TRICHODERMA HARZIANUM BIOFORMULATION FOR CONTROLLING ROOT AND ROOT ROT DISEASE AND STRENGTHENING PLANT GROWTH IN PUMPKIN.

Abstract

The invention relates to a granular bioformulation containing the isolate Trichoderma harzianum (NCBI accession number: MH351669) TRIC8, supplemented with wheat straw and sugar beet molasses, to prevent root rot disease caused by seed-borne Fusarium proliferatum in pumpkin seeds used in the production of snacks, confectionery and sweet foods, and to promote plant growth.

Description

1 TARIFF ROOT AND ROOT ROT DISEASE IN PUMPKIN (SNACK PUMPKIN) TRICHODERMA FOR CONTROL AND ENHANCE PLANT GROWTH. HARZIANUM BIOFORMULATION Technical Field to Which the Invention Relates The invention relates to pumpkin seeds used in the production of snacks, confectionery, and sweet foods. Root and collar rot caused by seed-borne Fusarium proliferatum To prevent disease and promote plant growth, Trichoderma harzianum (NCBI accession number: MH351669) Wheat straw and sugar containing TRIC8 isolate 10 It relates to a granular bioformulation with added beet molasses. State of the Art Soil-borne fungal diseases are a problem in crop cultivation, and This leads to significant crop losses. Soil-borne fungal agents vary by 15 By constantly adapting to soil conditions, they maintain their viability in the soil for many years. They are able to sustain themselves. They cause significant economic losses in plants. Crop rotation, the use of resistant varieties, and certain other methods can be used to control these types of pathogens. Fungicides, soil sterilants, and solarization are recommended, but not with sufficient success. This is not possible. There is a residue problem with soil fumigants on products. 20 By polluting the soil, water, and air, it negatively affects humans and other living beings. It is known to have an effect in that direction. Common fungal diseases in pumpkin cultivation include wilting, root rot, and stem rot. throat rot, pseudomildew, gray mold, powdery mildew, anthracnose and some leaf 25 and fruit spot diseases [1]. The severity of these diseases depends on the pathogen Virulence depends on the physical and chemical properties of the growth environment, soil moisture, temperature, and pH depends on nutritional factors and host sensitivity, as well as the soil environment. suitable for the formation and development of the fungus, but unsuitable for the host. The onset of disease is possible. Symptoms of root rot seen in the above-ground parts, 30 It is a result of a weakened root system and is usually caused by nutrient deficiencies or poor root systems. This manifests as developmental delays, chlorosis, wilting, or plant death. 2 In pumpkin cultivation, planting is crucial for preventing soil-borne fungal diseases. The root rot control is of great importance in cultivation areas. Multiple factors contribute to the disease. Growing conditions in Türkiye... Considering this, common soil-borne diseases that could negatively affect production Fungal species include Rhizoctonia spp., Sclerotinia spp., and Fusarium spp. (ranking 5th). However, within the scope of the irrigation project, there is a transition from dry farming to irrigated farming. With the increase in humidity in these regions, zoospores of Pythium and Phytophthora are carried in the water. Soil-borne fungi such as damping-off are also becoming widespread over time. soil-borne fungal diseases characterized by wilting, root rot, and root collar rot. Pathogens cause significant yield losses in the product. These pathogens are 10 They can remain alive in the soil for a long time in a resistant spore form. The disease... The most common symptoms in plants are the rotting of seeds in the soil before germination, and developing... root collar rot of seedlings, root collar rot of seedlings emerging from the soil surface It is caused by the thinning of the soil, resulting in it falling over and lying on the ground. Also, the soil... Leaf deformities may be observed in seedlings emerging from the surface. Diseased seedlings 15 softening and browning of the roots, especially in the bark tissue. This is observed in diseased fields, with clusters of collapsed plants, partial drying, and sparse growth. There are outbreaks. In severe infections, replanting in the field is necessary. This situation leads to loss of labor and production [2]. Seeds in snack pumpkin. When the fungal pathogen F. proliferatum of genetic origin is inoculated into the