Application of trichoderma harzianum and metabolite in aspects of plant root promotion and adversity stress resistance

Abstract

The invention discloses application of trichoderma harzianum and metabolite in aspects of plant root promotion and adversity stress resistance, and the preparation method comprises the following steps: inoculating the trichoderma harzianum into a PDB culture medium for culture, and inoculating a culture solution into a rice + catecholamine + trimethoprim culture medium for fermentation. The composition can stimulate the growth of plant root systems, has a relatively obvious promoting effect on crop germination potential and germination rate, promotes the growth of seedlings at the overground part of the crop, accelerates the growth of the top end of the crop, deepens the green color of leaves, enhances the saline-alkali stress resistance of rice, enhances the adversity stress resistance of virus resistance, high temperature resistance, low temperature resistance, drought resistance and waterlogging resistance of the crop, and improves the yield of the rice. Therefore, crops are resistant to adversity and root, and strong induced resistance and immune induced resistance are achieved. The trichoderma harzianum and metabolites prepared by the invention enhance the vitality of crops, are environment-friendly and pollution-free, and have wide application prospects.

Description

Technical Field

[0001] The present invention mainly relates to the "technical field of agricultural application of biological metabolites", and specifically relates to the use of Trichoderma harzianum and its metabolites in promoting plant root growth and resisting stress. Background Art

[0002] In agricultural production, improving the root growth of crops and enhancing their stress resistance are the keys to improving crop yield and quality. Traditional plant growth promotion and stress resistance technologies mainly rely on chemical fertilizers and pesticides. Although these methods can increase the growth rate and yield of crops in the short term, in the long run, they will pollute the soil and environment, affect the ecological balance, and may lead to increased drug resistance of crops, reducing the long-term effect of pesticides. In addition, some physical methods such as irrigation and covering are also used to improve the stress resistance of plants, but these methods often require a large amount of water resources and energy, with high costs, and are difficult to implement in arid and water-scarce areas. At the same time, these physical methods have limited effects on promoting the growth of plant roots and cannot fundamentally solve the problems of slow plant growth and weak stress resistance. In contrast, biotechnology provides a more environmentally friendly and sustainable solution. Using microorganisms and their metabolites to promote plant growth and enhance stress resistance is an emerging technical means. However, in existing biotechnology solutions, the effects of many microbial preparations are not stable and are sensitive to environmental conditions, restricting their wide application in different regions and different crops.

[0003] Research shows that the addition of specific antibacterial substances in the culture medium will cause changes in microbial metabolites. At present, there is little research on the promotion of metabolite changes of Trichoderma harzianum by trimethoprim. As a microorganism with broad biological activities, Trichoderma harzianum has great potential in promoting plant root growth and resisting stress. However, traditional cultivation methods of Trichoderma harzianum have the phenomenon of low disease-resistant and antibacterial metabolites. Therefore, developing a biotechnology that uses Trichoderma harzianum and its metabolites to promote plant root growth and enhance stress resistance is of great significance for improving crop yield and quality, protecting the environment and ecological balance. Summary of the Invention

[0004] Based on the existing technology, the technical problem to be solved by the present invention is to provide the application of Trichoderma harzianum and its metabolites in promoting plant root growth and resisting stress to solve the above problems.

[0005] The technical solution of the present invention:

[0006] A preparation method of Trichoderma harzianum and its metabolites, comprising the following preparation steps:

[0007] S1. Inoculate Trichoderma harzianum in a PDB medium for culture to obtain a Trichoderma harzianum culture solution;

[0008] S2, inoculating the culture fluid of Trichoderma harzianum into a solid culture medium containing rice, catecholamines and trimethoprim for fermentation, and centrifuging to obtain the supernatant to obtain a fermentation fluid;

[0009] S3, countercurrently eluting the fermented bacterial liquid to obtain a purified bacterial liquid;

[0010] S4, filtering the purified bacterial liquid, distilling under reduced pressure, concentrating, drying, and powdering to obtain finished powder.

[0011] Further, in step S2, the mass ratio of the rice, catecholamines and trimethoprim is 1:0.05-0.08:0.02-0.04, and the spore production of the solid culture medium after fermentation is greater than 1×10 8 Pieces / mL.

[0012] Further, in step S2, the culture temperature is 24°C-26°C, the humidity is 45%-75%, the pH value is 5.5-7.0, and the culture time is 6-8 hours; the centrifugal speed is 3500-4000r / min, and the centrifugal time is 15-20min.

