CN107338195B - Trichoderma for degrading organophosphorus pesticide and microbial inoculum thereof - Google Patents
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Abstract
The invention discloses Trichoderma for degrading organophosphorus pesticide and a microbial inoculum thereof, wherein the Trichoderma simens (Trichoderma simmonsii) T21-3 is preserved in the China general microbiological culture Collection center in 2016, 6 and 24 days, and the preservation number is CGMCC N0.12630. The microbial inoculum can degrade organophosphorus pesticide residues and prevent and control soil-borne diseases of various vegetables.
Description
Technical Field
The invention relates to trichoderma for degrading organophosphorus pesticide and a microbial inoculum thereof, belongs to the technical field of biology, utilizes a microbial method to degrade chemical pesticide residues, and is suitable for production and processing of green and pollution-free agricultural products in modern agricultural production.
Background
Pesticide pollution becomes one of the major agricultural ecological environment problems which are widely concerned at present, and poses serious threats to the sustainable development of modern agriculture and social economy, the agricultural ecological environment safety and the quality safety of agricultural products. The organophosphorus pesticide is used as a main pesticide in domestic and foreign agriculture, has the advantages of quick and efficient pesticide killing and low use cost, is widely used in modern agriculture, particularly in protected vegetable cultivation, and is the main body of vegetable pesticide residue exceeding standard. The pesticide residue is non-point source pollution, has low concentration and wide range, and adopts the traditional physical and chemical treatment method with great difficulty, high cost and secondary pollution. The treatment of pesticide residues with microorganisms is the safest, effective and inexpensive method to eliminate and detoxify high concentrations of pesticide residues.
Experts and scholars at home and abroad are dedicated to research the biodegradation technology of organophosphorus pesticides. The degrading bacteria are mainly enriched, domesticated and screened aiming at a specific organophosphorus pesticide from polluted media such as pesticide polluted soil or sludge of a pesticide factory, and the types of the degrading bacteria are mainly concentrated on bacteria, including pseudomonas, bacillus, flavobacterium, arthrobacter, acinetobacter, alcaligenes, ochrobactrum, brucella, enterobacter, rhodococcus and the like; the research on the degradation mechanism and the degrading enzyme gene of the organophosphorus pesticide is systematic. A great deal of research is also done in this aspect at home, a series of research results are obtained, and various species such as bacillus, pseudomonas putida, ochrobactrum, sphingomonas and the like apply for national invention patents. For example, Nanjing agriculture university separates and screens more than 500 microbial strains (mainly bacteria) for degrading chemical pesticides aiming at part of organophosphorus pesticides, organic nitrogen, organochlorine pesticides, bactericides, herbicides and the like, establishes a strain quality resource library with the largest microbial degradation of pesticides in China at present, and obtains national invented patents by 62 degrading strains and a fermentation process thereof. The degrading bacteria separated and screened at home and abroad have stronger degrading and repairing potential on one or more organophosphorus pesticides, but research results are generally in laboratory stage or plot test conditions, are less in actual product registration and popularization, and have single function, are not favorable for popularization and demonstration, and cannot meet the requirement of actual pollution control in soil at present.
Trichoderma has a special status in the field of agricultural production due to its application potential in the aspects of biocontrol ability, plant growth promotion, soil fertility improvement and the like, is a microbial species widely researched and applied, and has registration, popularization and application of microbial products (pesticides, fertilizers or decomposed microbial agents) obtained by a plurality of strains at home and abroad. Recent years of research at home and abroad shows that trichoderma has strong degradation or repair capability on pesticides (such as dichlorvos, chlorpyrifos, DDT, detelin, endosulfan and the like), polycyclic aromatic hydrocarbons, polychlorinated aromatic hydrocarbons, cyanides, heavy metals (such as cadmium, nickel, copper, aluminum and the like) and plastics and other pollutants, and also shows excellent application and development potential.
The development of greenhouse vegetables has been rapidly progressed since the middle of the 80 s of the last century. Due to perennial planting or continuous cropping, nutrient imbalance, serious pest and disease damage, frequent and excessive use of chemical fertilizers and pesticides, gradual deterioration of soil environment (such as soil secondary salinization, acidification and nutrient imbalance), long-term residue of various pesticides and microbial ecological imbalance.