seeds of susceptible varieties, 20 51.07% of the roots and root collar were affected when inoculated into the soil, a reduction of 43.75%. It causes decay. It is also inoculated from both soil and seed. When this is done, the number of leaves, shoot length, shoot age and This results in significant reductions in dry weight. Fusarium species are adapted to diverse climatic conditions around the world, except for the Antarctic continent. These are fungi that can be pathogenic [3]. Pathogenic Fusarium species are found on more than 100 host plants. It exhibits symptoms such as wilting or root rot and is resistant to spores. Their structures allow them to maintain their viability in the soil for a long time. Soil and seeds. These fungi, originating from seeds, can survive for 2 years, and in chlamydospore form, they can remain in the soil for many years (30). It can maintain its viability. Chemical control of soil-borne Fusarium species. It is quite difficult, and crop rotation significantly prevents the disease. When crop rotation is not practiced in the production area, soil-borne disease agents increase in population, activation of nematodes, organic matter in the soil 3 a decrease in quantity, disruption of the balance of nutrient elements in the soil, and There are numerous downsides, such as the increasing problem of toxic residues from pesticides. This can be the subject of discussion. Its broad spectrum of impact and environmentally protective properties. integrated protection against soil-borne pathogens, nematodes, and weeds. Solarization, a physical method included in pest control programs, is also among the 5 It is used to combat soil-borne Fusarium species. Solarization, It is the process of disinfecting soil using the sun's thermal energy. Toxic material. It is an environmentally friendly method that does not involve, is economical and easy to implement [4]. Solarization The process usually takes several weeks. This affects the agricultural production calendar. and leads to delays. In addition, the application is generally effective on the surface 10 Its effectiveness against pathogens deep in the soil is limited. In addition, sodium is used before planting to control soil-borne diseases. Seed disinfection with hypochlorite (1%) is important [2]. In addition, in nurseries hymexazole, captan, captan+pencycuron, hydroxyquinoline sulphate, azoxystrobin + metalaxyl-m + fludioxonil, fludioxonil + metalaxyl-m and propamocarb + fosetyl active 15 fungicides containing ethandinitrile, metam potassium and metam sodium as active ingredients. Fumigants are used in the form of soil application. Chemical control involves water and... The misuse of pesticides leads to soil pollution and disrupts ecosystem balance. It is damaging. To protect the biological balance disrupted by excessive chemical use, the environment... Research into alternative pest control methods that protect health is increasing day by day. It is gaining momentum day by day. Root bacteria, especially Pseudomonas fluorescens their ability to inhibit soil-borne fungal pathogens, competition against the pathogen, Induced Systemic Resistance (ISR) 25 stimulating the mechanism of producing siderophores, mycolytic enzymes, and antibiotics. Many researchers have reported that they have abilities [5-6]. Worldwide In numerous countries, Bacillus and Pseudomonas are used in the biological control of plant diseases. Commercial preparations of Streptomyces species are practically used for the control of some diseases. available for use [7]. Among these, the most commonly used are Bacillus 30 spp., Pseudomonas spp. Streptomyces spp. such as Agrobacterium radiobacter bacteria including Trichoderma spp., Candida oleophila, Gliocladium spp. and They are fungi such as Coniothyrium minitans and Ampelomyces quisqualis [8]. In our country Trichoderma harzianum Rifai is a licensed biological fungicide for root diseases. 