Description:TECHNICAL FIELD OF THE INVENTION:
The present invention relates to bio-fungicide and bio-stimulant composition comprising phytochemicals extracted from plants that is cost effective and environment friendly and process for preparation thereof.

BACKGROUND AND PRIOR ART:

Fungicides play a critical role in global agriculture by protecting crops from fungal pathogens that threaten food security, economic stability, and agricultural productivity. Fungal diseases, such as rusts, blights, mildews, and rots, can cause significant yield losses, estimated to reduce global crop production by 10–20% annually if left uncontrolled. In developing nations, where agriculture is a primary economic driver, fungal infections can devastate staple crops like rice, wheat, and maize, exacerbating food insecurity and poverty. Fungicides ensure consistent crop yields, maintain food quality, and support the livelihoods of millions of farmers worldwide. By preventing post-harvest losses and improving storage life, fungicides also contribute to reducing food waste, a critical factor in meeting the demands of a growing global population projected to reach 9.7 billion by 2050 (FAO, 2017). Additionally, fungicides enable sustainable intensification of agriculture by maximizing output on limited arable land, aligning with global efforts to balance productivity with environmental sustainability.
Uses of Fungicides in Global Agriculture
Fungicides are applied across various agricultural contexts to manage fungal diseases and enhance crop production.
Fungicides are used to control fungal pathogens affecting major crops.
Fungicides, such as carbendazim, are applied to seeds to prevent seed-borne fungal diseases, ensuring healthy germination and early plant growth.
Fungicides like propiconazole are used to prevent fungal spoilage during storage and transportation, extending the shelf life of perishable crops.
Copper compounds, such as copper oxychloride, are extensively used in organic farming to control fungal diseases in crops like grapes and tomatoes.
Fungicides with different modes of action (e.g., benzimidazoles, strobilurins) are used in rotation or mixtures to delay the development of fungicide-resistant fungal strains, ensuring long-term disease control.
Fungicides are applied to high-value crops to maintain aesthetic quality and prevent diseases supporting the horticultural industry.
Chemical fungicides are essential for protecting crops from fungal pathogens, ensuring global food security. However, their widespread and often uncontrolled use has led to significant environmental and health concerns, prompting the need for safer, sustainable alternatives. Research indicates that chemical fungicides, such as triazoles, strobilurins, and dithiocarbamates, cause soil and water pollution, harm non-target organisms, and contribute to health risks like cancer. Uncontrolled use, particularly in developing nations with limited regulations, exacerbates these issues, leading to residue buildup and fungicide-resistant fungal strains.
Environmental Hazards
While fungicides are indispensable for global agriculture, their widespread use, particularly of chemical fungicides like mancozeb, copper compounds, benzimidazoles, DMIs, and strobilurins, raises concerns about environmental and health impacts. These fungicides have half-lives ranging from days (e.g., mancozeb: 6–15 days in soil) to decades (e.g., copper compounds), leading to soil degradation, water contamination, and health risks like endocrine disruption and carcinogenicity.
The persistence of these fungicides, driven by their half-lives, leads to significant environmental impacts:
Soil Degradation
Water Contamination
Harms beneficial organisms
Bioaccumulation
Skin irritation, respiratory issues, or dermatitis due to direct exposure.
Chronic Health Effects
Fungicide Resistance
The half-lives of fungicides like mancozeb (6–15 days in soil), copper compounds (decades), carbendazim (25–180 days), propiconazole (70–214 days), and azoxystrobin (70–180 days) contribute to their environmental persistence, exacerbating soil degradation, water contamination, and health risks.
Uncontrolled fungicide use, especially in developing nations with limited regulations, intensifies these hazards. Indiscriminate application leads to residue buildup in food chains, environmental pollution, and soil infertility. For example, mancozeb overuse contaminates groundwater, affecting aquatic ecosystems and human health. This practice also drives fungicide resistance, further complicating disease management.
Need for Alternatives
Due to environmental persistence, toxicity, and environmental and health risks associated with chemical fungicides, coupled with resistance development, there is an urgent need for eco-friendly, effective, biodegradable and non-toxic alternatives like phytochemicals that protect ecosystems and human health while effectively managing fungal diseases.
Phytochemicals as Alternatives
Plants produce a diverse array of secondary metabolites, including phytochemicals, to protect themselves against biotic (pests, pathogens) and abiotic (drought, UV radiation, temperature) stresses. These compounds serve as direct defences by repelling or inhibiting attackers and as signalling molecules to trigger host defences responses, such as systemic acquired resistance (SAR) or induced systemic resistance (ISR).
Phytochemicals, such as alkaloids, flavonoids, and terpenoids from plants like neem (Azadirachta indica) and thyme (Thymus vulgaris), offer a sustainable alternative. These compounds are biodegradable, less toxic to non-target organisms, and effective against pathogens like Fusarium and Aspergillus without harmful residues. Their complex modes of action reduce resistance development, and their renewable nature enhances cost-effectiveness.
Phytochemicals offer a biodegradable, low-toxicity solution, aligning with global demands for sustainable agriculture. Phytochemicals mitigate environmental and health risks associated with chemical fungicides (e.g., mancozeb’s 6–15-day soil half-life, copper compounds’ decades-long accumulation) and toxicity.
US6482455B1 discloses a composition for the control of post-harvest pathologies of fruits and vegetables comprising active ingredient consisting of the association of thymol, eugenol and cinnamaldehyde, a secondary active ingredient consisting of the association of cuminaldehyde, geraniol, vanillin, borneol, menthol, anethole, terpineol, limonene and different glycosides contained in the essences of mustard and jasmine, a surface-active selected among the different families accepted as emulsifying food additives and an oligosaccharide selected among the mono-oligosaccharides, fructo-oligosaccharides, phyco-oligosaccharides proceeding from algae extracts or vegetable tissue.

2001/MUM/2014 discloses Cinnamaldehyde and eugenol as effective fungicide and bactericide for plants and also for protecting the crop against various pathogens, with special application to the cumin crop. The compounds cinnamaldehyde and eugenol not only prevents the crop against various diseases but is also helpful for the cumin crop growth and development.

US2024415129A1 discloses synergistic botanical based bioinsecticidal, bio-stimulant composition for controlling pests, increasing plant growth and crop yield by improving plant physiological process like photosynthesis and other essential metabolic process in plants/crops that is cost effective and environment friendly.

There is a growing trend towards eco-friendly agricultural practices that prioritize both safety and sustainability. This shift is crucial in addressing the negative impacts of chemical treatments on human health and the environment, promoting a more responsible approach to pest management. Thus, there exists a need in the art to provide improved compositions with fungicide activity that minimize the dependency on synthetic chemical compounds, are environmentally friendly and are safe for agriculture and humans thereby promote environmental sustainability.

The present inventors felt that there is a need to improve the prior art compositions to improve the bio-fungicide and bio-stimulant properties. Hence, the present inventors have come up with a synergistic bio-fungicide composition which is eco friendly, relatively inexpensive, bio-degradable, residue free, safe for agriculture and humans and minimizes the risk of resistance development. The phytochemicals and its composition thereof of the present invention are observed to be highly toxic to the fungal pathogens only and do not pose any risk to humans or the environment.

OBJECT OF THE INVENTION:

It is an object of the present invention to provide bio-fungicide composition, comprising phytochemicals useful for controlling and preventing the fungal pathogens and thereby protecting crops.

It is another object of the present invention to provide a process for preparation of said bio-fungicide composition.
SUMMARY OF THE INVENTION:

In accordance with the above, the present invention provides an effective, safe and environment friendly bio-fungicide and bio-stimulant composition comprising blend of phytochemicals selected from the group consisting of (i) (un)substituted or substituted alkaloids, (ii) (un)substituted or substituted terpenoids and cyclic ketones, (iii) (un)substituted or substituted, saturated or unsaturated aldehydes, ketones, amides, acids or esters; (iv) (un)substituted or substituted aryl or heteoaryl or cyclic or fused aryls; (v) (un)substituted or substituted heterocyclic compounds which may be fused; (vi) (un)substituted or substituted quinolones and isoquinolines; (vii) essential oils alone or combinations thereof.

In an aspect, the phytochemicals of the present invention may be in the form of salts, solvates, hydrates, isomers or its enantiomers.

In an aspect, the phytochemicals of the present invention as bio-fungicide and bio-stimulant comprises one or more of camphor, eugenol, citral, thymol, Piperine; Cuminaldehyde; Methyl chavicol; Carvacrol; D-limonene; Gingerol; β-Asarone; Menthol; Capsaicin; p-Cymene; Chavibetol; Thymoquinone; Berberine; Alpha-pinene; 1,8 cineole; Camphene; Cinnamic aldehyde; Mixture of Essential oils and oleoresin; Essential oils and Total turmerones in an amount ranging between 0.001 to 45.0 %.

In a main aspect, the present invention provides a bio-fungicide and bio-stimulant composition comprising two or more phytochemicals selected from the group consisting of:
Camphor at a concentration in a range of 0.001-35 %, more preferably 2.5 %;
Eugenol at a concentration in a range of 0.001-35 %, more preferably 13.5 %;
Citral at a concentration in a range of 0.001-25 %, more preferably 5.5 %;
Thymol at a concentration in a range of 0.001-45 %, more preferably 17.0 %;
Piperine at a concentration in a range of 0.001-25 %, more preferably 12.0 %;
Cuminaldehyde at a concentration in a range of 0.001-15 %, more preferably 6.5 %;
Methyl chavicol at a concentration in a range of 0.001-35 %, more preferably 8.5 %;
Carvacrol at a concentration in a range of 0.001-25 %, more preferably 7.5 %;
D-limonene at a concentration in a range of 0.001-45 %, more preferably 20.0 %;
Gingerol at a concentration in a range of 0.001-25 %, more preferably 4.5 %;
β-Asarone at a concentration in a range of 0.001-25 %, more preferably 8.5 %;
Menthol at a concentration in a range of 0.001-40 %, more preferably 5.0 %;
Capsaicin at a concentration in a range of 0.001-20 %, more preferably 3.5 %;
p-Cymene at a concentration in a range of 0.001-25 %, more preferably 6.0 %;
Chavibetol at a concentration in a range of 0.001-25 %, more preferably 5.5 %;
Thymoquinone at a concentration in a range of 0.001-25 %, more preferably 2.5 %;
Berberine at a concentration in a range of 0.001-40 %, more preferably 4.5 %;
Alpha-pinene at a concentration in a range of 0.001-40 %, more preferably 4.0 %;
1,8 cineole at a concentration in a range of 0.001-40 %, more preferably 3.0 %;
camphene at a concentration in a range of 0.001-20 %, more preferably 7.5 %;
Cinnamic aldehyde at a concentration in a range of 0.001-15 %, more preferably 5.5 %;
Mixture of Essential oils and oleoresin at a concentration in a range of 0.001-20 %, more preferably 5.0 %; and
Essential oils and Total turmerones at a concentration in a range of 0.001-20 %, more preferably 5.0 %;
together with agriculturally acceptable excipients or additives.

In another aspect, the present invention provides a bio-fungicide and bio-stimulant composition of said phytochemicals together with agriculturally acceptable excipients or additives in suitable amounts.
In yet another aspect, the agriculturally acceptable excipients or additives are selected from binders, diluents, surfactants, emulsifiers, carriers, lubricants, solvents, pH adjusters, colorants, essential oils, anti-caking agent, dispersing agent and the like, alone or mixtures thereof.

In yet another aspect, the emulsifier is selected from group consisting of Span 80, polysorbate 80, polysorbate 60, Gaur gum, ethoxylated castor oil, Polyorganosiloxane, soy lecithin, carrageenan, mono- and diglycerides, carboxymethylcellulose, and the like in a range of 0.001 – 10 %, more preferably 4.3 %.

In another aspect, the surfactant is selected from the group consisting of Sodium oleoyl amino fatty acid, Sodium dodecyl sulphate, Polyoxyl 35 hydrogenated castor oil, Sodium N methyl N-Oleyl taurate, Sodium alkyl naphthalene sulfonate and the like in a range of 0.1 % to 15 %, more preferably 5.0 %.
In another aspect, the solvents are selected from one or more water, Dimethyl sulfoxide (DMSO), Benzyl acetate, N-methyl pyrrolidinone, Diacetone alcohol, N-Ethyl-2 pyrrolidone (NEP) and the like in a range of 10.0- 55.0 %, more preferably 55.0 %.
In another aspect, the carriers are selected from at least one substantially water-miscible co-solvent, preferably selected from the group of N-rnethylpyrrolidinone; dimethylsulphoxide; dimethylfonnamide C9; methyl ethyl ketone, Ethylene Glycol Diacetate, dimethylisosorbide isophorone; acetophenone; cyclohexanone; Diacetone alcohol 1,3-dimethy1-2-imidazolidonone; ethylene, propylene, and butylene carbonates; lactate esters; Methyl oleate, dimethyl and diethylcarbonates; alkylglycol ethers; glycols, including propylene, carbapol 940 and biodiesel and the like in a range of 10.0- 55.0 %, more preferably 35.0 %.

In another aspect, the essential oils are selected from the group consisting of seed oil of Essential oil of Ferula asafoetida, Orange oil, camphor, thyme, clove, pepper, spearmint, citronella, cassia, orange oil, star anise, cedar wood, peppermint, ginger, turmeric and bay leaf and the like in a range of 0.1 to 10 %, more preferably 1.5 %.

In another aspect, the anti-caking agent is selected from fumed silica in a range of 0.001 to 18 %, more preferably 4.5 %. and dispersing agent is selected from sodium lignosulphonate in a range of 0.001 to 25 %, more preferably 5 %.

In another aspect, the composition of the present invention has a particle size in the range of 10-1000 nanometer, more specifically 1 - 100 nanometer.

In another aspect, the present invention provides a bio-fungicidal and bio-stimulant composition comprising:
Thymol in the range of 0.001 to 45 %;
Eugenol in the range of 0.001 to 35 %;
Piperine in the range of 0.001 to 25 %;
Sodium lignosulphonate in the range of 0.1 to 25 %;
Fumed silica in the range of 0.1 to 18 %;
Maltodextrin in the range of 35 to 95 %;
Sodium lauryl sulfate in the range of 0.001 to 20 %; and
Citric acid in the range of 0.001 to 18 %.
In another aspect, the present invention provides a bio-fungicidal and bio-stimulant composition comprising:
Thymol in the range of 3 to 45 %;
Sodium Lauryl sulphate in the range of 0.001 to 20 %;
Sodium Ligno sulphonate in the range of 0.001 to 15 %;
Fumed silica in the range of 0.001 to 18 %;
Polysorbate-80 in the range of 0.001 to 25 %;
Poly glycol based defoamer in the range of 0.001 to 25 %; and
China clay in the range of 25.0 to 85 %.

In another aspect, the present invention provides a bio-fungicidal and bio-stimulant composition comprising:
Piperine in the range of 0.001 to 25 %;
Total turmerones in the range of 0.001 to 20 %;
Gingerol in the range of 0.001 to 35 %;
Thymol in the range of 0.001 to 45 %;
Camphor in the range of 0.001 to 35 %;
Eugenol in the range of 0.001 to 35 %;
Sodium lignosulphonate in the range of 0.1 to 25 %;
Fumed silica in the range of 0.1 to 18 %;
Maltodextrin in the range of 35 to 95 %;
Sodium lauryl sulfate in the range of 0.001 to 20 %; and
Citric acid in the range of 0.001 to 18 %.

In another aspect, the present invention provides a bio-fungicidal and bio-stimulant composition comprising:
Thymol in the range of 0.001 to 45 %;
Cumin aldehyde in the range of 0.001 to 20 %;
Cinnamaldehyde in the range of 0.001 to 15 %;
Sodium lignosulphonate in the range of 0.1 to 25 %;
Fumed silica in the range of 0.1 to 18 %;
Cyclodextrins in the range of 35 to 95 %;
Sodium lauryl sulfate in the range of 0.001 to 20 %; and
Citric acid in the range of 0.001 to 18 %.

In another aspect, the present invention provides a bio-fungicidal and bio-stimulant composition comprising:
Thymol in the range of 0.001 to 45 %;
Piperine in the range of 0.001 to 25 %;
Eugenol in the range of 0.001 to 35 %;
Sodium lignosulphonate in the range of 0.001 to 25 %;
Fumed silica in the range of 0.001 to 18 %;
Lactose in the range of 35 to 95 %;
Sodium lauryl sulfate in the range of 0.001 to 20 %; and
Citric acid in the range of 0.001 to 18 %.

In another aspect, the present invention provides a bio-fungicidal and bio-stimulant composition comprising:
Piperine in the range of 0.001 to 25 %;
Eugenol in the range of 0.001 to 35 %;
Citral in the range of 0.001 to 25 %;
Sodium lignosulphonate in the range of 0.001 to 25 %;
Fumed silica in the range of 0.001 to 18 %;
China clay in the range of 35 to 95 %;
Sodium lauryl sulfate in the range of 0.001 to 20 %; and
Citric acid in the range of 0.001 to 18 %.

In another aspect, the present invention provides a bio-fungicidal and bio-stimulant composition comprising:
Menthol in the range of 0.001 to 40 %;
Total Turmerones in the range of 0.001 to 20 %;
Berberine in the range of 0.001 to 40 %;
Alpha pinene in the range of 0.001 to 40 %;
1,8 cineole in the range of 0.001 to 40 %;
Carvacrol in the range of 0.001 to 40 %;
eugenol in the range of 0.001 to 35 %;
Citral in the range of 0.001 to 25 %;
Sodium lignosulphonate in the range of 0.1 to 25 %;
Fumed silica in the range of 0.1 to 18 %;
China clay in the range of 35 to 95 %;
Sodium lauryl sulfate in the range of 0.001 to 20 %; and
Citric acid in the range of 0.001 to 18 %;

In another aspect, the present invention provides a bio-fungicidal and bio-stimulant composition comprising:
Thymol in the range of 0.001 to 45 %;
Eugenol in the range of 0.001 to 35 %;
Piperine in the range of 0.001 to 25 %;
Gaur gum in the range of 0.001 to 7.5 %;
ethoxylated castor oil in the range of 0.001 to 10 %;
Sodium oleoyl amino fatty acid in the range of 0.1 to 15 %;
Dimethyl sulfoxide (DMSO) in the range of 10.0 to 55 %; and
N-Ethyl-2 pyrrolidone (NEP) in the range of 10.0 to 55 %.

In another aspect, the present invention provides a bio-fungicidal and bio-stimulant composition comprising:
Piperine in the range of 0.001 to 25 %;
Methyl chavicol in the range of 0.001 to 35 %;
Camphor in the range of 0.001 to 35 %;
Gaur gum in the range of 0.001 to 7.5 %;
ethoxylated castor oil in the range of 0.001 to 10 %;
Sodium oleoyl amino fatty acid in the range of 0.1 to 15 %;
Benzyl acetate in the range of 10.0 to 35 %; and
Water in the range of 10.0 to 35 %;

In another aspect, the bio-fungicide and bio-stimulant composition of the present invention is in the form of powder and liquid formulation.

In an aspect, the present invention provides a process for preparation of the bio-fungicide and bio-stimulant powder composition comprising:
Dry blending the powdered phytochemicals and excipients or additives to form a pre-mix;
Gradually adding the surfactant and buffer into the pre-mix of step (i) and blending to form a homogenous intermediate powder blend;
Subjecting the intermediate powder blend of step (ii) to micronization to obtain micronized powder having fine particle size in the range of 1–10 µm;
Re-blending the micronized powder of step (iii) to obtain the final composition;
Passing the final composition obtained in step (iv) through a vibro-sifter equipped with a 100–120 mesh screen to ensure uniform particle distribution;
Recovering the final product.

In another aspect, the dry blending of step (i) is done by using any one of the following: V-Blender (Twin-Shell Blender), Double Cone Blender, Ribbon Blender (Horizontal Blender), Paddle Blender, Octagonal Blender, Bin Blender (IBC – Intermediate Bulk Container Blender), High-Shear Mixer (for wet powder mixing/granulation), Turbula Mixer / 3D Mixer, Ploughshare Mixer, Conical Screw Mixer (Nauta Mixer), more particularly ribbon blender and High-Shear Mixer.

In another aspect, the blending of step (ii) is done by using High-Shear Mixer and Sigma Mixer to attain homogeneity.

In yet another aspect, the micronization of step (iii) is done by jet milling, cryogenic milling and air classifier milling to achieve precise size reduction.

In an aspect, the present invention provides a process for preparation of the bio-fungicide and bio-stimulant liquid composition comprising:
Dissolving the emulsifier and surfactant in the solvent to make inert mixture and agitating with homogenizer until a uniform blend is formed;
Adding the phytochemicals to the blend formed in step (a) and homogenizing completely to make an emulsion concentration.
Stirring the mixture obtained in step (b) at 300-1000 RPM particularly at 350-800 RPM at 25-55°C followed by homogenizing the mix at a speed of 3700~27000 rpm to reduce the particle size;
Passing the homogenized mixture obtained in step (c) through in line shear pump with 1800 RPM to 5800 RPM to obtain the nano emulsion with particle size ranging from 10 to 1000 nano meter;
Passing the mixture obtained in step (d) through High-pressure homogenization to obtain the particle size below 100 nano meters;
Passing the mixture obtained in step (e) through sparkler filter having seven layers of membrane filters with pore size less than 1-2 micron to achieve desired filtration; and
Recovering the final product.

In another aspect, the high-pressure homogenization of step (e) is carried out at a speed of 4000 rpm to 10000 rpm and pressure up to 4,200 bar (60,000 psi).

In another aspect, the excipients in the above processes are selected from binders, diluents, surfactants, emulsifiers, carriers, lubricants, solvents, pH adjusters, colorants, essential oils, anti-caking agent, dispersing agent and the like, alone or mixtures thereof.

In another aspect, the bio-fungicide and bio-stimulant composition of the present invention may be used in conjunction with the known insecticidal or acaricidal active ingredient.

In yet another aspect, the phytochemicals of the present invention may be used with the active ingredient contained in other agents such as bactericides, nematocides, plant growth regulators, synergists, fertilizers, soil improvers, animal feeds and the like.

The composition of the present invention may be formulated into a known form such as wettable powder, granule, powder, tablet, emulsion, water-soluble agent, suspension, granule wettable powder, flowable agent, microcapsule, aerosol, propellant, spray, fogging agent, heating transpiration agent, smoking agent, baiting agent or the like.

In yet another aspect, the present invention provides a method for controlling fungal growth or killing fungal pathogens comprising applying said biofungicide composition to the affected parts of the plants in suitable amount thereof.

The composition of the present invention is useful for controlling fungal infections and improving yield of crop plants.