Disclosure of Invention
The invention aims to provide trichoderma and a microbial inoculum thereof for degrading organophosphorus pesticide aiming at the actual problems and requirements of organophosphorus pesticide residue in domestic facility agricultural soil, soil-borne plant diseases with increasingly serious diseases and the like. The trichoderma strain disclosed by the invention has better adaptability to the soil environment of facility agriculture, and has prevention and control effects on various common soil-borne diseases, and the developed trichoderma degrading microbial inoculum product is more beneficial to application and popularization.
the Trichoderma for degrading organophosphorus pesticide and the microbial inoculum thereof have the technical scheme that the Trichoderma strain for degrading organophosphorus pesticide adopted by the invention is Trichoderma simmondii (Trichoderma simmonsii) T21-3, is separated and screened from facility tomato soil in a laboratory, is stored in the common microorganism center of China Committee for culture Collection of microorganisms, and has the address of No. 3 of Ministry No. 1 of West Chen of south China, Beijing, the institute of microorganisms of the institute of China academy of sciences, 100101, the storage date of 2016 24 months and the storage number of CGMCC N0.12630, and the TEF-1 α of the strain is submitted to NCBI BlAST for similarity comparison, so that the Trichoderma is identified as Trichoderma simmondii (Trichoderma simmonsii), and the TEF-1 alpha gene sequence is submitted to a Genbank database with the accession number of KX 426072.
The trichoderma capable of degrading organophosphorus pesticide is applied to degrading organophosphorus pesticide residues in soil.
The organophosphorus pesticide is any one of methamidophos, chlorpyrifos, omethoate, parathion, isocarbophos and phoxim.
The trichoderma capable of degrading organophosphorus pesticide is applied to prevention and treatment of soil-borne diseases of vegetables.
The soil-borne disease is caused by at least one of the following bacteria: rhizoctonia solani (rhizoctonia solani), Fusarium oxysporum (Fusarium oxysporum), Pythium aphanidermatum (Pythium aphanidermatum), Fusarium solani (Fusarium solani), Verticillium dahliae (Verticillium dahliae), and botrytis cinerea (botrytisciprea).
Trichoderma reesei (Trichoderma simmonsii) T21-3 is prepared into Trichoderma agent, and the active ingredient of the Trichoderma agent is live Trichoderma conidium more than or equal to 3 multiplied by 108cfu/g。
The trichoderma preparation is wettable powder and consists of the following components in percentage by weight: 2-3% of trichoderma simmondii T21-3 conidium powder, 1-2% of sodium carboxymethylcellulose, 1-2% of xanthan gum, 1-2% of sodium fulvate, 5-10% of glucose, 75-85% of diatomite and less than 5% of water content.
The preparation method of the trichoderma agent comprises the following steps:
a. preparing a Trichoderma simmondii (Trichoderma simmonsii) T21-3 seed solution;
b. preparing a solid fermentation culture medium;
c. mixing the seed liquid obtained in the step a with the solid fermentation culture medium obtained in the step b according to the proportion of 1%, and fermenting to obtain a solid trichoderma simonii fermented product;
d. c, crushing the solid trichoderma simonii fermentation product obtained in the step c, and sieving the crushed product with a 60-mesh or 80-mesh sieve;
e. and d, mixing the diatomite, the xanthan gum, the sodium carboxymethylcellulose, the sodium fulvate and the glucose with the raw materials obtained in the step d, uniformly stirring and fully mixing to obtain a light green powder, namely the trichoderma preparation.