4 KRL-AG2 (T-22 Planter Box), Trichoderma harzianum Rifai KRL-AG1 (Rootshield Granules), Trichoderma harzianum Rifai Strain T22 (Trianum P), Trichoderma aspellerum race ICC 012+Trichoderma gamsii race ICC 080 (Remeider), Trichoderma viride (Bio-Cure F), Bacillus subtilis IAB / BS03 (Fungisei), Gliocladium virens strain GL21 (Soilgard 12 G), Bacillus amyloliquefaciens (B. subtilis) strain QST 713 5 (Serenade) is used. Of these, only Trianum P. is used on cotton seeds. some are applied, while others are applied to the soil. GreenMax Agro Tech, a company involved in the known state of the art, uses Trichoderma harzianum Bio Fertilizer Liquid with talc additive, plant growth enhancing 10 It is produced as follows. The content of this bioformulation is Trichoderma harzianum 1%. (w / w), adhesive - carboxymethyl cellulose (CMC, 1%) inactive ingredient (98%, w / w) and It consists of talc (63%). Bioformulation seed application (40 g / kg seed) (mixed in a ratio of 10 and dried in the shade for 24 hours), seedling dipping (2 kg / 50 l water, 10 (minutes) and soil application (5 kg bioformulation with 100 kg organic matter) (It is mixed and kept in the shade for 1 week before being applied to the soil.) It is used. This bioformulation is used for cold-resistant roots. It causes the formation of deep roots in corn and ornamental plants, and B. It is reported to be effective against cinerea. The limitations and inadequacies of current technological solutions, Cucurbitaceae Fusarium, a seed-borne pest, is found in plants belonging to the same family, especially in pumpkin seeds. Root rot disease caused by proliferatum affects the number of leaves and shoots. causing significant reductions in length, shoot fresh and dry weight. the fact that chemical pest control methods lead to water and soil pollution, 25 The misuse of pesticides disrupts ecosystem balance, solarization the process takes a long time and consequently causes delays in the production schedule the cause, also root rot caused by F. proliferatum such as the lack of a licensed fungicide or biopreparation against the disease. For these reasons, plants from the Cucurbitaceae family, especially snack plants, 30 Root rot in pumpkin caused by seed-borne Fusarium proliferatum. An improvement in disease prevention has become necessary. Brief Description and Objectives of the Invention The invention relates to the use of pumpkin seeds in the production of snacks, confectionery, and sweet foods. root rot disease caused by seed-borne Fusarium proliferatum to prevent and promote plant growth Trichoderma harzianum (NCBI access Wheat straw and sugar beet molasses containing TRIC8 isolate (number: MH351669) 5 An additive granular bioformulation is described. The aim of the invention is to develop a cultivar in plants of the Cucurbitaceae family, especially those used for snacking. Root rot in pumpkin caused by seed-borne Fusarium proliferatum. The goal is to prevent disease and promote plant growth. 10 One aim of the invention is to create a bioformulation that is low in production cost and easy to produce. The bioformulation that is the subject of the invention contains wheat straw and Sugar beet molasses is a readily available and low-cost ingredient. Furthermore, bioformulation directly to seed rather than large agricultural lands 15 Due to its applicability, it is used in small quantities. Repeated No application is needed. This reduces production costs. In addition, it also saves farmers money on their expenses. Because the bioformulation can be applied to the seed, the production in question is low. It is carried out at cost. 20 Another aim of the invention is to create a bioformulation that can maintain its viability for a long time. The aim is to provide the sugar beet molasses additive contained in the bioformulation in question. This substance allows T. harzianum conidia to easily attach to wheat straw and seeds. It ensures adhesion, thus the bioformulation adheres to the seed for 60 days (25). It retains its vitality. Explanation of the Figures Figure 1. 30 at 5 different doses of different bioformulations of Trichoderma harzianum. Rates of inhibition of Fusarium proliferatum colony growth (Doses; a: 0.25%, b: 0.50%, c: 0.75%, d: 1%, e: 2%). Figure 2. Different bioformulations prepared with Trichoderma harzianum (TRIC8). When applied in the prescribed doses, Fusarium proliferatum in PDA culture medium 6 Colony development (AE: Antagonist conidia suspension + CMC (TF1), FJ: Talc + CMC) (TF2), KO: Talc+CMC+CaCO3 (TF3), PT: Pesta granule (TF4), UY:Trichoderma harzianum (TRIC8) bioformulation (Invention), Zd: Maize straw (TF6), e: Control). (Gray-black areas represent bioformulations, and white areas represent Fusarium) (represents proliferatum) 5 Figure 3. The subject of the invention after inoculation of seeds with Fusarium proliferatum. Coating result with Trichoderma harzianum (TRIC8) bioformulation Disease severity in seedlings. Detailed Description of the Invention The invention relates to pumpkin seeds used in the production