DESC:Field of the invention
The invention relates to agricultural biotechnology and microbiology.
The present invention relates to synergistic bioformulation of Trichoderma to stimulate plant growth and Induced Systemic Resistance (ISR) responses to a wide spectrum of pathogens. The invention more particularly relates to a bioformulation comprising Trichoderma viride and Trichoderma harzianum. The invention also relates to a process of developing a synergistic bioformulation of Trichoderma for sustainable nutrition and disease management in plants.

Background of the invention
Currently there is a great demand for environment friendly biocontrol agents and plant nutrition compositions in the agricultural sector. Plant diseases caused by soil-borne and foliar pathogens is destructive for agriculture and spreads quickly causing huge loss of important crops. Several chemical pesticides have recently been banned globally due to their widespread use, which exacerbates soil degradation, kills non-target beneficial organisms, and causes yield stagnation, threatening food security and being harmful to the environment and human health. Chemical pesticides affect the ecosystem irreversible which is always undesirable. One alternative attempted more widely is to use biological control agents that exist in nature and control the plant pathogens. Many organisms that can be used in this way have been described in the state of the art. However, the use of said biocontrol agents has been limited, primarily because in the majority of the cases the level of control has not been comparable to that achieved with a very efficient chemical fungicide.

Trichoderma species are well-known biocontrol agents that have been widely used for their ability to suppress plant diseases and enhance plant growth. There are documents related to Trichoderma in a consortium. WO2019067379A1 provides method of producing Trichoderma fungi on an industrial scale. In specific embodiments, the subject invention provides methods of producing both a liquid Trichoderma-based product and a solid-state Trichoderma-based product from the same starting seed culture and inoculant.

WO2011099027A1; “Composition and method of preparation of bio fungicide based on Trichoderma viride for controlling soil borne diseases discloses composition and method of preparing a bio Fungicide containing Trichoderma viride with enzymes, fats and growth promoting molecules to control soil borne diseases like root rot, set rot, wilt, collar rot, red rot, rhizome rot, damping off, stem rot, and club rot.

The above-mentioned prior art references demonstrate the ongoing research and development in the field of Trichoderma for plant disease control and plant growth promotion.

Despite their potential benefits, the efficacy of Trichoderma in a consortium can vary depending on the composition, formulation, and application methods. Therefore, there is a need for improved formulations of Trichoderma in a consortium that exhibit enhanced efficacy, stability, and performance as an effective and sustainable alternative to chemical pesticides in crop protection.

In the existing system as given in Canadian Patent Application No. 1 106201- Granted Patent wherein the invention concerns a mycofungicidal product consisting of immunizing commensals (:IC) and cereal flour where the immunizing commensal is Scytalidium lignicola ATCC 16,675, Trichoderma polysporum rifai ATCC 20,475, Trichoderma viride sensu Bisby ATCC 20,476 or Trichoderma viride sensu Bisby strain CG BINAB.INRA T 030(Pasteur Institute, Paris). It is aimed also at uses of this product for the treatment of trees, wood and plants.

In the existing system as given in WIPO Patent Application No. PCT/US1988/002011- Granted Patent wherein the invention relates to A rhizosphere-competent $i(Trichoderma polysporum), (ATCC 20852) and a rhizosphere-competent $i(Trichoderma viride) (ATCC 20853) are described. They are respectively, mutants of rhyzosphere-incompetent $i(Trichoderma polysporum) ATCC i20475 and rhizosphere- incompetent $i(Trichoderma viride) ATCC 20476. The rhizosphere-competent strains are particularly effective biocontrol agents for plant and/or treeidiseases associated with various fungi e.g., Pythium spp., $i(Sclerotium), spp., $i(Rhizoctonia solani) and basidiomycetes such as $i(Chondrostereum purpureum).

Soil borne disease have until recently, received less attention than plant diseases affecting the shoot and foliage. However, this is not a reflection of their economic importance, but rather of the difficulties in investigating and detecting pathogens below soil level. Many soil borne diseases are stress related and it is in the tropics where crop growth is particularly limited by environmental stress, predisposing crops to infection by soil borne pathogen.

