Spreading oil (SO), Water soluble powder (SP), Water soluble tablet (ST), Ultra-low volume (ULV) suspension, Tablet (TB), Ultra-low volume (ULV) liquid, Water dispersible granules (WG), Wettable powder (WP), Water dispersible powder for slurry seed treatment (WS), Water dispersible tablet (WT), a mixed formulation of CS and SC (ZC), a mixed formulation of CS and SE (ZE), a mixed formulation of CS and EW (ZW). Preferably, the composition of the present invention is in form of suspension concentrate (SC).
In an embodiment of the present invention, the fungicidal composition comprises picoxystrobin in range from 8% to 20% (w/w) and validamycin in range from 4% to 20% (w/w).
In an embodiment, the compositions of the present invention may typically be produced by mixing the actives in the composition with an inert carrier and adding surfactants and other excipients and carriers as needed for various formulations. During application, a common excipient can be mixed with the composition.
In an embodiment, a method for controlling plant diseases, comprises applying the fungicidal composition of the present invention to agricultural and horticultural plants. The concentration of the fungicidal composition of the present invention cannot generally be defined, as it varies depending upon the crop plants to be treated, the application method, the type of the formulation, the dose, etc.
In accordance with the method of the present invention, other agricultural chemicals such as a fungicide, an insecticide, an acaricide, a nematicide, an antiviral agent, an attractant, a herbicide, a plant growth regulator and the like, can be used in combination, and such a case can sometimes show additional superior effect.
The present invention provides a fungicidal composition for agricultural and horticultural use in which the control effect of a plant disease, in particular, an effect for preventing and/or treating a plant disease, and a method for controlling a plant disease using the composition.
Wetting is the first stage of dispersion, in which the air surrounding the granular composition is substituted with water. Wetting of the fungicidal composition with water cannot occur if the surface tension of the liquid is very high. Hence, it is recommended to add a wetting agent to the fungicidal composition to facilitate the process of dispersion of the granules in the liquid. Accordingly, the composition of the present invention preferably contains a wetting agent suitable for use in the present invention are Ethoxylated TSP-phosphate ester, Amine salt of poly aryl phenyl ether phosphate, polyoxyethylenated alkylphenol, dialkyl naphthalene sulphonate sodium salt, alkyl naphthalene sulfonate (ANS) condensate blend, sodium ligno sulphonate, linear alcohol derivative and polyethylene glycol nonyl phenyl ether ammonium sulfate. Preferably, the wetting agent is present in a range from 0.2% to 1.5% (w/w).
It is generally observed that solid particles in a liquid undergo spontaneous aggregation to form lumps. Hence it is recommended to add a dispersant or a dispersing agent which prevents aggregation of solid particles and keeps them suspended in the fluid. One or more dispersing agents may be used in the synergistic composition of the present invention. Accordingly, the composition of the present invention preferably contains a dispersing agent suitable for use in the present invention are EO/PO polymeric dispersant, Ethoxylated TSP-phosphate ester, Polyoxy alkylene glycol butyl ether, Polyoxyethylene-Polyoxypropylene Block Polymer, Polyethylene glycol 2,4,6-tris(1-phenylethyl)phenyl ether sulfate ammonium salt, Methyl methacrylate graft copolymer, Sodium polyalkyl naphthalene sulfonate, Sodium salt of methyl naphthalene sulfonate, sodium ligno sulphonate, acrylate copolymer, phenol sulfonic acid-formaldehyde-polycondensation as sodium salt, sodium polycarboxylate, sodium methyl oleoyl taurate and sodium lauryl sulphate or combination thereof. Preferably, the dispersing agent is present in a range from 0.2% to 5% (w/w) in the compositions of the present invention.
Antifreezing agent is an additive which lowers the freezing point of a water-based liquid. An anti-freezing agent is used in suspension concentrate so as to prevent freezing of water. The suitable anti-freezing agent for the present invention is selected from the group comprising of Propylene glycol, Glycerin, and Mono ethylene glycol. Preferably, the anti-freeze agent is present in an amount of 1.0% (w/w).
Biocide is a chemical agent intended to destroy, render harmless, or control potentially harmful organism. The biocide suitable for use in the present invention is selected from the group comprising of 1,2-Benzisothiazol-3(2H)-one and formaldehyde. Preferably, the biocide is present in an amount of 0.1% (w/w).
It is essential in suspension concentrate to prevent the dispersed solid particles from sedimentation upon storage. The sedimentation may adversely affect efficacy of the composition during application. In order to maintain uniform distribution of active ingredient during shelf life, viscosity modifiers are used, like xanthan gum. Xanthan gum thickeners generate a supporting structure within the aqueous phase through controlled incompatibility with the water phase. Suitable viscosity modifiers for the purpose of the present invention are xanthan gum, Carboxymethyl cellulose, and polyvinyl pyrrolidine (PVP). Preferably, the viscosity modifier is present in an amount of 1.0% (w/w).
The composition of the suspension concentrate formulation, excipients, surfactant, and water, promotes the generation of foam in the presence of high shear equipment. Foam may adversely affect the efficiency of processing equipment and the bulk density of the formulation during packaging. Therefore, defoamers are incorporated into the formulation in order to eliminate foam formation during processing. Suitable defoamers for the present invention is Polydimethylsiloxane antifoam emulsion, present in an amount of 0.5% (w/w).
The base is liquid media and can be water or any organic/inorganic solvent in which the solid meets the insolubility criteria, under all temperature conditions. The base suitable for the present invention is distilled water. Preferably, the base is present in a range from 54% to 85% (w/w).
