Claims:CLAIMS

We Claim:

1. A botanical based bio-fungicidal composition for controlling the plant pathogenic fungus comprising synergistically effective combination of leaf and bark extract of Cinnamomum cassia orCinnamaldehyde extracted from botanical source such as Cinnamomum genus or chemically synthesized, bud extract of SyzygiumaromaticumorEugenol extracted from botanical source such as Syzygium genus or Ocimum genus or chemically synthesized, seed extract of CuminumcyminumorCuminaldehyde extracted from botanical source such as Cuminum genus or chemically synthesized, oil extracted from aerial parts of Thymus vulgaris orThymol extracted from botanical source such as Thymus genus or chemically synthesized, non-ionic emulsifier, anionic emulsifier and carrier oil at defined concentrations.

2. The botanical based bio-fungicidal composition as claimed in claim 1, wherein the leaf and bark extract of Cinnamomum cassiaorCinnamaldehyde extracted from botanical source such as Cinnamomum genus or chemically synthesizedranges from 0.001% to 40%; bud extract of SyzygiumaromaticumorEugenol extracted from botanical source such as Syzygium genus or Ocimum genus or chemically synthesizedranges from 0.001% to 25%; seed extract of CuminumcyminumorCuminaldehyde extracted from botanical source such as Cuminum genus or chemically synthesizedranges from 0.001% to 40% and oil extracted from aerial parts of Thymus vulgarisorThymol extracted from botanical source such as Thymus genus or chemically synthesizedranges from 0.001% to 45% respectively.

3. The botanical based bio-fungicidal composition as claimed in claim 1, wherein the non-ionic emulsifier ranges from 0.001% to 20%; anionic emulsifier ranges from 0.001% to 25% and carrier oil ranges from 0.001% to 75% respectively.

4. The botanical based bio-fungicidal composition as claimed in claim 1, wherein the carrier oil includes but not limited to seed oil of Anethumgraveolens, Pongamaiapinnata oil, Azadirachtaindica oil, castor oil, fish oil, Sesamumindicumoil, Brassica junceaoil and the like.

5. The botanical based bio-fungicidal composition as claimed in claim 1, wherein the said composition is used to control various categories of plant pathogenic fungus such as but not limited to Alternariaalternata (Leaf Spot), Lasiodioplodiatheobromae (Mango - Stem androt),Fusariumoxysporum (Gram wilt), Mycosphaerellafijensis (Banana- Sigatoka disease), Ceratocystisfimbriata(Pomegranate wilt disease) Colletotrichumgloeosporioides (Fruit rot) , Alternariasolani(Early Blight of Tomato) and Plasmoparaviticola (Downy Mildew of Grapes).

6. The botanical based bio-fungicidal composition as claimed in claim 1, wherein the said composition is useful for controlling plant pathogenic fungus of agricultural crops such as but not limited to olericulture, floriculture, horticulture, medicinal and economic plants.

7. The botanical based bio-fungicidal composition as claimed in claim 1, wherein the mode of application of said composition consist of spraying such as foliar sprays, applied to the plant shoots, used as drenching, or can even be used as spray for treating soil, or as seed treatment.

8. A method of preparation of botanical based bio-fungicidal composition for controlling the plant pathogenic fungus comprising:

a. Extracting 0.001 to 40% of leaf and bark extract of Cinnamomum cassiaor Cinnamaldehyde extracted from botanical source such as Cinnamomum genus or chemically synthesized, 0.001 to 25% of bud extract of SyzygiumaromaticumorEugenol extracted from botanical source such as Syzygium genus or Ocimum genus or chemically synthesized, 0.001 to 40% of seed extract of CuminumcyminumorCuminaldehyde extracted from botanical source such as Cuminum genus or chemically synthesizedand 0.001 to 45% of aerial parts of the Thymus vulgarisorThymol extracted from botanical source such as Thymus genus or chemically synthesizedby using plurality of extraction techniques;
b. mixing the obtained extracts at given proportion along with 0.001 to 25% of anionic emulsifier and 0.001 to 20% of non-ionic emulsifier;
c. blending the obtained mixture with 0.001 to 75% of carrier oils;
d. stirring the obtained mixture at 200RPM to 800 RPM at 350 C to 650 C in closed mixing vessel made up of stainless steel 316 grade with outer jacket for maintaining temperature, stirrer for continuous stirring, vessel homogenizer towards the bottom with the speed of 2800 RPM to reduce the particle size under very high pressure, sheer, turbulence, acceleration and impact to make more stable and effective;
e. passing the obtained mixture through in line shear pump with the speed of 1800RPM to 3600 RPM to reduce the particle size to nano, thereby achieving nano emulsion with particle size ranging from 10 to 900 nm;
f. passing the material thus obtained through the filter consisting seven layers of membrane filters with pore size less than 10 µm in order to achieve proper filtration; and
g. Finally, the product thus obtained is used for commercialization and field trails.

9. The method of preparation of botanical based bio-fungicide as claimed in claim 8, wherein the extraction techniques includes such as not limited to solvent extraction, oleoresin, steam distillation, super critical carbon-dioxide extraction and the like.

10. The method of preparation of botanical based bio-fungicide as claimed in claim 9, wherein the solvent used in the solvent extraction consist of methanol, Hexane, Acetone, Ethyl acetate, butanol and ethanol.

DATED THIS 4thday of June, 2020

GARGI AMOL PHADATARE (IN/PA/2209)
PATENT AGENT OF APPLICANT

, Description:
FORM – 2

THE PATENTS ACT, 1970
(39 of 1970)

&

THE PATENTS RULES, 2003

COMPLETE SPECIFICATION
(See section 10 and rule 13)

A BOTANICAL BASED BIO-FUNGICIDAL COMPOSITION AND MANUFACTURING METHOD THEREOF

KAY BEE BIO ORGANICS PVT LTD
An Indian Company of
At Post- Sastewadi, Khunte Road,
Taluka:Phaltan, District: Satara, State: Maharashtra,
Country: India, Pin Code: 415523

The following specification particularly describes the invention and the manner in which it is to be performed.

A) TECHNICAL FIELD
[0001] The present invention generally relates to the field of botanicalbased bio-fungicidal composition for controlling the plant pathogenic fungus. In particular the invention relates to botanicalbased bio-fungicidal composition based on a complex mixture of organic substances of various plant extracts. The present invention ismore particularly related to the botanicalbased bio-fungicidal compositions, method of making thebotanical basedbio-fungicidal compositions, and methods of applying such compositions to plants and to soil.

B) BACKGROUND OF THE INVENTION
[0002] Pesticides are chemical compounds used to kill the pests, that includes insects, rodents, fungi, bacteria and unwanted plants (weeds). The pests can cause damage to the crops which ultimately leads to decrease in food production, reduction in crop quality and increase in harvesting cost. Hence, the current practice to control the destructive pest is the application of pesticides containing synthetic chemical compounds.
[0003] However, the synthetic chemical pesticides are highly toxic and lethal for the pest, as well as the person who get exposed directly or indirectly if not handled with almost care. Therefore, in race of obtaining maximum productivity from crops, one is extensively using the chemical pesticides which results in detrimental environmental effects that are harmful to humans and other animals.
[0004] Initially the chemical pesticides were effective in controlling or killing the pests on crops. However, the repeated application of the chemical pesticides leads to the development of tolerance and resistance against the pests. Hence, the current scenario is that if any chemical pesticides lose its effectiveness, other synthetic chemicals that the pest have not developed resistance are been used. Hence the cycle of addition of constant new synthetic chemicals to control the pests leads to the release of toxic chemicals in the environment which are hazardous to humans and environment. Non selective way of action also makes the pesticides undesirable. Moreover, the synthetic chemicals can frequently persist in environment almost indefinitely.
[0005] Further, for numerous years the plant pathogenic fungi one of the major pests have caused devastating loses globally. Synthetic fungicides have effectively controlled them but their repeated use over decades have disturbed the natural biological systems, which often led to the development of resistance against the majority of compounds. Moreover, the pesticides also have undesired effects on non-target organisms, environment and on humans.
[0006] Hence, there is a need for an alternative measure which are safe to the eco-system, capable of bio-degradable, having multi-mechanism action and are less prone to the development of resistance. There is also aneed for an alternative fungicide which comprises reduced utilization of synthetic chemical compounds and are eco-friendly, safe for agriculture and humans.
[0007] The above-mentioned shortcomings, disadvantages and problemsare addressed herein and which will be understood by reading and studying thefollowing specification.