The present invention aims to provide a novel formulation of Trichoderma in a consortium of Trichoderma harzianum and Trichoderma viride for control of wide range of insect pest in wide range of crops, which offers distinct advantages over the existing prior art references. The present invention is an innovative combination of fungal based consortium containing Trichoderma viride and Trichoderma harzianum with enzymes, fats and growth promoting molecules to control soil borne diseases like root rot, set rot, wilt, collar rot, red rot, rizome rot, damping off, stem rot, club rot and alike.

Object of the invention
The main object of present invention is to provide a synergistic bioformulation of Trichoderma to stimulate plant growth and Induced Systemic Resistance (ISR) responses to a wide spectrum of pathogens.
It is a further object of the present invention to provide a bioformulation comprising consortia of Trichoderma viride and Trichoderma harzianum to effectively control the wide range of plant diseases and promoting plant growth.

It is a further object of the present invention to provide bioformulation comprising consortia of Trichoderma viride and Trichoderma harzianum along with adjuvants for better efficacy of the bioformulation against pest.

It is a further object of the present invention to provide a liquid, wettable powder & granule formulation comprising consortia of Trichoderma viride and Trichoderma harzianum along with adjuvants showing high viability and extended shelf life of microbes.

It is a further object of the present invention to provide a bioformulation providing high colony forming units of fungal microbial cultures.

It is a further object of the present invention to provide a process of developing a synergistic bioformulation of Trichoderma for sustainable nutrition and disease management in plants.

It is a further object of the present invention to provide a method of application of bioformulation of Trichoderma to be preventive or curative to control wide range of pest.

Summary of the invention
Accordingly, in an aspect of the present disclosure, the invention provides a synergistic bioformulation of Trichoderma to stimulate plant growth and Induced Systemic Resistance (ISR) responses to a wide spectrum of pathogens. The bioformulation comprises consortia of Trichoderma viride and Trichoderma harzianum to effectively control the wide range of plant diseases and promoting plant growth.

In an aspect of the present disclosure, the bioformulation comprises consortia of Trichoderma viride and Trichoderma harzianum along with adjuvants for better efficacy of the bioformulation against pest control. The bioformulation is a liquid, wettable powder & granule formulation comprising consortia of Trichoderma viride and Trichoderma harzianum along with adjuvants showing high viability and extended shelf life. The bioformulation demonstrates high colony forming units of fungal microbial cultures.

The bioformulation of the present invention is capable of controlling wide range of soil borne and foliar diseases. The fungal culture used in the bioformulation are provided as suspension concentrate formulation with high spore density using optimized media. The formulation comprises live viable cells which includes spores and mycelia, both of which contributes to the activity of formulation.

The process of developing the bioformulation of consortia of Trichoderma viride and Trichoderma harzianum involves standardization of carbon, nitrogen, macro elements, microelements and vitamins etc. to achieve desirable formulation.

The process provides suspension concentrate and wettable formulation of consortium of Trichoderma viride and Trichoderma harzianum, stable for at least 18 months and up to 36 months. The formulation is stable at low temperature, high temperature and room temperature for at least 18 months.

The bioformulation of the present invention shows synergistic effect of the combination of microbes in a consortium, thereby providing superior efficacy in controlling plant diseases and promoting plant growth. The bioformulation has established good efficacy in terms of pot and field trials on different crops.
Present invention involves the process of development of bioformulation of consortium of Trichoderma viride and Trichoderma harzianum and application of bioformulation on wide range of crops at different stages of development to cover wide range of pests in an economical, cost-effective and user friendly way.

The bioformulation is capable to control wide range of soil borne diseases like white mold, cottony rot, watery soft rot, stem rot, drop, crown rot (Sclerotinia sclerotiorum), fusarium wilt (Fusarium oxysporum) sheath blight of rice (Rhizoctonia solani) and foliar diseases such as Black sigatoka which is a leaf-spot disease of banana plants (Mycosphaerella fijiensis), leaf spot pomegranate (Cercospora punicae), leaf spot (Alternaria brassicicola) etc.