One kind or two or more kinds of other fungicides or pesticides, and synergists may be mixed with the agricultural and horticultural fungicidal composition of the present invention so long as they do not interfere with the effects of the present invention.
A method for controlling plant diseases, which comprises applying the fungicidal composition of the present invention to agricultural and horticultural plants, is also included in the present invention. The concentration of the fungicidal composition of the present invention cannot generally be defined, as it varies depending upon the crop plants to be treated, the application method, the type of the formulation, the dose, etc.
The fungicidal composition of the present invention, which has been formulated into a preparation, may be used such as spreading (spreading, spraying, misting, atomizing, grain diffusing or application on water surface), soil application (such as mixing or irrigation) or surface application (such as coating, dust coating or covering) in the form of a solution, a suspension, or an emulsion preferably, suspension concentrate.
Examples of useful plants to be treated in the fungicidal composition of the present invention include cereals, vegetables, root vegetables, potatoes, trees, grasses, and lawn. In this case, each part of these plants may be subjected to the treatment. Examples of the part of the plants include leaves, stems, flowers, buds, fruits, seeds, sprouts, roots, tubers, tuberous roots, shoots, and cuttings. It is also possible to subject the improved varieties/variants of these plants, and cultivars, mutants, hybrid bodies, or genetically modified bodies (GMO) to the treatment.
The fungicidal composition of the present invention can control various diseases generated upon cultivation of agricultural and horticultural crops, including flowers, lawn, and glasses by seed treatment, foliar spray, soil application, water application, or the like. Furthermore, the fungicidal composition of the present invention has an excellent fungicidal effect even on resistant fungi. Further, since the fungicidal composition exhibits the effect even when used at very low doses, it has an effect of preventing the emergence of new resistant fungi.
The fungicide composition for agricultural and horticultural use according to the present invention exhibits an excellent controlling effect against plant diseases even at a very low dosage and is free from concerns about phytotoxicity to useful plants. The fungicide composition for agricultural and horticultural use according to the present invention exhibits a remarkable synergistic plant disease control effect which cannot be predicted from the plant disease control effect obtained when using validamycin alone or picoxystrobin alone.
The present invention is more particularly described in the following examples that are intended as illustrations only, since numerous modifications and variations within the scope of the present invention will be apparent to those of skill in the art. Unless otherwise noted, all parts, percentages, and ratios reported in the following examples are on a weight basis and all reagent used in the example were obtained or are available from the chemical suppliers.
Examples:
Example 1: Preparation of composition of Validamycin and Picoxystrobin in suspension concentrate (SC) form.
The composition of Validamycin and Picoxystrobin in suspension concentrate (SC) form is prepared by following steps as below:
Step 1: Preparation of Aqueous Slurry
? Taking 10 parts by weight of propylene glycol (anti-freezing agent), in Vessel No. 1 (pre mixer). Adding 5 parts by weight of EO/PO Polymeric dispersant (dispersing agent/ wetting agent) and mixing well after 15 min adding 0.1 parts by weight of 1,2-Benzisothiazol-3(2H)-one (biocide) followed by 0.1 parts by weight of Polydimethylsiloxane antifoam emulsion (defoamer), xanthan gum (viscosity modifier) after 15 min, add 100 % of distilled water.
? Continuing the mixture for 20 minutes and then adding 3 parts by weight of validamycin (active) over the period of 15 min after that adding 7.5 parts by weight of picoxystrobin (active) over the period of 15 min.
? Continuing the mixture for homogenization (the mixture should be thoroughly homogenized) for 30 min.
Step 2: Wet Milling of Slurry
? Passing the wet slurry from Vessel No. 1 through Dyno-Mill at temperature less than 30 °C and collect material in Vessel No.2 (post-Mixer).
? Maintaining continuous flowrate of the Dyno mill to avoid the sedimentation.
? Submitting the sample to quality control department to check the particle size distribution (PSD). The PSD of the material should be d90 < 5 µ. If the d90 is higher than > 5 µ then grind the mixture further till the PSD value complies the specification.
? Collecting the sample to a vessel No. 2 under homogenization.
Step 3: Preparation of Gel Suspension
? Adding 10 parts by weight of Xanthan gum (viscosity modifier) in 4.0 parts by weight of Propylene glycol (anti-freezing agent) then add 0.1 part by weight 1,2-Benzisothiazol-3(2H)-one (biocide) to this slurry into 93.95 parts by weight of distilled water and stir the mixture until it gets lumps free homogeneous mixture.
? Keeping this gel as such for 4-5 hours to get a translucent homogeneous gel.
Step 4: Gel addition
? Adding required quantity of Gel (prepared above) to the wet slurry in Vessel No. 2 and stir the mixture for ~3h for homogenization.
? Submitting the sample for quality control for complete analysis.
? Packing the material in suitable bottle.
The process illustrated above may be used for preparation of other composition of Validamycin and Picoxystrobin in suspension concentrate (SC) form.
Example 2: Compositions of the present invention
The illustrative embodiments show the composition of Validamycin and Picoxystrobin in suspension concentrate (SC) form in different amount as follows:
Example-1: Validamycin 6% + Picoxystrobin 10% SC
S. No. Ingredients Function % (w/w)
1. Validamycin Fungicide 6.0
2. Picoxystrobin Fungicide 10.0
3. EO/PO Polymeric dispersant Dispersing agent 5.0
4. Propylene glycol Antifreezing agent 1.0
5. 1,2-Benzisothiazol-3(2H)-one Biocide 0.1
6. Xanthan gum Viscosity Modifier 1.0
7. Polydimethylsiloxane antifoam emulsion Defoamer 0.5
8. Distilled water Base 76.4
Total 100