C) OBJECTS OF THE INVENTION
[0008] The primary object of the present invention is to provide asynergistic botanicalbased bio-fungicidal composition which obviates the limitations as stated above.
[0009] Another object of the present invention is to provide an uniquebotanical based bio-fungicidal composition consisting of synergistically effective combination of leaf and bark extract of Cinnamomum cassia or Cinnamaldehyde extracted from botanical source such as Cinnamomum genus or chemically synthesized, bud extract of Syzygium aromaticumor Eugenol extracted from botanical source such as Syzygium genus or Ocimum genus or chemically synthesized, seed extract of Cuminumcyminum or Cuminaldehyde extracted from botanical source such as Cuminum genus or chemically synthesized, oil extracted from aerial parts of Thymus vulgaris or Thymol extracted from botanical source such as Thymus genus or chemically synthesized, non-ionic emulsifier, anionic emulsifier and carrier oil at defined concentrations.

[0010] Yet another object of the present invention is to provide a botanical based bio-fungicidal composition comprising of various plants extracts exhibiting synergism at specific concentration thereby controls the pest and protect agriculture, horticulture, olericulture and floricultural crops.

[0011] Yet another object of the present invention is to provide method of preparation of botanical based bio-fungicides and the mode of application of the bio-fungicides.

[0012] Yet another object of the present invention is to provide botanical based bio-fungicide comprising natural ingredients such as extracts of plants, used in combination or modified to be used as an alternative to synthetic chemical pesticides.

[0013] Yet another object of the present invention is to provide an eco-friendly, non-toxic botanical based bio-fungicide safe for agriculture and humans.

[0014] These and other objects and advantages of the present inventionwill become readily apparent from the following detailed description taken inconjunction with the accompanying drawings.

D) SUMMARY OF THE INVENTION
[0015] The present invention relates to botanical based bio-fungicidal composition for controlling the plant pathogenic fungus. In particular the invention relates to botanical based bio-fungicidal composition based on a complex mixture of organic substances of various plant extracts. The present invention ismore particularly related to the botanical based bio-fungicidal compositions, method of making the bio-fungicidal compositions, and methods of applying such compositions to plants and to soil to control the plant pathogens.

[0016] According to one embodiment of the present invention provides a botanical based bio-fungicidal composition for controlling the plant pathogenic fungus comprising synergistically effective combination of leaf and bark extract of Cinnamomum cassia or Cinnamaldehyde extracted from botanical source such as Cinnamomum genus or chemically synthesized, bud extract of Syzygium aromaticumor Eugenol extracted from botanical source such as Syzygium genus or Ocimum genus or chemically synthesized, seed extract of Cuminumcyminum or Cuminaldehyde extracted from botanical source such as Cuminum genus or chemically synthesized, oil extracted from aerial parts of Thymus vulgaris or Thymol extracted from botanical source such asThymus genus or chemically synthesized, non-ionic emulsifier, anionic emulsifier and carrier oil at defined concentrations.
[0017] According to one embodiment herein, the botanical based bio-fungicidal composition comprises by volume approximately from 0.001% to 40% of leaf and bark extract of Cinnamomum cassiaor 0.001 % to 40 % of Cinnamaldehyde extracted from botanical source such asCinnamomum genus or chemically synthesized, 0.001% to 25% of bud extract of Syzygium aromaticumor 0.001 % to 25 % of Eugenol extracted from botanical source such as Syzygiumgenus or Ocimum genus or chemically synthesized, 0.001% to 40% of seed extract of Cuminum cyminumor 0.001 % to 40 % of Cuminaldehyde extracted from botanical source such asCuminum genus or chemically synthesized, 0.001% to 45% of oil extracted from aerial parts of Thymus vulgarisor 0.001 % to 45 % of Thymol extracted from botanical source such asThymus genus or chemically synthesized 0.001% to 20% of non-ionic emulsifier, 0.001% to 25% of anionic emulsifier and 0.001% to 75% of carrier oilrespectively.
[0018] According to one embodiment of the present invention, the botanical based bio-fungicide is used against various categories of plant pathogenic fungus such as but not limited to Alternariaalternata (Leaf Spot), Lasiodioplodiatheobromae (Mango - Stem androt), Fusariumoxysporum (Gram wilt), Mycosphaerellafijensis (Banana- Sigatoka disease), Ceratocystis fimbriata (Pomegranate wilt disease) , Colletotrichumgloeosporioides (Fruit rot), Alternariasolani(Early Blight of Tomato) and Plasmoparaviticola (Downy Mildew of Grapes) .
[0019] According to one embodiment of the present invention, the botanical based bio-fungicide is useful for controlling plant pathogenic fungus of agricultural crops such as but not limited to olericulture, floriculture, horticulture, medicinal and economic plants.
[0020] According to one embodiment of the present invention, the mode of application of botanical based bio-fungicide incudes spraying such as foliar sprays, applied to the plant shoots, used as drenching, or can even be used as spray for treating soil, or as seed treatment.
[0021] According to one embodiment of the present invention, provides a method of preparation of botanical based bio-fungicide for controlling the plant pathogenic fungus comprising extracting 0.001 to 40% of leaf and bark extract of Cinnamomum cassiaor 0.001 % to 40 % of Cinnamaldehyde extracted from botanical source such as Cinnamomum genus or chemically synthesized, 0.001 to 25% of bud extract of Syzygium aromaticumor 0.001 % to 25 % of Eugenol extracted from botanical source such as Syzygium genus or Ocimum genus or chemically synthesized, 0.001 to 40% of seed extract of Cuminum cyminumor 0.001 % to 40 % of Cuminaldehyde extracted from botanical source such as Cuminum genus or chemically synthesizedand 0.001 to 45% of aerial parts of the Thymus vulgarisor 0.001 % to 45 % of Thymol extracted from botanical source such as Thymus genusby using plurality of extraction techniques; mixing the obtained extracts at given proportion along with 0.001 to 25% of anionic emulsifier and 0.001 to 20% of non-ionic emulsifier; followed by blending the obtained mixture with 0.001 to 75% of carrier oils; stirring the obtained mixture at 200 to 800 RPM at 350 C to 650 C in closed mixing vessel made up of stainless steel 316 grade with outer jacket for maintaining temperature, stirrer for continuous stirring, vessel homogenizer towards the bottom with the speed of 2800 RPM to reduce the particle size under very high pressure, sheer, turbulence, acceleration and impact to make more stable and effective; passing the obtained mixture through in line shear pump with the speed of 1800 RPM to 3600 RPM to reduce the particle size to nano, thereby achieving nano emulsion with particle size ranging from 10 to 900 nm. Further the material obtained is passed through the filter consisting seven layers of membrane filters with pore size less than 10 µm in order to achieve proper filtration. Finally, the product obtained is used for commercialization and field trails.
[0022] According to one embodiment of the present invention, the extraction techniques of the various parts of plurality of plant extracts are carried out by using various extraction techniques such as not limited to solvent extraction, oleoresin, steamdistillationand super critical carbon-dioxide extraction. Further, the solvent extraction includes various solvents such as but not limited to Methanol, Hexane, Acetone, Ethyl acetate, butanol and ethanol.
[0023] According to one embodiment of the present invention, the carrier oil used in the preparation of botanical based bio-fungicide includes but not limited to seed oil ofAnethumgraveolens, Pongamaiapinnataoil,Azadirachtaindica oil, castor oil, fish oil, Sesamumindicum oil and Brassica juncea oil.
[0024] According to one embodiment of the present invention, the botanical based bio-fungicide does not separate the phytochemicals which includes alkaloids, phytosterols, polyphenols, terpenoids, organosulfur compounds and glycosides. Hence a holistic approach of phytochemicals from plant to control the plant pathogenic fungus is thus employed. Hence, the plant extracts used as ingredients in composition are standardized, as well as Phyto profile is thus explored. Moreover, the composition is standardized for selected phytochemicals in order to assure the quality, as most of the botanicalbased pesticides lack the standardization.
[0025] According to one embodiment of the present invention, the in vitro and in vivo trials have been conducted in order to conform the potential outcome of the botanical based bio-fungicide to control the fungal attack, in competing with the leading commercial synthetic chemical fungicides which are toxic and not safe to the environment and humans.
[0026] These and other aspects of the embodiments herein will be betterappreciated and understood when considered in conjunction with the followingdescription and the accompanying drawings. It should be understood, however,that the following descriptions, while indicating the preferred embodiments andnumerous specific details thereof, are given by way of an illustration and not of alimitation. Many changes and modifications may be made within the scope ofthe embodiments herein without departing from the spirit thereof, and theembodiments herein include all such modifications.