[0013] Furthermore, in step S3, the inorganic salt used in the countercurrent elution is one of magnesium chloride, sodium chloride, magnesium sulfate, and sodium sulfate; the polar solvent is one of methanol, ethanol, and acetonitrile; the mass percentage of the inorganic salt in the eluent is less than 3%, the volume percentage of the polar solvent is 20%-75%, and the remainder is water; the volume ratio of the fermentation liquid to the eluent is 1:1-2.

[0014] Furthermore, in step S3, the elution temperature is 20-25°C, the flow rate of the fermentation liquid is 5-7 ml / min, and the flow rate of the eluent is 20-25 ml / min. In step S4, the pressure of the reduced pressure distillation is 0.08-1.0 MPa, and the temperature is 40°C-45°C.

[0015] Further, in step S4, the ingredients of the finished powder include 6-phosphate-glucose, acetylglucosamine, chitosan, free α-amino acids, β-amino acids, fatty acids, hydroxycinnamic acid, 6-pentyl-α-pyrone, 5,6-dihydro-6-pentyl-2H-pyran-2-one, isonitrile, khatzian lactone, collomycin, trichomycin, khatzianin, and gibberellic acid.

[0016] Compared with the prior art, the present invention has the following beneficial effects:

[0017] (1) This technical solution utilizes Trichoderma harzianum and its metabolites to resist Pythium ulminans, Pythium ultimum, Phytophthora capsici, Gibberella tritici, and Fusarium oxysporum, thereby enhancing the disease resistance of crops.

[0018] (2) Trichoderma harzianum and its metabolites can enhance plants' resistance to stress, have an obvious promoting effect on the germination potential and germination rate of rice, promote the growth of the roots and above-ground seedlings of individual wheat plants in the early stage, improve the apical growth of pepper, bitter gourd, and pea plants, deepen the green color of their leaves, and promote the absorption of nutrients by crops.

[0019] (3) The finished powder prepared by the method of the present invention can enable rice to resist saline-alkali stress and enhance the ability of rice roots to root under stress.

[0020] (4) The Trichoderma harzianum and its metabolites prepared by this technical solution enhance the stress resistance of crops to high temperature, low temperature, drought, and waterlogging, enable crops to root under stress, and have strong induced resistance and immune-induced resistance capabilities.

[0021] (5) This technical solution uses biotechnology, does not involve chemical additives, is environmentally friendly, pollution-free, and meets the requirements of sustainable development. The application of Trichoderma harzianum and its metabolites can be widely applied to a variety of plants and has broad application prospects. Description of the Drawings

[0022] Figure 1 : Finished powder

[0023] Figure 2 : Comparison chart of the antibacterial effect of the finished powder

[0024] Figure 3 : Comparison chart of the growth effect of maize seedling roots. The left figure shows the comparison data of root length and bud length under different treatments, and the right figure shows the control chart of root growth under different treatments. The annotations marked as 0.1, 0.2, 0.5, 1, 2, 5, and 10 mean that the dosage of the coated finished powder is 0.1 g / 100 Kg, 0.2 g / 100 Kg, 0.5 g / 100 Kg, 1 g / 100 Kg, 2 g / 100 Kg, 5 g / 100 Kg, and 10 g / 100 Kg

[0025] Figure 4 : Comparison chart of the growth effect of wheat under different treatments. The left figure shows the comparison data of root fresh weight and seedling fresh weight under different treatments, and the right figure shows the control chart of germination potential and germination rate under different treatments. The annotations marked as 0.1, 0.2, 0.5, 1, 2, and 5 mean that the dosage of the coated finished powder is 0.1 g / 100 Kg, 0.2 g / 100 Kg, 0.5 g / 100 Kg, 1 g / 100 Kg, 2 g / 100 Kg, and 5 g / 100 Kg

[0026] Figure 5:Comparison chart of the growth effects of different treatments on rice. The left figure shows the comparison data of the germination potential and germination rate of different treatments, and the right figure shows the control chart of the root system and seedling growth of the corresponding treatments. The annotations marked as 0.1, 0.2, 0.5, 1, 2, and 5 in the figure mean that the dosage of the coated finished product powder is 0.1 g / 100 Kg, 0.2 g / 100 Kg, 0.5 g / 100 Kg, 1 g / 100 Kg, 2 g / 100 Kg, and 5 g / 100 Kg