The preparation method of the trichoderma preparation specifically comprises the following steps:
a. inoculating Trichoderma simmondii (Trichoderma simmonsii) T21-3 strain into a PDA culture medium for activation, transferring the activated Trichoderma simsii T21-3 strain onto a PDA solid culture medium or a wheat grain culture medium, placing the PDA solid culture medium or the wheat grain culture medium in a constant-temperature culture box at 28 ℃ for culturing for 5-7 days, eluting the culture medium with sterile water after a large amount of conidia are generated to obtain Trichoderma simmondii seed solution, wherein the PDA culture medium is prepared by adding 200 g of peeled potatoes, 20 g of glucose and 15 g of agar powder into 1000mL of water and mixing; the wheat grain culture medium is prepared by soaking wheat grains in water, standing overnight, and performing moist heat sterilization at 121 deg.C;
b. preparing a solid fermentation culture medium: sterilizing a mixture of 40% of corn straws, 5% of wheat bran and the balance of water at 121 ℃ for 60 minutes to obtain a solid fermentation culture medium;
c. mixing the seed solution obtained in the step a with the solid fermentation culture medium obtained in the step b according to a proportion of 1%, uniformly stirring, covering with gauze or a sealing film, standing at a constant temperature of 25-28 ℃ for 7-10 days, allowing trichoderma conidium to reach 60-100 multiplied by 108/g, and naturally airing or drying after fermentation is finished to obtain a solid trichoderma simens fermentation product;
d. c, crushing the solid trichoderma simonii fermentation product obtained in the step c, and sieving the crushed solid trichoderma simonii fermentation product through a 60-mesh or 80-mesh sieve;
e. and d, mixing the diatomite, the xanthan gum, the sodium carboxymethylcellulose, the sodium fulvate and the glucose with the raw materials obtained in the step d, uniformly stirring and fully mixing to obtain a light green powder, namely the trichoderma preparation.
The trichoderma preparation is applied to degrading organophosphorus pesticide residues in soil and preventing and treating vegetable soil-borne diseases.
The invention has the beneficial effects that: the trichoderma simonsii T21-3 for degrading various organophosphorus pesticides is separated from a facility vegetable field which is cultivated and applied for many years, is easy to ferment, low in production cost, strong in adaptability to facility agricultural soil environment, environment-friendly, capable of degrading various organophosphorus pesticide residues such as methamidophos, chlorpyrifos, omethoate, parathion, isocarbophos and phoxim in soil and reducing residual hazard of pesticides in soil or vegetables; meanwhile, the soil conditioner has the effects of preventing and controlling various common soil-borne diseases caused by Rhizoctonia solani (Rhizoctonia solani), Fusarium oxysporum (Fusarium oxysporum), Pythium aphanidermatum (Pythium aphanidermatum), Fusarium solani (Fusarium solani), Verticillium dahliae (Verticillium dahliae) and Botrytis cinerea (Botrytis cinerea), reduces or replaces bactericides for use, and effectively regulates and controls a soil microflora. Meanwhile, the trichoderma degrading microbial inoculum product is more beneficial to application and popularization, and has high application and popularization values in the organic (green) vegetable cultivation process of modern facility agriculture.
The specific implementation mode is as follows:
for better understanding of the present invention, the technical solution of the present invention will be described in detail with specific examples, but the present invention is not limited thereto.
Example 1
Preparation of trichoderma agent for degrading organophosphorus pesticide
the Trichoderma reesei (Trichoderma sumingsii) T21-3 for degrading various organophosphorus pesticides is separated and screened from greenhouse tomato soil in a laboratory, is stored in the common microorganism center of China Committee for culture Collection of microorganisms, and has the address of No. 3 of No. 1 Hosth West Lu of Beijing south Kogyo of sunward area, the institute of microbiology of China academy of sciences, 100101, the storage date of 2016, 6 and 24 days, and the preservation number of CGMCC N0.12630. the TEF-1 α sequence of the Trichoderma reesei is submitted to NCBI BlAST for similarity comparison, and the Trichoderma reesei (Trichoderma sumingsii) is identified, and the TEF-1 α gene sequence is submitted to Genbank database, and has the accession number of KX 426072.
The preparation method of the trichoderma agent comprises the following steps:
a. inoculating a stored Trichoderma simmondii (Trichoderma simmonsii) T21-3 strain into a PDA (PDA dextrose agar) plate culture medium for activation, transferring the activated Trichoderma simmondii T21-3 strain onto a PDA solid culture medium in a 500mL triangular flask, placing the PDA solid culture medium in a constant-temperature incubator at 28 ℃ for culturing for 5 days, eluting the PDA solid culture medium with sterile water after a large amount of conidia are generated to obtain Trichoderma simmondii seed solution, wherein the PDA culture medium is prepared by mixing 200 g of peeled potatoes, 20 g of glucose and 15 g of agar powder with 1000mL of water;
b. preparing a solid fermentation culture medium: sterilizing a mixture of 40% of corn straws, 5% of wheat bran and the balance of water at 121 ℃ for 60 minutes to obtain a solid fermentation culture medium;
c. mixing the seed solution obtained in the step a with the solid fermentation culture medium obtained in the step b according to the proportion of 1%, uniformly stirring, covering with gauze or a sealing film, placing in a constant-temperature incubator at the temperature of 28 ℃ for static culture for 8 days, and allowing the conidia of the trichoderma to reach 90 multiplied by 108Per gram. After fermentation, the fermented product is naturally dried or air-driedDrying to obtain solid fermentation product of Trichoderma Simens (Trichoderma reesei) with conidium content of 150 × 108Per gram;
d. c, crushing the solid trichoderma simonii fermentation product obtained in the step c, and sieving the crushed solid trichoderma simonii fermentation product through a 60-mesh or 80-mesh sieve;
e. and (d) mixing 80% of diatomite, 2% of xanthan gum, 1% of sodium carboxymethylcellulose, 1% of sodium fulvate and 5% of glucose with 2% of the raw materials obtained in the step (d), uniformly stirring and fully mixing to obtain a light green powder, and controlling the water content to be below 5%, thus obtaining the trichoderma preparation.