E) BRIEF DESCRIPTION OF THE DRAWINGS
[0027] The other objects, features, and advantages will occur to thoseskilled in the art from the following description of the preferred embodiment andthe accompanying drawings in which:
[0028] FIG. 1 illustrates an in-vitro efficacy of botanical based bio-fungicide (referred in drawings as BB Bio-Fungicide) against Alternariasolani (Early Blight of Tomato), according to an embodimentof the present invention;
[0029] FIG. 2 illustrates a bio-assay efficacy of botanical based bio-fungicide (referred in drawings as BB Bio-Fungicide) against Plasmoparaviticola (Downy Mildew of Grapes) by leaf disc assay method, according to an embodiment of thepresent invention;
[0030] FIG. 3 illustrates a schematic view of after 10days of treatment, the bio-efficacy of botanical based bio-fungicide (referred in drawings as BB Bio-Fungicide) against Downy Mildew of Grapes by leaf disc assay method,according to an embodiment of the present invention;
[0031] FIG. 4 illustrates an in-vitro efficacy of botanical based bio-fungicide (referred in drawings as BB Bio-Fungicide) against Colletotrichumgloeosporioides(Leaf spot disease of chilli), according to anembodiment of the present invention;
[0032] FIG. 5 illustrates an in-vitro efficacy of botanical based bio-fungicide (referred in drawings as BB Bio-Fungicide) against Fusariumoxysporium, according to an embodiment of the presentinvention;
[0033] FIG. 6 illustrates an in-vitro efficacy of botanical based bio-fungicide (referred in drawings as BB Bio-Fungicide) against Mycospherellafijensis,according to one embodiment of the present invention;
[0034] FIG. 7 illustrates an in-vivo bio-efficacy of botanical based bio-fungicide (referred in drawings as BB Bio-Fungicide) against Downy mildew of Grapes, according to another embodiment of thepresent invention, wherein in the drawing Obs. are the observations recorded;
[0035] FIG. 8 illustrates an in-vivo bio-efficacy of botanical based bio-fungicide (referred in drawings as BB Bio-Fungicide) against Powdery mildew of Grapes,according to an embodiment of thepresent invention, wherein in the drawing Obs. are the observations recorded;
[0036] FIG. 9 illustrates an in-vivo bio-efficacy of botanical based bio-fungicide(referred in drawings as BB Bio-Fungicide) against Late blight of Potato, according to anembodiment of the present invention, wherein in the drawing Obs. are the observations recorded; and
[0037] FIG. 10 illustratesan in-vivo bio-efficacy of botanical based bio-fungicide (referred in drawings as BB Bio-Fungicide) against Early blight of Tomato, according to an embodiment of the presentinvention, wherein in the drawing Obs. are the observations recorded.
[0038] The specific features of the present invention are shownin some drawings and not in others. This is done for convenience only as eachfeature may be combined with any or all of the other features in accordance withthe present invention.