of snacks, confectionery, and sweet foods. root rot disease caused by seed-borne Fusarium proliferatum to prevent and promote plant growth Trichoderma harzianum (NCBI access Wheat straw and sugar beet molasses containing TRIC8 isolate (number: MH351669) 15 It relates to an additive granular bioformulation. The bioformulation that is the subject of the invention is F. proliferatum infestation of seedling roots and root collar of pumpkin seeds It prevents rot by 75.6%, and also increases seedling root length and shoot growth. It increases the length, seedling vigor index, seedling fresh and dry weight. The bioformulation described in this invention contains 2.0-3.0% Trichoderma harzianum (NCBI (Access number: MH351669) TRIC8 isolate, 37.0-41.0% wheat straw. and 57.0-60.0% sugar beet diluted with 8-12% distilled water. It contains molasses. In an application of the invention, the bioformulation subject to the invention, 2.3% Trichoderma harzianum (NCBI accession number: MH351669) TRIC8 25 The isolate was diluted with 39.1% wheat straw and 10% distilled water. It contains 58.6% sugar beet molasses. The method of preparing the bioformulation that is the subject of the invention; i. After wetting the ground wheat straw with distilled water, the excess water is removed by 30 and sugar beet molasses diluted with distilled water is added being done, ii. Sterilization of the resulting mixture in an autoclave and Trichoderma harzianum Addition of TRIC8 isolate from conidia suspension, 7 iii. Drying in a sterile environment by stirring daily, iv. After the drying process, in a sterile environment, through sterilized sieves transferring the mixture into sterile jars and sealing the lids with vacuum. It includes the steps involved in the process. Another application of the invention is the preparation of the bioformulation that is the subject of the invention. method; i. Soaking ground wheat straw in distilled water for 1.5-2.5 hours (250-350 ml) After soaking 100 g of straw (ml / 100 g), the excess water is removed and 140-160 ml / 100 g of straw is added. ml / 100 g straw diluted with 8-12% distilled water to sugar beet 10 the addition of molasses, ii. The resulting mixture is tested three times at 121°C for 20 minutes each, with a one-day interval between tests. autoclaving and sterilization of the conidia of Trichoderma harzianum TRIC8 isolate Adding 5-7 ml / 100 g of straw from the suspension (1 × 107 conidia / ml), iii. Store in a sterile environment at 24-26°C for 4-6 days, stirring daily. drying, iv. After the drying process, sterile medium with 0.5-1.4 mm pores in a sterile environment. passing through sieves, transferring into sterile jars and capping them vacuum sealing It includes the steps of the process. 20 In one application of the invention, the method of preparing the bioformulation that is the subject of the invention is as follows: i. ground wheat straw mixed with distilled water for 2 hours at a rate of 300 ml / 100 g After the straw is soaked, the excess water is removed and 150 ml / 100 g is added. straw diluted with 10% distilled water, sugar beet molasses added 25 being done, ii. The resulting mixture is tested three times at 121°C for 20 minutes each, with a one-day interval between tests. autoclaving and sterilization of the conidia of Trichoderma harzianum TRIC8 isolate Adding 6 ml / 100 g of straw from the suspension (1 × 107 conidia / ml), iii. Store in a sterile environment at 25°C for 5 days, stirring daily. drying, iv. After the drying process, sterile molds with 1 mm pores are placed in a sterile environment. passing through sieves, transferring into sterile jars and capping them vacuum sealing 8 It includes the steps involved in the process. In preparing the bioformulation that is the subject of this invention, ground wheat straw was processed for 2 hours. After being soaked in distilled water (300 ml / 100 g straw) throughout, the excess water was removed. and sugar beet molasses (10%) was added to it (150 ml / 100 g straw). The result is 5 The resulting mixture is sterilized three times in an autoclave at 121°C for 20 minutes each time, with a one-day interval between each sterilization. and from the conidia suspension of Trichoderma harzianum TRIC8 isolate (1 × 6 ml / 100 g of straw (107 conidia / ml) was added. Then, in a sterile environment at 25 °C. It was dried for 5 days by stirring it every day. As a result of this process... The antagonist was observed to remain alive. After the drying