Reference throughout this specification to “an aspect”, “another aspect” or similar language means that a particular feature, structure, or characteristic described in connection with the embodiment is included in at least one embodiment of the present invention. Thus, appearances of the phrase “in an embodiment”, “in another embodiment” and similar language throughout this specification may, but do not necessarily, all refer to the same embodiment.

The terms "comprises", "comprising", or any other variations thereof, are intended to cover a nonexclusive inclusion, such that a process or method that comprises a list of steps does not include only those steps but may include other steps not expressly listed or inherent to such process or method.

Unless otherwise defined, all terms, and especially any technical and/or scientific terms, used herein may be taken to have the same meaning as commonly understood by one having an ordinary skill in the art.

The terminology and structure employed herein is for describing, teaching and illuminating some embodiments and their specific features and elements and does not limit, restrict or reduce the spirit and scope of the claims or their equivalents.

It will be understood by those skilled in the art that the summary of the invention provided herein is exemplary and explanatory of the invention and are not intended to be restrictive thereof. Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by those skilled in the art to which this invention belongs. The composition, methods, and examples provided herein are only illustrative and not intended to be limiting.

Statement of the invention
The inventor provides a synergistic bioformulation Trichoderma viride and Trichoderma harzianum and process of preparing the same for plant growth and controlling various soil borne diseases like root rot, set rot, wilt, collar rot, red rot, rhizome rot, damping off, stem rot and club rot.

Detail description of the invention
It should be noted that the particular description and embodiments set forth in specification below are merely exemplary of the wide variety and arrangement of instructions which can be employed with the present invention. The present invention may be embodied in other specific form without departing from the spirit or essential characteristics thereof. All the feature disclosed in the complete specification may be replaced by similar other or alternative features performing similar or equivalent purposes. Thus, unless expressly stated otherwise, they all are within the scope of present invention.

Various modifications or substitutions are also possible without departing from the scope and spirit of present invention. Therefore it is to be understood that this specification has been described by way of the most preferred embodiment and purpose of illustration and not limitation.

Present disclosure provides a synergistic fungal based consortium comprising strains of Trichoderma for controlling broad spectrum of disease in various crops.

The present invention is an innovative combination of Trichoderma viride and Trichoderma harzianum and growth promoting molecules for plant growth and to control soil borne diseases like Fusarium wilt, Alternaria blight, Damping off, Sheath blight, blast black spot, sclerotinia, foot & seedling rot and mold.

The Trichoderma harzianum is accessed from vegetable field soil of Palwal, Faridabad, Haryana and deposited at IMTECH having MTCC 25013 dated 24.03.2015. The Trichoderma viridae is accessed from Baraut, Baghpat, Uttar Pradesh, India and deposited at IMTECH having MTCC 25012 dated 10.04.2015

The present invention also deals with the large scale production of this unique synergistic formulation by submerged fermentation in suspension concentrate, wettable powder & granular form for disease control in agriculture use.

Nucleus culture (Maintain culture)
Pure culture of fungus named Trichoderma viride and Trichoderma harzianum are inoculated aseptically on separate plates having 20 ml PAC-00 agar Media; such two to three plates are generally inoculated. Such inoculated plate is maintained in BOD incubator for at 26 ± 1 °C for 3 to 4 days. (Composition for one liter of PAC-00 Media contains 300 gm potato, 20 gm dextrose, 20 gm agar agar and 1000 ml distilled water).

Starter culture (In flask)
Cultures grown on plates are inoculated aseptically in a flask (1000 ml capacity) having 500 ml optimized broth media via cork borer (having 5 mm diameter cutting capacity), and allow to grow for at 26 ± 1 °C for at least 2 to 3 days. For superior growth, flask is put on shaker with agitation of media at 150-200 RPM. Such two, flasks are maintained.

Seed Fermenter (In small fermenter)
One (1) lit culture grown in step-2 is inoculated aseptically in small fermenter having 10 litre optimized growth media and allow it for further growth at 26 ± 1 °C for at least 2 to 3 days with agitation by motor at 100 -150 RPM.