F) DETAILED DESCRIPTION OF THE INVENTION
[0039] The present invention relates to botanical based bio-fungicidal composition for controlling the plant pathogenic fungus. In particular the invention relates to botanicalbased bio-fungicidal compositionbased on a complex mixture of organic substances of various plant extracts. The present invention ismore particularly related to the botanicalbased bio-fungicidal compositions, method of making the bio-fungicidal compositions, and methods of applying such compositions to plants and to soil to control the plant pathogens.
[0040] Before the present invention is disclosed and described, it is to be understood that this invention is not limited to the particular process steps and materials disclosed herein, as such process steps and materials may vary to some degree. It is also to be understood that the terminology used herein is used for the purpose of describing particular embodiments only and is not intended to be limiting as the scope of the present invention will be limited only by appended claims and equivalents thereof.
[0041] In order to more clearly and concisely describe and point out the subject matter of the claimed invention, the following definitions are provided for specific terms which are used in the following written description.
[0042] The term “pest” when used herein refers to insects, rodents, fungi, bacteria and unwanted plants (weeds) that are harmful to useful plants.
[0043] The term “botanical” when used herein refers to substance obtained from plants.
[0044] The term “fungicide” and “fungicidal” as employed in the specification and claims of the application is confined to the means, which adversely affects the existence, growth of the target organism. Such means may comprise a complete killing action, eradicate, arresting growth, inhibition, reducing in number or any combination thereof.
[0045] The term “emulsion” when used herein refers to a stable mixture of two or more immiscible held in suspension.
[0046] The term “emulsifier” when used herein refers to a substance that stabilizes an emulsion.
l[0047] The term “control” and “controlling” when used herein refers to alleviating or abrogating a disorder, disease, or condition, or one or more of the symptoms associated with the disorder, disease, or condition; or alleviating or eradicating the cause(s) of the disorder, disease, or condition itself.
[0048] The trademark “Folicur®” is owned by Bayer crop science refers to a fungicidal compound with broad-spectrum systemic action which can be used as a foliar spray or seed-dressing.
[0049] The trademark “Nativo®” is owned by Bayer crop science refers to systemic broad-spectrum fungicide with protective and curative action which offers not only a disease control but also improves quality and yield of crop.
[0050] The trademark “Kavach” is owned by Syngenta refers to broad spectrum contact fungicide and is highly effective against Anthracnose.
[0051] The trademark “Antracol®” is owned by Bayer crop science refers to broad spectrum contact fungicide with protective action.
[0052] The trademark “Bavistin” is owned by Biostadt India Limited refers to broad spectrum systemic fungicide containing effective against a wide range of pathogenic fungi and is highly specific in its control of important plant pathogens on a variety of crops, ornamental plants and plantation crops.
[0053] The trademark “Score” is owned by Syngenta refers to is a broad-spectrum systemic triazole fungicide that provides outstanding control of Leaf Rust and Leaf Spots in vegetables and ornamentals.
[0054] The trademark “Roko” is owned by Biostadt India Limited refers to broad spectrum systemic fungicide and has unique combination of Preventive, Curative and Systemic fungicidal properties.
[0055] The trademark “Kocide” is owned by Kocide LLC refers to copper-based fungicide/bactericide products.
[0056] The trademark “M-45” is owned by Indofil industries limited refers to broad spectrum fungicide which control diseases caused by all four major classes of plant pathogens in wide range of the cross.
[0057] The trademark “Revus” is owned by Syngenta refers to fungicide to control of Late blight in potatoes and Downy mildew in onions.
[0058] The trademark “Acrobat®” is owned by BASF refers to fungicide to control Downy mildew in Grapes.
[0059] The trademark “BenofitBenomyl” is owned by Agrosiaa refers to fungicide for effective control of wide range of disease in fruits, peanut, vegetables, food crops and ornamentals.
[0060] The trademark “Propiconazole” is owned by Tagros chemicals refers to triazole fungicide.
[0061] The trademark “Amistar” is owned by Syngenta refers to broad-spectrum fungicide to control wide range of disease in barley, maize, onions, peas, potatoes, sweetcorn, tomatoes and wheat.
[0062] The trademark “Alliate®” is owned by Bayer crop science refers to fungicide which protects against phytophthora, downy mildew and other diseases.
[0063] The trademark “Captan” is a general use fungicide used to control disease on ornamentals, fruits and vegetables.
[0064] The trademark “Blitox” is owned by Rallis India refers to fungicide for the control of diseases on field, garden and plantation crop.
[0065] According to one embodiment of the present invention provides a botanical based bio-fungicidal composition for controlling the plant pathogenic fungus comprising synergistically effective combination of leaf and bark extract of Cinnamomum cassiaorCinnamaldehyde extracted from botanical source such as Cinnamomum genus or chemically synthesized, bud extract of Syzygium aromaticumorEugenol extracted from botanical source such as Syzygium genus or Ocimum genus or chemically synthesized, seed extract of Cuminumcyminum or Cuminaldehyde extracted from botanical source such as Cuminum genus or chemically synthesized, oil extracted from aerial parts of Thymus vulgaris or Thymol extracted from botanical source such as Thymus genus or chemically synthesized, non-ionic emulsifier, anionic emulsifier and carrier oil at defined concentrations.
[0066] According to one embodiment herein, the botanical based bio-fungicidal composition comprises by volume approximately from 0.001% to 40% of leaf and bark extract of Cinnamomum cassiaor 0.001 % to 40 % of Cinnamaldehyde extracted from botanical source such as Cinnamomum genus or chemically synthesized, 0.001% to 25% of bud extract of Syzygium aromaticumor 0.001 % to 25 % of Eugenol extracted from botanical source such as Syzygium genus or Ocimum genus or chemically synthesized, 0.001% to 40% of seed extract of Cuminum cyminumor 0.001 % to 40 % of Cuminaldehyde extracted from botanical source such as Cuminum genus or chemically synthesized, 0.001% to 45% of oil extracted from aerial parts of Thymus vulgarisor 0.001 % to 45 % of Thymol extracted from botanical source such as Thymus genus or chemically synthesized 0.001% to 20% of non-ionic emulsifier, 0.001% to 25% of anionic emulsifier and 0.001% to 75% of carrier oilrespectively.
[0067] According to one embodiment of the present invention, the botanical based bio-fungicide is used against various categories of plant pathogenic fungus such as but not limited to Alternariaalternata (Leaf Spot), Lasiodioplodiatheobromae (Mango - Stem androt), Fusariumoxysporum (Gram wilt), Mycosphaerellafijensis (Banana- Sigatoka disease), Ceratocystis fimbriata (Pomegranate wilt disease) Colletotrichumgloeosporioides (Fruit rot), Alternariasolani(Early Blight of Tomato) and Plasmoparaviticola (Downy Mildew of Grapes) .
[0068] According to one embodiment of the present invention, the botanical based bio-fungicide is useful for controlling plant pathogenic fungus of agricultural crops such as but not limited to olericulture, floriculture, horticulture, medicinal and economic plants.
[0069] According to one embodiment of the present invention, the mode of application of botanical based bio-fungicide incudes spraying such as foliar sprays, applied to the plant shoots, used as drenching, or can even be used as spray for treating soil, or as seed treatment.
[0070] According to one embodiment of the present invention, provides a method of preparation of botanical based bio-fungicide for controlling the plant pathogenic fungus comprising extracting 0.001 to 40% of leaf and bark extract of Cinnamomum cassiaor 0.001 % to 40 % of Cinnamaldehyde extracted from botanical source such as Cinnamomum genus or chemically synthesized, 0.001 to 25% of bud extract of Syzygium aromaticumor 0.001 % to 25 % of Eugenol extracted from botanical source such as Syzygium genus or Ocimum genus or chemically synthesized, 0.001 to 40% of seed extract of Cuminum cyminumor 0.001 % to 40 % of Cuminaldehyde extracted from botanical source such as Cuminum genus or chemically synthesizedand 0.001 to 45% of aerial parts of the Thymus vulgarisor 0.001 % to 45 % of Thymol extracted from botanical source such as Thymus genusby using plurality of extraction techniques; mixing the obtained extracts at given proportion along with 0.001 to 25% of anionic emulsifier and 0.001 to 20% of non-ionic emulsifier; followed by blending the obtained mixture with 0.001 to 75% of carrier oils; stirring the obtained mixture at 200 to 800 RPM at 350 C to 650 C in closed mixing vessel made up of stainless steel 316 grade with outer jacket for maintaining temperature, stirrer for continuous stirring, vessel homogenizer towards the bottom with the speed of 2800 RPM to reduce the particle size under very high pressure, sheer, turbulence, acceleration and impact to make more stable and effective; passing the obtained mixture through in line shear pump with the speed of 1800 RPM to 3600 RPM to reduce the particle size to nano, thereby achieving nano emulsion with particle size ranging from 10 to 900 nm. Further the material obtained is passed through the filter consisting seven layers of membrane filters with pore size less than 10 µm in order to achieve proper filtration. Finally, the product obtained is used for commercialization and field trails.
[0071] According to one embodiment of the present invention, the extraction techniques of the various parts of plurality of plant extracts are carried out by using various extraction techniques such as not limited to solvent extraction, oleoresin, steam distillation and super critical carbon-dioxide extraction. Further, the solvent extraction includes various solvents such as but not limited to methanol, Hexane, Acetone, Ethyl acetate, butanol and ethanol.
[0072] According to one embodiment of the present invention, the carrier oil used in the preparation of botanical based bio-fungicide includes but not limited to Seed oil of Anethumgraveolens, Pongamaiapinnata oil, Azadirachtaindica oil, castor oil, fish oil, Sesamumindicumoil and Brassica junceaoil.
[0073] According to one embodiment of the present invention, the botanical based bio-fungicide does not separate the phytochemicals which includes alkaloids, phytosterols, polyphenols, terpenoids, organosulfur compounds and glycosides. Hence a holistic approach of phytochemicals from plant to control the plant pathogenic fungus is thus employed. Hence, the plant extracts used as ingredients in composition are standardized, as well as phyto profile is thus explored. Moreover, the composition is standardized for selected phytochemicals in order to assure the quality, as most of the botanical based pesticides lack the standardization.
[0074] According to one embodiment of the present invention, the in vitro and in vivo trials have been conducted in order to conform the potential outcome of the botanical based bio-fungicide to control the fungal attack, in competing with the leading commercial synthetic chemical fungicides which are toxic and not safe to the environment and humans.
[0075] According to one embodiment of the present invention, the FIG.1 illustrates an in-vitro efficacy of botanical based bio-fungicide against Alternariasolani (Early Blight of Tomato). Further the Table 1 provides the percentage of inhibition of botanical based bio-fungicide vs various fungicides available in the market. The Table 1 provides that botanical based bio fungicide offers 100% fungal inhibition after 7days of treatment at dosage of 0.25ml and 0.5 ml respectively when compared to other fungicides available in the market.
Table 1: In-vitro efficacy of botanical based bio-fungicide against Alternariasolani (Early Blight of Tomato)
Treat. No. Treatment Dose (per L) Mycelial Growth (mm) Per cent inhibition (%)
After 7th day
T1 Botanical based bio-fungicide 25% EC 0.25 ml 0 100
T2 Botanical basedbio-fungicide 25% EC 0.5 ml 0 100
T3 Folicur 25.9% EC 0.5 ml 0 100
T4 Nativo 75% WP 0.5 g 17.33 74.51
T5 Kavach 75% WP 1.25 g 0 100
T6 Antracol 70% WP 2.5 g 16.33 75.98
T7 Bavistin 50% WP 2 g 41.30 39.22
T8 Score 25% EC 0.5 ml 53.45 21.57
T9 Roko 70% WP 2 g 42.35 37.75
T10 Kocide 53.8 DF 2.5 g 18.35 37.04
T11 Control - 27 0

[0076] According to one embodiment of the present invention, the FIG.2 illustrates a bio-assay efficacy of botanical based bio-fungicide against Plasmoparaviticola (Downy Mildew of Grapes) by leaf disc assay method. Further the Table 2 provides the treatment efficacy of botanical based bio-fungicide vs various fungicides available in the market. Hence the Table 2 provides that botanical based bio fungicide offers 100% fungal inhibition after 10days of treatment at dosage of 0.5ml and 1 ml respectively when compared to other fungicides available in the market.
Table 2: Bio-assay efficacy of botanical based bio-fungicide against Plasmoparaviticola (Downy Mildew of Grapes) by leaf disc assay method

* Days after treatment, #Treatment efficacy

[0077] According to one embodiment of the present invention, the FIG. 3 illustrates a schematic view of leaves after 10days of treatment, the bio-efficacy of botanical based bio-fungicide against Downy Mildew of Grapes by leaf disc assay method. The Downy mildew development and each disc is assessed using a rating scale as given below:
Scale 0–4; where,
0 : No visible symptoms
1 : Infection but no sporulation
2 : Little sporulation, lesion size smaller than droplet size
3 : Sporulation, lesion size same as droplet size
4 : Sporulation, lesion size larger than droplet size
Treatment Efficacy:
Treatment efficacy = C – T X 100
T

Where,
C = Control
T = Treatment

[0078] According to one embodiment of the present invention, the FIG.4 illustrates an in-vitro efficacy of botanical based bio-fungicide against Colletotrichumgloeosporioides(Leaf spot disease of chilli). Further the Table 3 provides the percentage inhibition of botanical based bio-fungicide vs various fungicides available in the market. Hence the Table 3 provides that botanical based bio fungicide offers 100% fungal inhibition after 7days of treatment at dosage of 0.25ml and 0.5ml respectively when compared to other fungicides available in the market.
Table 3: In-vitro efficacy of botanical based bio-fungicide against Colletotrichumgloeosporioides(Leaf spot disease of chilli)

[0079] According to one embodiment of the present invention, the FIG.5 illustrates an in-vitro efficacy of botanical based bio-fungicide against Fusariumoxysporium. Further the Table 4 provides the percentage inhibition of botanical based bio-fungicide vs various fungicides available in the market. Hence the Table 4 provides that botanical based bio fungicide offers 100% fungal inhibition after 7days of treatment at dosage of 0.5ml respectively when compared to other fungicides available in the market.
Table 4: In-vitro efficacy of botanical based bio-fungicide against Fusariumoxysporium

[0080] According to one embodiment of the present invention, the FIG.6 illustrates an in-vitro efficacy of botanical based bio-fungicide against Mycospherellafijensis. Further the Table 5 provides the percentage inhibition of botanical based bio-fungicide vs various fungicides available in the market. Hence the Table 5 provides that botanical based bio fungicide offers 100% fungal inhibition after 7days of treatment at dosage of 0.25ml and 0.5ml respectively when compared to other fungicides available in the market.
Table 5: In-vitro efficacy of botanical based bio-fungicide against Mycospherellafijensis

[0081] According to one embodiment of the present invention, the FIG. 7 illustrates an in-vivo bio-efficacy of botanical based bio-fungicide against Downy mildew of Grapes. Further the Table 6 provides the yield obtained after 5 days of treatment usingbotanical based bio-fungicide vs various fungicides available in the market. Hence the Table 6 provides that botanical based bio fungicide offers 17.25(t./ha-1) yield after 5days of treatment at dosage of 0.75ml and 1.5ml respectively when compared to other fungicides available in the market.
Table 6: In-vivo bio-efficacy of botanical based bio-fungicide against Downy mildew of Grapes.