Description of Figures:
Figure 1: Graphical representation of Bio-efficacy of Bio-Fungicide and Synthetic Fungicide against Downy mildew disease of Grape.
Figure 2: Graphical representation of Bio-efficacy of Bio-Fungicide and Synthetic Fungicide against Rust disease of Wheat.
Figure 3: Bio-efficacy of Bio-Fungicide and Synthetic Fungicide on Powdery mildew disease of Chilli.
Figure 4: Bio-efficacy of Bio-Fungicide and Synthetic Fungicide against Rhizome Rot of Ginger.
Figure 5: Graphical representation of Bio-efficacy of Bio-Fungicide and Synthetic Fungicide against blast disease of Paddy.
Figure 6: Graphical representation of the in vitro effect of bio-fungicides on growth and inhibition of Pyricularia oryzae.
Figure 7: Graphical representation of the in vitro effect of bio-fungicides on growth and inhibition of Colletotrichum capsici.
Figure 8: Graphical representation of the in vitro effect of bio-fungicides on growth and inhibition of Phytophthora infestans.
Figure 9: Graphical representation of the in vitro effect of bio-fungicides on growth and inhibition of Rhizoctonia bataticola.
Figure 10: Graphical representation of the in vitro effect of bio-fungicides on growth and inhibition of Fusarium oxysporum f. sp. ciceris.

DETAILED DESCRIPTION OF THE INVENTION:

The invention will now be described in detail in connection with certain preferred and
optional embodiments, so that various aspects thereof may be more fully understood and
appreciated.

For the purpose of present invention the term ‘phytochemicals’, ‘phyto ingredient’ and ‘active ingredients’ are used interchangeably and mean phytochemicals extracted from plants useful as bio-fungicides and bio-stimulants.
For the purpose of present invention the term ‘composition’ and ‘formulation’ are used interchangeably and mean the bio-fungicide and bio-stimulant composition comprising phytochemicals extracted from plants.
The phytochemicals—camphor, eugenol, citral, thymol, piperine, cuminaldehyde, methyl chavicol, essential oil mixtures, turmerones, carvacrol, D-limonene, gingerol, β-asarone, menthol, capsaicin, p-cymene, chavibetol, thymoquinone, berberine, alpha-pinene, 1,8-cineole, and camphene—play dual roles in plant defences. They directly repel pests and inhibit pathogens while triggering host defence mechanisms like SAR and ISR, enhancing resistance to biotic and abiotic stresses.
In an embodiment, the present invention provides an effective, safe and environment friendly bio-fungicide and bio-stimulant composition comprising blend of phytochemicals selected from the group consisting of (i) (un)substituted or substituted alkaloids, (ii) (un)substituted or substituted terpenoids and cyclic ketones, (iii) (un)substituted or substituted, saturated or unsaturated aldehydes, ketones, amides, acids or esters; (iv) (un)substituted or substituted aryl or heteoaryl or cyclic or fused aryls; (v) (un)substituted or substituted heterocyclic compounds which may be fused; (vi) (un)substituted or substituted quinolones and isoquinolines; (vii) essential oils alone or combinations thereof.

In another embodiment, the phytochemicals of the present invention may be in the form of salts, solvates, hydrates, isomers or its enantiomers.

In an embodiment, the phytochemicals of the present invention comprise;
Sr. No. Phyto Ingredient Structure of Phyto Ingredient Molecular Formula Molecular Weight
1. Camphor
C10H16O
152.23
2. Eugenol
C10H12O2
164.20
3. Citral
C10H16O
152.23
4. Thymol
C10H14O
150.22
5. Piperine
C17H19NO3
285.34
6. Cuminaldehyde
C10H12O
148.20
7. Methyl chavicol
C10H12O
148.20
12. Carvacrol
C10H14O
150.22
13. D-limonene
C10H16
136.23
14. Gingerol

C17H26O4
294.39
15. β-Asarone
C12H16O3

208.25
16. Menthol
C10H20O

156.27
17. Capsaicin

C18H27NO3 .

305.41
18. p-Cymene
C10H14
134.22
22. Chavibetol
C10H12O2
164.20
23. Thymoquinone C10H12O2

164.20
24. Berberine

C20H18NO4+
336.4
26. Alpha-pinene
C10H16

136.23
27. 1,8 cineole
C10H18O
154.25
28. camphene
C10H16
136.23
29. Cinnamic aldehyde C9H8O 132.16

In another embodiment, the phytochemicals of the present invention as bio-fungicides and bio-stimulant comprises one or more of camphor, eugenol, citral, thymol, Piperine; Cuminaldehyde; Methyl chavicol; Carvacrol; D-limonene; Gingerol; β-Asarone; Menthol; Capsaicin; p-Cymene; Chavibetol; Thymoquinone; Berberine; Alpha-pinene; 1,8 cineole; Camphene; Cinnamic aldehyde; Mixture of Essential oils and oleoresin; Essential oils and Total turmerones in an amount ranging between 0.001 to 45 %.

In another embodiment, the phytochemicals of the present invention may be in the form of salts, solvates, hydrates, isomers or its enantiomers.

In a preferred embodiment, the present invention provides a bio-fungicide and bio-stimulant composition comprising two or more phytochemicals selected from the group consisting of:
Camphor at a concentration in a range of 0.001-35 %, more preferably 2.5 %;
Eugenol at a concentration in a range of 0.001-35 %, more preferably 13.5 %;
Citral at a concentration in a range of 0.001-25 %, more preferably 5.5 %;
Thymol at a concentration in a range of 0.001-45 %, more preferably 17.0 %;
Piperine at a concentration in a range of 0.001-25 %, more preferably 12.0 %;
Cuminaldehyde at a concentration in a range of 0.001-15 %, more preferably 6.5 %;
Methyl chavicol at a concentration in a range of 0.001-35 %, more preferably 8.5 %;
Carvacrol at a concentration in a range of 0.001-25 %, more preferably 7.5 %;
D-limonene at a concentration in a range of 0.001-45 %, more preferably 20.0 %;
Gingerol at a concentration in a range of 0.001-25 %, more preferably 4.5 %;
β-Asarone at a concentration in a range of 0.001-25 %, more preferably 8.5 %;
Menthol at a concentration in a range of 0.001-40 %, more preferably 5.0 %;
Capsaicin at a concentration in a range of 0.001-20 %, more preferably 3.5 %;
p-Cymene at a concentration in a range of 0.001-25 %, more preferably 6.0 %;
Chavibetol at a concentration in a range of 0.001-25 %, more preferably 5.5 %;
Thymoquinone at a concentration in a range of 0.001-25 %, more preferably 2.5 %;
Berberine at a concentration in a range of 0.001-40 %, more preferably 4.5 %;
Alpha-pinene at a concentration in a range of 0.001-40 %, more preferably 4.0 %;
1,8 cineole at a concentration in a range of 0.001-40 %, more preferably 3.0 %;
camphene at a concentration in a range of 0.001-20 %, more preferably 7.5 %;
Cinnamic aldehyde at a concentration in a range of 0.001-15 %, more preferably 5.5 %;
Mixture of Essential oils and oleoresin at a concentration in a range of 0.001-20 %, more preferably 5.0 %; and
Essential oils and Total turmerones at a concentration in a range of 0.001-20 %, more preferably 5.0 %;
together with agriculturally acceptable excipients or additives.

In another embodiment, the present invention provides a bio-fungicide and bio-stimulant composition of said phytochemicals together with agriculturally acceptable excipients or additives in suitable amounts.
In yet another embodiment, the agriculturally acceptable excipients or additives are selected from binders, diluents, surfactants, emulsifiers, carriers, lubricants, solvents, pH adjusters, colorants, essential oils, anti-caking agent, dispersing agent and the like, alone or mixtures thereof.

In yet another embodiment, the emulsifier is selected from group consisting of Span 80, polysorbate 80, polysorbate 60, Gaur gum, ethoxylated castor oil, Polyorganosiloxane, soy lecithin, carrageenan, mono- and diglycerides, carboxymethylcellulose, and the like in a range of 0.001 – 10 %, more preferably 4.3 %.

In another embodiment, the surfactant is selected from the group consisting of sodium lauryl sulfate, Polysorbate 80, polyethylene glycol derivatives, and sodium lauryl sulfate, Sodium oleoyl amino fatty acid, Sodium dodecyl sulphate, Polyoxyl 35 hydrogenated castor oil, Sodium N methyl N-Oleyl taurate, Sodium alkyl naphthalene sulfonate and the like in a range of 0.1 % to 15 %, more preferably 5.0 %.
In another embodiment, the solvents are selected from one or more water, Dimethyl sulfoxide (DMSO), Benzyl acetate, N-methyl pyrrolidinone, Diacetone alcohol, N-Ethyl-2 pyrrolidone (NEP) and the like in a range of 10.0- 55.0 %, more preferably 55.0 %.

In another embodiment, the carriers are selected from at least one substantially water-miscible co-solvent, preferably selected from the group of maltodextrin, Cyclodextrins, China clay, N-rnethylpyrrolidinone; dimethylsulphoxide; dimethylfonnamide C9; methyl ethyl ketone, Ethylene Glycol Diacetate, dimethylisosorbide isophorone; acetophenone; cyclohexanone; Diacetone alcohol 1,3-dimethy1-2-imidazolidonone; ethylene, propylene, and butylene carbonates; lactate esters; Methyl oleate, dimethyl and diethylcarbonates; alkylglycol ethers; glycols, including propylene, carbapol 940 and biodiesel and the like in a range of 10.0 - 55.0 %, more preferably 35.0 %.

In another embodiment, the essential oils are selected from the group consisting of seed oil of Essential oil of Ferula asafoetida, Orange oil, camphor, thyme, clove, pepper, spearmint, citronella, cassia, orange oil, star anise, cedar wood, peppermint, ginger, turmeric and bay leaf and the like in a range of 0.1 to 10 %, more preferably 1.5 %.

In another embodiment, the anti-caking agent is selected from fumed silica in a range of 0.001 to 18 %, more preferably 4.5 %. and dispersing agent is selected from sodium lignosulphonate in a range of 0.001 to 25 %, more preferably 5 %.

In another embodiment, the composition of the present invention has a particle size in the range of 10-1000 nanometer, more specifically 1 - 100 nanometer.

In another embodiment, the present invention provides a bio-fungicide and bio-stimulant composition comprising:
Thymol in the range of 0.001 to 45 %;
Eugenol in the range of 0.001 to 35 %;
Piperine in the range of 0.001 to 25 %;
Sodium lignosulphonate in the range of 0.1 to 25 %;
Fumed silica in the range of 0.1 to 18 %;
Maltodextrin in the range of 35 to 95 %;
Sodium lauryl sulfate in the range of 0.001 to 20 %; and
Citric acid in the range of 0.001 to 18 %.

In another embodiment, the present invention provides a bio-fungicide and bio-stimulant composition comprising:
Thymol in the range of 3 to 45 %;
Sodium Lauryl sulphate in the range of 0.001 to 20 %;
Sodium Ligno sulphonate in the range of 0.001 to 15 %;
Fumed silica in the range of 0.001 to 18 %;
Polysorbate-80 in the range of 0.001 to 25 %;
Poly glycol based defoamer in the range of 0.001 to 25 %; and
China clay in the range of 25.0 to 85 %.

In yet another embodiment, the present invention provides a bio-fungicide and bio-stimulant composition comprising:
Piperine in the range of 0.001 to 25 %;
Total turmerones in the range of 0.001 to 20 %;
Gingerol in the range of 0.001 to 35 %;
Thymol in the range of 0.001 to 45 %;
Camphor in the range of 0.001 to 35 %;
Eugenol in the range of 0.001 to 35 %;
Sodium lignosulphonate in the range of 0.1 to 25 %;
Fumed silica in the range of 0.1 to 18 %;
Maltodextrin in the range of 35 to 95 %;
Sodium lauryl sulfate in the range of 0.001 to 20 %; and
Citric acid in the range of 0.001 to 18 %.

In another embodiment, the present invention provides a bio-fungicide and bio-stimulant composition comprising:
Thymol in the range of 0.001 to 45 %;
Cumin aldehyde in the range of 0.001 to 20 %;
Cinnamaldehyde in the range of 0.001 to 15 %;
Sodium lignosulphonate in the range of 0.1 to 25 %;
Fumed silica in the range of 0.1 to 18 %;
Cyclodextrins in the range of 35 to 95 %;
Sodium lauryl sulfate in the range of 0.001 to 20 %; and
Citric acid in the range of 0.001 to 18 %.

In yet another embodiment, the present invention provides a bio-fungicide and bio-stimulant composition comprising:
Thymol in the range of 0.001 to 45 %;
Piperine in the range of 0.001 to 25 %;
Eugenol in the range of 0.001 to 35 %;
Sodium lignosulphonate in the range of 0.001 to 25 %;
Fumed silica in the range of 0.001 to 18 %;
Lactose in the range of 35 to 95 %;
Sodium lauryl sulfate in the range of 0.001 to 20 %; and
Citric acid in the range of 0.001 to 18 %.

In another embodiment, the present invention provides a bio-fungicide and bio-stimulant composition comprising:
Piperine in the range of 0.001 to 25 %;
Eugenol in the range of 0.001 to 35 %;
Citral in the range of 0.001 to 25 %;
Sodium lignosulphonate in the range of 0.001 to 25 %;
Fumed silica in the range of 0.001 to 18 %;
China clay in the range of 35 to 95 %;
Sodium lauryl sulfate in the range of 0.001 to 20 %; and
Citric acid in the range of 0.001 to 18 %.

In another embodiment, the present invention provides a bio-fungicide and bio-stimulant composition comprising:
Menthol in the range of 0.001 to 40 %;
Total Turmerones in the range of 0.001 to 20 %;
Berberine in the range of 0.001 to 40 %;
Alpha pinene in the range of 0.001 to 40 %;
1,8 cineole in the range of 0.001 to 40 %;
Carvacrol in the range of 0.001 to 40 %;
eugenol in the range of 0.001 to 35 %;
Citral in the range of 0.001 to 25 %;
Sodium lignosulphonate in the range of 0.1 to 25 %;
Fumed silica in the range of 0.1 to 18 %;
China clay in the range of 35 to 95 %;
Sodium lauryl sulfate in the range of 0.001 to 20 %; and
Citric acid in the range of 0.001 to 18 %.

In yet another embodiment, the present invention provides a bio-fungicidal and bio-stimulant composition comprising:
Thymol in the range of 0.001 to 45 %;
Eugenol in the range of 0.001 to 35 %;
Piperine in the range of 0.001 to 25 %;
Gaur gum in the range of 0.001 to 7.5 %;
ethoxylated castor oil in the range of 0.001 to 10 %;
Sodium oleoyl amino fatty acid in the range of 0.1 to 15 %;
Dimethyl sulfoxide (DMSO) in the range of 10.0 to 55 %; and
N-Ethyl-2 pyrrolidone (NEP) in the range of 10.0 to 55 %.

In another embodiment, the present invention provides a bio-fungicidal and bio-stimulant composition comprising:
Piperine in the range of 0.001 to 25 %;
Methyl chavicol in the range of 0.001 to 35 %;
Camphor in the range of 0.001 to 35 %;
Gaur gum in the range of 0.001 to 7.5 %;
ethoxylated castor oil in the range of 0.001 to 10 %;
Sodium oleoyl amino fatty acid in the range of 0.1 to 15 %;
Benzyl acetate in the range of 10.0 to 35 %; and
Water in the range of 10.0 to 35 %.

In another embodiment, the present invention provides a bio-fungicide and bio-stimulant composition comprising said phytochemicals with agriculturally acceptable excipients or additives with a synergistic biological action that efficiently controls a wide range of plant fungal pathogens.

In another embodiment, the bio-fungicide and bio-stimulant composition of the present invention is in the form of powder and liquid formulation.

In an embodiment, the present invention provides a process for preparation of the bio-fungicide and bio-stimulant powder composition comprising:
Dry blending the powdered phytochemicals and excipients or additives to form a pre-mix;
Gradually adding the surfactant and buffer into the pre-mix of step (i) and blending to form a homogenous intermediate powder blend;
Subjecting the intermediate powder blend of step (ii) to micronization to obtain micronized powder having fine particle size in the range of 1–10 µm;
Re-blending the micronized powder of step (iii) to obtain the final composition;
Passing the final composition obtained in step (iv) through a vibro-sifter equipped with a 100–120 mesh screen to ensure uniform particle distribution;
Recovering the final product.

In another embodiment, the dry blending of step (i) is done by using any one of the following: V-Blender (Twin-Shell Blender), Double Cone Blender, Ribbon Blender (Horizontal Blender), Paddle Blender, Octagonal Blender, Bin Blender (IBC – Intermediate Bulk Container Blender), High-Shear Mixer (for wet powder mixing/granulation), Turbula Mixer / 3D Mixer, Ploughshare Mixer, Conical Screw Mixer (Nauta Mixer), more particularly ribbon blender and High-Shear Mixer.

In another embodiment, the blending of step (ii) is done by using High-Shear Mixer and Sigma Mixer to attain homogeneity.

In yet another embodiment, the micronization of step (iii) is done by jet milling, cryogenic milling and air classifier milling to achieve precise size reduction.

In an embodiment, the present invention provides a process for preparation of the bio-fungicide and bio-stimulant liquid composition comprising:
Dissolving the emulsifier and surfactant in the solvent to make inert mixture and agitating with homogenizer until a uniform blend is formed;
Adding the phytochemicals to the blend formed in step (a) and homogenizing completely to make an emulsion concentrate.
Stirring the emulsion concentrate obtained in step (b) at 300-1000 RPM particularly at 350-800 RPM at 25-55°C followed by homogenizing the mix at a speed of 3700~27000 rpm to reduce the particle size;
Passing the homogenized mixture obtained in step (c) through in line shear pump with 1800 RPM to 5800 RPM to obtain the nano emulsion with particle size ranging from 10 to 1000 nano meter;
Passing the mixture obtained in step (d) through High-pressure homogenization to obtain the particle size below 100 nano meters;
Passing the mixture obtained in step (e) through sparkler filter having seven layers of membrane filters with pore size less than 1-2 micron to achieve desired filtration; and
Recovering the final product.

In another embodiment, the high-pressure homogenization of step (e) is carried out at a speed of 4000 rpm to 10000 rpm and pressure up to 4,200 bar (60,000 psi).

In another embodiment, the excipients or additives in both the processes are selected from binders, diluents, surfactants, emulsifiers, carriers, lubricants, solvents, pH adjusters, colorants, essential oils, anti-caking agent, dispersing agent and the like, alone or mixtures thereof.

In another embodiment, the emulsifier in the process is selected from group consisting of Span 80, polysorbate 80, polysorbate 60, Gaur gum, ethoxylated castor oil, Polyorganosiloxane, soy lecithin, carrageenan, mono- and diglycerides, carboxymethylcellulose, and the like alone or mixtures thereof.

In another embodiment, the surfactants in the process are selected from the group consisting of sodium lauryl sulfate, Polysorbate 80, polyethylene glycol derivatives, and sodium lauryl sulfate, Sodium oleoyl amino fatty acid, Sodium dodecyl sulphate, Polyoxyl 35 hydrogenated castor oil, Sodium N methyl N-Oleyl taurate, Sodium alkyl naphthalene sulfonate and the like alone or mixtures thereof.

In another embodiment, the buffer in the process is selected from group consisting of Citric acid and the like.
In another embodiment, the solvents used in the process are selected from one or more water, Dimethyl sulfoxide (DMSO), Benzyl acetate, N-methyl pyrrolidinone, Diacetone alcohol, N-Ethyl-2 pyrrolidone (NEP) and the like alone or mixtures thereof.
In another embodiment, the carriers used in the process are selected from at least one substantially water-miscible co-solvent, preferably selected from the group of maltodextrin, Cyclodextrins, China clay, N-rnethylpyrrolidinone; dimethylsulphoxide; dimethylfonnamide C9; methyl ethyl ketone, Ethylene Glycol Diacetate, dimethylisosorbide isophorone; acetophenone; cyclohexanone; Diacetone alcohol 1,3-dimethy1-2-imidazolidonone; ethylene, propylene, and butylene carbonates; lactate esters; Methyl oleate, dimethyl and diethylcarbonates; alkylglycol ethers; glycols, including propylene, carbapol 940 and biodiesel and the like alone or mixtures thereof.

In another embodiment, the essential oils used in the process are selected from the group consisting of seed oil of Essential oil of Ferula asafoetida, Orange oil, camphor, thyme, clove, pepper, spearmint, citronella, cassia, orange oil, star anise, cedar wood, peppermint, ginger, turmeric and bay leaf and the like alone or mixtures thereof.

In another embodiment, the anti-caking agent is selected from fumed silica and dispersing agent is selected from sodium lignosulphonate.

In another embodiment, the further quantification of Phyto ingredient obtained in both the processes is carried out by using various instruments but not limited to liquid chromatography-mass spectrometry, gas chromatography-mass spectrometry, High pressure liquid chromatograph, gas chromatography, spectroscopy compound and the like.
In an embodiment, the particle size of the composition/formulation is reduced to the nano range for improved efficacy and penetration. The reduction in the particle size in the nano range is achieved by employing different modules such as a filter press, shear pump, homogenizer, and high pressure homogenization (pressure max up to 45,000 PSI) to create a nano formulation with an average particle size range of 10 to 1000 nm. Nano formulations are stable and may be obtained without the use of any additives or chemicals. Nanoparticles are obtained in the present invention employing mechanical methods with a temperature control system to maintain heat sensitive substances.

In another embodiment, the composition is provided at a concentration ranging between 0.5 to 1.0 ml /L or 0.5 to 1.0 gm /L.

In another embodiment, the bio-fungicide and bio-stimulant composition of the present invention exhibits synergism at specific concentration and thereby controls the plant fungal pathogens and protects agriculture, horticulture, olericulture and floricultural crops.

The present inventors have compared the in vitro and in vivo bio-efficacy of the present composition (Embodiments 1 to 9) with the bio-efficacy of individual phytochemicals and observed that the activity of the present Embodiments 1 to 9 were significantly better than the bio-efficacy shown by individual phytochemicals. The results are illustrated in Examples 12 to 21.

In another embodiment, the bio-fungicide and bio-stimulant composition of the present invention was effective against broad group of fungal pathogens within 48 -72 hrs of application with low risk of resistance development and most importantly safe for environment and mammalian.

In another embodiment, the bio-fungicide and bio-stimulant composition of the present invention comprises of a number of plant base Phyto ingredient which also significantly boost the plant growth with respect of height, growth, chlorophyll content, increase in flower and fruit number and subsequently the yield and most importantly being originated from plants they are easily uptake by plants.