[0027] Figure 6 :Comparison chart of the growth trends of different treatments on bitter gourd. In the figure, Experimental Group 10000, Experimental Group 20000, and Experimental Group 30000 mean that the finished product powder is diluted 10000 times, 20000 times, and 30000 times with water

[0028] Figure 7 :Comparison chart of the growth trends of different treatments on peppers and peas. The left figure shows the comparison data of peppers with different dilution multiples, and the right figure shows the growth control chart of peas with different dilution multiples. 10000 times, 20000 times, and 30000 times in the figure mean that the finished product powder is diluted 10000 times, 20000 times, and 30000 times with water

[0029] Figure 8 :Comparison chart of the growth trends of the adventitious roots of different treatments on rice under saline-alkali stress. In the figure, A: Alginate oligosaccharide is diluted 1 million times; B: PHA (polyhydroxyalkanoate) + trehalose 2 g / 100 ml is diluted 700 times; C: The finished product powder prepared in Example 3 is diluted 50000 times; D: The finished product powder prepared in Example 3 is diluted 200000 times; E: PHA is diluted 700 times; F: PHA + ectoine (8:2) is diluted 700 times; G: PHA + ectoine (9:1) is diluted 700 times; CK1: Non-stress control; CK2: Saline-alkali stress control Detailed implementation methods

[0030] To better understand the technical content of the present invention, specific examples are provided below to further illustrate the present invention

[0031] Unless otherwise specified, the experimental methods used in the embodiments of the present invention are all conventional methods, and the reagents used in the embodiments can all be obtained through commercial channels

[0032] Example 1

[0033] The preparation of Trichoderma harzianum and its metabolites includes the following steps

[0034] Step (1): Inoculate Trichoderma harzianum in a PDB medium and culture it to obtain a Trichoderma harzianum culture solution

[0035] Step (2): Inoculate the Trichoderma harzianum culture solution onto a solid medium of rice, catecholamine, and trimethoprim for fermentation culture. The mass ratio of rice, catecholamine, and trimethoprim is 1:0.05:0.02. The culture temperature is 24°C, the humidity is 45%, the pH value is 5.5, and the culture time is 6 hours. After culturing, perform centrifugation at a rotational speed of 3500 r / min for 15 minutes. After centrifugation, take the supernatant to obtain the fermented bacterial liquid;

[0036] Step (3): Perform countercurrent elution on the fermented bacterial liquid. The inorganic salt used in the countercurrent elution is magnesium chloride with a mass percentage content of 2.5%, the polar solvent is acetonitrile with a volume percentage content of 45%, and the balance is water. The volume ratio of the fermented bacterial liquid to the eluent is 1:1.5. The elution temperature is 20°C, the flow rate of the fermented bacterial liquid is 5 mL / min, and the flow rate of the eluent is 20 mL / min. After countercurrent elution, obtain the purified bacterial liquid;

[0037] Step (4): Filter the purified bacterial liquid, perform vacuum distillation. The pressure of the vacuum distillation is 0.08 MPa, the temperature is 40°C. After heating and concentration, dry it and spray powder to obtain the finished product powder.

[0038] Example 2

[0039] The preparation of Trichoderma harzianum and its metabolites includes the following steps:

[0040] Step (1): Inoculate Trichoderma harzianum into a PDB medium for culture to obtain a Trichoderma harzianum culture solution;

[0041] Step (2): Inoculate the Trichoderma harzianum culture solution onto a solid medium of rice, catecholamine, and trimethoprim for fermentation culture. The mass ratio of rice, catecholamine, and trimethoprim is 1:0.06:0.02. The culture temperature is 26°C, the humidity is 55%, the pH value is 6.0, and the culture time is 8 hours. After culturing, perform centrifugation at a rotational speed of 4000 r / min for 20 minutes. After centrifugation, take the supernatant to obtain the fermented bacterial liquid;

[0042] Step (3): Perform countercurrent elution on the fermented bacterial liquid. The inorganic salt used in the countercurrent elution is sodium chloride with a mass percentage content of 2%, the polar solvent is acetonitrile with a volume percentage content of 40%, and the balance is water. The volume ratio of the fermented bacterial liquid to the eluent is 1:1.2. The elution temperature is 25°C, the flow rate of the fermented bacterial liquid is 7 mL / min, and the flow rate of the eluent is 25 mL / min. After countercurrent elution, obtain the purified bacterial liquid;

[0043] Step (4): Filter the purified bacterial liquid, perform vacuum distillation. The pressure of the vacuum distillation is 1 MPa, the temperature is 45°C. After heating and concentration, dry it and spray powder to obtain the finished product powder.