process, it was stored in a sterile environment for 10 minutes. Passed through sterilized sieves with a mesh size of 1 mm and into sterile jars. The products have been collected and their lids have been vacuum-sealed. The shelf life of the bioformulation is 150 at +4°C. It's been a day. To control root rot disease in pumpkin seeds and improve plant growth 15 To strengthen, clean seeds that have not been mechanically damaged are light. moistened (so that 1 / 3 of the seed's surface is wet) and placed on a clean area. The bioformulation is applied to the seeds at a dose of 1 g / 100 g seed. It is sprinkled, mixed until the surface is completely covered with the bioformulation, and It is dried in a clean environment for 15 minutes. 20 Firstly, the bioformulation that is the subject of the invention is pathogenic snack pumpkin (Cucurbita pepo). Fusarium proliferatum, which causes root and root collar disease in plants Its ability to inhibit colony growth has been determined. For this purpose, T. harzianum TRIC8 Six different bioformulations were made with the isolate, these were given at 5 different doses of Potato Dextrose 25 It was mixed into the culture medium. Experiments were conducted with 5 replicates. 6 samples among the bioformulations, the bioformulation prepared with wheat (Figures 1 and 2) It has been observed that this method inhibits colony growth to the greatest extent. Doses Among the findings, 1% and 2% doses of the bioformulation related to the invention were used to inhibit the colony of the pathogen. inhibiting its development at the highest rates, 94.13% and 94.16% respectively. 30 that they were capable of it and that there wasn't a huge difference between these two doses It has been observed that other applications are more economical. A 1% dose of the bioformulation was selected. In the second stage, the aforementioned The study investigated whether the bioformulation had a negative impact on seedling development. 9 In tests, bioformulation was applied to seeds of the pathogen-sensitive variety while they were slightly moist. After applying the mixture (1 g / 100 g seed) and ensuring a good mixture, it is left in a clean environment for 15 minutes. Dried for a few minutes, then placed in petri dishes containing moist blotting paper. (13 petri dishes, 8 seeds in each petri dish) were placed. No treatment was applied. Seeds were evaluated as a control. The bioformulation of our invention, 5 When applied to pumpkin seeds, it negatively affects germination and seedling development. There was no effect (Table 1). Also, control – (No intervention was applied). (seedlings developed from seed) root length, shoot length, seedling The strength index, wet weight and dry weight were 1.20, 1.16, 1.18, 1.39 and 1.07 respectively. It has increased by a factor of 10. Table 1. Coated with Trichoderma harzianum (TRIC8) bioformulation. germination rates of seeds compared to untreated seeds and resulting seedling development. In the third stage, the bioformulation of the invention was tested against the pathogen F. proliferatum. in its presence, the ability to prevent disease severity and promote plant growth. They were examined. In these tests, seeds of the susceptible variety were found to contain the pathogen F. After being inoculated with proliferatum, coated with the bioformulation of the invention (1 20 (g / 100 g seeds) dried in a clean environment for 15 minutes, then moistened inside. petri dishes containing blotting paper (13 petri dishes, 8 seeds in each petri dish) It was placed there for the purpose of comparison during the trials in question. TRIC8 isolate conidia suspension (1 × 107 conidia / ml + Tween 20, 0.1%) prepared (without bioformulation) and seeds were placed in this 25 for 6 hours. It was shaken in suspension. Only seeds contaminated with the pathogen were positive. The control was evaluated as (Control +). The pathogen was present in 48.08% of the seedlings. This has led to increased disease severity. However, the bioformulation of the invention... When applied to pathogen-inoculated seeds, disease severity was 11.73%, with TRIC8. When only conidia suspension was applied, the percentage was 14.04% (Figure 3). The invention... While the bioformulation of TRIC8 prevented disease severity by 75.6%, only TRIC8 did not. The blocking rate is lower (70.8%) in conidia suspension application. When plant growth parameters are taken into consideration, bioformulation is preferred. In the application, all