In another embodiment, the synergistic fungicidal activity of the phytochemicals—camphor, eugenol, citral, thymol, piperine, cuminaldehyde, methyl chavicol, mixture of essential oils and oleoresin, essential oils and total turmerones, carvacrol, D-limonene, gingerol, β-asarone, menthol, capsaicin, p-cymene, chavibetol, thymoquinone, berberine, alpha-pinene, 1,8-cineole, and camphene was evaluated in comparison to their individual activities.
The bio-fungicide and bio-stimulant composition of the present invention is useful for organic and residue free grower.

The in vitro and in vivo bio-efficacy studies conducted on numerous crops demonstrated that the bio-efficacy is consistent and stable against various fungal pathogens.

In another embodiment, the composition of the present invention is useful for controlling fungal infection in plants and improving yield of crop plants.

In another embodiment, the composition of the present invention exhibits biostimulant activity and thereby improves plant growth and crop yield.

In an embodiment, the composition of the present invention may be applied by sprinkler application, sprayer application or drip application, more preferably by sprayer application such as foliar sprays, sprays to be applied to plants shoots and the like.

In yet another embodiment, the composition of the present invention is effective against fungal pathogens such as Plasmopara viticola, Puccinia graminis, Leveillula taurica, Pythium aphanidermatum, Pyricularia oryzae, Colletotrichum capsici, Phytophthora infestans, Rhizoctonia bataticola, Fusarium oxysporum f. sp. ciceris, Fusarium oxysporum, Fusarium graminearum, Fusarium solani, Fusarium verticillioides, Magnaporthe oryzae, Magnaporthe grisea, Puccinia graminis f. sp. tritici, Puccinia striiformis f. sp. tritici, Puccinia triticina, Uromyces appendiculatus, Botrytis cinerea, Colletotrichum gloeosporioides, Colletotrichum lindemuthianum, Alternaria solani, Alternaria alternata, Alternaria brassicae, Sclerotinia sclerotiorum, Sclerotium rolfsii, Rhizoctonia solani, Rhizoctonia bataticola, Macrophomina phaseolin, Phytophthora infestan, Phytophthora capsica, Phytophthora palmivora, Peronospora destructor, Hyaloperonospora parasitica, Pseudoperonospora cubensis, Albugo candida, Erysiphe graminis f. sp. tritici, Oidium spp., Cercospora arachidicola, Cercospora zeae-maydis, Mycosphaerella fijiensis, Venturia inaequalis, Guignardia citricarpa, Neonectria ditissima, Cryphonectria parasitica, Monilinia fructicola, and the like.
In an embodiment, the present invention provides a method for controlling fungal pathogens or preventing fungal infections in plants comprising applying said phytochemical composition to the plant or to the affected parts of the plants in suitable amount thereof.
In the present invention, water is used as a carrier for water-based formulations, and for wettable powder formulations, one of the ingredients is chosen from the list below: lactose, anhydrous lactose, dextrose, calcium bentonite powder, sodium bentonite powder, white carbon, kaolin, precipitated calcium carbonate, cornflour, powdered sugar, potter's clay, and the like.

In another embodiment, the composition of the present invention may be used in conjunction with the known insecticidal or acaricidal active ingredient.
In yet another embodiment, the phytochemicals of the present invention may be used with the active ingredient contained in other agents such as fungicides, bactericide, nematicide, plant growth regulators, synergists, fertilizers, soil improvers, animal feeds and the like.
The phytochemicals/ingredients of the present invention provide synergistic effects when combined. They work together to disrupt the vital fungal cellular processes, hindering growth, and preventing reproduction.
In another embodiment, the essential oils and botanical concretes of the present invention enhance the fungicidal action of the active ingredients.
The bio-fungicide and bio-stimulant composition of the present invention may be formulated into a known form such as wettable powder, granule, powder, tablet, emulsion, water-soluble agent, suspension, granule wettable powder, flowable agent, microcapsule, aerosol, propellant, spray, fogging agent, heating transpiration agent, smoking agent, baiting agent or the like.
In an embodiment, the composition of the present invention is useful for controlling fungal infections and improving yield of crop plants.
In yet another embodiment, the present invention provides a method for controlling or preventing fungal infections in plants comprising applying said phytochemical composition to the plant or to the affected parts of the plants in suitable amount thereof.
In an embodiment, the phytochemicals of the present invention are safe, non-toxic to humans and environment while being highly potent towards the fungal pathogens.
In another embodiment, the phytochemical composition of the present invention is applied to the plants/crops at all developmental stages in suitable dosages.

The composition of the present invention has multiple advantages/benefits such as –
The composition of the present invention aids in managing plant fungal pathogens and protects agriculture, horticulture, olericulture and floricultural crops.
It promotes the growth of plants and increases the yield of crops.
It is environmentally friendly, relatively inexpensive, safe for both agriculture and humans, and reduces the risk of resistance development.
The compositions of the present invention are easily degradable, exhibit low toxicity, and are relatively safe for humans and other animals.
It is safe for the environment, biodegradable, free of residues, and therefore reduces the chemical residue on our planet.

The following example, which includes preferred embodiments, will serve to illustrate the
practice of this invention, it being understood that the particulars shown are by way of example and for purpose of illustrative discussion of preferred embodiments of the invention.

Examples:
Example 1: Phytochemical ingredients, including their purity, fungicidal properties like modes of action, and target sites:
Table 1
Sr. No. Phyto ingredient Botanical Source Purity range of Phyto ingredient Fungicide Based on Mode of Action Fungal Target Site
1. Camphor Cinnamomum camphora 45-100% Contact Fungicides
Protective Fungicides
Multi-site Fungicides
Contact Fungicides
Fungal cell walls and membranes
Disruption of fungal cell membrane integrity
Direct toxicity to fungal hyphae on contact
Protective Fungicides
Inhibition of spore germination and early mycelial growth
Formation of a protective barrier on plant tissues
Blocking fungal penetration into host cells
Multi-site Fungicides
Fungal cell membrane integrity (disruption)
Multiple biochemical pathways simultaneously to prevent resistance
2. Eugenol Syzygium aromaticum 50-100% Contact Fungicides
Protective Fungicides
Curative Fungicides
Multi-site Fungicides Contact Fungicides
Fungal spore surfaces
Fungal cell walls and membranes
Disruption of fungal cell membrane integrity
Inhibition of spore germination on plant surfaces
Direct toxicity to fungal hyphae on contact
Protective Fungicides
Prevention of spore adhesion to plant surfaces
Inhibition of spore germination and early mycelial growth
Curative Fungicides
Early-stage fungal mycelium inside plant tissues
Inhibition of fungal enzymatic pathways
Disruption of fungal metabolism and growth after infection onset
Multi-site Fungicides
Fungal cell membrane integrity (disruption)
Enzyme systems involved in respiration and metabolism
3. Citral Ociumum gratissimum 50-100% Contact Fungicides
Protective Fungicides
Curative Fungicides
Multi-site Fungicides Contact Fungicides
Fungal spore surfaces
Fungal cell walls and membranes
Disruption of fungal cell membrane integrity
Protective Fungicides
Prevention of spore adhesion to plant surfaces
Inhibition of spore germination and early mycelial growth
Curative Fungicides
Early-stage fungal mycelium inside plant tissues
Inhibition of fungal enzymatic pathways
Multi-site Fungicides
Fungal cell membrane integrity (disruption)
Enzyme systems involved in respiration and metabolism
4. Thymol Thymus vulgaris 50-100% Contact Fungicides
Protective Fungicides
Curative Fungicides
Multi-site Fungicides Contact Fungicides
Fungal spore surfaces
Disruption of fungal cell membrane integrity
Inhibition of spore germination on plant surfaces
Direct toxicity to fungal hyphae on contact
Protective Fungicides
Prevention of spore adhesion to plant surfaces
Inhibition of spore germination and early mycelial growth
Formation of a protective barrier on plant tissues
Blocking fungal penetration into host cells
Prevention of fungal colonization at initial stages
Curative Fungicides
Early-stage fungal mycelium inside plant tissues
Inhibition of fungal enzymatic pathways
Disruption of fungal metabolism and growth after infection onset
Halting spore germination post-infection
Multi-site Fungicides
Protein and nucleic acid synthesis
Oxidative stress induction inside fungal cells
Multiple biochemical pathways simultaneously to prevent resistance
5. Piperine Piper longum 50-100% Contact Fungicides
Protective Fungicides
Curative Fungicides
Multi-site Fungicides Contact Fungicides
Disruption of fungal cell membrane integrity
Inhibition of spore germination on plant surfaces
Direct toxicity to fungal hyphae on contact
Protective Fungicides
Prevention of spore adhesion to plant surfaces
Inhibition of spore germination and early mycelial growth
Curative Fungicides
Inhibition of fungal enzymatic pathways
Interference with fungal nutrient uptake
Halting spore germination post-infection
Multi-site Fungicides
Fungal cell membrane integrity (disruption)
Enzyme systems involved in respiration and metabolism
6. Cuminaldehyde Cuminum cyminum 50-100% Contact Fungicides
Protective Fungicides
Curative Fungicides
Multi-site Fungicides Contact Fungicides
Fungal spore surfaces
Fungal cell walls and membranes
Disruption of fungal cell membrane integrity
Inhibition of spore germination on plant surfaces
Direct toxicity to fungal hyphae on contact
Protective Fungicides
Prevention of spore adhesion to plant surfaces
Inhibition of spore germination and early mycelial growth
Formation of a protective barrier on plant tissues
Blocking fungal penetration into host cells
Prevention of fungal colonization at initial stages
Curative Fungicides
Early-stage fungal mycelium inside plant tissues
Inhibition of fungal enzymatic pathways
Disruption of fungal metabolism and growth after infection onset
Interference with fungal nutrient uptake
Halting spore germination post-infection
Multi-site Fungicides
Fungal cell membrane integrity (disruption)
Enzyme systems involved in respiration and metabolism
Protein and nucleic acid synthesis
Oxidative stress induction inside fungal cells
Multiple biochemical pathways simultaneously to prevent resistance
7. Methyl chavicol Ocimum sanctum 50-100% Contact Fungicides
Protective Fungicides
Curative Fungicides
Multi-site Fungicides Contact Fungicides
Disruption of fungal cell membrane integrity
Inhibition of spore germination on plant surfaces
Direct toxicity to fungal hyphae on contact
Protective Fungicides
Formation of a protective barrier on plant tissues
Blocking fungal penetration into host cells
Prevention of fungal colonization at initial stages
Curative Fungicides
Early-stage fungal mycelium inside plant tissues
Inhibition of fungal enzymatic pathways
Disruption of fungal metabolism and growth after infection onset
Multi-site Fungicides
Fungal cell membrane integrity (disruption)
Enzyme systems involved in respiration and metabolism
Protein and nucleic acid synthesis
Multiple biochemical pathways simultaneously to prevent resistance
8. Mixture of Essential oils and oleoresin Ferula asafoetida 5 – 45% Contact Fungicides
Protective Fungicides
Curative Fungicides
Multi-site Fungicides Contact Fungicides
Fungal spore surfaces
Fungal cell walls and membranes
Disruption of fungal cell membrane integrity
Protective Fungicides
Prevention of spore adhesion to plant surfaces
Inhibition of spore germination and early mycelial growth
Formation of a protective barrier on plant tissues
Curative Fungicides
Disruption of fungal metabolism and growth after infection onset
Interference with fungal nutrient uptake
Halting spore germination post-infection
Multi-site Fungicides
Protein and nucleic acid synthesis
Oxidative stress induction inside fungal cells
Multiple biochemical pathways simultaneously to prevent resistance
9. Essential oils and Total turmerones Curcuma longa 10 – 60% Contact Fungicides
Protective Fungicides
Curative Fungicides
Multi-site Fungicides Contact Fungicides
Fungal spore surfaces
Fungal cell walls and membranes
Protective Fungicides
Prevention of spore adhesion to plant surfaces
Inhibition of spore germination and early mycelial growth
Curative Fungicides
Early-stage fungal mycelium inside plant tissues
Halting spore germination post-infection
Multi-site Fungicides
Fungal cell membrane integrity (disruption)
Enzyme systems involved in respiration and metabolism
Protein and nucleic acid synthesis
10.
Carvacrol
Origanum vulgare 50-100% Contact Fungicides
Protective Fungicides
Curative Fungicides
Multi-site Fungicides Contact Fungicides
Fungal spore surfaces
Fungal cell walls and membranes
Protective Fungicides
Prevention of spore adhesion to plant surfaces
Blocking fungal penetration into host cells
Prevention of fungal colonization at initial stages
Curative Fungicides
Early-stage fungal mycelium inside plant tissues
Interference with fungal nutrient uptake
Halting spore germination post-infection
Multi-site Fungicides
Fungal cell membrane integrity (disruption)
Enzyme systems involved in respiration and metabolism
Protein and nucleic acid synthesis
Oxidative stress induction inside fungal cells
Multiple biochemical pathways simultaneously to prevent resistance
11.
D-limonene
Citrus sinensis 50-100% Contact Fungicides
Protective Fungicides
Curative Fungicides
Multi-site Fungicides Contact Fungicides
Disruption of fungal cell membrane integrity
Inhibition of spore germination on plant surfaces
Direct toxicity to fungal hyphae on contact
Protective Fungicides
Formation of a protective barrier on plant tissues
Blocking fungal penetration into host cells
Prevention of fungal colonization at initial stages
Curative Fungicides
Early-stage fungal mycelium inside plant tissues
Interference with fungal nutrient uptake
Halting spore germination post-infection
Multi-site Fungicides
Protein and nucleic acid synthesis
Oxidative stress induction inside fungal cells
Multiple biochemical pathways simultaneously to prevent resistance
12. Gingerol Zingiber officinale 5 - 55 % Contact Fungicides
Protective Fungicides
Curative Fungicides
Multi-site Fungicides Contact Fungicides
Fungal spore surfaces
Fungal cell walls and membranes
Disruption of fungal cell membrane integrity
Inhibition of spore germination on plant surfaces
Direct toxicity to fungal hyphae on contact
Protective Fungicides
Prevention of spore adhesion to plant surfaces
Inhibition of spore germination and early mycelial growth
Formation of a protective barrier on plant tissues
Blocking fungal penetration into host cells
Prevention of fungal colonization at initial stages
Curative Fungicides
Early-stage fungal mycelium inside plant tissues
Inhibition of fungal enzymatic pathways
Halting spore germination post-infection
Multi-site Fungicides
Fungal cell membrane integrity (disruption)
Enzyme systems involved in respiration and metabolism
Protein and nucleic acid synthesis
13. β-Asarone Acorus calamus 50-100% Contact Fungicides
Protective Fungicides
Curative Fungicides
Multi-site Fungicides Contact Fungicides
Fungal spore surfaces
Fungal cell walls and membranes
Direct toxicity to fungal hyphae on contact
Protective Fungicides
Prevention of spore adhesion to plant surfaces
Inhibition of spore germination and early mycelial growth
Blocking fungal penetration into host cells
Prevention of fungal colonization at initial stages
Curative Fungicides
Early-stage fungal mycelium inside plant tissues
Interference with fungal nutrient uptake
Halting spore germination post-infection
Multi-site Fungicides
Fungal cell membrane integrity (disruption)
Enzyme systems involved in respiration and metabolism
Protein and nucleic acid synthesis
Oxidative stress induction inside fungal cells
Multiple biochemical pathways simultaneously to prevent resistance
14. Menthol Mentha piperita 50-100% Contact Fungicides
Protective Fungicides
Curative Fungicides
Multi-site Fungicides Contact Fungicides
Fungal cell walls and membranes
Disruption of fungal cell membrane integrity
Inhibition of spore germination on plant surfaces
Protective Fungicides
Prevention of spore adhesion to plant surfaces
Inhibition of spore germination and early mycelial growth
Blocking fungal penetration into host cells
Curative Fungicides
Early-stage fungal mycelium inside plant tissues
Inhibition of fungal enzymatic pathways
Disruption of fungal metabolism and growth after infection onset
Multi-site Fungicides
Fungal cell membrane integrity (disruption)
Enzyme systems involved in respiration and metabolism
Protein and nucleic acid synthesis
Multiple biochemical pathways simultaneously to prevent resistance
15. Capsaicin Capsicum frutescens 5-30 % Contact Fungicides
Protective Fungicides
Curative Fungicides
Multi-site Fungicides Contact Fungicides
Fungal spore surfaces
Direct toxicity to fungal hyphae on contact
Protective Fungicides
Prevention of spore adhesion to plant surfaces
Inhibition of spore germination and early mycelial growth
Formation of a protective barrier on plant tissues
Curative Fungicides
Early-stage fungal mycelium inside plant tissues
Inhibition of fungal enzymatic pathways
Multi-site Fungicides
Fungal cell membrane integrity (disruption)
Multiple biochemical pathways simultaneously to prevent resistance
16. p-Cymene Trachyspermum ammi 65 – 100 % Contact Fungicides
Protective Fungicides
Curative Fungicides
Multi-site Fungicides Contact Fungicides
Fungal cell walls and membranes
Disruption of fungal cell membrane integrity
Inhibition of spore germination on plant surfaces
Direct toxicity to fungal hyphae on contact
Protective Fungicides
Prevention of spore adhesion to plant surfaces
Inhibition of spore germination and early mycelial growth
Blocking fungal penetration into host cells
Prevention of fungal colonization at initial stages
Curative Fungicides
Inhibition of fungal enzymatic pathways
Disruption of fungal metabolism and growth after infection onset
Interference with fungal nutrient uptake
Halting spore germination post-infection
Multi-site Fungicides
Protein and nucleic acid synthesis
Oxidative stress induction inside fungal cells
Multiple biochemical pathways simultaneously to prevent resistance
17. Chavibetol Piper betle 50-100% Contact Fungicides
Protective Fungicides
Curative Fungicides
Multi-site Fungicides Contact Fungicides
Fungal cell walls and membranes
Disruption of fungal cell membrane integrity
Direct toxicity to fungal hyphae on contact
Protective Fungicides
Prevention of spore adhesion to plant surfaces
Inhibition of spore germination and early mycelial growth
Blocking fungal penetration into host cells
Prevention of fungal colonization at initial stages
Curative Fungicides
Early-stage fungal mycelium inside plant tissues
Inhibition of fungal enzymatic pathways
Halting spore germination post-infection
Multi-site Fungicides
Fungal cell membrane integrity (disruption)
Enzyme systems involved in respiration and metabolism
Multiple biochemical pathways simultaneously to prevent resistance
18. Thymoquinone Nigella sativa 50-100% Contact Fungicides
Protective Fungicides
Curative Fungicides
Multi-site Fungicides Contact Fungicides
Fungal cell walls and membranes
Disruption of fungal cell membrane integrity
Direct toxicity to fungal hyphae on contact
Protective Fungicides
Prevention of spore adhesion to plant surfaces
Inhibition of spore germination and early mycelial growth
Curative Fungicides
Early-stage fungal mycelium inside plant tissues
Inhibition of fungal enzymatic pathways
Halting spore germination post-infection
Multi-site Fungicides
Enzyme systems involved in respiration and metabolism
Protein and nucleic acid synthesis
Oxidative stress induction inside fungal cells
Multiple biochemical pathways simultaneously to prevent resistance
19. Berberine Berberis aristata 50-100% Contact Fungicides
Protective Fungicides
Curative Fungicides
Multi-site Fungicides Contact Fungicides
Fungal cell walls and membranes
Disruption of fungal cell membrane integrity
Inhibition of spore germination on plant surfaces
Direct toxicity to fungal hyphae on contact
Protective Fungicides
Prevention of spore adhesion to plant surfaces
Inhibition of spore germination and early mycelial growth
Blocking fungal penetration into host cells
Prevention of fungal colonization at initial stages
Curative Fungicides
Early-stage fungal mycelium inside plant tissues
Inhibition of fungal enzymatic pathways
Interference with fungal nutrient uptake
Halting spore germination post-infection
Multi-site Fungicides
Fungal cell membrane integrity (disruption)
Enzyme systems involved in respiration and metabolism
Protein and nucleic acid synthesis
Multiple biochemical pathways simultaneously to prevent resistance
20. Alpha-pinene Rosmarinus officinalis 50-100% Contact Fungicides
Protective Fungicides
Curative Fungicides
Multi-site Fungicides Contact Fungicides
Fungal cell walls and membranes
Disruption of fungal cell membrane integrity
Inhibition of spore germination on plant surfaces
Protective Fungicides
Prevention of spore adhesion to plant surfaces
Inhibition of spore germination and early mycelial growth
Prevention of fungal colonization at initial stages
Curative Fungicides
Early-stage fungal mycelium inside plant tissues
Inhibition of fungal enzymatic pathways
Interference with fungal nutrient uptake
Halting spore germination post-infection
Multi-site Fungicides
Fungal cell membrane integrity (disruption)
Enzyme systems involved in respiration and metabolism
Protein and nucleic acid synthesis
Multiple biochemical pathways simultaneously to prevent resistance
21. 1,8 cineole Rosmarinus officinalis 50-100% Contact Fungicides
Protective Fungicides
Curative Fungicides
Multi-site Fungicides Contact Fungicides
Fungal cell walls and membranes
Disruption of fungal cell membrane integrity
Inhibition of spore germination on plant surfaces
Direct toxicity to fungal hyphae on contact
Protective Fungicides
Prevention of spore adhesion to plant surfaces
Inhibition of spore germination and early mycelial growth
Formation of a protective barrier on plant tissues
Blocking fungal penetration into host cells
Prevention of fungal colonization at initial stages
Curative Fungicides
Early-stage fungal mycelium inside plant tissues
Inhibition of fungal enzymatic pathways
Disruption of fungal metabolism and growth after infection onset
Halting spore germination post-infection
Multi-site Fungicides
Enzyme systems involved in respiration and metabolism
Protein and nucleic acid synthesis
Multiple biochemical pathways simultaneously to prevent resistance
22. camphene Rosmarinus officinalis 50-100% Contact Fungicides
Protective Fungicides
Curative Fungicides
Multi-site Fungicides Contact Fungicides
Fungal cell walls and membranes
Disruption of fungal cell membrane integrity
Inhibition of spore germination on plant surfaces
Protective Fungicides
Prevention of spore adhesion to plant surfaces
Formation of a protective barrier on plant tissues
Blocking fungal penetration into host cells
Prevention of fungal colonization at initial stages
Curative Fungicides
Early-stage fungal mycelium inside plant tissues
Inhibition of fungal enzymatic pathways
Halting spore germination post-infection
Multi-site Fungicides
Fungal cell membrane integrity (disruption)
Enzyme systems involved in respiration and metabolism
Protein and nucleic acid synthesis
Multiple biochemical pathways simultaneously to prevent resistance
23. Cinnamic aldehyde Cinnamomum cassia 50-100% Contact Fungicides
Protective Fungicides
Curative Fungicides
Multi-site Fungicides Contact Fungicides
Fungal spore surfaces
Fungal cell walls and membranes
Disruption of fungal cell membrane integrity
Inhibition of spore germination on plant surfaces
Direct toxicity to fungal hyphae on contact
Protective Fungicides
Prevention of spore adhesion to plant surfaces
Inhibition of spore germination and early mycelial growth
Curative Fungicides
Early-stage fungal mycelium inside plant tissues
Inhibition of fungal enzymatic pathways
Disruption of fungal metabolism and growth after infection onset
Multi-site Fungicides
Fungal cell membrane integrity (disruption)
Enzyme systems involved in respiration and metabolism