Optimum process parameters for optimum growth in minimum time is the important criteria for the production of maximum cell biomass of consortium of Trichoderma viride with Trichoderma harzianum strains by submerged culture. The inoculated medium is maintained under substantially constant agitation and aeration at a temperature from about 28° C. to 35° C. The pH of the medium is not monitored during the incubation period. The initial pH was kept at 6.0-7.5. The culture is grown until the density of the cell reached from the inoculant is about 2×1010 CFU per ml. The optimum growth is attained at incubation of period of 24-96 hours. Aeration optimized for shake flask condition is 150-200 RPM.

The medium comprises a carbon source, a nitrogen source and other micro and macro nutrient sources like magnesium sulphate, manganese sulphate, Dipotassium hydrogen phosphate, Potassium dihydrogen phosphate, ferric chloride, zinc Sulphate, cobalt chloride, and Boric Acid.

The carbon source is preferably sources like Jaggery, fructooligosaccharide, molasses, and glucose, glycerol, sucrose, maltose, fructose or mixtures thereof. The nitrogen source is preferably yeast extract, beef extract, peptone, soya meal, oat meal and bengal gram.

The essential micro and macro nutrients for the optimum production of Trichoderma viride with Trichoderma harzianum are supplied from sources like magnesium sulphate, Calcium chloride, Dipotassium hydrogen phosphate, Potassium di-hydrogen phosphate, manganese sulphate, ferric chloride, zinc sulphate, cobalt chloride.

The essential vitamins are supplied like riboflavin, thiamine and pantothenic acid for the optimum production of Trichoderma viride with Trichoderma harzianum.

Large scale fermenter
The present invention also describes a process of large scale production of synergistic consortium of Trichoderma viride and Trichoderma harzianum in submerged culture using fermenter from volume ranging from 50 litres to 6000 litres. The pre-inoculum of the Trichoderma viride and Trichoderma harzianum are prepared in a suitable liquid medium under continuous aeration using BOD orbital shaker. For inoculation of fermenter media for large scale production of consortium of Trichoderma viride and Trichoderma harzianum as liquid inoculum size of 0.5- 2.5 % can be used. The fermenter medium contains water as solvent or base in which previously optimized media components are added sequentially and mixed uniformly.

Industrial or commercial fermenter are sterilized differently using additional utilities under pressure and steam. The temperature range of 120-125oC at 15 lbs. is maintained for 15-30 minutes for sterilization of entire vessel and other connected pipelines and filters along with media. This process of sterilization is known as full vessel sterilization in place (FSIP). Proper sterilization of the vessel along with the media will ensure zero microbial presence in the fermenter media which shall be ready for inoculation with inoculum of desired percentage for running a commercial batch.

Fermentation media and raw material:
The medium comprises a carbon source, a nitrogen source and other micro and macro nutrient sources like magnesium sulphate, manganese sulphate, Dipotassium hydrogen phosphate, Potassium dihydrogen phosphate, ferric chloride, copper sulphate & calcium chloride.
The carbon sources used are Jaggery, molasses, fructooligosaccharides, glucose, glycerol, sucrose, maltose & fructose or mixtures thereof. The nitrogen source is preferably yeast extract, meat extract, oat meal, peptone, soya flour and bengal gram.
The essential micro and macro nutrients for the optimum production of Trichoderma viride with Trichoderma harzianum are supplied from sources like magnesium sulphate, Dipotassium hydrogen phosphate, Potassium di-hydrogen phosphate, manganese sulphate, ferric chloride, zinc sulphate, cobalt chloride.
The essential vitamins are supplied like riboflavin, thiamine and pantothenic acid for the optimum production of Trichoderma viride with Trichoderma harzianum.
The pre-inoculum is added to the sterilized media in the fermenter.

Submerged fermentation is done in fermenter with optimized parameters as follows:
Process Time: 24-96 h
Temperature: 28 to 35° C
Agitation: 25-80 RPM (Rotation per minute)
Aeration: 0.2-1.0 VVM (Vessel Volume per minute)
After attaining optimum growth, the fermented broth is mixed with preservatives before harvesting.