Example 2
Preparation of trichoderma agent for degrading organophosphorus pesticide
The preparation method of the microbial inoculum comprises the following steps:
the difference between this embodiment and embodiment 1 is: in the step a, activating trichoderma strains, transferring the activated trichoderma strains to a wheat solid culture medium, and culturing for 7d at room temperature; in step c, after inoculation, the mixture is placed under the condition of room temperature (23-28 ℃) for static culture for 10 days, and the conidia of the trichoderma reaches 80 multiplied by 108Per gram; in step e, 75% of diatomite and 8% of glucose are mixed with the raw material obtained in step d.
Example 3
Test for degrading organophosphorus pesticide by using trichoderma in liquid shake flask condition
The method comprises the following steps: inoculating a stored Trichoderma simmondii (Trichoderma simmonsii) T21-3 strain into a PDA plate culture medium for activation, transferring the activated strain onto a PDA solid culture medium in a 500mL triangular flask, placing the PDA solid culture medium in a constant-temperature incubator at 28 ℃ for 5 days, eluting by using sterile water after a large amount of conidia are generated, and diluting to obtain a Trichoderma simmondii seed solution (1.0 multiplied by 108 CFU/mL);
step two: adding six pesticides of methamidophos, chlorpyrifos, omethoate, parathion, isocarbophos and phoxim into a sterilized 250mL triangular flask filled with 50mL of inorganic salt liquid culture medium (MSM) according to the concentration content of 100mg/L, and respectively inoculating 1mL of trichoderma spore seed liquid (1.0 multiplied by 108CFU/mL) of each strain, wherein the MSM inorganic salt liquid culture medium is K2HPO41.50g，KH2PO40.50g，MgSO4·7H2O 0.50g，(NH4)2SO40.50g，FeSO4·7H2O 0.002g，CaCl20.04g，NaCl 0.5g，H2O 1000ml，pH7.0；
Step three: the treatment with a suspension of spores of non-inoculated wood mould was used as a control. Each treatment is repeated for 3 times, the mixture is placed at 30 ℃ and 140rpm for culturing for 5 days, samples are taken, and the organophosphorus pesticide residue is detected by using a GC-ECD method, and the results are shown in Table 1.
TABLE 1 degradation of organophosphorus pesticides by Trichoderma under liquid shake flask conditions
Example 4
Bacteriostasis test of trichoderma in the invention on several common vegetable soil-borne pathogens
The method comprises the following steps: respectively inoculating preserved Trichoderma T21-3 and several soil-borne pathogenic fungi such as Rhizoctonia solani (Rhizoctonia solani), Fusarium oxysporum (Fusarium oxysporum), Pythium aphanidermatum (Pythium aphanidermatum), Fusarium solani (Fusarium solani), Verticillium dahliae (Verticillium dahliae) and Botrytis cinerea (Botrytis cinerea) on PDA medium for activation for 2 d;
step two: picking lawn from the edges of trichoderma and each pathogenic fungus colony with sterilized punch with diameter of 5mm, respectively transferring onto PDA plate with diameter of 9cm (spacing of 6cm), culturing at 25 deg.C, repeating for 3 times without treatment, and comparing with control of inoculating pathogenic fungus only;
step three: and when the control pathogenic bacteria colony grows to the trichoderma inoculation point or the trichoderma colony grows to the pathogenic bacteria inoculation point, measuring the radius of each colony and calculating the inhibition rate. The results are shown in Table 2.