Example 2 - Extraction and purification methods for the listed Phytoingredients:
Table 2
Ingredient Method Type Solvent Parameters Description
Camphor Steam Distillation Extraction Water (steam) Temp: 100°C, Pressure: Atmospheric, Time: 4-6 h Steam passes through Cinnamomum camphora leaves/wood, condensing camphor (yield ~0.5%)
Supercritical CO₂ Extraction Extraction CO₂ Temp: 45°C, Pressure: 25 MPa, Time: 2-3 h High-yield extraction (4.63%) from camphor tree leaves
Sublimation Purification None Temp: 50-60°C, Pressure: Low (~0.1 atm) Camphor sublimes and recrystallizes, achieving high purity.
Eugenol Steam Distillation Extraction Water (steam) Temp: 100°C, Pressure: Atmospheric, Time: 3-5 h Extracts eugenol from clove buds (Syzygium aromaticum), yield 15-20%.
Solvent Extraction Extraction Ethanol Temp: 50-60°C, Pressure: Atmospheric, Time: 4-6 h Ethanol dissolves eugenol from cloves, followed by evaporation.
Fractional Distillation Purification None Temp: 248-254°C (boiling point), Pressure: Atmospheric or vacuum Separates eugenol from clove oil components.
Citral Steam Distillation Extraction Water (steam) Temp: 100°C, Pressure: Atmospheric, Time: 3-4 h Extracts citral from lemongrass (Cymbopogon flexuosus), yield ~1-2%
Hydrotropic Extraction Extraction Sodium salicylate (aqueous) Temp: 50-60°C, Pressure: Atmospheric, Time: 2-3 h Enhances citral yield from lemongrass
Column Chromatography Purification Hexane:Ethyl acetate (9:1) Temp: Ambient, Pressure: Atmospheric Separates citral using silica gel based on polarity.
Thymol Steam Distillation Extraction Water (steam) Temp: 100°C, Pressure: Atmospheric, Time: 3-5 h Extracts thymol from thyme (Thymus vulgaris), yield ~1-3%.
Solvent Extraction Extraction Ethanol Temp: 50-60°C, Pressure: Atmospheric, Time: 4-6 h Ethanol extracts thymol from plant material.
Crystallization Purification Ethanol Temp: 0-5°C (cooling), Pressure: Atmospheric Recrystallizes thymol for high purity.
Piperine Soxhlet Extraction Extraction Ethanol Temp: 78°C (ethanol boiling point), Pressure: Atmospheric, Time: 6-8 h Extracts piperine from black pepper (Piper nigrum), yield ~5%
Ultrasound-Assisted Extraction Extraction Ethanol Temp: 40-50°C, Pressure: Atmospheric, Ultrasound: 20 kHz, Time: 30-60 min Enhances piperine yield (up to 5.8 mg/g)
Recrystallization Purification Ethanol Temp: 0-10°C (cooling), Pressure: Atmospheric Purifies piperine by recrystallization.
Cuminaldehyde
Steam Distillation Extraction Water (steam) Temp: 100°C, Pressure: Atmospheric, Time: 3-4 h Extracts cuminaldehyde from cumin seeds (Cuminum cyminum).
Fractional Distillation Purification None Temp: 235-240°C (boiling point), Pressure: Atmospheric or vacuum Separates cuminaldehyde based on boiling point.
Methyl Chavicol Steam Distillation Extraction Water (steam) Temp: 100°C, Pressure: Atmospheric, Time: 3-4 h Extracts methyl chavicol from basil
Fractional Distillation Purification None Temp: 215-220°C (boiling point), Pressure: Atmospheric Isolates methyl chavicol from essential oil.
Mixture of Essential Oils and Oleoresin Supercritical CO₂ Extraction Extraction CO₂ Temp: 40-50°C, Pressure: 20-30 MPa, Time: 2-3 h Extracts complex mixtures from spices
Solvent Extraction Extraction Hexane Temp: 50-60°C, Pressure: Atmospheric, Time: 4-6 h Extracts oils and oleoresins, followed by solvent evaporation.
Column Chromatography Purification Hexane: Ethyl acetate (8:2) Temp: Ambient, Pressure: Atmospheric Separates components based on polarity.
Essential Oils and Total Turmerones Supercritical CO₂ Extraction Extraction CO₂ Temp: 40-50°C, Pressure: 25-30 MPa, Ti me: 2-3 h High-quality extraction from turmeric
Column Chromatography Purification Hexane: Ethyl acetate (9:1) Temp: Ambient, Pressure: Atmospheric Isolates turmerones from essential oil mixture.
Carvacrol Steam Distillation Extraction Water (steam) Temp: 100°C, Pressure: Atmospheric, Time: 3-4 h Extracts carvacrol from oregano
Crystallization Purification Ethanol Temp: 0-5°C (cooling), Pressure: Atmospheric Purifies solid carvacrol.
D-Limonene Cold Pressing Extraction None Temp: Ambient, Pressure: Mechanical, Time: 1-2 h Extracts D-limonene from citrus peels (Citrus spp.), yield ~1-5%
Steam Distillation Extraction Water (steam) Temp: 100°C, Pressure: Atmospheric, Time: 3-4 h Alternative for D-limonene extraction.
Fractional Distillation Purification None Temp: 175-178°C (boiling point), Pressure: Atmospheric Isolates D-limonene from citrus oil.
Gingerol Solvent Extraction Extraction Ethanol Temp: 50-60°C, Pressure: Atmospheric, Time: 4-6 h Extracts gingerol from ginger (Zingiber officinale)
HPLC Purification Methanol:Water (70:30) Temp: Ambient, Pressure: High (HPLC system) High-precision purification of gingerol.
β-Asarone Steam Distillation Extraction Water (steam) Temp: 100°C, Pressure: Atmospheric, Time: 3-4 h Extracts β-asarone from calamus roots (Acorus calamus).
Column Chromatography Purification Hexane:Ethyl acetate (9:1) Temp: Ambient, Pressure: Atmospheric Isolates β-asarone from essential oil.
Menthol Steam Distillation Extraction Water (steam) Temp: 100°C, Pressure: Atmospheric, Time: 3-5 h Extracts menthol from peppermint (Mentha piperita).
Crystallization Purification Ethanol Temp: 0-5°C (cooling), Pressure: Atmospheric Purifies menthol crystals.
Capsaicin Solvent Extraction Extraction Ethanol Temp: 50-60°C, Pressure: Atmospheric, Time: 4-6 h Extracts capsaicin from chili peppers (Capsicum spp.).
HPLC Purification Methanol:Water (80:20) Temp: Ambient, Pressure: High (HPLC system) High-purity capsaicin isolation.
p-Cymene Steam Distillation Extraction Water (steam) Temp: 100°C, Pressure: Atmospheric, Time: 3-4 h Extracts p-cymene from thyme or cumin.
Fractional Distillation Purification None Temp: 177-178°C (boiling point), Pressure: Atmospheric Isolates p-cymene from essential oil.
Chavibetol Steam Distillation Extraction Water (steam) Temp: 100°C, Pressure: Atmospheric, Time: 3-4 h Extracts chavibetol from betel leaves (Piper betle).
Column Chromatography Purification Hexane:Ethyl acetate (8:2) Temp: Ambient, Pressure: Atmospheric Purifies chavibetol from essential oil.
Thymoquinone Solvent Extraction Extraction Hexane Temp: 50-60°C, Pressure: Atmospheric, Time: 4-6 h Extracts thymoquinone from black cumin (Nigella sativa).
Recrystallization Purification Ethanol Temp: 0-10°C (cooling), Pressure: Atmospheric Purifies thymoquinone.
Berberine Acid-Base Extraction Extraction Acidic water (HCl), Ethanol Temp: 50-60°C, Pressure: Atmospheric, Time: 4-6 h Selective extraction from barberry (Berberis spp.) using pH changes.
Recrystallization Purification Ethanol Temp: 0-10°C (cooling), Pressure: Atmospheric Purifies berberine alkaloid.
Alpha-Pinene Steam Distillation Extraction Water (steam) Temp: 100°C, Pressure: Atmospheric, Time: 3-4 h Extracts alpha-pinene from pine (Pinus spp.).
Fractional Distillation Purification None Temp: 155-156°C (boiling point), Pressure: Atmospheric Isolates alpha-pinene from pine oil.
1,8-Cineole Steam Distillation Extraction Water (steam) Temp: 100°C, Pressure: Atmospheric, Time: 3-4 h Extracts 1,8-cineole from eucalyptus (Eucalyptus spp.).
Fractional Distillation Purification None Temp: 174-176°C (boiling point), Pressure: Atmospheric Isolates 1,8-cineole from eucalyptus oil.
Camphene Steam Distillation Extraction Water (steam) Temp: 100°C, Pressure: Atmospheric, Time: 3-4 h Extracts camphene from pine (Pinus spp.).
Fractional Distillation Purification None Temp: 159-160°C (boiling point), Pressure: Atmospheric Isolates camphene from pine oil.
Cinnamic Aldehyde Steam Distillation Extraction Water (steam) Temp: 100°C, Pressure: Atmospheric, Time: 3-4 h Extracts cinnamic aldehyde from cinnamon (Cinnamomum zeylanicum).
Fractional Distillation Purification None Temp: 248-252°C (boiling point), Pressure: Vacuum Isolates cinnamic aldehyde to avoid thermal degradation.
Note: In Examples 3 to 11 below, all the raw materials, including active phytochemicals and excipients, were verified for identity, purity, and compliance with predefined specifications and each component was accurately weighed according to the batch formulation protocol.
Example 3: Composition of Phytochemical-Based Bio fungicide (Embodiment 1) in Water-Soluble Powder (WSP) Formulation
Table 3
Sr No. Ingredient Actual Percent
Percent Range
1. Thymol 5.0 % 0.001 to 45 %
2. Eugenol 3.0 % 0.001 to 35 %
3. Piperine 1.5 % 0.001 to 25 %
4. Sodium lignosulphonate 8.0 % 0.1 to 25 %
5. Fumed silica 1.0 % 0.1 to 18 %
6. Maltodextrin 70.0 % 35 to 95 %
7. Sodium lauryl sulfate 2.0 % 0.001 to 20 %
8. Citric acid (buffer) 0.5 % 0.001 to 18 %

Preparation of Phytochemical-Based Bio-fungicide (Embodiment 1) in Water-Soluble Powder (WSP) Formulation:
The process for preparing Bio-fungicide (Embodiment 1) comprises the steps of:
Primary Dry Blending of Powdered Ingredients: The active ingredients ( Thymol, Eugeno, and piperine), maltodextrin (carrier), fumed silica (anti-caking agent), and sodium lignosulphonate (dispersing agent) were subjected to initial dry mixing by using any one of the following V-Blender (Twin-Shell Blender), Double Cone Blender, Ribbon Blender (Horizontal Blender), Paddle Blender, Octagonal Blender, Bin Blender (IBC – Intermediate Bulk Container Blender), High-Shear Mixer (for wet powder mixing/granulation), Turbula Mixer / 3D Mixer, Ploughshare Mixer, Conical Screw Mixer (Nauta Mixer), more precisely ribbon blender and High-Shear Mixer for min 10–15 minutes. This step ensured a uniform distribution of ingredients before the addition of liquid excipients.
Surfactants such as sodium lauryl sulfate in powder and citric acid as buffer in powder were gradually introduced into the pre-mix. The blend was processed until a homogeneous coating of dry particles was achieved by using High-Shear Mixer and Sigma Mixer for 15–20 minutes or until homogeneity was attained.
Particle Size Reduction was achieved by intermediate blend was subjected to micronization to achieve a fine particle size, typically in the range of 1–10 µm. Jet milling, cryogenic milling and air classifier milling were employed for precise size reduction. For heat-sensitive actives like Thymol, eugenol or piperine, cryogenic milling was preferred to preserve integrity.
Final Homogenization: Post-micronization, the powder was reblended to ensure rehomogenization of actives and excipients in tumbler blender or V-Blender more precisely in tumbler blender for 20-35 mins. This step enhanced consistency across the final formulation.
Final Sieving: The final product was passed through a vibro-sifter equipped with a 100–120 mesh screen to eliminate agglomerates, improve powder flow, and ensure uniform particle distribution.
Product recovered from step 5 was further quantified for the active phyto constituent by using various instruments but not limited to High pressure liquid chromatograph, gas chromatography, spectroscopy compounds etc.
The product obtained in step 6 was also tested for bio-efficacy against pests on different crops in vitro and in vivo, and it is compared to other chemically synthesised fungicides.
The resultant product obtained was effective against broad group of fungal pathogens within 48 -72 hrs of application with low risk of resistance development and safe for environment and mammalian.

Example 4: Composition of Phytochemical-Based Bio fungicide (Embodiment 2) in Water-Soluble Powder (WSP) Formulation:
Table 4
Sr No. Ingredient Actual Percent
Percent Range
1. Thymol 15.0 % 3 to 45 %
2. Sodium Lauryl sulphate 10.0 % 0.001 to 20 %
3. Sodium Ligno sulphonate 5.0 % 0.001 to 15 %
5. Fumed silica 10.0 % 0.001 to 18 %
6. Polysorbate-80 12.0 % 0.001 to 25 %
7. Poly glycol based defoamer 2.0 % 0.001 to 25 %
8. China clay 46.0 % 25.0 to 85 %

Preparation of Phytochemical-Based Bio fungicide (Embodiment 2) in Water-Soluble Powder (WSP) Formulation:
The process for preparing the Bio-fungicide (Embodiment 2) comprises the steps of:
Primary Dry Blending of Powdered Ingredients: The active ingredients (Thymol), China clay (carrier), fumed silica (anti-caking agent), and sodium lignosulphonate (dispersing agent) were subjected to initial dry mixing by using any one of the following V-Blender (Twin-Shell Blender), Double Cone Blender, Ribbon Blender (Horizontal Blender), Paddle Blender, Octagonal Blender, Bin Blender (IBC – Intermediate Bulk Container Blender), High-Shear Mixer (for wet powder mixing/granulation), Turbula Mixer / 3D Mixer, Ploughshare Mixer, Conical Screw Mixer (Nauta Mixer), more precisely ribbon blender and High-Shear Mixer for min 10–15 minutes. This step ensured a uniform distribution of ingredients before the addition of liquid excipients.
Surfactants such as Polysorbate 80, polyethylene glycol derivatives, and sodium lauryl sulfate (in powder or paste form, depending on grade) were gradually introduced into the pre-mix. The blend was processed until a homogeneous coating of dry particles was achieved by using High-Shear Mixer and Sigma Mixer for 15–20 minutes or until homogeneity was attained.
Particle Size Reduction was achieved by intermediate blend was subjected to micronization to achieve a fine particle size, typically in the range of 1–10 µm. Jet milling, cryogenic milling and air classifier milling were employed for precise size reduction. For heat-sensitive actives like Thymol, cryogenic milling was preferred to preserve integrity.
Final Homogenization, Post-micronization, the powder was reblended to ensure rehomogenization of actives and excipients in tumbler blender or V-Blender more precisely in tumbler blender for 20-35 mins. This step enhanced consistency across the final formulation.
Final Sieving: The final product was passed through a vibro-sifter equipped with a 100–120 mesh screen to eliminate agglomerates, improve powder flow, and ensure uniform particle distribution.
Product recovered from step 5 was further quantified for the active phyto constituent by using various instruments but not limited to High pressure liquid chromatograph, gas chromatography, spectroscopy compounds etc.
The product obtained in step 6 was also tested for bio-efficacy against pests on different crops in vitro and in vivo, and it is compared to other chemically synthesised fungicides.
The resultant product obtained was effective against broad group of fungal pathogen within 48 -72 hrs of application with low risk of resistance development and safe for environment and mammalian.

Example 5: Composition of Phytochemical-Based Bio fungicide (Embodiment 3) in Water-Soluble Powder (WSP) Formulation:
Table 5
Sr No. Ingredient Actual Percent
Percent Range
1. Piperine 3.0 % 0.001 to 25 %
2. Total turmerones 3.0 % 0.001 to 20 %
3. Gingerol 1.5 % 0.001 to 35 %
4. Thymol 1.5 % 0.001 to 45 %
5. camphor 1.5 % 0.001 to 35 %
6. eugenol 1.5 % 0.001 to 35 %
7. Sodium lignosulphonate 8.0 % 0.1 to 25 %
8. Fumed silica 1.0 % 0.1 to 18 %
9. Maltodextrin 76.5 % 35 to 95 %
10. Sodium lauryl sulfate 2.0 % 0.001 to 20 %
11. Citric acid (buffer) 0.5 % 0.001 to 18 %

Process of Preparation of Phytochemical-Based Bio fungicide (Embodiment 3) in Water-Soluble Powder (WSP) Formulation:
The process for preparing Bio-fungicide (Embodiment 3) comprises the steps:
Primary Dry Blending of Powdered Ingredients: The active ingredients ( piperine, Total turmerones, gingerol, thymol, camphor and eugenol ), maltodextrin (carrier), fumed silica (anti-caking agent), and sodium lignosulphonate (dispersing agent) were subjected to initial dry mixing by using any one of the following V-Blender (Twin-Shell Blender), Double Cone Blender, Ribbon Blender (Horizontal Blender), Paddle Blender, Octagonal Blender, Bin Blender (IBC – Intermediate Bulk Container Blender), High-Shear Mixer (for wet powder mixing/granulation), Turbula Mixer / 3D Mixer, Ploughshare Mixer, Conical Screw Mixer (Nauta Mixer), more precisely ribbon blender and High-Shear Mixer for min 10–15 minutes. This step ensured a uniform distribution of ingredients before the addition of liquid excipients.
Surfactants such as Sodium lauryl sulfate and citric acid as buffer in powder were gradually introduced into the pre-mix. The blend wasprocessed until a homogeneous coating of dry particles was achieved by using High-Shear Mixer and Sigma Mixer for 15–20 minutes or until homogeneity was attained.
Particle Size Reduction was achieved by intermediate blend was subjected to micronization to achieve a fine particle size, typically in the range of 1–10 µm. Jet milling, cryogenic milling and air classifier milling were employed for precise size reduction. For heat-sensitive actives like cryogenic milling is preferred to preserve integrity.
Final Homogenization: Post-micronization, the powder was reblended to ensure rehomogenization of actives and excipients in tumbler blender or V-Blender more precisely in tumbler blender for 20-35 mins. This step enhanced consistency across the final formulation.
Final Sieving: The final product was passed through a vibro-sifter equipped with a 100–120 mesh screen to eliminate agglomerates, improve powder flow, and ensure uniform particle distribution.
Product recovered from step 5 was further quantified for the active phyto constituent by using various instruments but not limited to High pressure liquid chromatograph, gas chromatography, spectroscopy compounds etc.
The product obtained in step 6 was also tested for bio-efficacy against pests on different crops in vitro and in vivo, and it is compared to other chemically synthesised fungicides.
The resultant product obtained was effective against broad group of fungal pathogens within 48 -72 hrs of application with low risk of resistance development and safe for environment and mammalian.

Example 6: Composition of Phytochemical-Based Bio fungicide (Embodiment 4) in Water-Soluble Powder (WSP) Formulation:
Table 6
Sr No. Ingredient Actual Percent
Percent Range
1. Thymol 10.0 % 0.001 to 45 %
2. Cumin aldehyde 6.5 % 0.001 to 20 %
3. Cinnamaldehyde 5.5 % 0.001 to 15 %
4. Sodium lignosulphonate 8.0 % 0.1 to 25 %
5. Fumed silica 1.0 % 0.1 to 18 %
6. Cyclodextrins 66.5 % 35 to 95 %
7. Sodium lauryl sulfate 2.0 % 0.001 to 20 %
8. Citric acid (buffer) 0.5 % 0.001 to 18 %

Preparation of Phytochemical Based Bio-fungicide (Embodiment 4):
The process for preparing Bio-fungicide (Embodiment 4) comprises the steps of:
Primary Dry Blending of Powdered Ingredients: The active ingredients ( Thymol, Cumin aldehyde, and Cinnamaldehyde ), Cyclodextrins (carrier), fumed silica (anti-caking agent), and sodium lignosulphonate (dispersing agent) were subjected to initial dry mixing by using any one of the following V-Blender (Twin-Shell Blender), Double Cone Blender, Ribbon Blender (Horizontal Blender), Paddle Blender, Octagonal Blender, Bin Blender (IBC – Intermediate Bulk Container Blender), High-Shear Mixer (for wet powder mixing/granulation), Turbula Mixer / 3D Mixer, Ploughshare Mixer, Conical Screw Mixer (Nauta Mixer), more precisely ribbon blender and High-Shear Mixer for min 10–15 minutes. This step ensured a uniform distribution of ingredients before the addition of liquid excipients.
Surfactants such as Sodium lauryl sulfate in powder and citric acid as buffer in powder were gradually introduced into the pre-mix. The blend was processed until a homogeneous coating of dry particles was achieved by using High-Shear Mixer and Sigma Mixer for 15–20 minutes or until homogeneity was attained.
Particle Size Reduction was achieved by intermediate blend was subjected to micronization to achieve a fine particle size, typically in the range of 1–10 µm. Jet milling, cryogenic milling and air classifier milling were employed for precise size reduction. For heat-sensitive actives like cryogenic milling is preferred to preserve integrity.
Final Homogenization: Post-micronization, the powder was reblended to ensure rehomogenization of actives and excipients in tumbler blender or V-Blender more precisely in tumbler blender for 20-35 mins. This step enhanced consistency across the final formulation.
Final Sieving: The final product was passed through a vibro-sifter equipped with a 100–120 mesh screen to eliminate agglomerates, improve powder flow, and ensure uniform particle distribution.
Product recovered from step 5 was further quantified for the active phyto constituent by using various instruments but not limited to High pressure liquid chromatograph, gas chromatography, spectroscopy compounds etc.
The product obtained in step 6 was also tested for bio-efficacy against pests on different crops in vitro and in vivo, and it is compared to other chemically synthesised fungicides.
The resultant product obtained product was effective against broad group of fungal pathogens within 48 -72 hrs of application with low risk of resistance development and safe for environment and mammalian.