development parameters are higher than Control (+) 5 It has been observed (Table 2). The bioformulation of the invention is effective even in the presence of the pathogen. germination, root length, shoot length, seedling vigor index, seedling dryness and It increased wet weight (by 2.07, 1.37, 1.38, 2.85, 1.60, and 1.16 times, respectively). Table 2. Trichoderma harzianum 10 after Fusarium proliferatum inoculation. Germination rates and formation of seeds coated with (TRIC8) bioformulation seedling development. In the fourth stage, the bioformulation of the invention is applied to seeds of the pathogen-sensitive variety. After application (1 g / 100 g seed), the pathogen (Fusarium proliferatum) Planted in contaminated soil and monitored for 30 days to observe disease severity and some development. The parameters have been determined. Experiments were conducted with 5 replicates. The subject matter... For comparison purposes during the experiments, the conidia of the TRIC8 isolate were 20 A suspension (1 × 10⁷ conidia / ml + Tween 20, 0.1%) was also prepared. (non-bioformulated version) and seeds are suspended in this mixture for 6 hours. It has been shaken. At the end of this period, the bioformulation of the invention reduced the disease severity. It was effective in reducing the number of patients by 86.5% (Table 3). Also, the control group (+) (Only (plants developed from pathogen-infected seeds) increased the number of leaves (1.2 times), 25 shoot length (1.23 times), shoot age (1.56 times) and dry weight (1.35 times) It has increased the disease. Seed application with the non-bioformulated version of TRIC8. 11 It was found to have a lesser effect on severity (82.7%), but improvement It has resulted in an increase in its parameters. Table 3. Trichoderma in soil contaminated with the pathogen (Fusarium proliferatum). 30 days after sowing of seeds treated with harzianum (TRIC8) bioformulation 5 then disease severity and some plant growth parameters. Industrial Applicability of the Invention The invention relates to pumpkin seeds used in the production of snacks, confectionery, and sweet foods. root rot disease caused by seed-borne Fusarium proliferatum to prevent and promote plant growth Trichoderma harzianum (NCBI access Wheat straw and sugar beet molasses containing TRIC8 isolate (number: MH351669) 15 It relates to an additive granular bioformulation and is suitable for industrial application. The invention is not limited to the above descriptions, and a person skilled in the field can easily make further discoveries. It can demonstrate different applications of the invention. These are the claims and demands of the invention. It should be evaluated within the scope of the protection granted. 20 12 REFERENCES [1] Snowdon, A.L. (2010). A color atlas of post-harvest diseases and disorders fruits and vegetables, Vol: 2, Vegetables, Manson Publishing, United Kingdom, 12- 51 p. [2] Kurt, Ş. 2012. Bitki fungal hastalıkları. Akademisyen Kitabevi Yayın Dağıtım ve 5 Pazarlama Ltd. Şti. Ankara, 214s [3] Stoner, M.F. 1981. Ecology of Fusarium in noncultivated soils. (P.E. Nelson, T.A. Toussoun and R.J. COOK, eds.) Fusarium diseases, biology, and taxonomy. The Pennsylvania State University Pres, University Park., 276-286p. [4] Katan, J. 1996. Soil solarization: Integrated control aspects. In: Hall, R. [Ed.] 10 Principles and Practice of Managing Soilborne Plant Pathogens. APS Press, St. Paul, MN, USA. pp. 250-278. [5] Meyer, J.M., Azelvandre, P., Georges, C. 1992. Iron metabolism in Pseudomonas: salicylic acid, a siderophore of Pseudomonas fluorescens CHAO. Biofactors 4: 23-27. 15 [6] Sneh, B., Dupler, M., Elad, Y., Baker, R. 1984. Clamidospore germination of Fusarium oxysporum f.sp. cucumerinum as affected by fluorescent and lytic bacteria from fusarium-suppresive soil. Phytopathology, 74: 1115-1124. [7] Janisiewicz, W., Korsten, L. 2002. Biological control of post-harvest diseases of fruits. Annual Review of Phytopathology, 40: 411-441. 20 [8] Fravel, D.R. 2000. Commercial biocontrol products, available for use against plant pathogens. http: / / www.oardc.ohio-state.edu 30

Claims

13 REQUESTS 1. Root disease caused by seed-borne Fusarium proliferatum in pumpkin seeds. to prevent rot disease and promote plant growth It is a bioformulation and its characteristic is; Trichoderma harzianum (NCBI access) TRIC8 isolate (number: MH351669) mixed with wheat straw and distilled water 5 It contains diluted sugar beet molasses.