[0082] According to one embodiment of the present invention, the FIG. 8 illustrates an in-vivo bio-efficacy of botanical based bio-fungicide against Powdery mildew of Grapes. Further the Table 7 provides the yield obtained after 7 days of treatment using botanical based bio-fungicide vs various fungicides available in the market. Hence the Table 7 provides that botanical based bio fungicide offers 17.25(t./ha-1) yield after 7days of treatment at dosage of 0.75ml and 1.5ml respectively when compared to other fungicides available in the market.
Table 7: In-vivo bio-efficacy of botanical based bio-fungicide against Powdery mildew of Grapes

[0083] According to one embodiment of the present invention, the FIG. 9 illustrates a bio-efficacy of botanical based bio-fungicide against Late blight of Potato. Further the Table 8 provides the yield obtained after 7 days of treatment using botanical based bio-fungicide vs various fungicides available in the market.
Table 8: bio-efficacy of botanical based bio-fungicide against Late blight of Potato

[0084] According to one embodiment of the present invention, the FIG. 10 illustrates an in-vivo bio-efficacy of botanical based bio-fungicide against Early blight of Tomato. Further the Table 9 provides the yield obtained after 7 days of treatment using botanical based bio-fungicide vs various fungicides available in the market. Hence the Table 9 provides that botanical based bio fungicide offers 67(t./ha-1) yield after 7days of treatment at dosage of 0.75ml and 1.5ml respectively when compared to other fungicides available in the market.
Table 9: In-vivo bio-efficacy of botanical based bio-fungicide against Early blight of Tomato

[0085] The present invention may be more clearly understood with reference to the following examples of the invention which are given by way of example only. One has to consider that the following examples are included to demonstrate certain non-limiting aspects of the invention. It should be appreciated by those of skill in the art that the techniques disclosed in the examples which follow represent techniques discovered by the inventor to function well in the practice of the invention. However, those of skilled in the art should, in light of the present disclosure, appreciate that many changes can be made in the specific embodiments which are disclosed and still obtain a like or similar result without departing from the spirit and scope of the invention.
[0086] Example 1
Representative botanical-based bio-fungicide composition 1 and the corresponding method of preparation
An exemplary composition 1 in accordance with the principle of the present invention comprises
Sr No. Ingredient Actual Percent Percent Range
1. Cinnamomum cassia 25 0.00 1 to 40 %
2. Syzygium aromaticum 10 0.001 to 25 %
3. Thymus vulgaris 20 0.001 to 45 %
4. Non-Ionic Emulsifier 5 0.001 to 20 %
5. Anionic Emulsifier 5 0.001 to 25 %
6. Carrier oil 35 0.001 to 75 %

Method of preparation of botanical based bio-fungicide composition 1 comprises the following steps:
1. Extracting leaf and bark extract of Cinnamomum cassiaorCinnamaldehyde extracted from botanical source such as Cinnamomum genus or chemically synthesizedof percentage range about 0.001% to 40% by using various extraction techniques such as but not limited to solvent (methanol, hexane, acetone, ethyl acetate, butanol, ethanol) extract, oleoresin, steam distillation or super critical carbon dioxide extraction.
2. Similarly extracting bud extract of Syzygium aromaticumorEugenol extracted from botanical source such as Syzygium genus or Ocimum genus or chemically synthesizedof percentage range about 0.001% to 25% and aerial parts of Thymus vulgaris or Thymol extracted from botanical source such as Thymus genus or chemically synthesizedof percentage range of 0.001% to 45% by using various extraction techniques such as but not limited to solvent (methanol, hexane, acetone, ethyl acetate, butanol, ethanol) extract, oleoresin, steam distillation or super critical carbon dioxide extraction.
3. Mixing the obtained extracts with anionic emulsifier in percentage range of 0.001% to 25% and non-ionic emulsifier in percentage range 0.001% to 20% respectively.
4. Further blending the obtained mixture with carrier oil such as but limited to seed oil of Anethumgraveolens, Pongamaiapinnataoil, Azadirachtaindica oil, castor oil, fish oil, Sesamumindicumoil or Brassica junceaoil in a percentage range 0.001% to 75%.
5. Stirring the obtained mixture at 200RPM to 800RPM speed and subjecting to 350C to 650C temperature in closed mixing vessel, wherein the vessel is made up of stainless steel 316 grade consisting of outer jacket for maintaining temperature, connected with stirrer such as blade stirrer, propeller stirrer, turbine stirrer, anchor stirrer or universal stirrer for continuous stirring of the mixture. Further the vessel also comprises homogenizer towards the bottom which runs at a speed of 2800 RPM is attached to reduce the particle size under very high pressure, sheer, turbulence, acceleration and impact to make the particles more stable and effective.
6. Further the obtained mixture is then subjected to line shear pump working at a speed of 1800 RPM to 3600 RPM in order to reduce the particle size of the mixture to nano, so as to achieve the nano emulsion with particle size ranging from 10 nm to 900 nm.
7. After shearing the material is then passed through the filter consisting of seven layers of membrane filters with pore size less than 10 µm to achieve proper filtration.
8. Finally, the product is recovered and is used for commercialization and field trails.

[0087] Example 2
Representative botanical-based bio-fungicidecomposition 2 and the corresponding method of preparation
Sr No. Ingredient Actual Percent Percent Range
1. Cinnamomum cassia 25 0.00 1 to 40 %
2. Syzygium aromaticum 5 0.001 to 25 %
3. Cuminum cyminum 25 0.001 to 45 %
4. Non-Ionic Emulsifier 5 0.001 to 20 %
5. Anionic Emulsifier 5 0.001 to 25 %
6. Carrier oil 35 0.001 to 75 %

Method of preparation of botanical based bio-fungicide composition 2 comprises the following steps:
1. Extracting leaf and bark extract of Cinnamomum cassiaorCinnamaldehyde extracted from botanical source such as Cinnamomum genus or chemically synthesizedof percentage range about 0.001% to 40% by using various extraction techniques such as but not limited to solvent (methanol, hexane, acetone, ethyl acetate, butanol, ethanol) extract, oleoresin, steam distillation or super critical carbon dioxide extraction.
2. Similarly extracting bud extract of Syzygium aromaticumorEugenol extracted from botanical source such as Syzygium genus or Ocimum genus or chemically synthesizedof percentage range about 0.001% to 25% and seed extract of Cuminumcyminum or Cuminaldehyde extracted from botanical source such as Cuminum genus or chemically synthesizedof percentage range of 0.001% to 45% by using various extraction techniques such as but not limited to solvent (methanol, hexane, acetone, ethyl acetate, butanol, ethanol) extract, oleoresin, steam distillation or super critical carbon dioxide extraction.
3. Mixing the obtained extracts with anionic emulsifier in percentage range of 0.001% to 25% and non-ionic emulsifier in percentage range 0.001% to 20% respectively.
4. Further blending the obtained mixture with carrier oil such as but limited to seed oil of Anethumgraveolens, Pongamaiapinnataoil, Azadirachtaindica oil, castor oil, fish oil, Sesamumindicumoil or Brassica junceaoil in a percentage range 0.001% to 75%.
5. Stirring the obtained mixture at 200RPM to 800RPM speed and subjecting to 350C to 650C temperature in closed mixing vessel, wherein the vessel is made up of stainless steel 316 grade consisting of outer jacket for maintaining temperature, connected with stirrer such as blade stirrer, propeller stirrer, turbine stirrer, anchor stirrer or universal stirrer for continuous stirring of the mixture. Further the vessel also comprises homogenizer towards the bottom which runs at a speed of 2800 RPM is attached to reduce the particle size under very high pressure, sheer, turbulence, acceleration and impact to make the particles more stable and effective.
6. Further the obtained mixture is then subjected to line shear pump working at a speed of 1800 RPM to 3600 RPM in order to reduce the particle size of the mixture to nano, so as to achieve the nano emulsion with particle size ranging from 10 nm to 900 nm.
7. After shearing the material is then passed through the filter consisting of seven layers of membrane filters with pore size less than 10 µm to achieve proper filtration.
8. Finally, the product is recovered and is used for commercialization and field trails.
[0088] Example 3
Representative botanical-basedbio-fungicidecomposition 3 and the corresponding method of preparation
An exemplary composition 3 in accordance with the principle of the present invention comprises
Sr No. Ingredient Actual Percent Percent Range
1. Cinnamomum cassia 25 0.00 1 to 40 %
2. Syzygium aromaticum 15 0.001 to 25 %
3. Non-Ionic Emulsifier 5 0.001 to 20 %
4. Anionic Emulsifier 5 0.001 to 25 %
5. Carrier oil 50 0.001 to 75 %