Example 7: Composition of Phytochemical Based Bio-fungicide (Embodiment 5) in Water-Soluble Powder (WSP) Formulation:
Table 7
Sr No. Ingredient Actual Percent
Percent Range
1. Thymol 30.0 % 0.001 to 45 %
2. Piperine 7.0 % 0.001 to 25 %
3. eugenol 3.5 % 0.001 to 35 %
4. Sodium lignosulphonate 8.0 % 0.001 to 25 %
5. Fumed silica % 0.001 to 18 %
6. Lactose 48.0 % 35 to 95 %
7. Sodium lauryl sulfate 2.0 % 0.001 to 20 %
8. Citric acid (buffer) 0.5 % 0.001 to 18 %

Preparation of Phytochemical Based Bio-fungicide (Embodiment 5):
The process for preparing Bio-fungicide (Embodiment 5) comprises the steps of:

Primary Dry Blending of Powdered Ingredients: The active ingredients ( Thymol, Piperine, and eugenol), lactose (carrier), fumed silica (anti-caking agent), and sodium lignosulphonate (dispersing agent) were subjected to initial dry mixing by using any one of the following V-Blender (Twin-Shell Blender), Double Cone Blender, Ribbon Blender (Horizontal Blender), Paddle Blender, Octagonal Blender, Bin Blender (IBC – Intermediate Bulk Container Blender), High-Shear Mixer (for wet powder mixing/granulation), Turbula Mixer / 3D Mixer, Ploughshare Mixer, Conical Screw Mixer (Nauta Mixer), more precisely ribbon blender and High-Shear Mixer for min 10–15 minutes. This step ensured a uniform distribution of ingredients before the addition of liquid excipients.
Surfactants such as sodium lauryl sulfate and citric acid as buffer in powder were gradually introduced into the pre-mix. The blend was processed until a homogeneous coating of dry particles was achieved by using High-Shear Mixer and Sigma Mixer for 15–20 minutes or until homogeneity was attained.
Particle Size Reduction was achieved by intermediate blend was subjected to micronization to achieve a fine particle size, typically in the range of 1–10 µm. Jet milling, cryogenic milling and air classifier milling were employed for precise size reduction. For heat-sensitive actives cryogenic milling is preferred to preserve integrity.
Final Homogenization: Post-micronization, the powder was reblended to ensure rehomogenization of actives and excipients in tumbler blender or V-Blender more precisely in tumbler blender for 20-35 mins. This step enhanced consistency across the final formulation.
Final Sieving: The final product was passed through a vibro-sifter equipped with a 100–120 mesh screen to eliminate agglomerates, improve powder flow, and ensure uniform particle distribution.
Product recovered from step 5 was further quantified for the active phyto constituent by using various instruments but not limited to High pressure liquid chromatograph, gas chromatography, spectroscopy compounds etc.
The product obtained in step 6 was also tested for bio-efficacy against pests on different crops in vitro and in vivo, and it is compared to other chemically synthesised fungicides.
The resultant product btained was effective against broad group of fungal pathogens within 48 -72 hrs of application with low risk of resistance development and safe for environment and mammalian.

Example 8: Composition of Phytochemical Based Bio-fungicide (Embodiment 6) in Water-Soluble Powder (WSP) Formulation:
Table 8
Sr No. Ingredient Actual Percent
Percent Range
1. Piperine 10.0 % 0.001 to 25 %
2. Eugenol 6.5 % 0.001 to 35 %
3. Citral 5.5 % 0.001 to 25 %
4. Sodium lignosulphonate 8.0 % 0.001 to 25 %
5. Fumed silica 1.0 % 0.001 to 18 %
6. China clay 66.5 % 35 to 95 %
7. Sodium lauryl sulfate 2.0 % 0.001 to 20 %
8. Citric acid (buffer) 0.5 % 0.001 to 18 %

Preparation of Bio-fungicide (Embodiment 6):
The process for preparing Bio-fungicide (Embodiment 6) comprises the steps of:
Primary Dry Blending of Powdered Ingredients: The active ingredients ( Piperine, eugenol and Citral), China clay (carrier), fumed silica (anti-caking agent), and sodium lignosulphonate (dispersing agent) were subjected to initial dry mixing by using any one of the following V-Blender (Twin-Shell Blender), Double Cone Blender, Ribbon Blender (Horizontal Blender), Paddle Blender, Octagonal Blender, Bin Blender (IBC – Intermediate Bulk Container Blender), High-Shear Mixer (for wet powder mixing/granulation), Turbula Mixer / 3D Mixer, Ploughshare Mixer, Conical Screw Mixer (Nauta Mixer), more precisely ribbon blender and High-Shear Mixer for min 10–15 minutes. This step ensured a uniform distribution of ingredients before the addition of liquid excipients.
Surfactants such as Sodium lauryl sulfate in powder and citric acid as buffer in powder were gradually introduced into the pre-mix. The blend was processed until a homogeneous coating of dry particles was achieved by using High-Shear Mixer and Sigma Mixer for 15–20 minutes or until homogeneity was attained.
Particle Size Reduction was achieved by intermediate blend was subjected to micronization to achieve a fine particle size, typically in the range of 1–10 µm. Jet milling, cryogenic milling and air classifier milling were employed for precise size reduction. For heat-sensitive actives cryogenic milling was preferred to preserve integrity.
Final Homogenization: Post-micronization, the powder was reblended to ensure rehomogenization of actives and excipients in tumbler blender or V-Blender more precisely in tumbler blender for 20-35 mins. This step enhanced consistency across the final formulation.
Final Sieving: The final product was passed through a vibro-sifter equipped with a 100–120 mesh screen to eliminate agglomerates, improve powder flow, and ensure uniform particle distribution.
Product recovered from step 5 was further quantified for the active phyto constituent by using various instruments but not limited to High pressure liquid chromatograph, gas chromatography, spectroscopy compounds etc.
The product obtained in step 6 was also tested for bio-efficacy against pests on different crops in vitro and in vivo, and it is compared to other chemically synthesised fungicides.
The resultant product btained was effective against broad group of fungal pathogens within 48 -72 hrs of application with low risk of resistance development and safe for environment and mammalian.

Example 9: Composition of Phytochemical-Based Bio fungicide (Embodiment 7) in Water-Soluble Powder (WSP) Formulation:
Table 9
Sr No. Ingredient Actual Percent
Percent Range
1. Menthol 5.0 % 0.001 to 40 %
2. Total Turmerones 5.0 % 0.001 to 20 %
3. Berberine 4.5 % 0.001 to 40 %
4. Alpha pinene 4.0 % 0.001 to 40 %
5. 1,8 cineole 3.0 % 0.001 to 40 %
6. Carvacrol 1.5 % 0.001 to 40 %
7. eugenol 1.5 % 0.001 to 35 %
8. Citral 1.5 % 0.001 to 25 %
9. Sodium lignosulphonate 8.0 % 0.1 to 25 %
10. Fumed silica 1.0 % 0.1 to 18 %
11. China clay 62.5 % 35 to 95 %
12. Sodium lauryl sulfate 2.0 % 0.001 to 20 %
13. Citric acid (buffer) 0.5 % 0.001 to 18 %

Preparation of Bio-fungicide (Embodiment 7):
The process for preparing Bio-fungicide (Embodiment 7) comprises the steps of:
Primary Dry Blending of Powdered Ingredients: The active ingredients (Menthol, Total Turmerones, Berberine, Alpha pinene, Carvacrol, eugenol and Citral), China clay (carrier), fumed silica (anti-caking agent), and sodium lignosulphonate (dispersing agent) were subjected to initial dry mixing by using any one of the following V-Blender (Twin-Shell Blender), Double Cone Blender, Ribbon Blender (Horizontal Blender), Paddle Blender, Octagonal Blender, Bin Blender (IBC – Intermediate Bulk Container Blender), High-Shear Mixer (for wet powder mixing/granulation), Turbula Mixer / 3D Mixer, Ploughshare Mixer, Conical Screw Mixer (Nauta Mixer), more precisely ribbon blender and High-Shear Mixer for min 10–15 minutes. This step ensured a uniform distribution of ingredients before the addition of liquid excipients.
Surfactants such as Sodium lauryl sulfate in powder and citric acid as buffer in powder were gradually introduced into the pre-mix. The blend was processed until a homogeneous coating of dry particles was achieved by using High-Shear Mixer and Sigma Mixer for 15–20 minutes or until homogeneity was attained.
Particle Size Reduction was achieved by intermediate blend was subjected to micronization to achieve a fine particle size, typically in the range of 1–10 µm. Jet milling, cryogenic milling and air classifier milling were employed for precise size reduction. For heat-sensitive actives cryogenic milling was preferred to preserve integrity.
Final Homogenization: Post-micronization, the powder was reblended to ensure rehomogenization of actives and excipients in tumbler blender or V-Blender more precisely in tumbler blender for 20-35 mins. This step enhanced consistency across the final formulation.
Final Sieving: The final product was passed through a vibro-sifter equipped with a 100–120 mesh screen to eliminate agglomerates, improve powder flow, and ensure uniform particle distribution.
Product recovered from step 5 was further quantified for the active phyto constituent by using various instruments but not limited to High pressure liquid chromatograph, gas chromatography, spectroscopy compounds etc.
The product obtained in step 6 was also tested for bio-efficacy against pests on different crops in vitro and in vivo, and it is compared to other chemically synthesised fungicides.
The resultant product btained was effective against broad group of fungal pathogens within 48 -72 hrs of application with low risk of resistance development and safe for environment and mammalian.

Example 10: Composition of Phytochemical-Based Bio fungicide (Embodiment 8) in Water-Soluble Powder (WSP) Formulation:
Table 10
Sr No. Ingredient Actual Percent Percent Range
1. Thymol 17.0 % 0.001 to 45 %
2. Eugenol 13.5 % 0.001 to 35 %
3. Piperine 2.5 % 0.001 to 25 %
4. Gaur gum (Emulsifier) 3.2% 0.001 to 7.5 %
5. ethoxylated castor oil (Emulsifier) 4.3 % 0.001 to 10 %
6. Sodium oleoyl amino fatty acid (Surfactant) 5.0 % 0.1 to 15 %
7. Dimethyl sulfoxide (DMSO) 24.4 % 10.0 to 55 %
8. N-Ethyl-2 pyrrolidone (NEP) 30.1 % 10.0 to 55 %

Preparation of Bio-fungicide (Embodiment 8):
The process for preparing Bio-fungicide (Embodiment 8) comprises the steps of:
The emulsifier and surfactant was dissolved in dimethyl sulfoxide and N-Ethyl-2 pyrrolidone to make inert mixture and agitated with homogenizer until the uniform blend is formed.
The active ingredients, thymol, eugenol and piperine in given quantity were added to the blend formed in step no. 1 and homogenized completely to make emulsion concentration.
The emulsion concentrate prepared in step no. 2 was stirred at 300 to 800 rpm at 25 to 55 degrees Celsius in close mixing vessel made up of stainless steel 316 grade with outer jacket for maintaining temperature, the mixing vessel was connected with stirrer (blade stirrer, propeller stirrer, turbine stirrer, anchor stirrer, universal stirrer) for continues stirring. Towards the bottom of vessel homogenizer with the speed of 3700~27000 rpm was attached to reduce the particle size under very high pressures, sheer, turbulence, acceleration and impact, to make them more stable and effective.
The blend prepared in step no. 3 was passed through in line shear pump with 1800 RPM to 5800 RPM to reduce particle size in nano, so as to achieve nano emulsion with particle size ranging from 10 to 900 nano meter.
Further to obtain nano particles base formulation the blend formed in step no. 4 was further passed through High-pressure homogenization (homogenization at speed of 4000 rpm to 10000 rpm and pressure max up to 4,200 bar (60,000 psi)) to reduce the particle size below 100 nano meters.
After passing through high pressure homogenizer the material was passed through sparkler filter having seven layers of membrane filters with pore size less than 1-2 micron to achieve proper filtration.
Product recovered from step no. 6 was further quantified for the active phyto constituent by using various instruments but not limited to High pressure liquid chromatograph, gas chromatography, spectroscopy compounds etc.
The product obtained in step no. 7 was also tested for bio-efficacy against pests on different crops in vitro and in vivo, and it is compared to other chemically synthesisedfungicides.
The product obtained was effective against broad group of pests within 48 -72 hrs of application with low risk of resistance development and safe for environment and mammalian.
Example 11: Composition of Phytochemical-Based Bio fungicide (Embodiment 9) in Water-Soluble Powder (WSP) Formulation:
Table 11
Sr No. Ingredient Actual Percent Percent Range
1. Piperine 12.0 % 0.001 to 25 %
2. Methyl chavicol 8.5 % 0.001 to 35 %
3. Camphor 2.5 % 0.001 to 35 %
4. Gaur gum (Emulsifier) 3.2% 0.001 to 7.5 %
5. ethoxylated castor oil (Emulsifier) 4.3 % 0.001 to 10 %
6. Sodium oleoyl amino fatty acid (Surfactant) 5.0 % 0.1 to 15 %
7. Benzyl acetate 9.5 % 10.0 to 35 %
8. water 55.0 % 10.0 to 35 %

Preparation of Bio-fungicide (Embodiment 9):
The process for preparing Bio-fungicide (Embodiment 9) comprises the steps of:
The emulsifier and surfactant was dissolved in benzyl acetate and water to make inert mixture and agitated with homogenizer until the uniform blend was formed.
The active ingredients, piperine, Methyl chavicol and camphor in given quantity were added to the blend formed in step no. 1 and homogenized completely to make emulsion concentration.
The emulsion concentrate prepared in step no.2 was stirred at 300 to 800 rpm at 25 to 55 degrees Celsius in close mixing vessel made up of stainless steel 316 grade with outer jacket for maintaining temperature, the mixing vessel is connected with stirrer (blade stirrer, propeller stirrer, turbine stirrer, anchor stirrer, universal stirrer) for continues stirring. Towards the bottom of vessel homogenizer with the speed of 3700~27000 rpm was attached to reduce the particle size under very high pressures, sheer, turbulence, acceleration and impact, to make them more stable and effective.
The blend prepared in step no. 3 was passed through in line shear pump with 1800 RPM to 5800 RPM to reduce particle size in nano, so as to achieve nano emulsion with particle size ranging from 10 to 900 nano meter.
Further to obtain nano particles base formulation the blend formed in step no. 4 was further passed through High-pressure homogenization (homogenization at speed of 4000 rpm to 10000 rpm and pressure max up to 4,200 bar (60,000 psi)) to reduce the particle size below 100 nano meters.
After passing through high pressure homogenizer the material was passed through sparkler filter having seven layers of membrane filters with pore size less than 1-2 micron to achieve proper filtration.
Product recovered from step no. 6 was further quantified for the active phyto constituent by using various instruments but not limited to High pressure liquid chromatograph, gas chromatography, spectroscopy compounds etc.
The product obtained in step no. 7 was also tested for bio-efficacy against pests on different crops in vitro and in vivo, and it is compared to other chemically synthesised fungicides.
The product obtained was effective against broad group of pests within 48 -72 hrs of application with low risk of resistance development and safe for environment and mammalian.
Example 12: Bio-efficacy of present Bio-Fungicide and Synthetic Fungicide against Downy Mildew Disease of Grape (Plasmopara viticola)
Material and Method:`
A field experiment carried out to study the bio-efficacy of present Bio-Fungicide and synthetic fungicide in Grape was conducted at Kay-Bee Bio-Organics Pvt. Ltd’s farm in Maharashtra. The experiment comprising of twenty two treatments was laid out in Randomized Block Design (RBD) with three replication. A susceptible variety Super Sonaka was used in the experiments with spacing’s of 2 X 2 m in plot size of 10 X 10 m. Recommended agronomic practices were followed for raise the Grape orchid. To record disease severity, five plants from each plot were selected. Up to 10 leaves of the tagged plants from top to bottom were observed. To record disease severity, a rating scale of 0-5 was used to score selected plants from each treatment, avoiding border rows. The data were calculated by using standard statistical methods.

Per cent Disease Index (PDI)=(Summation of all numerical ratings )/(Total number of leaves examined X maximum ratings) X 100
0 No visible symptoms apparent
1 A few minute lesions to about 10% of the total leaf area is blighted and usually confined to the 2 bottom leaves
2 Leaves on about 25% of the total area are infected
3 Leaves on about 50% of the total area are infected
4 Leaves on about 75% of the total area are infected
5 Leaves on whole vine are blighted and dead

Results:
Bio-efficacy of Bio-Fungicide and Synthetic Fungicide against Downy Mildew Disease of Grape are depicted in Table 12 and Figure 1.
Phyto-tonic effects of Bio-Fungicide and Synthetic Fungicide on Grape orchid are depicted in Table 13.
Table 12
Sr. No. Treatment No. Dose gm/ml/lit. of water Pre spray PDI PDI after 1st spray PDI after 2nd spray PDC* Over Control
3rd Day 5th Day 3rd Day 5th Day
1 Thymol 1 28.12 24.68 21.45 17.44 15.42 77.87
2 Camphor 1 27.14 25.32 22.09 18.21 16.04 76.98
3 Eugenol 1 27.43 24.43 21.22 17.22 15.23 78.14
4 Citral 1 28.88 26.12 23.32 19.00 17.43 74.98
5 Cumin aldehyde 1 26.78 24.25 20.95 16.87 14.88 78.64
6 Methyl chavicol 1 28.53 25.68 22.38 18.35 16.20 76.75
7 Cinnamaldehyde 1 27.65 25.98 22.86 18.66 16.87 75.79
8 Embodiment :1 1 26.31 22.65 19.35 15.61 12.78 81.66
9 Embodiment :2 1 25.03 22.00 18.77 15.03 11.77 83.11
10 Embodiment :3 1 25.78 22.78 19.53 15.91 12.96 81.40
11 Embodiment :4 1 26.23 23.13 19.92 16.15 13.32 80.88
12 Embodiment :5 1 24.19 22.10 18.87 15.13 12.19 82.50
13 Embodiment :6 1 25.32 22.95 19.72 16.04 13.05 81.27
14 Embodiment :7 1 26.32 23.31 17.11 16.32 13.41 80.75
15 Embodiment :8 1 25.65 22.28 19.03 15.24 12.36 82.26
16 Embodiment :9 1 26.23 22.42 19.17 15.4 12.56 81.97
17 Cymoxanil 8% + Mancozeb 64% WP 3 gm 25.56 23.96 20.45 16.45 14.15 79.69
18 Amisulbrom 20% SC 0.75 ml 27.34 24.87 21.65 17.56 15.66 77.52
19 Dimethomorph 50% WP 1.3 gm 27.43 24.1 20.78 16.66 14.68 78.93
20 Cyazofamid a.i - 34.5%, 0.4 ml 27.76 25.86 22.76 18.56 16.48 76.35
21 Propineb 70% WP 3 gm 28.43 25.09 21.93 17.89 15.88 77.21
22 Control - 26.07 34.63 41.57 57.07 69.67 0.00
SEm (±) 0.94 0.71 0.29 1.25 0.57
CD 5% NS NS 0.89 3.62 1.70
*Per cent disease Control
Table 13
Sr. No. Treatment No. Dose gm/ml/lit. of water Inter-nodal length (cm) Number of bunches Bunch weight (g) Berry length (mm) Berry diameter (mm) Fruit yield per vine (kg)

1 Thymol 1 6.81 32.29 336.59 19.32 18.76 9.34
2 Camphor 1 6.90 31.25 338.00 19.34 18.04 10.64
3 Eugenol 1 5.88 32.40 329.67 19.65 18.90 9.65
4 Citral 1 6.72 32.15 342.34 20.29 18.81 10.75
5 Cumin aldehyde 1 7.05 32.48 327.86 20.45 18.98 10.54
6 Methyl chavicol 1 6.98 31.50 339.65 19.77 18.30 10.82
7 Cinnamaldehyde 1 6.18 32.02 330.50 20.11 18.56 9.85
8 Embodiment :1 1 7.60 33.21 351.20 21.25 19.53 11.48
9 Embodiment :2 1 7.85 33.92 361.28 21.49 19.76 11.78
10 Embodiment :3 1 7.58 33.12 350.78 21.21 19.45 11.45
11 Embodiment :4 1 7.45 33.03 350.53 21.05 19.43 11.36
12 Embodiment :5 1 7.80 33.76 358.76 21.40 19.70 11.73
13 Embodiment :6 1 7.54 32.98 348.44 20.98 19.29 11.25
14 Embodiment :7 1 7.42 33.62 347.12 20.90 19.40 11.17
15 Embodiment :8 1 7.71 33.48 356.43 21.37 19.65 11.65
16 Embodiment :9 1 7.65 33.35 353.23 21.34 19.61 11.55
17 Cymoxanil 8% + Mancozeb 64% WP 3 gm 7.10 31.96 313.21 20.02 18.37 10.38
18 Amisulbrom 20% SC 0.75 ml 7.33 32.77 334.77 20.81 19.05 11.04
19 Dimethomorph 50% WP 1.3 gm 6.45 32.70 345.56 20.76 19.21 10.95
20 Cyazofamid a.i - 34.5%, 0.4 ml 7.28 32.63 344.58 19.87 18.48 11.11
21 Propineb 70% WP 3 gm 6.32 31.77 331.23 20.78 19.14 10.10
22 Control 5.89 29.57 325.67 16.40 15.37 8.57
SEm (±) 0.24 0.12 4.25 0.41 0.61 0.37
CD 5% 0.71 NS 12.68 1.24 1.82 1.12

Conclusion:
Results of the trial revealed that an application of Embodiment: 2 @ 1 ml/l recorded maximum per cent disease control 83.11 followed by Embodiment: 5 @ 1 ml/l 82.50 in grapes against downy mildew disease than the rest of treatments.
All the Bio-Fungicide reported minimum percent disease index than the control and all the Embodiments reported minimum percent disease index than synthetic fungicides.
The Embodiment 2 @ 1 ml/l recorded maximum fruit yield 11.78 kg/vine followed Embodiment 5 @ 1 ml/l 11.73 kg/vine than the rest of treatments.
All tested Embodiments demonstrated significantly superior efficacy compared to individual phytochemicals, chemical standards, and the control treatment.
All the Embodiments reported maximum yield as well as morphological improvement than the synthetic fungicides and control.