Example-2: Validamycin 4% + Picoxystrobin 8% SC
S. No. Ingredients Function % (w/w)
1. Validamycin Fungicide 4.0
2. Picoxystrobin Fungicide 8.0
3. Ethoxylated TSP-phosphate ester Wetting/Dispersing agent 0.2
4. Polyoxyalkylene Glycol Butyl Ether Dispersing agent 0.2
5. Propylene glycol Antifreezing agent 1.0
6. 1,2-Benzisothiazol-3(2H)-one Biocide 0.1
7. Xanthan gum Viscosity Modifier 1.0
8. Polydimethylsiloxane antifoam emulsion Defoamer 0.5
9. Distilled water Base 85.0
Total 100

Example-3: Validamycin 10% + Picoxystrobin 12% SC
S. No. Ingredients Function % (w/w)
1. Validamycin Fungicide 10.0
2. Picoxystrobin Fungicide 12.0
3. Polyethylene glycol 2,4,6-tris(1-phenylethyl)phenyl ether sulfate ammonium salt Dispersing agent 0.2
4. Amine salt of poly aryl phenyl ether phosphate Wetting agent 0.2
5. propane-1,2-diol Antifreezing agent 1.0
6. 1,2-Benzisothiazol-3(2H)-one Biocide 0.1
7. Xanthan gum Viscosity Modifier 1.0
8. Polydimethylsiloxane antifoam emulsion Defoamer 0.5
9. Distilled water Base 75.0
Total 100