The preservatives comprise Starch, Sodium chloride and Polyvinylpyrrolidone (PVP) at different concentration of:
Sodium chloride: 5- 10 %
Starch: 1.0: 10.0 %
PVP: 1.0-3.0 %
The pre-determined process parameters for submerged fermentation comprises Process Time: 24 h to 96 h, Temperature: 28 °C to 35 °C, Agitation: 25 RPM to 80 RPM (Rotation per minute); Aeration: 0.2 VVM to-1.0 VVM (Vessel Volume per minute). The optimum viable cell count of the microbial consortium in the liquid formulation is in the range of 2×109 CFU/ml to 2 × 1010 CFU/ml.

The Fermented broth is harvested and bottled in different capacity of 100 ml to 5 litres HDPE container.
The formulation can be formulated into powder as well as liquid formulation using following components and mass percentage thereof:
Liquid formulation
Harvested broth 90-95%
Adjuvant 0.02-0.5 %
Sticker 1.0-3.0 %

The adhesive used in the liquid formulation is Polyvinylpyrrolidone.

The adjuvant used in the formulation is polysorbate 80.

The sticker used in the liquid formulation is vegetable oil.

Powder formulation
Harvested broth 15-30%
Adjuvant 0.05-1.5 %
Carrier 60-80%

The adjuvant used in the powder formulation is Poly ethylene glycol.

The carrier used in the powder formulation is talc.
The optimum viable cell count of the microbial consortium in the powder formulation is in the range of 2×108 CFU/ml to 2 × 109 CFU/gm.
In a non-limiting example, the bioformulation of the present disclosure is effective in controlling 36 fungal and 09 bacterial plant pathogens (Table 1).

Table 1: Bioefficacy study data
S. No. Pathogens screened Growth of Pathogen in control (in cm) Radial growth of Pathogen in present consortium (in cm) Reduction in growth (%)
Soil borne pathogens
1 Fusarium oxysporum f.sp. capsica 3.00 1.47 51.00
2 Fusarium oxysporum f.sp. vasinfectum 3.00 1.70 65.33
3 Fusarium solani 2.45 1.45 60.81
4 Fusarium oxysporum fsp. Cumini 2.30 1.30 73.47
5 Fusarium moniliformae 2.80 1.45 48.21
6 Fusarium oxysporum f.sp. zingiberi 2.57 1.40 45.52
7 Rhizoctonia solani 2.35 1.40 40.42
8 Fusarium oxysporum 2.10 1.55 66.19
Foliar pathogens
9 Alternaria alternata 2.47 1.15 53.44
10 Curvularia sp. 2.45 1.25 48.97
11 Venturia inequalis 2.50 1.75 30.00
12 Phomopsis vexans 1.82 0.97 46.70
13 Magnaporthe grisae 2.25 1.37 39.11
14 Coniella granati 3.00 1.70 43.33
15 Alternaria brassicae 2.37 1.22 48.52
16 Alternaria brassicicola 2.32 1.37 40.94
17 Cercospora brassicicola 2.17 1.30 40.09
18 Dreschlera maydis 2.35 1.30 44.68
19 Mycospharella fijiensis 1.92 1.17 40.00
20 Alternaria porri 2.30 1.37 40.43
21 Colletotrichum gloeosporoides 2.50 1.30 48.00
22 Alternaria jacinthicola 2.1 1.1 47.61
23 Colletotrichum falcatum 1.92 1.02 46.87
24 Alternaria macrospora 1.80 1.02 43.33
25 Alternaria solani 1.82 0.82 54.94
26 Colletotrichum coccodes 2.07 1.17 43.47
27 Alternaria tenuissima 2.20 1.12 49.09
28 Gaeumannomyces graminis tritici 2.40 1.15 52.08
29 Bipolaris sorokiniana 1.67 1.12 32.93
30 Pythium aphanidermatum 3.00 1.60 46.66
31 Curvularia clavata 1.55 1.07 30.96
32 Michrodochium nivale 1.25 0.50 60.00
33 Alternaria padwickii 1.92 1.10 42.70
34 Bipolaris oryzae 2.42 0.97 59.91
Postharvest pathogens
35 Botrytis cineria 3.00 1.30 56.66
36 Aspergillus niger 3.00 1.77 41.00
NC= No control; *C= Control; *NR= No result
Conclusion: Sanjeevni Plus formulation was found effective in controlling 36 fungal 09 bacterial plant pathogens.
In another example, the shelf life study was conducted on the formulation for 24 months and 36 months with an intermediate period of 3 months. The shelf life data are presented in the Table 2 and Table 3.
The shelf life studies shows that the shelf life of the liquid formulation is more than 36 months and for powder formulation is 24 months. The shelf life of the liquid was tested at every 3 months interval for 36 months. While the shelf life of the powder formulation was tested at every 3 months interval for 24 months. The shelf life data for liquid and powder formulation are presented in table 2 and 3 below, respectively.