Example 13: Bio-efficacy of present Bio-Fungicide and Synthetic Fungicide against Rust (Puccinia graminis) of Wheat
Material and Method
A field experiment carried out to study the bio-efficacy of present Bio-Fungicide and Synthetic fungicide in Wheat was conducted at Kay-Bee Bio-Organics Pvt. Ltd’s. farm in Maharashtra. The experiment comprising of twenty two treatments was laid out in Randomized Block Design (RBD) with three replications. A susceptible variety Netravati was used in the experiments in plot size of 5.00 m x 3.80 m2. Recommended agronomic practices were followed for raise the Wheat crop. To record disease severity, ten plants from each plot were selected. Up to 10 leaves of the tagged plants from top to bottom were observed. To record disease severity, A rating scale of 0-100% was used to score selected plants from each treatment, avoiding border rows. The data were calculated by using standard statistical methods.

Per cent Disease Severity (PDI)=(Sum of individual plant severity percentages)/(Total number of plants assessed)
Rust Severity Class Approximate % Leaf/Stem Area Covered Description / Notes
0 0% No visible infection
T Trace (≈0.1–1%) Few small pustules scattered
5 5% Small but distinct rust patches
10 10% About one-tenth of surface covered
20 20% About one-fifth of surface covered
40 40% About two-fifths of surface covered
60 60% More than half of surface covered
100 100% Entire leaf covered

Results:
Bio-efficacy of Bio-Fungicide and Synthetic Fungicide against Rust (Puccinia graminis) disease of Wheat are depicted in Table 14 and Figure 2.
Phyto-tonic effects of Bio-Fungicide and Synthetic Fungicide on Wheat crop are depicted in Table 15.
Table 14
Sr. No. Treatment No. Dose gm/ml/lit. of water Pre spray PDI PDS after 1st spray PDS after 2nd spray PDC* Over Control
3rd Day 5th Day 3rd Day 5th Day
1 Thymol 1 20.75 20.47 20.05 19.57 19.03 63.07
2 Camphor 1 20.32 20.13 19.87 18.35 17.87 65.32
3 Eugenol 1 19.58 19.47 18.67 17.53 14.17 72.50
4 Citral 1 19.34 18.37 16.87 15.23 14.00 72.83
5 Cumin aldehyde 1 20.67 19.57 19.43 18.57 16.43 68.12
6 Methyl chavicol 1 21.24 20.57 19.87 17.63 14.65 71.57
7 Cinnamaldehyde 1 19.78 19.07 18.37 17.27 15.87 69.20
8 Embodiment :1 1 20.45 18.97 17.07 15.87 13.00 74.77
9 Embodiment :2 1 21.30 19.70 15.67 12.33 9.76 81.06
10 Embodiment :3 1 21.98 20.17 18.87 16.37 12.57 75.61
11 Embodiment :4 1 20.29 20.07 17.57 15.69 11.97 76.77
12 Embodiment :5 1 21.43 20.26 17.23 13.71 9.87 80.85
13 Embodiment :6 1 20.78 20.58 19.57 16.83 12.87 75.02
14 Embodiment :7 1 19.67 19.17 18.57 14.27 12.00 76.71
15 Embodiment :8 1 20.37 19.98 16.97 13.90 11.05 78.56
16 Embodiment :9 1 20.68 20.01 17.02 14.02 11.78 77.14
17 Carbendazim 12% + Mancozeb 63% WP 1 g 21.25 20.27 19.83 17.87 15.67 69.59
18 Azoxystrobin 18.2% + Difenoconazole 11.4% SC 1 g 19.69 17.87 17.03 16.47 13.47 73.86
19 Difenoconazole 3% WS 2 g 20.22 19.43 18.77 17.67 16.87 67.26
20 Tebuconazole 50%+ Trifloxystrobin 25% w/w WG (75 WG) 1 g 19.28 18.47 15.57 14.83 13.03 74.71
21 Propiconazole 25% EC 1 ml 20.43 19.57 17.00 16.53 14.57 71.73
22 Control _ 20.37 27.76 33.60 39.93 51.53 0.00
SEm (±) 0.48 0.41 0.87 0.63 1.05
CD 5% NS NS 2.60 1.89 3.14
*Per cent disease Control
Table 15
Sr. No. Treatment No. Dose gm/ml/lit. of water Plant Height (cm) Effective tillers m-2 Spike length (cm) No. of grains spike-1 Grain yield q ha-1 Straw yield q ha-1

1 Thymol 1 96.49 283.47 8.96 38.97 29.57 86.97
2 Camphor 1 96.27 282.43 8.45 38.73 29.83 86.43
3 Eugenol 1 97.28 290.08 9.01 40.97 30.86 88.73
4 Citral 1 99.37 291.43 9.13 40.53 31.06 89.67
5 Cumin aldehyde 1 97.48 285.37 9.00 39.20 29.43 87.62
6 Methyl chavicol 1 98.14 289.30 9.10 39.63 30.50 87.73
7 Cinnamaldehyde 1 98.01 274.60 9.05 39.43 29.97 87.41
8 Embodiment :1 1 98.12 291.47 9.13 41.43 31.36 90.73
9 Embodiment :2 1 104.58 306.93 9.53 42.53 32.16 94.24
10 Embodiment :3 1 100.27 300.37 9.31 41.57 31.60 91.76
11 Embodiment :4 1 101.37 304.57 9.17 42.10 31.47 92.48
12 Embodiment :5 1 101.49 306.27 9.37 42.13 32.11 93.58
13 Embodiment :6 1 99.37 292.27 9.15 41.20 31.47 92.04
14 Embodiment :7 1 98.57 299.57 9.17 41.23 31.68 91.13
15 Embodiment :8 1 102.35 304.33 9.27 42.10 32.03 93.16
16 Embodiment :9 1 101.37 301.07 9.23 42.07 31.94 93.05
17 Carbendazim 12% + Mancozeb 63% WP 1 g 98.25 287.53 9.07 39.87 30.32 87.73
18 Azoxystrobin 18.2% + Difenoconazole 11.4% SC 1 g 97.58 281.40 9.13 41.00 31.22 89.46
19 Difenoconazole 3% WS 2 g 97.29 283.27 8.97 39.03 29.59 88.28
20 Tebuconazole 50%+ Trifloxystrobin 25% w/w WG (75 WG) 1 g 98.05 296.37 9.10 41.07 31.13 90.21
21 Propiconazole 25% EC 1 ml 97.49 289.03 9.07 40.07 30.74 89.02
22 Control 94.36 276.53 8.56 36.00 27.37 83.62
SEm (±) 0.96 1.87 0.57 0.87 0.58 0.75
CD 5% 2.88 5.61 NS 2.61 1.73 2.25

Conclusion:
Results of the trial revealed that an application of Embodiment: 2 @ 1 ml/l recorded maximum per cent disease control 81.06 followed by Embodiment: 5 @ 1 ml/l 80.85 in Wheat against rust disease than the rest of treatments.
All the Bio-Fungicide reported minimum percent disease index than the control and all the Embodiments reported minimum percent disease index than synthetic fungicides.
The Embodiment 2 @ 1 ml/l recorded maximum fruit yield 32.16 q/ha followed Embodiment 5 @ 1 ml/l 32.11q/ha than the rest of treatments.
All the Embodiments reported maximum yield as well as morphological improvement than the synthetic fungicides and control.

Example 14: Bio-efficacy of present Bio-Fungicide and Synthetic Fungicide against Powdery Mildew (Leveillula taurica) disease of Chilli
Material and Method
A field experiment carried out to study the bio-efficacy of present Bio-Fungicide and Synthetic fungicide in Chilli was conducted at Kay-Bee Bio-Organics Pvt. Ltd’s. farm at Phaltan, Maharashtra. The experiment comprising of twenty-two treatments was laid out in Randomized Block Design (RBD) with three replications was used. A susceptible variety Pusa Jwala was used in the experiments with spacings of 60 X 45 cm in plot size of 2.40 X 3.15 m. Recommended agronomic practices were followed raise the Chilli crop. To record disease severity, A rating scale of 0-5 was used to score selected plants from each treatment, avoiding border rows . The grades were assigned numerical ratings proportional to the diseased area. The data were calculated by using standard statistical methods.
Per cent Disease Index (%)=(Summation of all numerical ratings )/(Total number of leaves examined X maximum ratings) X 100

0 No visible symptoms apparent
1 A few minute lesions to about 10% of the total leaf area is blighted and usually confined to the 2 bottom leaves
2 Leaves on about 25% of the total area are infected
3 Leaves on about 50% of the total area are infected
4 Leaves on about 75% of the total area are infected
5 Leaves on whole vine are blighted and dead

Results:
Bio-efficacy of Bio-Fungicide and Synthetic Fungicide against Powdery mildew disease of Chilli are depicted in Table 16 and Figure 3.
Phyto-tonic effect of Bio-Fungicide and Synthetic Fungicide on Chilli Crop is depicted in Table 17.
Table 16
Sr. No. Treatment No. Dose gm/ml/lit. of water Pre spray PDI PDI after 1st spray PDI after 2nd spray PDC* Over Control
3rd Day 5th Day 3rd Day 5th Day
1 Thymol 1 25.11 22.70 19.98 17.00 16.76 75.73
2 Camphor 1 23.89 21.80 18.65 15.78 15.34 77.79
3 Eugenol 1 25.67 21.88 20.75 17.65 17.66 74.43
4 Citral 1 23.65 21.44 17.80 14.60 14.00 79.73
5 Cumin aldehyde 1 25.88 23.12 20.22 17.23 17.00 75.39
6 Methyl chavicol 1 24.67 21.66 18.21 14.96 14.76 78.63
7 Cinnamaldehyde 1 24.89 21.77 20.27 17.32 17.23 75.05
8 Embodiment :1 1 24.54 20.11 16.74 13.21 10.45 84.87
9 Embodiment :2 1 22.13 19.27 16.30 12.47 9.23 86.64
10 Embodiment :3 1 23.40 20.45 16.84 13.45 10.65 84.58
11 Embodiment :4 1 23.78 20.66 16.92 13.67 11.21 83.77
12 Embodiment :5 1 23.48 19.32 16.34 12.56 9.37 86.43
13 Embodiment :6 1 22.67 20.78 17.00 13.73 11.45 83.42
14 Embodiment :7 1 23.14 20.91 17.32 13.88 11.78 82.94
15 Embodiment :8 1 22.46 19.56 16.48 12.78 9.56 86.16
16 Embodiment :9 1 24.66 19.66 16.60 12.84 9.74 85.90
17 Carbendazim 12% + Mancozeb 63% WP 1.5 gm 25.12 22.32 19.14 16.34 16.21 76.53
18 Azoxystrobin 18.2% + Difenoconazole 11.4% SC 1 ml 24.65 21.00 17.45 14.00 12.45 81.97
19 Tebuconazole 10% + Sulphur 65% WG 2.5 gm 24.18 22.12 18.90 16.00 15.76 77.18
20 Tebuconazole 50%+ Trifloxystrobin 25% w/w WG (75 WG) 0.5 gm 24.98 22.45 19.21 16.78 16.44 76.20
21 Azoxystrobin 4.7% + Mancozeb 59.7% + Tebuconazole 5.6% WG 4 gm 24.48 21.32 17.54 14.32 13.56 80.37
22 Control - 22.33 31.03 49.37 58.00 69.07 0.00
SEm (±) 0.56 0.86 0.92 1.43 1.53
CD 5% NS 2.57 2.74 4.30 4.60
*Per cent disease Control
Table 17
Sr. No. Treatment No. Dose gm/ml/lit. of water Plant Height (cm) Number of Branches Days to flower initiation No. of fruits per plant Fruit yield per plant (g/plant) Fruit yield per ha. (t/ha.)

1 Thymol 1 71.20 5.99 55.25 62.89 369.52 14.04
2 Camphor 1 68.44 6.21 54.96 60.12 366.78 13.94
3 Eugenol 1 74.87 5.80 55.18 64.34 356.98 12.98
4 Citral 1 74.12 6.43 54.30 62.45 371.40 14.66
5 Cumin aldehyde 1 73.88 5.55 55.04 63.29 335.76 13.76
6 Methyl chavicol 1 75.14 6.40 54.70 61.11 362.65 14.27
7 Cinnamaldehyde 1 70.65 5.69 54.41 62.22 327.65 13.21
8 Embodiment :1 1 80.34 7.38 57.00 66.96 403.43 16.08
9 Embodiment :2 1 82.60 8.23 58.33 67.47 427.59 16.87
10 Embodiment :3 1 79.66 7.11 56.84 66.26 398.77 16.00
11 Embodiment :4 1 79.35 6.76 56.70 66.78 400.78 15.70
12 Embodiment :5 1 81.97 8.07 57.76 67.23 420.70 16.65
13 Embodiment :6 1 78.21 6.98 56.53 66.08 395.76 15.93
14 Embodiment :7 1 78.56 6.66 56.28 66.18 392.12 15.83
15 Embodiment :8 1 81.23 7.89 57.50 67.01 415.67 16.45
16 Embodiment :9 1 80.87 7.50 57.23 65.89 408.77 16.21
17 Carbendazim 12% + Mancozeb 63% WP 1.5 gm 76.21 6.48 55.40 65.21 381.40 14.88
18 Azoxystrobin 18.2% + Difenoconazole 11.4% SC 1 ml 73.16 6.18 54.53 65.77 341.60 15.32
19 Tebuconazole 10% + Sulphur 65% WG 2.5 gm 77.57 6.56 56.02 61.89 385.60 15.04
20 Tebuconazole 50%+ Trifloxystrobin 25% w/w WG (75 WG) 0.5 gm 75.87 6.03 54.40 65.58 348.12 13.65
21 Azoxystrobin 4.7% + Mancozeb 59.7% + Tebuconazole 5.6% WG 4 gm 75.82 6.12 55.89 61.48 375.00 13.50
22 Control 62.30 4.90 53.50 54.57 298.99 11.58
SEm (±) 1.11 0.25 2.35 0.89 0.61 0.37
CD 5% 3.32 NS NS 2.67 1.8 1.12

Conclusion
Results of the trial revealed that an application of Embodiment: 2 @ 1 ml/l recorded maximum per cent disease control 86.64 followed by Embodiment: 5 @ 1 ml/l 86.43 in Chilli against Powdery Mildew disease than the rest of treatments.
All the Bio-Fungicide reported minimum percent disease index than the control and all the Embodiments reported minimum percent disease index than synthetic fungicides.
The Embodiment 2 @ 1 ml/l recorded maximum fruit yield 16.87 t/ha followed Embodiment 5 @ 1 ml/l 16.65 t/ha than the rest of treatments.
All tested Embodiments demonstrated significantly superior efficacy compared to individual phytochemicals, chemical standards, and the control treatment.
All the Embodiments reported maximum yield as well as morphological improvement than the synthetic fungicides and control.
Example 15: Bio-efficacy of present Bio-Fungicide and Synthetic fungicide against Rhizome Rot (Pythium aphanidermatum) of Ginger
Material and Method
A field experiment carried out to study the bio-efficacy of present Bio-Fungicide and synthetic fungicide against rhizome rot in Ginger was conducted at Kay-Bee Bio-Organics Pvt. Ltd’s. farm Satara, Maharashtra, India during 2023-2024. The experiment comprising of twenty-two treatments was laid out in Randomized Block Design (RBD) with three replications. The trial was laid in a standard plot size of 3 X 1 m2 in ridges and furrows method and rhizomes were planted in 30 X 20 cm apart. Recommended agronomic practices were followed to raise the Ginger crop. The percent disease incidence were recorded and calculated by using following formula. The collected data were analyzed by using standard statistical methods

Per cent Disease Incidence (%)=(Number of infected plant)/(Total number of Plant)*100
Results:
Bio-efficacy of Bio-Fungicide and Synthetic fungicide against Rhizome Rot (Pythium aphanidermatum) of Ginger are depicted in Table 18 and Figure 4.
Phyto-tonic effect of Bio-Fungicide and Synthetic fungicide on Ginger Crop is depicted in Table 19.

Table 18
Sr. No. Treatment No. Dose gm/ml/lit. of water Pre-drench PDI PDI after 1st drench PDI after 2nd drench PDC* Over Control
3rd Day 5th Day 3rd Day 5th Day
1 Thymol 1 20.43 19.87 18.50 16.87 16.43 62.29
2 Camphor 1 18.33 17.57 16.83 12.53 11.93 72.62
3 Eugenol 1 19.43 19.53 17.47 15.23 15.43 64.59
4 Citral 1 20.73 18.57 16.40 14.35 13.83 68.26
5 Cumin aldehyde 1 21.53 19.43 17.40 15.27 12.77 70.69
6 Methyl chavicol 1 21.67 18.40 16.47 15.47 14.73 66.19
7 Cinnamaldehyde 1 19.47 18.67 17.54 16.43 14.53 66.65
8 Embodiment :1 1 17.40 16.43 15.20 13.42 10.03 76.98
9 Embodiment :2 1 20.57 18.43 13.50 10.70 7.80 82.10
10 Embodiment :3 1 19.43 17.63 14.47 11.83 9.78 77.55
11 Embodiment :4 1 17.67 16.87 15.63 12.17 9.97 77.12
12 Embodiment :5 1 21.53 19.57 14.23 12.87 8.63 80.19
13 Embodiment :6 1 19.00 17.57 15.43 13.57 10.87 75.05
14 Embodiment :7 1 19.43 17.43 14.86 12.27 10.43 76.06
15 Embodiment :8 1 18.93 17.87 15.43 12.60 8.87 79.64
16 Embodiment :9 1 17.40 16.67 14.80 13.50 9.03 79.27
17 Thiophanate Methyl 70 % 2.0 gm 19.87 17.07 15.73 13.57 11.97 72.53
18 Metalaxyl 4%+Manconzeb 64% 3.0 gm 21.50 20.73 18.83 15.37 13.53 68.95
19 Mancozeb 63% + Carbendazim 12 % 1.5 gm 16.93 16.73 14.35 13.73 12.27 71.84
20 Fosetyl Al 80 WP (80% w/w) 2.0 gm 20.23 18.47 15.30 14.73 11.17 74.36
21 Copper Oxychloride 50 % WP 2.0 gm 18.83 18.13 16.67 14.73 14.47 66.79
22 Control - 18.43 22.93 27.87 35.53 43.57 0.00
SEm (±) 0.48 0.97 0.47 0.75 0.34
CD 5% NS 2.90 1.41 2.26 1.02
*Per cent disease Control
Table 17
Sr. No. Treatment No. Dose gm/ml/lit. of water Plant Height (cm) No. of pseudostem/ clump No. of leaves/ clump Yield (t/ha.)

1 Thymol 1 80.34 4.17 61.47 57.89
2 Camphor 1 89.71 4.43 65.73 58.43
3 Eugenol 1 79.60 4.43 62.50 56.41
4 Citral 1 86.37 4.37 64.93 58.87
5 Cumin aldehyde 1 84.39 4.57 64.75 57.93
6 Methyl chavicol 1 81.37 4.23 63.33 58.72
7 Cinnamaldehyde 1 80.46 4.20 63.70 57.62
8 Embodiment :1 1 91.43 4.83 67.23 61.03
9 Embodiment :2 1 95.49 5.67 70.53 65.64
10 Embodiment :3 1 93.47 5.00 67.57 62.31
11 Embodiment :4 1 92.03 5.00 67.53 61.48
12 Embodiment :5 1 94.13 5.63 69.47 64.82
13 Embodiment :6 1 89.37 4.97 67.27 60.29
14 Embodiment :7 1 92.84 4.77 66.75 61.72
15 Embodiment :8 1 95.21 5.07 69.73 63.48
16 Embodiment :9 1 94.47 5.63 68.47 62.78
17 Thiophanate Methyl 70 % 2.0 gm 85.37 4.47 66.77 59.37
18 Metalaxyl 4%+Manconzeb 64% 3.0 gm 82.44 4.40 64.37 59.48
19 Mancozeb 63% + Carbendazim 12 % 1.5 gm 83.50 4.47 66.47 57.98
20 Fosetyl Al 80 WP (80% w/w) 2.0 gm 87.87 4.57 67.03 60.09
21 Copper Oxychloride 50 % WP 2.0 gm 82.49 4.33 62.48 58.49
22 Control 73.84 3.77 55.45 49.12
SEm (±) 0.78 0.28 0.89 0.74
CD 5% 2.36 0.85 2.68 2.22

Conclusion
Results of the trial revealed that an application of Embodiment: 2 @ 1 ml/l recorded maximum per cent disease control 82.10 followed by Embodiment: 5 @ 1 ml/l 80.19 in Ginger against Rhizome Rot (Pythium aphanidermatum) than the rest of treatments.
All the Bio-Fungicide reported minimum percent disease index than the control and all the Embodiments reported minimum percent disease index than synthetic fungicides.
The Embodiment 2 @ 1 ml/l recorded maximum rhizome yield 65.64 t/ha followed by Embodiment 5 @ 1 ml/l 64.82 t/ha than the rest of treatments.
All tested Embodiments demonstrated significantly superior efficacy compared to individual phytochemicals, chemical standards, and the control treatment.
All the Embodiments reported maximum yield as well as morphological improvement than the synthetic fungicides and control.

Example 16: Bio-efficacy of present Bio-Fungicide and Synthetic Fungicide against Blast (Pyricularia oryzae) of Paddy
Material and Method
The field experimental trial was conducted to study the bio-efficacy of different bio-fungicides and synthetic fungicides against Blast of Paddy during kharif season at Kay Bee Research and Development Farm, Maharashtra. The experiment was laid out by using randomized block design with three replication and twenty two treatments following spacing’s of 20 cm and 15 cm between rows and plants respectively. Each treatment consisted of an area of 3.5 × 5 m. Recommended agronomic practices were followed to raise the paddy crop. Observations on blast disease were taken on randomly selected ten hills or plants per plot. Two sprays were taken & disease intensity was recorded as per standard method use before and after the application of fungicides. To record disease severity, a rating scale of 0-9 was used to score selected plants from each treatment, avoiding border rows (IRRI, 1996). The yield parameters were recorded at harvesting stage. The data were analyzed by applying standard statistical methods.