Example-4: Validamycin 12% + Picoxystrobin 14% SC
S. No. Ingredients Function % (w/w)
1. Validamycin Fungicide 12.0
2. Picoxystrobin Fungicide 14.0
3. Polyoxyethylene-Polyoxypropylene Block Polymer Dispersing agent 1.5
4. Amine salt of poly aryl phenyl ether phosphate Wetting agent 1.5
5. propane-1,2-diol Antifreezing agent 1.0
6. 1,2-Benzisothiazol-3(2H)-one Biocide 0.1
7. Xanthan gum Viscosity Modifier 1.0
8. Polydimethylsiloxane antifoam emulsion Defoamer 0.5
9. Distilled water Base 68.4
Total 100

Example-5: Validamycin 20% + Picoxystrobin 20% SC
S. No. Ingredients Function % (w/w)
1. Validamycin Fungicide 20.0
2. Picoxystrobin Fungicide 20.0
3. Polyoxyethylenated alkylphenol Wetting agent 1.5
4. Methyl methacrylate graft copolymer Dispersing agent 1.5
5. propane-1,2-diol Antifreezing agent 1.0
6. 1,2-Benzisothiazol-3(2H)-one Biocide 0.1
7. Xanthan gum Viscosity Modifier 1.0
8. Polydimethylsiloxane antifoam emulsion Defoamer 0.5
9. Distilled water Base 54.4
Total 100

Example 3: Efficacy study for combination of Validamycin and Picoxystrobin in suspension concentrate (SC) form
Field Bio-efficacy trials: Pomegranate Anthracnose, Bacterial leaf spot.
SYNERGY STUDIES:
After calculating % disease control, the synergism was calculated by below formula:
The synergistic fungicidal action of the inventive mixtures calculated by Colby’s formula as follows:
Appropriate analysis of plant response to fungicide combination is critical in determining the type of activity observed. The most widely used model is one Gowing* derived and Colby** modified. Gowing described a mathematical formula for calculating the predicting response values for pesticide mixtures. He suggested the expected (E) percent inhibition of growth induced by pesticide A plus pesticide B is as follows, *(Jerry Flint et al, 1988)***
If A1 = the percent inhibition of growth by pesticide A at given rate
B1 = the percent inhibition of growth by pesticide B at given rate then,

When the percentage of pesticidal control observed for the combination is greater than the expected percentage, there is a synergistic effect. (Ratio of O/E > 1, means synergism observed.)
Reference:
*Gowing, D. P. 1960. Comments on tests of fungicide mixtures. Weeds 8:379–391.
**Colby, S. R. 1967. Calculating synergistic and antagonistic responses of fungicide combinations. Weeds 15:20–22
*** Jerry Flint et al, 1988. Analyzing Fungicide Interactions: A Statistical Treatment of Colby's Method. Weed Technology 2: 304-309

Trial 1: Bio-efficacy against pomegranate anthracnose
The anthracnose is caused by the fungal pathogen Colletotrichum gloeosporioides. This is potentially devasting disease and the fungus cause wide range of symptoms depending upon tissue attack and weather conditions can infect pomegranate foliage at any stage of crop development. There is a small regular or irregular black or brown coloured spots appear on the leaves, stem, flowers & fruits which turn later as dark brown spots. The outbreak of this disease occurs during August-September. Frequent rainfall, high humidity about 50-80 % and the temperature range between 25-30°C favours this disease.
The field trial was conducted to evaluate the efficacy of innovative mixtures of Picoxystrobin and Validamycin against Colletotrichum gloeosporioides fungus in pomegranate crop. Trial was conducted with randomized block design with net plot size of 5m x 6m. Pomegranate crop was raised with all standard agronomic practices. Spraying was done with manual operated backpack knapsack sprayer with 500 – 1000 L of water spray volume per hectare at 45 to 50 days after pruning.
The visual observations were recorded for % disease control from ten plants per plot. The observations were recorded at before spraying, 7 DAA (Days after application) and 14 DAA (Days after application).
Table 6: Anthracnose disease control in pomegranate:
Compositions Dose (gram active per hectare) Percent disease control –Anthracnose.
07 DAA 14 DAA
Validamycin 6% + Picoxystrobin 10% SC 120 90 80
Validamycin 45 45 25
Picoxystrobin 75 75 65
SC – Suspension concentrates, and DAA - Days after application.
The trial results show excellent efficacy of Validamycin 6% + Picoxystrobin 10% SC combinations against Anthracnose disease of Pomegranate. The solo application of two active ingredients tested here, were also not able to provide satisfactory control of anthracnose disease. The combination of Validamycin 6% + Picoxystrobin 10% SC found very promising against Pomegranate anthracnose in terms of efficacy as well as residual control.