TABLE 2 test of the confronting of Trichoderma on several soil-borne phytopathogenic fungi
Example 5
The trichoderma preparation of the invention is used for degradation test of organophosphorus pesticide in soil
The method comprises the following steps: sieving soil (from tomato vegetable field of Shouguang facility) with 20 mesh sieve, treating with two groups, wherein group A is sterilized at 121 deg.C for 120 min, and group B is not sterilized; respectively adding organophosphorus pesticides such as methamidophos, chlorpyrifos, omethoate, parathion, isocarbophos, phoxim and the like according to the amount of 100mg/Kg, uniformly mixing and adsorbing for 24 hours;
step two: filling the pesticide-treated soil into 18cm × 15cm × 15cm plastic test pots, wherein each pot contains 1kg of soil;
step three: adding the trichoderma preparation prepared in the example 1 into the treatment in the step two according to the inoculation amount of 5 percent, and taking the treatment without adding trichoderma as a reference; setting 3 basins for each treatment;
step four: randomly placing the treated group and the control group into a constant temperature climate chamber (the temperature is 28 +/-2 ℃), managing according to conventional potting conditions, continuously culturing for 2 weeks, and taking a soil sample to determine the residual quantity of the organophosphorus pesticide by adopting a GC-ECD method. The results are shown in Table 3.
TABLE 3 degradation of organophosphorus pesticides in soil by Trichoderma
Example 6
Prevention and treatment test of common tomato soil-borne diseases by trichoderma agent
The method comprises the following steps: and (4) treating the soil. The facility vegetable field (Wuxiliu village, Jitai town of Shouguang), applying organic fertilizer according to conventional operation, deep turning, and making ridges (adopting a mode of large and small rows and small ridges with a height of 70-80 cm, 40 cm and 10-15 cm). Dividing the west to east into 8 equal sections, treating 2, 4 and 6 sections, and comparing 3, 5 and 7 sections; in the treatment interval, the trichoderma preparation (5 Kg/mu) prepared in the embodiment 1 is added with 25Kg of moist fine soil or organic fertilizer to be uniformly stirred, and is uniformly spread and raked; the control interval was not loaded with the trichoderma preparation of the present invention.
Step two: and (5) transplanting. Planting two lines in each ridge, transplanting tomato seedlings, and simultaneously processing the ridges to irrigate roots at the dosage of 5 Kg/mu.
Step three: disease investigation and data analysis. Sampling at 5 points in each interval in the greenhouse, investigating 10 plants at each point, counting the incidence rate of common soil-borne diseases of the tomatoes, and calculating the control effect; statistical analysis is carried out on the calculated data by SPSS software, and the difference significance between the processing mean values is tested; the results are shown in Table 4.
TABLE 4 controlling Effect of Trichoderma preparation of the present invention on common soil-borne diseases of tomato
Example 7
Test for controlling common soil-borne diseases of cucumbers by trichoderma agent
The method comprises the following steps: and (4) treating the soil. In the facility vegetable field (the depressed village of Taoist Tokyo, shou Guang city), applying organic fertilizer according to the conventional operation, deep turning, and making ridges (by adopting a large-row mode, namely a large row of 80 cm and a small row of 40 cm). Dividing the west to east into 8 equal sections, treating 2, 4 and 6 sections, and comparing 3, 5 and 7 sections; in the treatment interval, the trichoderma preparation (5 Kg/mu) prepared in the embodiment 1 is added with 25Kg of moist fine soil or organic fertilizer to be uniformly stirred, and is uniformly spread and raked; the control interval was not loaded with the trichoderma preparation of the present invention.
Step two: and (5) transplanting. Planting two lines (the row spacing is 25-30 cm) in each ridge, transplanting cucumber seedlings, and simultaneously processing the ridges to irrigate roots at the dosage of 5 Kg/mu.
Step three: disease investigation and data analysis. Sampling at 5 points in each interval in the greenhouse, investigating 10 plants at each point, counting the incidence rate of common soil-borne diseases of the cucumbers, and calculating the control effect; statistical analysis is carried out on the calculated data by SPSS software, and the difference significance between the processing mean values is tested; the results are shown in Table 5.