2. It is a bioformulation according to Claim 1, characterized by a concentration of 2.0-3.0%. Trichoderma harzianum (NCBI accession number: MH351669) TRIC8 isolate, With 37.0-41.0% wheat straw and 8-12% distilled water It contains 57.0-60.0% diluted sugar beet molasses. 10 3. It is a bioformulation according to Claim 2, characterized by a concentration of 2.3%. Trichoderma harzianum (NCBI accession number: MH351669) TRIC8 isolate, Diluted with 39.1% wheat straw and 10% distilled water. It contains 58.6% sugar beet molasses.

4. Root rot caused by seed-borne Fusarium proliferatum in snack pumpkins. 15 to prevent rot disease and promote plant growth It is a method of preparing bioformulations, and its characteristic feature is; i. the excess after the ground wheat straw is moistened with distilled water the water is removed and sugar beet is diluted with distilled water. the addition of molasses, 20 ii. Sterilization of the resulting mixture in an autoclave and Trichoderma Addition of conidia suspension of harzianum TRIC8 isolate, iii. Drying in a sterile environment by stirring daily, iv. After the drying process, in a sterile environment, through sterilized sieves the mixture is transferred to sterile jars and sealed with a vacuum 25 minutes. closing It includes the steps of the process.

5. It is a method according to claim 4, and its characteristic is; i. ground wheat straw is mixed with distilled water for 1.5-2.5 hours at a rate of 250- After soaking 350 ml / 100 g of straw, the excess water should be removed and it should be left for 30 minutes. into 140-160 ml / 100 g of straw with 8-12% distilled water Addition of diluted sugar beet molasses, ii. The resulting mixture is tested three times at 121°C for 20 minutes each, with a one-day interval between tests. sterilization in an autoclave and Trichoderma harzianum TRIC8 isolate 14 Add 5-7 ml / 100 g of straw from conidia suspension (1 × 107 conidia / ml) being done, iii. Store in a sterile environment at 24-26°C, stirring daily for 4-6 days. drying for a period of time, iv. After the drying process, in a sterile environment, to a pore size of 0.5-1.4 mm 5 passing through sterilized sieves and transferring into sterile jars and lids are closed by vacuum It includes the steps of the process.

6. It is a method according to claim 5, and its characteristic is; i. ground wheat straw mixed with distilled water for 2 hours at a rate of 300 ml / 100 10 After the straw is soaked, the excess water is removed and 150 is added. ml / 100 g straw sugar beet diluted with 10% distilled water the addition of molasses, ii. The resulting mixture is tested three times at 121°C for 20 minutes each, with a one-day interval between tests. autoclaving and sterilization of Trichoderma harzianum TRIC8 isolate 15 Add 6 ml / 100 g of straw from conidia suspension (1 × 107 conidia / ml) being done, iii. Store in a sterile environment at 25°C, stirring daily for 5 days. drying, iv. After the drying process, 20 with 1 mm pores in a sterile environment. passing through sterilized sieves, transferring into sterile jars, and lids closing with vacuum It includes the steps of the process.

7. A bioformulation prepared by a method according to any of claims 4-6. 30

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