Table 2: Shelf life data of liquid formulation
S. No Months of study Optimized Media PDB (Broth)
1 3 Month 2 x 1010 2 x 108
2 6 Month 2 x 1010 2 x 105
3 9 Month 2 x 1010 2 x 105
4 12 Month 2 x 1010 2 x 104
5 15 Month 9 x 109 2 x 103
6 18 Month 6 x 109 2 x 103
7 21 Month 6 x 109 2 x 103
8 24 Month 5 x 109 Nil
9 27 Month 4 x 109 Nil
10 30 Month 2 x 109 Nil
11 33 Month 2 x 109 Nil
12 36 Month 2 x 109 Nil

Table 3: Shelf life data of powder formulation
S.no Months of study Optimized Media +Talc PDB (Broth)+ Talc
1 3 Month 2 x 109 1 x 108
2 6 Month 2 x 109 2 x 106
3 9 Month 2 x 109 3 x 105
4 12 Month 9 x 108 1 x 104
5 15 Month 7 x 108 1 x 103
6 18 Month 5 x 108 Nil
7 21 Month 2 x 108 Nil
8 24 Month 2 x 108 Nil
Uniqueness in the formulation and process of manufacturing:
1. The synergistic consortium of Trichoderma viride and Trichoderma harzianum contains viable cell count of minimum of 2×109 CFU/ml and maximum of 2 × 1010 CFU/ml.
2. The production and mass multiplication to attain the high CFU count is only 24-96 hrs of batch run.
3. The shelf life of liquid bioformulation is 18-36 months and for powder and granular formulation is 18 months to 24 months
4. The synergistic bioformulation of Trichoderma is containing adjuvant (Sticker, Wetting agent) for better efficacy and rain fastness. The adjuvant is an organic adjuvant in an amount of 1-5%.
5. The synergistic bioformulation contains UV Protectant for better stability of spore after foliar application.
UV protectant 1-5%
6. The bioformulation is highly effective in controlling diseases like Fusarium wilt, Alternaria blight, Damping off, Sheath blight, blast black spot, sclerotinia, foot & seedling rot and mold.
7. The bioformulationn can be applied by spraying.
8. Suspension concentrate formulation with high spore density using optimized media comprises live viable cells which includes spores and mycelia, both of which contributes to the activity of formulation.
9. The bioformulation can be applied to different types of crops like fruits, paddy, cotton, tomato, vegetables, tea, coffee, cardamom, aromatic and medicinal crops.

So accordingly, the present invention provides an improved, highly stable, nontoxic and environment friendly bio-formulation for the effective utilization of biocontrol agents for sustainable agriculture in pilot scale and commercial scale, said bio-formulation effective for both agricultural and non-agricultural crops, said bio-formulation comprising Trichoderma viride with Trichoderma harzianum as consortium, along with an optimized media for the growth of the Trichoderma viride with Trichoderma harzianum under submerged fermentation and optimum preservatives added in the formulations to obtain a shelf life for more than two years, such that all the ingredients are working together in synergistic manner to obtain the desired efficiency.
,CLAIMS:I/We Claim:
1. A bioformulation for the effective control of insects and pests, said bio-formulation comprises Trichoderma viride and Trichoderma harzianum as consortium having viable cell count in the range from 2 × 109 CFU/ml to 2 × 1010 CFU/ml in an optimized media, preservatives, and adjuvants, such that all the ingredients are working together in synergistic manner for sustainable nutrition, growth and disease management in plants.