Method of preparation of botanical based bio-fungicide composition 3 comprises the following steps:
1. Extracting leaf and bark extract of Cinnamomum cassiaorCinnamaldehyde extracted from botanical source such as Cinnamomum genus or chemically synthesizedof percentage range about 0.001% to 40% by using various extraction techniques such as but not limited to solvent (methanol, hexane, acetone, ethyl acetate, butanol, ethanol) extract, oleoresin, steam distillation or super critical carbon dioxide extraction.
2. Similarly extracting bud extract of Syzygium aromaticumorEugenol extracted from botanical source such as Syzygium genus or Ocimum genus or chemically synthesizedof percentage range about 0.001% to 25% by using various extraction techniques such as but not limited to solvent (methanol, hexane, acetone, ethyl acetate, butanol, ethanol) extract, oleoresin, steam distillation or super critical carbon dioxide extraction.
3. Mixing the obtained extracts with anionic emulsifier in percentage range of 0.001% to 25% and non-ionic emulsifier in percentage range 0.001% to 20% respectively.
4. Further blending the obtained mixture with carrier oil such as but limited to seed oil of Anethumgraveolens, Pongamaiapinnataoil, Azadirachtaindica oil, castor oil, fish oil, Sesamumindicumoil or Brassica junceaoil in a percentage range 0.001% to 75%.
5. Stirring the obtained mixture at 200RPM to 800RPM speed and subjecting to 350C to 650C temperature in closed mixing vessel, wherein the vessel is made up of stainless steel 316 grade consisting of outer jacket for maintaining temperature, connected with stirrer such as blade stirrer, propeller stirrer, turbine stirrer, anchor stirrer or universal stirrer for continuous stirring of the mixture. Further the vessel also comprises homogenizer towards the bottom which runs at a speed of 2800 RPM is attached to reduce the particle size under very high pressure, sheer, turbulence, acceleration and impact to make the particles more stable and effective.
6. Further the obtained mixture is then subjected to line shear pump working at a speed of 1800 RPM to 3600 RPM in order to reduce the particle size of the mixture to nano, so as to achieve the nano emulsion with particle size ranging from 10 nm to 900 nm.
7. After shearing the material is then passed through the filter consisting of seven layers of membrane filters with pore size less than 10 µm to achieve proper filtration.
8. Finally, the product is recovered and is used for commercialization and field trails.
[0089] Example 4
Representative botanical-basedbio-fungicidecomposition 4 and the corresponding method of preparation
An exemplary composition 4 in accordance with the principle of the present invention comprises
Sr No. Ingredient Actual Percent Percent Range
1. Syzygium aromaticum 5 0.001 to 25 %
2. Cuminum cyminum 20 0.001 to 40 %
3. Thymus vulgaris 15 0.001 to 45 %
4. Non-Ionic Emulsifier 5 0.001 to 20 %
5. Anionic Emulsifier 5 0.001 to 25 %
6. Carrier oil 25 0.001 to 75 %

Method of preparation of botanical based bio-fungicide composition 4 comprises the following steps:
1. Extracting bud extract of Syzygium aromaticumorEugenol extracted from botanical source such as Syzygium genus or Ocimum genus or chemically synthesizedof percentage range about 0.001% to 25% by using various extraction techniques such as but not limited to solvent (methanol, hexane, acetone, ethyl acetate, butanol, ethanol) extract, oleoresin, steam distillation or super critical carbon dioxide extraction.
2. Similarly extracting the aerial parts of Thymus vulgaris or Thymol extracted from botanical source such as Thymus genus or chemically synthesized percentage range about 0.001% to 45% and seed extract of Cuminumcyminumor Cuminaldehyde extracted from botanical source such as Cuminum genus or chemically synthesizedof percentage range of 0.001% to 40% by using various extraction techniques such as but not limited to solvent (methanol, hexane, acetone, ethyl acetate, butanol, ethanol) extract, oleoresin, steam distillation or super critical carbon dioxide extraction.
3. Mixing the obtained extracts with anionic emulsifier in percentage range of 0.001% to 25% and non-ionic emulsifier in percentage range 0.001% to 20% respectively.
4. Further blending the obtained mixture with carrier oil such as but limited to seed oil of Anethumgraveolens, Pongamaiapinnataoil, Azadirachtaindica oil, castor oil, fish oil, Sesamumindicumoil or Brassica junceaoil in a percentage range 0.001% to 75%.
5. Stirring the obtained mixture at 200RPM to 800RPM speed and subjecting to 350C to 650C temperature in closed mixing vessel, wherein the vessel is made up of stainless steel 316 grade consisting of outer jacket for maintaining temperature, connected with stirrer such as blade stirrer, propeller stirrer, turbine stirrer, anchor stirrer or universal stirrer for continuous stirring of the mixture. Further the vessel also comprises homogenizer towards the bottom which runs at a speed of 2800 RPM is attached to reduce the particle size under very high pressure, sheer, turbulence, acceleration and impact to make the particles more stable and effective.
6. Further the obtained mixture is then subjected to line shear pump working at a speed of 1800 RPM to 3600 RPM in order to reduce the particle size of the mixture to nano, so as to achieve the nano emulsion with particle size ranging from 10 nm to 900 nm.
7. After shearing the material is then passed through the filter consisting of seven layers of membrane filters with pore size less than 10 µm to achieve proper filtration.
8. Finally, the product is recovered and is used for commercialization and field trails.
[0090] Example 5
Representative botanical-based bio-fungicidecomposition 5 and the corresponding method of preparation
An exemplary composition5 in accordance with the principle of the present invention comprises
Sr No. Ingredient Actual Percent Percent Range
1. Cuminum cyminum 25 0.001 to 40 %
2. Thymus vulgaris 20 0.001 to 45 %
3. Non-Ionic Emulsifier 8 0.001 to 20 %
4. Anionic Emulsifier 7 0.001 to 25 %
5. Carrier oil 40 0.001 to 75 %

Method of preparation of botanical based bio-fungicide composition 5 comprises the following steps:
1. Extracting seed extract of Cuminumcyminum or Cuminaldehyde extracted from botanical source such as Cuminum genus or chemically synthesizedof percentage range about 0.001% to 40% by using various extraction techniques such as but not limited to solvent (methanol, hexane, acetone, ethyl acetate, butanol, ethanol) extract, oleoresin, steam distillation or super critical carbon dioxide extraction.
2. Similarly extracting the aerial parts of Thymus vulgaris or Thymol extracted from botanical source such as Thymus genus or chemically synthesized percentage range about 0.001% to 45% by using various extraction techniques such as but not limited to solvent (methanol, hexane, acetone, ethyl acetate, butanol, ethanol) extract, oleoresin, steam distillation or super critical carbon dioxide extraction.
3. Mixing the obtained extracts with anionic emulsifier in percentage range of 0.001% to 25% and non-ionic emulsifier in percentage range 0.001% to 20% respectively.
4. Further blending the obtained mixture with carrier oil such as but limited to seed oil of Anethumgraveolens, Pongamaiapinnataoil, Azadirachtaindica oil, castor oil, fish oil, Sesamumindicumoil or Brassica junceaoil in a percentage range 0.001% to 75%.
5. Stirring the obtained mixture at 200RPM to 800RPM speed and subjecting to 350C to 650C temperature in closed mixing vessel, wherein the vessel is made up of stainless steel 316 grade consisting of outer jacket for maintaining temperature, connected with stirrer such as blade stirrer, propeller stirrer, turbine stirrer, anchor stirrer or universal stirrer for continuous stirring of the mixture. Further the vessel also comprises homogenizer towards the bottom which runs at a speed of 2800 RPM is attached to reduce the particle size under very high pressure, sheer, turbulence, acceleration and impact to make the particles more stable and effective.
6. Further the obtained mixture is then subjected to line shear pump working at a speed of 1800 RPM to 3600 RPM in order to reduce the particle size of the mixture to nano, so as to achieve the nano emulsion with particle size ranging from 10 nm to 900 nm.
7. After shearing the material is then passed through the filter consisting of seven layers of membrane filters with pore size less than 10 µm to achieve proper filtration.
8. Finally, the product is recovered and is used for commercialization and field trails.
[0090] Example 6
Representative botanical-basedbio-fungicidecomposition 6 and the corresponding method of preparation
An exemplary composition 6 in accordance with the principle of the present invention comprises
Sr No. Ingredient Actual Percent Percent Range
1. Cinnamomum cassia 30 0.00 1 to 40 %
2. Syzygium aromaticum 10 0.001 to 25 %
4. Thymus vulgaris 20 0.001 to 45 %
5. Non-Ionic Emulsifier 8 0.001 to 20 %
6. Anionic Emulsifier 7 0.001 to 25 %
7. Carrier oil 25 0.001 to 75 %