Table 7: Percent Anthracnose disease control at 14 DAA
Compositions Dose
(g or ml/ha) % Anthracnose disease control
Expected Actual
Validamycin 45 25
Picoxystrobin 75 65
Validamycin 6% + Picoxystrobin 10% SC 120 73.91 80
Ratio of O/E 1.08

The results in table 7 clearly demonstrates synergy between Picoxystrobin and Validamycin in controlling Pomegranate Anthracnose disease. The large difference between the observed and the expected efficacy clearly demonstrates the synergistic effect of the combination.

Trial 2: Bio-efficacy against Pomegranate Bacterial leaf spot
Bacterial blight caused by Xanthomonas axonopodis pv. punicae is a major disease of pomegranate. Appearance of one to several small water soaked, dark-coloured irregular spots on leaves resulting in premature defoliation under severe cases. The pathogen also infects stem and branches causing girdling and cracking symptoms. Spots on fruits were dark brown irregular slightly raised with oily appearance, which split open with L-shaped cracks under severe cases.
Primary source of inoculum is infected cuttings and secondary source of inoculum spreads through wind splashed rains. The increase in day temperature (38.6°C) and afternoon relative humidity of 30.4% along with cloudy weather and intermittent rainfall favoured the disease initiation and further spread of the disease.
The field trial was conducted to evaluate the efficacy of innovative mixtures of Picoxystrobin and Validamycin against Xanthomonas axonopodis pv. punicae in Pomegranate crop. Trial was conducted with randomized block design with net plot size of 5m x 6m. Pomegranate crop was raised with all standard agronomic practices. Spraying was done with manual operated backpack knapsack sprayer with 500 L of water spray volume per hectare at 45 days after transplanting.
The visual observations were recorded for % disease control from ten hills per plot. The observations were recorded at before spraying, 7 DAA (Days after application) and 14 DAA (Days after application).
Table 8: Bacterial leaf spot disease control in Pomegranate
Compositions Dose (gram active per hectare) Percent disease control –Bacterial leaf spot.
07 DAA 14 DAA
Validamycin 6% + Picoxystrobin 10% SC 120 90 80
Validamycin 45 75 50
Picoxystrobin 75 50 30
SC – Suspension concentrates, and DAA - Days after application.

The trial results show excellent efficacy of Validamycin 6% + Picoxystrobin 10% SC combinations against Bacterial leaf spot disease of Pomegranate. The solo application of two active ingredients tested here, were also not able to provide satisfactory control of Bacterial leaf spot disease. The combination of Validamycin 6% + Picoxystrobin 10% SC found very promising against Pomegranate Bacterial leaf spot in terms of efficacy as well as residual control.

Table 9: Percent Bacterial leaf spot disease control at 14 DAA
Compositions Dose
(g or ml/ha) % Bacterial leaf spot disease control
Expected Actual
Validamycin 45 50
Picoxystrobin 75 30
Validamycin 6% + Picoxystrobin 10% SC 120 65.15 80.2
Ratio of O/E 1.23

The results in table 9 clearly demonstrates synergy between Picoxystrobin and Validamycin in controlling Pomegranate Bacterial leaf spot disease. The large difference between the observed and the expected efficacy clearly demonstrates the synergistic effect of the combination.