TABLE 5 prevention and treatment effects of Trichoderma preparation of the present invention on common soil-borne diseases of cucumber
Claims (7)
1. The trichoderma for degrading organophosphorus pesticide is trichoderma simens (trichoderma sammonsii) T21-3, which is preserved in China general microbiological culture Collection center (CGMCC) at 24 months 6 in 2016, with the preservation number: CGMCC N0.12630.
2. Use of the organophosphorus pesticide-degrading trichoderma as claimed in claim 1 for degrading organophosphorus pesticide residues in soil; the organophosphorus pesticide is any one of methamidophos, chlorpyrifos, omethoate, parathion, isocarbophos and phoxim.
3. The use of a trichoderma fungus degrading organophosphorus pesticide according to claim 1 for controlling soil-borne diseases of vegetables;
the soil-borne disease is caused by at least one of the following bacteria: rhizoctonia solani (Rhizoctonia solani), Fusarium oxysporum (Fusarium oxysporum), Pythium aphanidermatum (Pythium aphanidermatum), Fusarium solani (Fusarium solani), Verticillium dahliae (Verticillium dahliae), and botrytis cinerea (botrytiscipinea).
4. The use according to claim 2 or 3, wherein Trichoderma reesei (Trichoderma Simonisii) T21-3 is prepared into Trichoderma preparation, and the Trichoderma preparation effective component is live Trichoderma conidium ≥ 3 x 108cfu/g。
5. The trichoderma preparation for trichoderma capable of degrading organophosphorus pesticides according to claim 1, wherein the trichoderma preparation is wettable powder and comprises the following components in parts by weight: 2-3% of trichoderma simmondii T21-3 conidium powder, 1-2% of sodium carboxymethylcellulose, 1-2% of xanthan gum, 1-2% of sodium fulvate, 5-10% of glucose, 75-85% of diatomite and less than 5% of water content.
6. The trichoderma preparation of claim 5, comprising the steps of:
a. preparing a Trichoderma simens T21-3 seed solution;
b. preparing a solid fermentation culture medium;
c. mixing the seed solution obtained in the step a with the solid fermentation culture medium obtained in the step b according to the proportion of 1%, and fermenting to obtain a solid trichoderma simonii T21-3 fermentation product;
d. c, crushing the fermentation product of the trichoderma simmondii T21-3 obtained in the step c, and sieving the crushed fermentation product with a 60-or 80-mesh sieve;
e. and d, mixing the diatomite, the xanthan gum, the sodium carboxymethylcellulose, the sodium fulvate and the glucose with the raw materials obtained in the step d, uniformly stirring and fully mixing to obtain a light green powder, namely the trichoderma preparation.
7. The trichoderma preparation of claim 6, comprising the steps of:
a. inoculating Trichoderma simens T21-3 strain into a PDA culture medium for activation, transferring the activated strain onto a PDA solid culture medium or a wheat grain culture medium, culturing the strain in a constant-temperature incubator at 28 ℃ for 5-7 d, eluting the strain with sterile water after a large amount of conidia are generated to obtain Trichoderma simens T21-3 seed liquid, wherein the PDA culture medium is prepared by mixing 200 g of peeled potatoes, 20 g of glucose and 15 g of agar powder with 1000mL of water; the wheat grain culture medium is prepared by soaking wheat grains in water, standing overnight, and performing moist heat sterilization at 121 deg.C;
b. preparing a solid fermentation culture medium: sterilizing a mixture of 40% of corn straws, 5% of wheat bran and the balance of water at 121 ℃ for 60 minutes to obtain a solid fermentation culture medium;
c. mixing the seed solution obtained in the step a with the solid fermentation culture medium obtained in the step b according to a proportion of 1%, uniformly stirring, covering with gauze or a sealing film, standing and culturing at a constant temperature of 25-28 ℃ for 7-10 days, wherein conidia of trichoderma reaches 60-100 multiplied by 108Drying the solid fermentation product in the air or in the air after fermentation is finished to obtain the solid fermentation product of the trichoderma simens;
d. c, crushing the solid trichoderma simonii fermentation product obtained in the step c, and sieving the crushed solid trichoderma simonii fermentation product through a 60-mesh or 80-mesh sieve;
e. and d, mixing the diatomite, the xanthan gum, the sodium carboxymethylcellulose, the sodium fulvate and the glucose with the raw materials obtained in the step d, uniformly stirring and fully mixing to obtain a light green powder, namely the trichoderma preparation.
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