2. The bioformulation as claimed in claim 1, wherein the preservative comprises Sodium chloride, Starch and Polyvinylpyrrolidone at different concentration in the range of: Sodium chloride 5-10 %; Starch- 1.0 10.0 %; PVP-1.0-3.0%.

3. The bioformulation as claimed in claim 1, wherein the optimized media comprises a carbon source, a nitrogen source and other micro and macro nutrient sources, wherein said carbon source is selected from Jaggery, molasses, and glucose, glycerol, sucrose, maltose, fructose, or any combination thereof; said nitrogen source is selected from yeast extract, beef extract, peptone, soya meal and Bengal gram, or any combination thereof; said essential micro and macro nutrients are selected from magnesium sulphate, Dipotassium hydrogen phosphate, Potassium di-hydrogen phosphate, manganese sulphate, or any combination thereof.

4. The bioformulation as claimed in claim 1, the bioformulation is in liquid form, granular form, suspension concentrate form or powder form.

5. The bioformulation as claimed in claim 1, wherein the adjuvants are selected from plant growth promoter molecules and UV protectants.

6. The bioformulation as claimed in claim 1, wherein the adjuvants are stickers, spreaders or surfactants.
7. The bioformulation as claimed in claim 1, wherein shelf life of the bioformulation in liquid form is up to 36 months and in powder form, suspension concentrate form and in granular form is up to 24 months at room temperature.

8. A process for preparing the bioformulation as claimed in claim 1, wherein said process comprises:
- Preparing an optimized liquid media comprising carbon source, nitrogen source and other micro and macro nutrient sources;
- preparing a pre-inoculum of Trichoderma viride and Trichoderma harzianum in the optimized liquid media under specific process parameters using BOD orbital shaker, said specific process parameters are incubation period of 24-96 hours and aeration at 150-200 RPM to obtain inoculum of consortium,
- using the inoculum of size of 0.5 to 5 % for inoculation of fermenter media for submerged fermentation under specific process parameters to obtain fermented broth with optimum viable call count of consortium,
- adding preservatives to the fermented broth followed by harvesting to obtain bioformulation, and bottling in different capacity of HDPE container.

9. The process as claimed in claim 8, wherein said optimized process parameters are: Process Time- 24-96 hrs.; Temperature 28 to 35° C; Agitation 25-80; RPM (Rotation per minute) and Aeration 0.2-1.0 VVM (Vessel Volume per minute).

10. The process as claimed in claim 8, wherein said preservatives comprises Sodium chloride, Starch and PVP at different concentration in the range of: Sodium chloride 5-10 %; Starch- 1.0 10.0 %; PVP-1.0-3.0%.
11. The process as claimed in claim 8, wherein the optimum viable count in the bioformulation is in the range of 2 × 109 CFU/ml to 2 × 1010 CFU/ml.

12. The process as claimed in claim 8, wherein the shelf life of bioformulation is in the rage of 24 months to 36 months at room temperature.

13. The process as claimed in claim 8, wherein the bioformulation in liquid form comprises Harvested broth in the range of 90-95%, Polyvinylpyrrolidone in the range of 0.02%-0.5 %, vegetable oil in the range of in the range of 1.0%-3.0%, polysorbate 80 in the range of 1.0%-3.0%; and in powder form comprises Harvested broth in the range of 15%-30%, Poly ethylene glycol in the range of 0.05%-1.5 % and Carrier in the range of 60-80% and talc.

14. The process as claimed in claim 8, wherein the bioformulation further comprises adjuvants selected from stickers, spreaders or surfactants for better efficacy of the formulation against broad spectrum of disease management of crops.