Per cent Disease Index (PDI)=(Summation of all numerical ratings )/(Total number of leaves examined X maximum ratings) X 100

0 No infection
1 Vertical spread of the lesions up to 20% of plant height
3 Vertical spread of the lesions up to 21-30% of plant height
5 Vertical spread of the lesions up to 31-45% of plant height
7 Vertical spread of the lesions up to 46-65% of plant height
9 Vertical spread of the lesions up to 66-100% of plant height

Results:
Bio-efficacy of Bio-Fungicide and Synthetic Fungicide against blast (Pyricularia oryzae) of Paddy are depicted in Table 20 and Figure 5.
Phyto-tonic effect of Bio-Fungicide and Synthetic Fungicide on Paddy Crop is depicted in Table 21.
Table 20
Sr. No. Treatment No. Dose gm/ml/lit. of water Pre spray PDI PDI after 1st spray PDI after 2nd spray PDC* Over Control
3rd Day 5th Day 3rd Day 5th Day
1 Thymol 1 31.16 29.14 28.83 24.60 21.97 62.46
2 Camphor 1 29.89 28.76 28.12 24.20 21.45 63.35
3 Eugenol 1 29.85 28.87 27.76 23.40 20.14 65.59
4 Citral 1 31.15 28.85 28.34 24.34 21.48 63.30
5 Cumin aldehyde 1 30.18 28.58 27.56 22.76 19.95 65.91
6 Methyl chavicol 1 29.78 28.65 27.57 23.66 20.21 65.47
7 Cinnamaldehyde 1 30.12 28.98 28.75 24.56 21.87 62.63
8 Embodiment :1 1 28.65 28.08 25.66 18.63 16.34 72.08
9 Embodiment :2 1 29.33 26.37 22.80 17.53 11.47 80.40
10 Embodiment :3 1 30.14 28.44 26.12 18.97 17.18 70.65
11 Embodiment :4 1 30.88 29.44 26.86 19.66 18.65 68.14
12 Embodiment :5 1 28.54 26.38 23.56 20.18 13.32 77.24
13 Embodiment :6 1 29.68 26.45 24.68 20.67 19.05 67.45
14 Embodiment :7 1 30.05 27.32 25.96 20.74 19.66 66.41
15 Embodiment :8 1 29.18 27.77 24.76 18.54 14.48 75.26
16 Embodiment :9 1 29.88 27.96 25.12 18.10 15.88 72.87
17 Carbendazim 12% + Mancozeb 63% WP 1 gm 30.15 28.33 28.12 23.45 20.34 65.25
18 Azoxystrobin 18.2% + Difenoconazole 11.4% SC 1 ml 29.60 28.12 26.98 22.12 19.67 66.39
19 Hexaconazole 5% EC 2 ml 30.42 28.28 27.21 22.45 19.88 66.03
20 Tebuconazole 50%+ Trifloxystrobin 25% w/w WG (75 WG) 0.4 g 30.65 28.57 27.99 24.22 20.66 64.70
21 Azoxystrobin 4.7% + Mancozeb 59.7% + Tebuconazole 5.6% WG 4 gm 31.15 28.57 27.43 23.56 19.94 65.93
22 Control 26.97 30.53 36.53 49.70 58.53 0.00
SEm (±) 0.28 0.73 1.02 0.98 1.25
CD 5% NS NS 3.06 2.96 3.76
*Per cent disease Control

Table 21
Sr. No. Treatment No. Dose gm/ml/lit. of water Plant Height (cm) Effective tillers m-2 Panicle length (cm) Grains panicle-1 Grain yield qt. ha-1 Straw yield qt. ha-1

1 Thymol 1 67.21 7.35 19.35 112.66 38.34 53.12
2 Camphor 1 75.67 7.29 21.66 112.56 46.74 56.77
3 Eugenol 1 71.43 8.12 19.08 113.54 41.20 52.95
4 Citral 1 77.33 8.32 22.02 120.45 47.32 57.00
5 Cumin aldehyde 1 68.77 7.20 21.77 119.34 39.21 52.10
6 Methyl chavicol 1 72.22 7.77 19.45 114.54 45.88 55.67
7 Cinnamaldehyde 1 76.39 7.53 21.48 118.12 40.31 51.67
8 Embodiment :1 1 81.76 9.12 23.56 123.67 51.03 62.00
9 Embodiment :2 1 85.48 11.00 24.56 128.34 53.36 64.50
10 Embodiment :3 1 80.78 10.34 22.87 126.56 50.80 61.45
11 Embodiment :4 1 79.56 9.78 23.40 126.44 49.67 60.34
12 Embodiment :5 1 84.31 12.32 24.42 127.28 52.38 63.45
13 Embodiment :6 1 81.34 10.23 23.04 125.45 48.40 60.70
14 Embodiment :7 1 80.21 9.54 22.67 124.28 49.32 59.89
15 Embodiment :8 1 82.68 11.76 24.07 127.12 51.34 62.35
16 Embodiment :9 1 82.08 11.34 23.77 126.78 51.20 62.10
17 Carbendazim 12% + Mancozeb 63% WP 1 gm 78.37 7.65 22.10 120.52 46.72 54.88
18 Azoxystrobin 18.2% + Difenoconazole 11.4% SC 1 ml 78.59 8.88 22.56 121.66 48.50 59.21
19 Hexaconazole 5% EC 2 ml 73.18 8.45 20.57 117.56 42.48 57.89
20 Tebuconazole 50%+ Trifloxystrobin 25% w/w WG (75 WG) 0.4 g 69.45 7.44 20.05 116.00 43.55 53.48
21 Azoxystrobin 4.7% + Mancozeb 59.7% + Tebuconazole 5.6% WG 4 gm 74.45 8.41 22.24 121.66 47.53 58.48
22 Control 65.48 7.02 17.34 106.45 36.10 48.54
SEm (±) 1.03 0.37 2.35 1.43 0.97 0.64
CD 5% 3.09 1.1 7.04 4.27 2.9 1.93

Conclusion
Results of the trial revealed that an application of Embodiment: 2 @ 1 ml/l recorded maximum per cent disease control 80.40% followed by Embodiment: 5 @ 1 ml/l 77.24 in paddy against blast disease than the rest of treatments.
All the Bio-Fungicide reported minimum percent disease index than the control and all the Embodiments reported minimum percent disease index than synthetic fungicides.
The Embodiment 2 @ 1 ml/l recorded maximum grain yield 53.36 qt/ha. followed Embodiment 5 @ 1 ml/l 52.38 qt/ha. than the rest of treatments.
All tested Embodiments demonstrated significantly superior efficacy compared to individual phytochemicals, chemical standards, and the control treatment.
All the Embodiments reported maximum yield as well as morphological improvement than the synthetic fungicides and control.

Example 17: In vitro Bio-efficacy of present Bio-Fungicides (Embodiment) and Synthetic Fungicides against Blast (Pyricularia oryzae) of Rice
Material and Method:
Pathogen Studied : Pyricularia oryzae (Collected culture from Plant Pathology section)
Technique Used : Poison Food Technique
Media used : Potato Dextrose Agar medium (For Fungus growing)
No. of Treatments : 47
Replications : 3
Micropipette : Require for taking accurate volume of Fungicides as per recommendations.
Weighing Balance : Require for taking accurate volume of Fungicides as per recommendations
Antibiotic Zone Scale : Require for measuring accurate zone
Observation Recorded : After three Days Application
Statistical Design Used : Completely Randomized Design
Calculations : Zone of inhibition (mm)

Results:-
In vitro effect of bio-fungicides on growth and inhibition of Pyricularia oryzae is provided in Table 22 and figure 6.
Table 22
Sr. No. Treatment No. Dose ml or gm/lit Mean colony Diameter (mm) Per cent Inhibition of growth (%)
1 Thymol 1 19.02 74.53
2 Camphor 1 18.21 75.62
3 Eugenol 1 20.76 72.20
4 Gingerol 1 19.49 73.90
5 Carvacrol 1 20.87 72.05
6 Piperine 1 20.46 72.60
7 Citral 1 21.02 71.85
8 Cumin aldehyde 1 22.47 69.91
9 1,8, cineole 1 23.84 68.08
10 Thymoquinone 1 22.49 69.88
11 β-Asarone 1 25.91 65.31
12 Alpha-pinene 1 19.63 73.71
13 Chavibetol 1 21.03 71.84
14 Berberine 1 18.98 74.58
15 Essential oils and total turmerones 1 19.01 74.54
16 ρ-Cymene 1 18.85 74.76
17 Mixture of Essential oils and oleoresin 1 21.78 70.84
18 Camphene 1 21.02 71.85
19 Menthol 1 25.86 65.37
20 D-limonene 1 19.95 73.29
21 Capsaicin 1 21.02 71.85
22 Methyl chavicol 1 17.63 76.39
23 Cinnamaldehyde 1 17.84 76.11
24 Embodiment :1 0.5 18.39 75.37
25 Embodiment :1 1 15.92 78.68
26 Embodiment :2 0.5 13.78 81.55
27 Embodiment :2 1 9.51 87.27
28 Embodiment :3 0.5 21.78 70.84
29 Embodiment :3 1 19.79 73.50
30 Embodiment :4 0.5 17.71 76.29
31 Embodiment :4 1 18.03 75.86
32 Embodiment :5 0.5 13.47 81.96
33 Embodiment :5 1 11.89 84.08
34 Embodiment :6 0.5 21.75 70.88
35 Embodiment :6 1 19.76 73.54
36 Embodiment :7 0.5 22.13 70.37
37 Embodiment :7 1 18.9 74.69
38 Embodiment :8 0.5 14.92 80.02
39 Embodiment :8 1 14.81 80.17
40 Embodiment :9 0.5 15.13 79.74
41 Embodiment :9 1 12.76 82.91
42 Kasugamycin 5% + Copper Oxychloride 45% WP 1.5 21.78 70.84
43 Picoxystrobin 6.78% + Tricyclazole 20.33% w/w SC 2 19.78 73.51
44 Pyraclostrobin 100g/l CS 2 22.89 69.35
45 Azoxystrobin 18.2% + Difenoconazole 11.4% SC 1 23.49 68.55
46 Tricyclazole 18 % + Mancozeb 62 % WP 1.0 gm 23.47 68.57
47 Control 74.68

Conclusions:
All tested Embodiments demonstrated significantly superior efficacy compared to individual phytochemicals, chemical standards, and the control treatment.
In vitro bio-efficacy study revealed that Embodiment 2 @ 1.0ml/lit showed maximum zone of inhibition 9.51 (mm) followed by Embodiment 5 @ 1.0ml/lit and Embodiment 9 @ 1 ml/lit against Pyricularia oryzae.

Example 18: In vitro Bio-efficacy of present Bio-Fungicides (Embodiment) and Synthetic Fungicides against Anthracnose (Colletotrichum capsici) of Chilli
Material and Methodology:
Pathogen Studied : Colletotrichum capsici (Collected culture from Plant Pathology section)
Technique Used : Poison Food Technique
Media used : Potato Dextrose Agar medium (For Fungus growing)
No. of Treatments : 47
Replications : 3
Micropipette : Require for taking accurate volume of Fungicides as per recommendations.
Weighing Balance : Require for taking accurate volume of Fungicides as per recommendations
Antibiotic Zone Scale : Require for measuring accurate zone
Observation Recorded : After three Days Application
Statistical Design Used : Completely Randomized Design
Calculations : Zone of inhibition (mm)
Results:-
In vitro effect of bio-fungicides on growth and inhibition of Colletotrichum capsici is provided in Table 23 and figure 7.
Table 23
Sr. No. Treatment No. Dose ml or gm/lit Mean colony Diameter (mm) Per cent Inhibition of growth (%)
1 Thymol 1 17.63 77.91
2 Camphor 1 17.53 78.04
3 Eugenol 1 16.66 79.13
4 Gingerol 1 21.66 72.86
5 Carvacrol 1 22.47 71.85
6 Piperine 1 23.33 70.77
7 Citral 1 15.43 80.67
8 Cumin aldehyde 1 16.33 79.54
9 1,8, cineole 1 25.15 68.49
10 Thymoquinone 1 24.43 69.39
11 β-Asarone 1 23.15 71.00
12 Alpha-pinene 1 21.57 72.98
13 Chavibetol 1 23.17 70.97
14 Berberine 1 21.13 73.53
15 Essential oils and total turmerones 1 21.67 72.85
16 ρ-Cymene 1 23.17 70.97
17 Mixture of Essential oils and oleoresin 1 23.13 71.02
18 Camphene 1 22.66 71.61
19 Menthol 1 23.17 70.97
20 D-limonene 1 23.53 70.52
21 Capsaicin 1 22.13 72.28
22 Methyl chavicol 1 17.13 78.54
23 Cinnamaldehyde 1 16.17 79.74
24 Embodiment :1 0.5 19.53 75.53
25 Embodiment :1 1 17.66 77.88
26 Embodiment :2 0.5 13.55 83.02
27 Embodiment :2 1 9.33 88.31
28 Embodiment :3 0.5 19.83 75.16
29 Embodiment :3 1 17.83 77.66
30 Embodiment :4 0.5 20.37 74.48
31 Embodiment :4 1 18.57 76.74
32 Embodiment :5 0.5 14.23 82.17
33 Embodiment :5 1 11.13 86.06
34 Embodiment :6 0.5 21.23 73.40
35 Embodiment :6 1 19.35 75.76
36 Embodiment :7 0.5 21.59 72.95
37 Embodiment :7 1 18.63 76.66
38 Embodiment :8 0.5 15.57 80.49
39 Embodiment :8 1 12.44 84.41
40 Embodiment :9 0.5 15.96 80.01
41 Embodiment :9 1 12.67 84.13
42 Carbendazim 12% + Mancozeb 63% WP 1.5 23.53 70.52
43 Azoxystrobin 18.2% + Difenoconazole 11.4% SC 1 17.63 77.91
44 Tebuconazole 10% + Sulphur 65% WG 2.5 22.37 71.97
45 Tebuconazole 50%+ Trifloxystrobin 25% w/w WG (75 WG) 0.5 21.69 72.83
46 Captan 70% + Hexaconazole 5% WP 2 22.66 71.61
47 Control 79.82

Conclusion :
All tested Embodiments demonstrated significantly superior efficacy compared to individual phytochemicals, chemical standards, and the control treatment.
In vitro bio-efficacy study revealed that Embodiment 2 @ 1.0ml/lit showed maximum zone of inhibition 9.33 (mm) followed by Embodiment 5 @ 1.0ml/lit and Embodiment 8 @ 1 ml/lit against Colletotrichum capsici.
Example 19: In vitro Bio-efficacy of present Bio-Fungicides (Embodiment) and Synthetic Fungicides against Late Blight (Phytophthora infestans) of Potato
Material and Methodology:
Pathogen Studied : Phytophthora infestans (Collected culture from Plant Pathology section)
Technique Used : Poison Food Technique
Media used : Potato Dextrose Agar medium (For Fungus growing)
No. of Treatments : 47
Replications : 3
Micropipette : Require for taking accurate volume of Fungicides as per recommendations.
Weighing Balance : Require for taking accurate volume of Fungicides as per recommendations
Antibiotic Zone Scale : Require for measuring accurate zone
Observation Recorded : After three Days Application
Statistical Design Used : Completely Randomized Design
Calculations : Zone of inhibition (mm)
Results:-
In vitro effect of bio-fungicides on growth and inhibition of Phytophthora infestans is provided in Table 24 and figure 8.
Table 24
Sr. No. Treatment No. Dose ml or gm/lit Mean colony Diameter (mm) Per cent Inhibition of growth (%)
1 Thymol 1 15.49 80.81
2 Camphor 1 18.45 77.15
3 Eugenol 1 15.76 80.48
4 Gingerol 1 20.46 74.66
5 Carvacrol 1 21.57 73.28
6 Piperine 1 21.2 73.74
7 Citral 1 14.55 81.98
8 Cumin aldehyde 1 15.53 80.76
9 1,8, cineole 1 24.19 70.04
10 Thymoquinone 1 21.53 73.33
11 β-Asarone 1 24.07 70.18
12 Alpha-pinene 1 20.76 74.28
13 Chavibetol 1 22.5 72.13
14 Berberine 1 21.13 73.83
15 Essential oils and total turmerones 1 20.43 74.69
16 ρ-Cymene 1 21.75 73.06
17 Mixture of Essential oils and oleoresin 1 22.15 72.56
18 Camphene 1 21.97 72.79
19 Menthol 1 24.97 69.07
20 D-limonene 1 20.5 74.61
21 Capsaicin 1 21.57 73.28
22 Methyl chavicol 1 16.5 79.56
23 Cinnamaldehyde 1 18.17 77.49
24 Embodiment :1 0.5 17.53 78.29
25 Embodiment :1 1 16.67 79.35
26 Embodiment :2 0.5 12.53 84.48
27 Embodiment :2 1 8.76 89.15
28 Embodiment :3 0.5 20.5 74.61
29 Embodiment :3 1 18.15 77.52
30 Embodiment :4 0.5 19.65 75.66
31 Embodiment :4 1 17.25 78.63
32 Embodiment :5 0.5 14.53 82.00
33 Embodiment :5 1 10.25 87.30
34 Embodiment :6 0.5 22.23 72.46
35 Embodiment :6 1 18.25 77.39
36 Embodiment :7 0.5 20.65 74.42
37 Embodiment :7 1 17.6 78.20
38 Embodiment :8 0.5 14.15 82.47
39 Embodiment :8 1 13.43 83.36
40 Embodiment :9 0.5 14.65 81.85
41 Embodiment :9 1 11.53 85.72
42 Carbendazim 12% + Mancozeb 63% WP 3.5 20.43 74.69
43 Azoxystrobin 11% & Tebuconazole 18.3% w/w SC 1.5 19.67 75.63
44 Cymoxanil 8% + Mancozeb 64% WP 3 21.23 73.70
45 Chlorothalonil 75% WP 1.5 22.67 71.92
46 Captan 70% + Hexaconazole 5% WP 2 23.47 70.93
47 Control 80.73

Conclusion:
All tested Embodiments demonstrated significantly superior efficacy compared to individual phytochemicals, chemical standards, and the control treatment.
In vitro bio-efficacy study revealed that Embodiment 2 @ 1.0ml/lit showed maximum zone of inhibition 8.76 (mm) followed by Embodiment 5 @ 1.0ml/lit and Embodiment 9 @ 1 ml/lit against Phytophthora infestans.
Example 20: In vitro Bio-efficacy of present Bio-Fungicides (Embodiment) and Synthetic Fungicides against Root Rot (Rhizoctonia bataticola) of Cotton
Material and Methodology:
Pathogen Studied : Rhizoctonia bataticola (Collected culture from Plant Pathology section)
Technique Used : Poison Food Technique
Media used : Potato Dextrose Agar medium (For Fungus growing)
No. of Treatments : 47
Replications : 3
Micropipette : Require for taking accurate volume of Fungicides as per recommendations.
Weighing Balance : Require for taking accurate volume of Fungicides as per recommendations
Antibiotic Zone Scale : Require for measuring accurate zone
Observation Recorded : After three Days Application
Statistical Design Used : Completely Randomized Design
Calculations : Zone of inhibition (mm)
Results:-
In vitro effect of bio-fungicides on growth and inhibition of Rhizoctonia bataticola is provided in Table 25 and figure 9.
Table 25
Sr. No. Treatment No. Dose ml or gm/lit Mean colony Diameter (mm) Per cent Inhibition of growth (%)
1 Thymol 1 20.03 75.88
2 Camphor 1 19.45 76.57
3 Eugenol 1 21.23 74.43
4 Gingerol 1 20.88 74.85
5 Carvacrol 1 22.46 72.95
6 Piperine 1 21.89 73.64
7 Citral 1 22.03 73.47
8 Cumin aldehyde 1 23.41 71.81
9 1,8, cineole 1 24.43 70.58
10 Thymoquinone 1 23.33 71.90
11 β-Asarone 1 26.82 67.70
12 Alpha-pinene 1 20.21 75.66
13 Chavibetol 1 22.87 72.46
14 Berberine 1 19.47 76.55
15 Essential oils and total turmerones 1 19.47 76.55
16 ρ-Cymene 1 19.84 76.11
17 Mixture of Essential oils and oleoresin 1 21.82 73.72
18 Camphene 1 21.08 74.61
19 Menthol 1 21.87 73.66
20 D-limonene 1 21.87 73.66
21 Capsaicin 1 22.41 73.01
22 Methyl chavicol 1 18.98 77.14
23 Cinnamaldehyde 1 18.81 77.35
24 Embodiment :1 0.5 19.52 76.49
25 Embodiment :1 1 16.66 79.93
26 Embodiment :2 0.5 14.21 82.89
27 Embodiment :2 1 10.59 87.25
28 Embodiment :3 0.5 22.84 72.49
29 Embodiment :3 1 20.05 75.85
30 Embodiment :4 0.5 18.42 77.82
31 Embodiment :4 1 19.85 76.09
32 Embodiment :5 0.5 14.81 82.16
33 Embodiment :5 1 12.78 84.61
34 Embodiment :6 0.5 22.19 73.27
35 Embodiment :6 1 20.87 74.86
36 Embodiment :7 0.5 23.77 71.37
37 Embodiment :7 1 19.33 76.72
38 Embodiment :8 0.5 15.85 80.91
39 Embodiment :8 1 15.93 80.81
40 Embodiment :9 0.5 16.78 79.79
41 Embodiment :9 1 13.03 84.31
42 Thiophanate Methyl 70 % 2.0 gm 22.61 72.77
43 Metalaxyl 4%+Manconzeb 64% 3.0 gm 20.22 75.65
44 Mancozeb 63% + Carbendazim 12 % 1.5 gm 23.72 71.43
45 Fosetyl Al 80 WP (80% w/w) 2.0 gm 24.03 71.06
46 Copper Oxychloride 50 % WP 2.0 gm 24.96 69.94
47 Control 83.03

Conclusion:
All tested Embodiments demonstrated significantly superior efficacy compared to individual phytochemicals, chemical standards, and the control treatment.
In vitro bio-efficacy study revealed that Embodiment 2 @ 1.0ml/lit showed maximum zone of inhibition 10.59 (mm) followed by Embodiment 5 @ 1.0ml/lit and Embodiment 9 @ 1 ml/lit against Rhizoctonia bataticola.
Example 21: In vitro Bio-efficacy of present Bio-Fungicides (Embodiment) and Synthetic Fungicides against Wilt (Fusarium oxysporum f. sp. ciceris) of Gram
Material and Methodology:
Pathogen Studied : Fusarium oxysporum f. sp. ciceris (Collected culture from Plant Pathology section)
Technique Used : Poison Food Technique
Media used : Potato Dextrose Agar medium (For Fungus growing)
No. of Treatments : 47
Replications : 3
Micropipette : Require for taking accurate volume of Fungicides as per recommendations.
Weighing Balance : Require for taking accurate volume of Fungicides as per recommendations
Antibiotic Zone Scale : Require for measuring accurate zone
Observation Recorded : After three Days Application
Statistical Design Used : Completely Randomized Design
Calculations : Zone of inhibition (mm)
Results:-
In vitro effect of bio-fungicides on growth and inhibition of Fusarium oxysporum f. sp. ciceris is provided in Table 26 and figure 10.
Table 26
Sr. No. Treatment No. Dose ml or gm/lit Mean colony Diameter (mm) Per cent Inhibition of growth (%)
1 Thymol 1 21 75.55
2 Camphor 1 18.57 78.38
3 Eugenol 1 19.91 76.82
4 Gingerol 1 19.58 77.20
5 Carvacrol 1 21.27 75.24
6 Piperine 1 20.58 76.04
7 Citral 1 20.09 76.61
8 Cumin aldehyde 1 22.01 74.37
9 1,8, cineole 1 24.15 71.88
10 Thymoquinone 1 22.63 73.65
11 β-Asarone 1 25.94 69.80
12 Alpha-pinene 1 20.24 76.43
13 Chavibetol 1 20.47 76.17
14 Berberine 1 21.56 74.90
15 Essential oils and total turmerones 1 18.64 78.30
16 ρ-Cymene 1 20.54 76.09
17 Mixture of Essential oils and oleoresin 1 20.24 76.43
18 Camphene 1 23.96 72.10
19 Menthol 1 22.39 73.93
20 D-limonene 1 22.33 74.00
21 Capsaicin 1 21.11 75.42
22 Methyl chavicol 1 20.97 75.59
23 Cinnamaldehyde 1 19.84 76.90
24 Embodiment :1 0.5 20.21 76.47
25 Embodiment :1 1 17.82 79.25
26 Embodiment :2 0.5 15.84 81.56
27 Embodiment :2 1 9.92 88.45
28 Embodiment :3 0.5 18.24 78.76
29 Embodiment :3 1 17.51 79.61
30 Embodiment :4 0.5 20.02 76.69
31 Embodiment :4 1 17.87 79.19
32 Embodiment :5 0.5 16.57 80.71
33 Embodiment :5 1 13.76 83.98
34 Embodiment :6 0.5 19.81 76.94
35 Embodiment :6 1 18.12 78.90
36 Embodiment :7 0.5 18.1 78.93
37 Embodiment :7 1 16.21 81.13
38 Embodiment :8 0.5 17 80.21
39 Embodiment :8 1 16.48 80.81
40 Embodiment :9 0.5 15.46 82.00
41 Embodiment :9 1 12.92 84.96
42 Thiophanate Methyl 70 % 2.0 gm 23.76 72.34
43 Metalaxyl 4%+Manconzeb 64% 3.0 gm 20.02 76.69
44 Mancozeb 63% + Carbendazim 12 % 1.5 gm 22.48 73.83
45 Fosetyl Al 80 WP (80% w/w) 2.0 gm 19.97 76.75
46 Copper Oxychloride 50 % WP 2.0 gm 23.12 73.08
47 Control 85.89

Conclusion:
All tested Embodiments demonstrated significantly superior efficacy compared to individual phytochemicals, chemical standards, and the control treatment.
In vitro bio-efficacy study revealed that Embodiment 2 @ 1.0ml/lit showed maximum zone of inhibition 9.92 (mm) followed by Embodiment 9 @ 1.0ml/lit and Embodiment 5 @ 1 ml/lit against Fusarium oxysporum f. sp. ciceris.