[0044] Example 3

[0045] The preparation of Trichoderma harzianum and its metabolites includes the following steps:

[0046] Step (1): Inoculate Trichoderma harzianum into a PDB medium for cultivation to obtain a Trichoderma harzianum culture solution.

[0047] Step (2): Inoculate the Trichoderma harzianum culture solution into a solid medium of rice, catecholamine, and trimethoprim for fermentation cultivation. The mass ratio of rice, catecholamine, and trimethoprim is 1:0.08:0.04. The cultivation temperature is 25°C, the humidity is 70%, the pH value is 7.0, and the cultivation time is 7 hours. After cultivation, perform centrifugation at a rotational speed of 3800 r / min for 18 min. After centrifugation, take the supernatant to obtain a fermented bacterial solution.

[0048] Step (3): Perform countercurrent elution on the fermented bacterial solution. The inorganic salt used in the countercurrent elution is magnesium chloride with a mass percentage content of 2.5%, the polar solvent is acetonitrile with a volume percentage content of 45%, and the balance is water. The volume ratio of the fermented bacterial solution to the eluent is 1:1.5. The elution temperature is 23°C, the flow rate of the fermented bacterial solution is 6 mL / min, and the flow rate of the eluent is 23 mL / min. After countercurrent elution, obtain a purified bacterial solution.

[0049] Step (4): Filter the purified bacterial solution, perform vacuum distillation at a pressure of 0.5 MPa and a temperature of 43°C, heat and concentrate, and then dry and spray powder to obtain a finished product powder.

[0050] Experiment 1: Determination of antibacterial activity

[0051] Dilute the finished product powder prepared in Example 3 of the present invention with water, add it to a culture medium to pour plates, inoculate pathogenic bacteria, observe the growth of pathogenic bacteria, record and analyze the growth data of pathogenic bacteria, draw an antibacterial spectrum, and clarify the inhibitory effect of the sample on pathogenic bacteria.

[0052] Test strains: Pathogenic bacteria preserved by Hanxing Biological Laboratory (Pythium myriotylum, Pythium ultimum, Phytophthora capsici, Gibberella zeae, Fusarium oxysporum); Types of culture media: PDA, PDB; Experimental equipment: autoclave, shaker, Erlenmeyer flasks (250 mL), pipettes (100 μL, 1000 μL, 5000 μL), pipette tips (200 μL, 1000 μL), centrifuge tubes (10 mL, 5 mL), disposable spreading rods, disposable petri dishes. First, perform secondary culture and purification (6 - 8 d) on all strains (Pythium myriotylum, Pythium ultimum, Phytophthora capsici, Gibberella zeae, Fusarium oxysporum). Select the edges of colonies with consistent growth of pathogenic bacteria to punch 5 - mm bacterial discs for standby. Set up PDA sample - containing plates with sample dilution multiples of 10, 100, 1000, and 10000 times. Inoculate the pathogenic - bacteria discs into the centers of the sample - containing plates with different sample contents, record and observe the colony growth conditions, analyze the experimental data, calculate the initial bacteriostatic concentration of the sample, and draw the bacteriostatic spectrum.

[0053] Dilute the finished powder prepared in Example 2 by 30, 80, 100, 500, 800, and 1000 times, coat it on PDA plates, observe the growth of colonies on the plates, pick and isolate the strains and inoculate them into PDB medium. Set the shaker at 37 °C and 180 r / min, and take them out for standby after 24 h. Double - streak the obtained target - bacteria liquid 25 mm away from the center of the PDA plate, inoculate a pathogenic - bacteria disc in the center of the plate, and use the plate inoculated with pathogenic bacteria without streaking as the control group. Incubate at a constant temperature of 27 °C for 4 d, record the data, and observe the size and morphology of the pathogenic - bacteria colonies.

[0054] The experimental results show that when the finished powder is diluted 30 times, the growth of pathogenic bacteria is completely inhibited; when diluted 500 times, the inhibitory effect on three strains of pathogenic bacteria can reach 50%; when the dilution multiple is greater than 1000, there is no bacteriostatic effect. The data and analysis results are shown in Tables 1 and 2. Through double - streak confrontation culture, it is observed that the pathogenic bacteria (oomycetes, fungi) are inhibited, but there is no obvious bacteriostatic zone between the target bacteria and oomycetes.