Field Bio-efficacy trials: Rice Sheath blight, Bacterial leaf blight
Trial 1: Bio-efficacy against rice sheath blight
The sheath blight is caused by the fungal pathogen Rhizoctonia solani. This is potentially devasting disease can infect rice foliage at any stage of crop development. Its major threat to many crops and can also affect the crop stand and yield. Rhizoctonia solani fungus is cosmopolitan, polyphagous, widely distributed in tropical, subtropical, and temperate regions and for its development required hot and humid environment. The fungus spreads in the field by growing its runner hyphae from tiller to tiller, from leaf to leaf, and from plant to plant, resulting in a circular pattern of damage. The infection spreads most quickly when susceptible varieties are grown under favourable conditions such as warm temperature (28 to 32°C), high humidity (95% or above), and dense stands with a heavily developed canopy.
The field trial was conducted to evaluate the efficacy of innovative mixtures of Picoxystrobin and Validamycin against Rhizoctonia solani fungus in rice crop. Trial was conducted with randomized block design with net plot size of 5m x 6m. Rice crop was raised with all standard agronomic practices. Spraying was done with manual operated backpack knapsack sprayer with 500 L of water spray volume per hectare at 45 days after transplanting.
The visual observations were recorded for % disease control from ten hills per plot. The observations were recorded at before spraying, 7 DAA (Days after application) and 14 DAA (Days after application).
Table 10: Sheath blight disease control in rice
Compositions Dose (gram active per hectare) Percent disease control –Sheath blight.
07 DAA 14 DAA
Validamycin 6% + Picoxystrobin 10% SC 120 90.4 85.9
Validamycin 45 50 40
Picoxystrobin 75 70 60
SC – Suspension concentrates, and DAA - Days after application.

The trial results show excellent efficacy of Validamycin 6% + Picoxystrobin 10% SC combinations against sheath blight disease of rice. The solo application of two active ingredients tested here, were also not able to provide satisfactory control of sheath blight disease. The combination of Validamycin 6% + Picoxystrobin 10% SC found very promising against rice sheath blight in terms of efficacy as well as residual control.

Table 11: Percent sheath blight disease control at 14 DAA
Compositions Dose
(g or ml/ha) % Sheath blight disease control
Expected Actual
Validamycin 45 40
Picoxystrobin 75 60
Validamycin + Picoxystrobin 16% SC 120 76.24 85.5
Ratio of O/E 1.12

The results in table 11 clearly demonstrates synergy between Picoxystrobin and Validamycin in controlling rice sheath blight disease. The large difference between the observed and the expected efficacy clearly demonstrates the synergistic effect of the combination.

Trial 2: Bio-efficacy against rice bacterial leaf blight
Bacterial blight is caused by Xanthomonas oryzae pv. oryzae. This is potentially devasting disease and it causes wilting of seedlings and yellowing and drying of leaves. The disease is most likely to develop in areas that have weeds and stubbles of infected plants. It can occur in both tropical and temperate environments, particularly in irrigated and rainfed lowland areas. In general, the disease favour’s temperatures at 25-34°C, with relative humidity above 70%. It is commonly observed when strong winds and continuous heavy rains occur, allowing the disease-causing bacteria to easily spread through ooze droplets on lesions of infected plants. Bacterial blight can be severe in susceptible rice varieties under high nitrogen fertilization.
The field trial was conducted to evaluate the efficacy of innovative mixtures of Picoxystrobin and Validamycin against Xanthomonas oryzae pv. oryzae in rice crop. Trial was conducted with randomized block design with net plot size of 5m x 6m. Rice crop was raised with all standard agronomic practices. Spraying was done with manual operated backpack knapsack sprayer with 500 L of water spray volume per hectare at 45 days after transplanting.
The visual observations were recorded for % disease control from ten hills per plot. The observations were recorded at before spraying, 7 DAA (Days after application) and 14 DAA (Days after application).
Table 12: Bacterial leaf blight disease control in rice
Compositions Dose (gram active per hectare) Percent disease control –Bacterial leaf blight.
07 DAA 14 DAA
Validamycin 6% + Picoxystrobin 10% SC 120 90 85.7
Validamycin 45 75 60
Picoxystrobin 75 45 30
SC – Suspension concentrates, and DAA - Days after application.