Method of preparation of botanical based bio-fungicide composition 6 comprises the following steps:
1. Extracting leaf and bark extract of Cinnamomum cassiaorCinnamaldehyde extracted from botanical source such as Cinnamomum genus or chemically synthesizedof percentage range about 0.001% to 40% by using various extraction techniques such as but not limited to solvent (methanol, hexane, acetone, ethyl acetate, butanol, ethanol) extract, oleoresin, steam distillation or super critical carbon dioxide extraction.
2. Similarly extracting bud extract of Syzygium aromaticumorEugenol extracted from botanical source such as Syzygium genus or Ocimum genus or chemically synthesizedof percentage range about 0.001% to 25% and aerial parts of Thymus vulgaris or Thymol extracted from botanical source such as Thymus genus or chemically synthesizedof percentage range of 0.001% to 45% by using various extraction techniques such as but not limited to solvent (methanol, hexane, acetone, ethyl acetate, butanol, ethanol) extract, oleoresin, steam distillation or super critical carbon dioxide extraction.
3. Mixing the obtained extracts with anionic emulsifier in percentage range of 0.001% to 25% and non-ionic emulsifier in percentage range 0.001% to 20% respectively.
4. Further blending the obtained mixture with carrier oil such as but limited to seed oil of Anethumgraveolens, Pongamaiapinnataoil, Azadirachtaindicaoil, castor oil, fish oil, Sesamumindicumoil or Brassica junceaoil in a percentage range 0.001% to 75%.
5. Stirring the obtained mixture at 200RPM to 800RPM speed and subjecting to 350C to 650C temperature in closed mixing vessel, wherein the vessel is made up of stainless steel 316 grade consisting of outer jacket for maintaining temperature, connected with stirrer such as blade stirrer, propeller stirrer, turbine stirrer, anchor stirrer or universal stirrer for continuous stirring of the mixture. Further the vessel also comprises homogenizer towards the bottom which runs at a speed of 2800 RPM is attached to reduce the particle size under very high pressure, sheer, turbulence, acceleration and impact to make the particles more stable and effective.
6. Further the obtained mixture is then subjected to line shear pump working at a speed of 1800 RPM to 3600 RPM in order to reduce the particle size of the mixture to nano, so as to achieve the nano emulsion with particle size ranging from 10 nm to 900 nm.
7. After shearing the material is then passed through the filter consisting of seven layers of membrane filters with pore size less than 10 µm to achieve proper filtration.
8. Finally, the product is recovered and is used for commercialization and field trails.
[0091] Example 7
Representative botanical-based bio-fungicidecomposition 7 and the corresponding method of preparation
An exemplary composition 7 in accordance with the principle of the present invention comprises
Sr No. Ingredient Actual Percent Percent Range
1. Cinnamomum cassia 25 0.00 1 to 40 %
2. Syzygium aromaticum 15 0.001 to 25 %
3. Cuminum cyminum 20 0.001 to 40 %
4. Non-Ionic Emulsifier 8 0.001 to 20 %
5. Anionic Emulsifier 7 0.001 to 25 %
6. Carrier oil 25 0.001 to 75 %

Method of preparation of botanical based bio-fungicide composition 7 comprises the following steps:
1. Extracting leaf and bark extract of Cinnamomum cassiaorCinnamaldehyde extracted from botanical source such as Cinnamomum genus or chemically synthesizedof percentage range about 0.001% to 40% by using various extraction techniques such as but not limited to solvent (methanol, hexane, acetone, ethyl acetate, butanol, ethanol) extract, oleoresin, steam distillation or super critical carbon dioxide extraction.
2. Similarly extracting bud extract of Syzygium aromaticumorEugenol extracted from botanical source such as Syzygium genus or Ocimum genus or chemically synthesizedof percentage range about 0.001% to 25% and seed extract of Cuminumcyminum or Cuminaldehyde extracted from botanical source such as Cuminum genus or chemically synthesizedof percentage range of 0.001% to 40% by using various extraction techniques such as but not limited to solvent (methanol, hexane, acetone, ethyl acetate, butanol, ethanol) extract, oleoresin, steam distillation or super critical carbon dioxide extraction.
3. Mixing the obtained extracts with anionic emulsifier in percentage range of 0.001% to 25% and non-ionic emulsifier in percentage range 0.001% to 20% respectively.
4. Further blending the obtained mixture with carrier oil such as but limited to seed oil of Anethumgraveolens, Pongamaiapinnataoil, Azadirachtaindica oil, castor oil, fish oil, Sesamumindicumoil or Brassica junceaoil in a percentage range 0.001% to 75%.
5. Stirring the obtained mixture at 200RPM to 800RPM speed and subjecting to 350C to 650C temperature in closed mixing vessel, wherein the vessel is made up of stainless steel 316 grade consisting of outer jacket for maintaining temperature, connected with stirrer such as blade stirrer, propeller stirrer, turbine stirrer, anchor stirrer or universal stirrer for continuous stirring of the mixture. Further the vessel also comprises homogenizer towards the bottom which runs at a speed of 2800 RPM is attached to reduce the particle size under very high pressure, sheer, turbulence, acceleration and impact to make the particles more stable and effective.
6. Further the obtained mixture is then subjected to line shear pump working at a speed of 1800 RPM to 3600 RPM in order to reduce the particle size of the mixture to nano, so as to achieve the nano emulsion with particle size ranging from 10 nm to 900 nm.
7. After shearing the material is then passed through the filter consisting of seven layers of membrane filters with pore size less than 10 µm to achieve proper filtration.
8. Finally, the product is recovered and is used for commercialization and field trails.
[0092] Example 8
Representative botanical based bio-fungicidecomposition 8 and the corresponding method of preparation
An exemplary composition 8 in accordance with the principle of the present invention comprises
Sr No. Ingredient Actual Percent Percent Range
1. Cinnamomum cassia 30 0.00 1 to 40 %
2. Non-Ionic Emulsifier 8 0.001 to 20 %
3. Anionic Emulsifier 7 0.001 to 25 %
4. Carrier oil 55 0.001 to 75 %

Method of preparation of botanical based bio-fungicide composition 8 comprises the following steps:
1. Extracting leaf and bark extract of Cinnamomum cassiaorCinnamaldehyde extracted from botanical source such as Cinnamomum genus or chemically synthesizedof percentage range about 0.001% to 40% by using various extraction techniques such as but not limited to solvent (methanol, hexane, acetone, ethyl acetate, butanol, ethanol) extract, oleoresin, steam distillation or super critical carbon dioxide extraction.
2. Mixing the obtained extract with anionic emulsifier in percentage range of 0.001% to 25% and non-ionic emulsifier in percentage range 0.001% to 20% respectively.
3. Further blending the obtained mixture with carrier oil such as but limited to seed oil of Anethumgraveolens, Pongamaiapinnataoil, Azadirachtaindica oil, castor oil, fish oil, Sesamumindicumoil or Brassica junceaoil in a percentage range 0.001% to 75%.
4. Stirring the obtained mixture at 200RPM to 800RPM speed and subjecting to 350C to 650C temperature in closed mixing vessel, wherein the vessel is made up of stainless steel 316 grade consisting of outer jacket for maintaining temperature, connected with stirrer such as blade stirrer, propeller stirrer, turbine stirrer, anchor stirrer or universal stirrer for continuous stirring of the mixture. Further the vessel also comprises homogenizer towards the bottom which runs at a speed of 2800 RPM is attached to reduce the particle size under very high pressure, sheer, turbulence, acceleration and impact to make the particles more stable and effective.
5. Further the obtained mixture is then subjected to line shear pump working at a speed of 1800 RPM to 3600 RPM in order to reduce the particle size of the mixture to nano, so as to achieve the nano emulsion with particle size ranging from 10 nm to 900 nm.
6. After shearing the material is then passed through the filter consisting of seven layers of membrane filters with pore size less than 10 µm to achieve proper filtration.
7. Finally, the product is recovered and is used for commercialization and field trails.
[0093] Example 9
Representative botanical based bio-fungicidecomposition 9 and the corresponding method of preparation
An exemplary composition 9 in accordance with the principle of the present invention comprises
Sr No. Ingredient Actual Percent Percent Range
1. Cuminum cyminum 25 0.001 to 40 %
2. Non-Ionic Emulsifier 10 0.001 to 20 %
3. Anionic Emulsifier 10 0.001 to 25 %
4. Carrier oil 55 0.001 to 75 %