Although the invention has been described in detail in the foregoing for the purpose of illustration, it is to be understood that such detail is solely for that purpose and that variations can be made therein by those skilled in the art without departing from the spirit and scope of the invention except as it may be limited by the claims.
, Claims:1. A bio-fungicide and bio-stimulant composition comprising two or more phytochemicals selected from the group consisting of:
i. Camphor at a concentration in a range of 0.001-35 %, more preferably 2.5 %;
ii. Eugenol at a concentration in a range of 0.001-35 %, more preferably 13.5 %;
iii. Citral at a concentration in a range of 0.001-25 %, more preferably 5.5 %;
iv. Thymol at a concentration in a range of 0.001-45 %, more preferably 17.0 %;
v. Piperine at a concentration in a range of 0.001-25 %, more preferably 12.0 %;
vi. Cuminaldehyde at a concentration in a range of 0.001-15 %, more preferably 6.5 %;
vii. Methyl chavicol at a concentration in a range of 0.001-35 %, more preferably 8.5 %;
viii. Carvacrol at a concentration in a range of 0.001-25 %, more preferably 7.5 %;
ix. D-limonene at a concentration in a range of 0.001-45 %, more preferably 20.0 %;
x. Gingerol at a concentration in a range of 0.001-25 %, more preferably 4.5 %;
xi. β-Asarone at a concentration in a range of 0.001-25 %, more preferably 8.5 %;
xii. Menthol at a concentration in a range of 0.001-40 %, more preferably 5.0 %;
xiii. Capsaicin at a concentration in a range of 0.001-20 %, more preferably 3.5 %;
xiv. p-Cymene at a concentration in a range of 0.001-25 %, more preferably 6.0 %;
xv. Chavibetol at a concentration in a range of 0.001-25 %, more preferably 5.5 %;
xvi. Thymoquinone at a concentration in a range of 0.001-25 %, more preferably 2.5 %;
xvii. Berberine at a concentration in a range of 0.001-40 %, more preferably 4.5 %;
xviii. Alpha-pinene at a concentration in a range of 0.001-40 %, more preferably 4.0 %;
xix. 1,8 cineole at a concentration in a range of 0.001-40 %, more preferably 3.0 %;
xx. camphene at a concentration in a range of 0.001-20 %, more preferably 7.5 %;
xxi. Cinnamic aldehyde at a concentration in a range of 0.001-15 %, more preferably 5.5 %;
xxii. Mixture of Essential oils and oleoresin at a concentration in a range of 0.001-20 %, more preferably 5.0 %; and
xxiii. Essential oils and Total turmerones at a concentration in a range of 0.001-20 %, more preferably 5.0 %;
together with agriculturally acceptable excipients or additives.
2. The composition, as claimed in Claim 1, wherein said phytochemicals may be in the form of salts, solvates, hydrates, isomers or its enantiomers.
3. The composition, as claimed in Claim 1, wherein said agriculturally acceptable excipients or additives are selected from binders, diluents, surfactants, emulsifiers, carriers, lubricants, solvents, pH adjusters, colorants, essential oils, anti-caking agent, dispersing agent and the like alone or mixtures thereof.
4. The composition, as claimed in Claim 3, wherein said emulsifier is selected from group consisting of Span 80, polysorbate 80, polysorbate 60, Gaur gum, ethoxylated castor oil, Polyorganosiloxane, soy lecithin, carrageenan, mono- and diglycerides, carboxymethylcellulose, and the like in a range of 0.001 – 10 %, more preferably 4.3 %.
5. The composition, as claimed in Claim 3, wherein said surfactant is selected from the group consisting of sodium lauryl sulfate, Polysorbate 80, polyethylene glycol derivatives, and sodium lauryl sulfate, Sodium oleoyl amino fatty acid, Sodium dodecyl sulphate, Polyoxyl 35 hydrogenated castor oil, Sodium N methyl N-Oleyl taurate, Sodium alkyl naphthalene sulfonate and the like in a range of 0.1 % to 15 %, more preferably 5.0 %.
6. The composition, as claimed in Claim 3, wherein said solvents are selected from one or more water, Dimethyl sulfoxide (DMSO), Benzyl acetate, N-methyl pyrrolidinone, Diacetone alcohol, N-Ethyl-2 pyrrolidone (NEP) and the like in a range of 10.0- 55.0 %, more preferably 55.0 %.
7. The composition, as claimed in Claim 3, wherein said carriers are selected from at least one substantially water-miscible co-solvent, preferably selected from the group of maltodextrin, Cyclodextrins, China clay, N-rnethylpyrrolidinone; dimethylsulphoxide; dimethylfonnamide C9; methyl ethyl ketone, Ethylene Glycol Diacetate, dimethylisosorbide isophorone; acetophenone; cyclohexanone; Diacetone alcohol 1,3-dimethy1-2-imidazolidonone; ethylene, propylene, and butylene carbonates; lactate esters; Methyl oleate, dimethyl and diethylcarbonates; alkylglycol ethers; glycols, including propylene, carbapol 940 and biodiesel and the like in a range of 10.0- 55.0 %, more preferably 35.0 %.
8. The composition, as claimed in Claim 3, wherein said essential oils are selected from the group consisting of seed oil of Essential oil of Ferula asafoetida, Orange oil, camphor, thyme, clove, pepper, spearmint, citronella, cassia, orange oil, star anise, cedar wood, peppermint, ginger, turmeric and bay leaf and the like in a range of 0.1 to 10 %, more preferably 1.5 %.
9. The composition, as claimed in Claim 3, wherein said anti-caking agent is selected from fumed silica in a range of 0.001 to 18 %, more preferably 4.5 % and dispersing agent is selected from sodium lignosulphonate in a range of 0.001 to 25 %, more preferably 5 %.
10. The composition, as claimed in Claim 1, wherein said composition has a particle size in the range of 10-1000 nanometer.
11. The composition, as claimed in Claim 10, wherein said composition has a particle size in the range of 1 - 100 nanometer.
12. The composition, as claimed in any one of the preceding claims 1 to 11, wherein said composition comprises:
i. Thymol in the range of 0.001 to 45 %;
ii. Eugenol in the range of 0.001 to 35 %;
iii. Piperine in the range of 0.001 to 25 %;
iv. Sodium lignosulphonate in the range of 0.1 to 25 %;
v. Fumed silica in the range of 0.1 to 18 %;
vi. Maltodextrin in the range of 35 to 95 %;
vii. Sodium lauryl sulfate in the range of 0.001 to 20 %; and
viii. Citric acid in the range of 0.001 to 18 %.
13. The composition, as claimed in any one of the preceding claims 1 to 11, wherein said composition comprises:
i. Thymol in the range of 3 to 45 %;
ii. Sodium Lauryl sulphate in the range of 0.001 to 20 %;
iii. Sodium Ligno sulphonate in the range of 0.001 to 15 %;
iv. Fumed silica in the range of 0.001 to 18 %;
v. Polysorbate-80 in the range of 0.001 to 25 %;
vi. Poly glycol based defoamer in the range of 0.001 to 25 %; and
vii. China clay in the range of 25.0 to 85 %.
14. The composition, as claimed in any one of the preceding claims 1 to 11, wherein said composition comprises:
i. Piperine in the range of 0.001 to 25 %;
ii. Total turmerones in the range of 0.001 to 20 %;
iii. Gingerol in the range of 0.001 to 35 %;
iv. Thymol in the range of 0.001 to 45 %;
v. Camphor in the range of 0.001 to 35 %;
vi. Eugenol in the range of 0.001 to 35 %;
vii. Sodium lignosulphonate in the range of 0.1 to 25 %;
viii. Fumed silica in the range of 0.1 to 18 %;
ix. Maltodextrin in the range of 35 to 95 %;
x. Sodium lauryl sulfate in the range of 0.001 to 20 %; and
xi. Citric acid in the range of 0.001 to 18 %.
15. The composition, as claimed in any one of the preceding claims 1 to 11, wherein said composition comprises:
i. Thymol in the range of 0.001 to 45 %;
ii. Cumin aldehyde in the range of 0.001 to 20 %;
iii. Cinnamaldehyde in the range of 0.001 to 15 %;
iv. Sodium lignosulphonate in the range of 0.1 to 25 %;
v. Fumed silica in the range of 0.1 to 18 %;
vi. Cyclodextrins in the range of 35 to 95 %;
vii. Sodium lauryl sulfate in the range of 0.001 to 20 %; and
viii. Citric acid in the range of 0.001 to 18 %.
16. The composition, as claimed in any one of the preceding claims 1 to 11, wherein said composition comprises:
i. Thymol in the range of 0.001 to 45 %;
ii. Piperine in the range of 0.001 to 25 %;
iii. Eugenol in the range of 0.001 to 35 %;
iv. Sodium lignosulphonate in the range of 0.001 to 25 %;
v. Fumed silica in the range of 0.001 to 18 %;
vi. Lactose in the range of 35 to 95 %;
vii. Sodium lauryl sulfate in the range of 0.001 to 20 %; and
viii. Citric acid in the range of 0.001 to 18 %.
17. The composition, as claimed in any one of the preceding claims 1 to 11, wherein said composition comprises:
i. Piperine in the range of 0.001 to 25 %;
ii. Eugenol in the range of 0.001 to 35 %;
iii. Citral in the range of 0.001 to 25 %;
iv. Sodium lignosulphonate in the range of 0.001 to 25 %;
v. Fumed silica in the range of 0.001 to 18 %;
vi. China clay in the range of 35 to 95 %;
vii. Sodium lauryl sulfate in the range of 0.001 to 20 %; and
viii. Citric acid in the range of 0.001 to 18 %.
18. The composition, as claimed in any one of the preceding claims 1 to 11, wherein said composition comprises:
i. Menthol in the range of 0.001 to 40 %;
ii. Total Turmerones in the range of 0.001 to 20 %;
iii. Berberine in the range of 0.001 to 40 %;
iv. Alpha pinene in the range of 0.001 to 40 %;
v. 1,8 cineole in the range of 0.001 to 40 %;
vi. Carvacrol in the range of 0.001 to 40 %;
vii. eugenol in the range of 0.001 to 35 %;
viii. Citral in the range of 0.001 to 25 %;
ix. Sodium lignosulphonate in the range of 0.1 to 25 %;
x. Fumed silica in the range of 0.1 to 18 %;
xi. China clay in the range of 35 to 95 %;
xii. Sodium lauryl sulfate in the range of 0.001 to 20 %; and
xiii. Citric acid in the range of 0.001 to 18 %.
19. The composition, as claimed in any one of the preceding claims 1 to 11, wherein said composition comprises:
i. Thymol in the range of 0.001 to 45 %;
ii. Eugenol in the range of 0.001 to 35 %;
iii. Piperine in the range of 0.001 to 25 %;
iv. Gaur gum in the range of 0.001 to 7.5 %;
v. ethoxylated castor oil in the range of 0.001 to 10 %;
vi. Sodium oleoyl amino fatty acid in the range of 0.1 to 15 %;
vii. Dimethyl sulfoxide (DMSO) in the range of 10.0 to 55 %; and
viii. N-Ethyl-2 pyrrolidone (NEP) in the range of 10.0 to 55 %.
20. The composition, as claimed in any one of the preceding claims 1 to 11, wherein said composition comprises:
i. Piperine in the range of 0.001 to 25 %;
ii. Methyl chavicol in the range of 0.001 to 35 %;
iii. Camphor in the range of 0.001 to 35 %;
iv. Gaur gum in the range of 0.001 to 7.5 %;
v. ethoxylated castor oil in the range of 0.001 to 10 %;
vi. Sodium oleoyl amino fatty acid in the range of 0.1 to 15 %;
vii. Benzyl acetate in the range of 10.0 to 35 %; and
viii. Water in the range of 10.0 to 35 %.
21. The composition, as claimed in claim 1, wherein said composition is in the form of powder and liquid formulation.
22. A process for preparation of the bio-fungicide and bio-stimulant powder composition comprising:
i. Dry blending the powdered phytochemicals and excipients or additives to form a pre-mix;
ii. Gradually adding the surfactant and buffer into the pre-mix of step (i) and blending to form a homogenous intermediate powder blend;
iii. Subjecting the intermediate powder blend of step (ii) to micronization to obtain micronized powder having fine particle size in the range of 1–10 µm;
iv. Re-blending the micronized powder of step (iii) to obtain the final composition;
v. Passing the final composition obtained in step (iv) through a vibro-sifter equipped with a 100–120 mesh screen to ensure uniform particle distribution;
vi. Recovering the final product.
23. The process as claimed in Claim 22, wherein the dry blending of step (i) is done by using any one of the following: V-Blender (Twin-Shell Blender), Double Cone Blender, Ribbon Blender (Horizontal Blender), Paddle Blender, Octagonal Blender, Bin Blender (IBC – Intermediate Bulk Container Blender), High-Shear Mixer (for wet powder mixing/granulation), Turbula Mixer / 3D Mixer, Ploughshare Mixer, Conical Screw Mixer (Nauta Mixer), more particularly ribbon blender and High-Shear Mixer.
24. The process as claimed in Claim 22, wherein the blending of step (ii) is done by using High-Shear Mixer and Sigma Mixer to attain homogeneity.
25. A process for preparation of the bio-fungicide and bio-stimulant liquid composition comprising:
a. Dissolving the emulsifier and surfactant in the solvent to make inert mixture and agitating with homogenizer until a uniform blend is formed;
b. Adding the phytochemicals to the blend formed in step (a) and homogenizing completely to make an emulsion concentration.
c. Stirring the mixture obtained in step (b) at 300-1000 RPM particularly at 350-800 RPM at 25-55°C followed by homogenizing the mix at a speed of 3700~27000 rpm to reduce the particle size;
d. Passing the homogenized mixture obtained in step (c) through in line shear pump with 1800 RPM to 5800 RPM to obtain the nano emulsion with particle size ranging from 10 to 1000 nano meter;
e. Passing the mixture obtained in step (d) through High-pressure homogenization to obtain the particle size below 100 nano meters;
f. Passing the mixture obtained in step (e) through sparkler filter having seven layers of membrane filters with pore size less than 1-2 micron to achieve desired filtration; and
g. Recovering the final product.
26. The process as claimed in Claim 25, wherein high-pressure homogenization of step (e) is carried out at a speed of 4000 rpm to 10000 rpm and pressure up to 4,200 bar (60,000 psi).
27. The process as claimed in Claim 22 and 25, wherein the excipients in the process are selected from binders, diluents, surfactants, emulsifiers, carriers, lubricants, solvents, pH adjusters, colorants, essential oils, anti-caking agent, dispersing agent and the like alone or mixtures thereof.
28. The process as claimed in Claim 22 and 25, wherein said surfactants are selected from the group consisting of sodium lauryl sulfate, Polysorbate 80, polyethylene glycol derivatives, and sodium lauryl sulfate, Sodium oleoyl amino fatty acid, Sodium dodecyl sulphate, Polyoxyl 35 hydrogenated castor oil, Sodium N methyl N-Oleyl taurate, Sodium alkyl naphthalene sulfonate and the like alone or mixtures thereof.
29. The process as claimed in Claim 22 and 25, wherein said buffer are selected from group consisting of Citric acid, and the like.
30. The process as claimed in Claim 22 and 25, wherein said emulsifier used in the process is selected from group consisting of Span 80, polysorbate 80, polysorbate 60, Gaur gum, ethoxylated castor oil, Polyorganosiloxane, soy lecithin, carrageenan, mono- and diglycerides, carboxymethylcellulose and the like alone or mixtures thereof.
31. The process as claimed in Claim 22 and 25, wherein said solvents used in the process are selected from one or more water, Dimethyl sulfoxide (DMSO), Benzyl acetate, N-methyl pyrrolidinone, Diacetone alcohol, N-Ethyl-2 pyrrolidone (NEP) and the like alone or mixtures thereof.
32. The process as claimed in Claim 22 and 25, wherein said carriers used in the process are selected from at least one substantially water-miscible co-solvent, preferably selected from the group of maltodextrin, cyclodextrins, china clay, N-rnethylpyrrolidinone; dimethylsulphoxide; dimethylfonnamide C9; methyl ethyl ketone, Ethylene Glycol Diacetate, dimethylisosorbide isophorone; acetophenone; cyclohexanone; Diacetone alcohol 1,3-dimethy1-2-imidazolidonone; ethylene, propylene, and butylene carbonates; lactate esters; Methyl oleate, dimethyl and diethylcarbonates; alkylglycol ethers; glycols, including propylene, carbapol 940 and biodiesel and the like or mixtures thereof.
33. The process as claimed in Claim 22 and 25, wherein said essential oils used in the process are selected from the group consisting of seed oil of Essential oil of Ferula asafoetida, Orange oil, camphor, thyme, clove, pepper, spearmint, citronella, cassia, orange oil, star anise, cedar wood, peppermint, ginger, turmeric and bay leaf and the like or mixtures thereof.
34. The process, as claimed in Claim 22 and 25, wherein said anti-caking agent is selected from fumed silica and dispersing agent is selected from sodium lignosulphonate.
35. The process as claimed in Claim 22 and 25, wherein further purification of Phyto ingredient is carried out by using various instruments but not limited to liquid chromatography-mass spectrometry, gas chromatography-mass spectrometry, High pressure liquid chromatograph, gas chromatography, spectroscopy compound and the like.
36. The composition, as claimed in any one of the preceding claims, wherein said composition is provided at a concentration ranging between 0.5 to 1.0 ml /L or 0.5 to 1.0 gm /L.
37. The composition, as claimed in any one of the preceding claims, wherein said composition may be used in conjunction with the known insecticidal or acaricidal active ingredient.
38. The composition, as claimed in any one of the preceding claims, wherein said phytochemicals may be used with the active ingredient contained in other agents such as insecticides, bactericide, nematicide, plant growth regulators, synergists, fertilizers, soil improvers, animal feeds and the like.
39. The composition, as claimed in any one of the preceding claims, wherein the composition may be in the form of wettable powder, granule, powder, tablet, emulsion, water-soluble agent, suspension, granule wettable powder, flowable agent, microcapsule, aerosol, propellant, spray, fogging agent, heating transpiration agent, smoking agent, baiting agent or the like.
40. The composition, as claimed in any one of the preceding claims, wherein said composition may be applied by sprinkler application, sprayer application or drip application, more preferably by sprayer application such as foliar sprays, sprays to be applied to plants shoots and the like.
41. The composition, as claimed in any one of the preceding claims, wherein said composition is useful for controlling and preventing fungal pathogens and improving yield of crop plants.
42. The composition, as claimed in any one of the preceding claims, wherein said composition is effective against fungal pathogens such as Plasmopara viticola, Puccinia graminis, Leveillula taurica, Pythium aphanidermatum, Pyricularia oryzae, Colletotrichum capsici, Phytophthora infestans, Rhizoctonia bataticola, Fusarium oxysporum f. sp. ciceris, Fusarium oxysporum, Fusarium graminearum, Fusarium solani, Fusarium verticillioides, Magnaporthe oryzae, Magnaporthe grisea, Puccinia graminis f. sp. tritici, Puccinia striiformis f. sp. tritici, Puccinia triticina, Uromyces appendiculatus, Botrytis cinerea,Colletotrichum gloeosporioides, Colletotrichum lindemuthianum, Alternaria solani, Alternaria alternata, Alternaria brassicae, Sclerotinia sclerotiorum, Sclerotium rolfsii, Rhizoctonia solani, Rhizoctonia bataticola, Macrophomina phaseolin, Phytophthora infestan, Phytophthora capsica, Phytophthora palmivora, Peronospora destructor, Hyaloperonospora parasitica, Pseudoperonospora cubensis, Albugo candida, Erysiphe graminis f. sp. tritici, Oidium spp., Cercospora arachidicola, Cercospora zeae-maydis, Mycosphaerella fijiensis, Venturia inaequalis, Guignardia citricarpa, Neonectria ditissima, Cryphonectria parasitica, Monilinia fructicola, and the like.
43. A method for controlling or preventing fungal infections in plants comprising applying said composition to the plant or to the affected parts of the plants in suitable amount thereof.