The trial results show excellent efficacy of Validamycin 6% + Picoxystrobin 10% SC combinations against bacterial leaf blight disease of rice. The solo application of two active ingredients tested here, were also not able to provide satisfactory control of bacterial leaf blight disease. The combination of Validamycin 6% + Picoxystrobin 10% SC found very promising against rice bacterial leaf blight in terms of efficacy as well as residual control.
Table 13: Percent bacterial leaf blight disease control at 14 DAA
Compositions Dose
(g or ml/ha) % Bacterial leaf blight disease control
Expected Actual
Validamycin 45 60
Picoxystrobin 75 30
Validamycin 6% + Picoxystrobin 10% SC 120 72.18 85.43
Ratio of O/E 1.18

The results in table 13 clearly demonstrates synergy between Picoxystrobin and Validamycin in controlling rice bacterial leaf blight disease. The large difference between the observed and the expected efficacy clearly demonstrates the synergistic effect of the combination.
The combinations of the present invention have several advantages over solo application of either Picoxystrobin and Validamycin and, likewise, over any solo application of these fungicides. The combination of the present invention shows enhanced fungicidal action in comparison with the fungicidal action of Picoxystrobin and Validamycin against undesirable fungal pathogens. Moreover, the combinations of the invention show a persistent fungicidal activity, even under difficult weathering conditions. The combinations are generally non-toxic or of low toxicity against mammals.
Although the present invention has been described and illustrated with respect to preferred embodiments and a preferred use thereof, it is not to be so limited since modifications and changes can be made therein which are within the full scope of the invention.
,CLAIMS:We claim:
1. A fungicidal composition comprising as its active ingredient validamycin present in a range from 4% to 20% (w/w), picoxystrobin present in a range from 8% to 20% (w/w) and excipients.
2. The fungicidal composition as claimed in claim 1, wherein the excipients are selected from the group comprising a wetting agent, a dispersing agent, an anti-freezing agent, a biocide, a viscosity modifier, a defoamer and a base.
3. The fungicidal composition as claimed in claim 2, wherein the wetting agent is selected from the group comprising Ethoxylated TSP-phosphate ester, Amine salt of poly aryl phenyl ether phosphate, polyoxyethylenated alkylphenol, dialkyl naphthalene sulphonate sodium salt, alkyl naphthalene sulfonate (ANS) condensate blend, sodium ligno sulphonate, linear alcohol derivative and polyethylene glycol nonyl phenyl ether ammonium sulfate present in an amount of 0.2 to 1.5% (w/w).
4. The fungicidal composition as claimed in claim 2, wherein the dispersing agent is selected from the group comprising EO/PO polymeric dispersant, Ethoxylated TSP-phosphate ester, Polyoxy alkylene glycol butyl ether, Polyoxyethylene-Polyoxypropylene Block Polymer, Polyethylene glycol 2,4,6-tris(1-phenylethyl)phenyl ether sulfate ammonium salt, Methyl methacrylate graft copolymer, Sodium polyalkyl naphthalene sulfonate, Sodium salt of methyl naphthalene sulfonate, sodium ligno sulphonate, acrylate copolymer, phenol sulfonic acid-formaldehyde-polycondensation as sodium salt, sodium polycarboxylate, sodium methyl oleoyl taurate and sodium lauryl sulphate or combination thereof present in an amount of 0.2 to 5% (w/w).
5. The fungicidal composition as claimed in claim 2, wherein the anti-freezing agent is selected from propylene glycol, glycerin, and mono ethylene glycol, present in an amount of 1.0% (w/w).
6. The fungicidal composition as claimed in claim 2, wherein the biocide is selected from 1,2-Benzisothiazol-3(2H)-one and formaldehyde, present in an amount of 0.1% (w/w).
7. The fungicidal composition as claimed in claim 2, wherein the viscosity modifier is selected from the group comprising of xanthan gum, carboxymethyl cellulose, and polyvinyl pyrrolidine (PVP), present in an amount of 1.0% (w/w).
8. The fungicidal composition as claimed in claim 2, wherein the defoamer is Polydimethylsiloxane antifoam emulsion, present in an amount of 0.5% (w/w).
9. The fungicidal composition as claimed in claim 2, wherein the base is water present in an amount of 54% to 85% (w/w).
10. The fungicidal composition as claimed in claim 1, wherein the composition is formulated as a suspension concentrate.