Method of preparation of botanical based bio-fungicide composition 9 comprises the following steps:
1. Extracting seed extract of Cuminumcyminum or Cuminaldehyde extracted from botanical source such as Cuminum genus or chemically synthesized of percentage range about 0.001% to 40% by using various extraction techniques such as but not limited to solvent (methanol, hexane, acetone, ethyl acetate, butanol, ethanol) extract, oleoresin, steam distillation or super critical carbon dioxide extraction.
2. Mixing the obtained extract with anionic emulsifier in percentage range of 0.001% to 25% and non-ionic emulsifier in percentage range 0.001% to 20% respectively.
3. Further blending the obtained mixture with carrier oil such as but limited to seed oil of Anethumgraveolens, Pongamaiapinnataoil, Azadirachtaindica oil, castor oil, fish oil, Sesamumindicumoil or Brassica junceaoil in a percentage range 0.001% to 75%.
4. Stirring the obtained mixture at 200RPM to 800RPM speed and subjecting to 350C to 650C temperature in closed mixing vessel, wherein the vessel is made up of stainless steel 316 grade consisting of outer jacket for maintaining temperature, connected with stirrer such as blade stirrer, propeller stirrer, turbine stirrer, anchor stirrer or universal stirrer for continuous stirring of the mixture. Further the vessel also comprises homogenizer towards the bottom which runs at a speed of 2800 RPM is attached to reduce the particle size under very high pressure, sheer, turbulence, acceleration and impact to make the particles more stable and effective.
5. Further the obtained mixture is then subjected to line shear pump working at a speed of 1800 RPM to 3600 RPM in order to reduce the particle size of the mixture to nano, so as to achieve the nano emulsion with particle size ranging from 10 nm to 900 nm.
6. After shearing the material is then passed through the filter consisting of seven layers of membrane filters with pore size less than 10 µm to achieve proper filtration.
7. Finally, the product is recovered and is used for commercialization and field trails.
[0094] Example 10
Representative botanical based bio-fungicide composition10 and the corresponding method of preparation
An exemplary composition 10 in accordance with the principle of the present invention comprises
Sr No. Ingredient Actual Percent Percent Range
4. Thymus vulgaris 30 0.001 to 45 %
5. Non-Ionic Emulsifier 10 0.001 to 20 %
6. Anionic Emulsifier 7 0.001 to 25 %
7. Carrier oil 53 0.001 to 75 %

Method of preparation of botanical based bio-fungicide composition 10 comprises the following steps:
1. Extracting aerial parts of Thymus vulgaris or Thymol extracted from botanical source such as Thymus genus or chemically synthesized of percentage range about 0.001% to 45% by using various extraction techniques such as but not limited to solvent (methanol, hexane, acetone, ethyl acetate, butanol, ethanol) extract, oleoresin, steam distillation or super critical carbon dioxide extraction.
2. Mixing the obtained extract with anionic emulsifier in percentage range of 0.001% to 25% and non-ionic emulsifier in percentage range 0.001% to 20% respectively.
3. Further blending the obtained mixture with carrier oil such as but limited to seed oil of Anethumgraveolens, Pongamaiapinnataoil, Azadirachtaindica oil, castor oil, fish oil, Sesamumindicumoil or Brassica junceaoil in a percentage range 0.001% to 75%.
4. Stirring the obtained mixture at 200RPM to 800RPM speed and subjecting to 350C to 650C temperature in closed mixing vessel, wherein the vessel is made up of stainless steel 316 grade consisting of outer jacket for maintaining temperature, connected with stirrer such as blade stirrer, propeller stirrer, turbine stirrer, anchor stirrer or universal stirrer for continuous stirring of the mixture. Further the vessel also comprises homogenizer towards the bottom which runs at a speed of 2800 RPM is attached to reduce the particle size under very high pressure, sheer, turbulence, acceleration and impact to make the particles more stable and effective.
5. Further the obtained mixture is then subjected to line shear pump working at a speed of 1800 RPM to 3600 RPM in order to reduce the particle size of the mixture to nano, so as to achieve the nano emulsion with particle size ranging from 10 nm to 900 nm.
6. After shearing the material is then passed through the filter consisting of seven layers of membrane filters with pore size less than 10 µm to achieve proper filtration.
7. Finally, the product is recovered and is used for commercialization and field trails.

[0095] Example 11
Representative botanical based bio-fungicidecomposition 11 and the corresponding method of preparation
An exemplary composition 11 in accordance with the principle of the present invention comprises
Sr No. Ingredient Actual Percent Percent Range
1. Syzygium aromaticum 30 0.001 to 50 %
2. Non-Ionic Emulsifier 10 0.001 to 25 %
3. Anionic Emulsifier 7 0.001 to 25 %
4. Carrier oil 53 0.001 to 80 %

Method of preparation of botanical based bio-fungicide composition 11 comprises the following steps:
1. Extracting bud extract of Syzygium aromaticumorEugenol extracted from botanical source such as Syzygium genus or Ocimum genus or chemically synthesized of percentage range about 0.001% to 50% by using various extraction techniques such as but not limited to solvent (methanol, hexane, acetone, ethyl acetate, butanol, ethanol) extract, oleoresin, steam distillation or super critical carbon dioxide extraction.
2. Mixing the obtained extract with anionic emulsifier in percentage range of 0.001% to 25% and non-ionic emulsifier in percentage range 0.001% to 25% respectively.
3. Further blending the obtained mixture with carrier oil such as but limited to seed oil of Anethumgraveolens, Pongamaiapinnataoil, Azadirachtaindica oil, Ricinuscommunis oil, fish oil, Sesamumindicum oil, or Brassica junceaoil in a percentage range 0.001% to 80%.
4. Stirring the obtained mixture at 200RPM to 800RPM speed and subjecting to 350C to 650C temperature in closed mixing vessel, wherein the vessel is made up of stainless steel 316 grade consisting of outer jacket for maintaining temperature, connected with stirrer such as blade stirrer, propeller stirrer, turbine stirrer, anchor stirrer or universal stirrer for continuous stirring of the mixture. Further the vessel also comprises homogenizer towards the bottom which runs at a speed of 2800 RPM is attached to reduce the particle size under very high pressure, sheer, turbulence, acceleration and impact to make the particles more stable and effective.
5. Further the obtained mixture is then subjected to line shear pump working at a speed of 1800 RPM to 3600 RPM in order to reduce the particle size of the mixture to nano, so as to achieve the nano emulsion with particle size ranging from 10 nm to 900 nm.
6. After shearing the material is then passed through the filter consisting of seven layers of membrane filters with pore size less than 10 µm to achieve proper filtration.
7. Finally, the product is recovered and is used for commercialization and field trails.

[0096] From the foregoing discussion, it is apparent that the botanical based bio-fungicide composition prepared from the various plant extracts have a holistic defense approach to control the plant pathogenic fungus. However, the risk of resistance development of botanic bio-fungicide against the pathogenic fungus is very low when compared to the synthetic chemical fungicides. Therefore, the botanical based bio-fungicide composition thus prepared is safe and non-toxic for the environment, bio-degradable and residue free.

[0097] The foregoing description of the specific embodiments will sofully reveal the general nature of the embodiments herein that others can, byapplying current knowledge, readily modify and/or adapt for various applicationssuch specific embodiments without departing from the generic concept, and,therefore, such adaptations and modifications should and are intended to becomprehended within the meaning and range of equivalents of the disclosedembodiments.

[0098] It is to be understood that the phraseology or terminologyemployed herein is for the purpose of description and not of limitation.Therefore, while the embodiments herein have been described in terms ofpreferred embodiments, those skilled in the art will recognize that theembodiments herein can be practiced with modifications.