[0055] Table 1 Colony growth data of Sclerotinia sclerotiorum, Gibberella zeae, and Pythium ultimum

[0056]

[0057]

[0058] Table 2 Bacteriostatic concentrations of Sclerotinia sclerotiorum, Gibberella zeae, and Pythium ultimum

[0059] Pathogenic bacteria EC50 EC75 EC95 Pythium ultimum 0.0040 0.0071 0.0165 Sclerotinia sclerotiorum 0.0014 0.0134 0.3441 Gibberella zeae 0.0406 0.0956 0.3282

[0060] Experiment 2: Root - promoting experiments on corn, wheat, and rice

[0061] Field experiments were conducted using water agar to sow maize (maize variety Xianyu 335), wheat (wheat variety Bainong 307), and rice (rice variety Longgeng 31), with 10 plants of each crop planted in each treatment. The treatments set for maize were: the control treatment was normal cultivation CK without adding any reagents, the treatment with coating by Cheng 46# film-forming agent, and the treatment with Cheng 46# film-forming agent plus different doses of the finished product powder (the finished product powder prepared in Example 3), and different doses of the finished product powder were used to wrap the maize roots ( Figure 3 ); the treatments set for wheat were: the control treatment for wheat was normal cultivation CK without adding any reagents, the treatment with Bigasai, and the treatment with Bigasai + different concentrations of the finished product powder ( Figure 4 ); the treatments set for rice were: the control treatment for rice was without adding any reagents, the treatment with Suliang added, and the treatment with Suliang + different concentrations of the finished product powder ( Figure 5 ). Concentration gradients were set, and the details can be seen in Figures 3 - 5 . The annotation meanings marked as 0.1, 0.2, 0.5, 1, 2, 5, and 10 were that the dosages of the coated finished product powder were 0.1 g / 100 Kg, 0.2 g / 100 Kg, 0.5 g / 100 Kg, 1 g / 100 Kg, 2 g / 100 Kg, 5 g / 100 Kg, and 10 g / 100 Kg, Figures 3 - 5 and the digital markings in

[0062] all had this meaning.

[0063] Experiment 3: Growth promotion and disease prevention experiment

[0064] Commercially available peppers, bitter gourds, and peas were selected for field experiments. The finished product powder (the finished product powder prepared in Example 3) at three concentrations, namely diluted 10,000 times, diluted 20,000 times, and diluted 30,000 times, was used to irrigate the roots of peppers, bitter gourds, and peas once, and a control group (without irrigating the roots with the finished product powder and normal cultivation) was set up, and the experimental results were observed 11 days later ( Figures 6 - 7 ).

[0065] The test results show that: the finished product powder was diluted into three concentrations, 10,000 times, 20,000 times, and 30,000 times. After observation and comparison, after the experimental group irrigated the roots of peppers with the finished product powder diluted 20,000 times, the growth of the pepper tops was vigorous, new young leaves grew at the tops, the leaves were green, and they blossomed again, while dead plants appeared in the control group. After the experimental group irrigated the roots of bitter gourds with the finished product powder diluted 30,000 times, the growth of the bitter gourd tops was vigorous, the vines were significantly thick, the leaves were thick and large, and the number of blossoms increased. After the experimental group irrigated the roots of peas with the finished product powder diluted 10,000 times, the root system of the peas developed well and there were no diseases.

[0066] Experiment 4: Rice Salt-alkali Stress Resistance and Adverse Condition Rooting Experiment

[0067] The rice variety Longgeng 31 was selected for a pot experiment. The pH value of the cultivation soil was 8.0 and the EC value was 13. The cultivation lasted for 11 days in total. A total of 9 treatments, namely A, B, C, D, E, F, G, CK1, and CK2, were set up for root irrigation, and the number of root irrigation times was 1 time. Among them, treatment A was the brown algae oligosaccharide diluted 1,000,000 times; treatment B was PHA (polyhydroxyalkanoate) + trehalose 2 g / 100 ml diluted 700 times; treatment C was the finished product powder prepared in Example 3 diluted 50,000 times; treatment D was the finished product powder prepared in Example 3 diluted 200,000 times; treatment E was PHA diluted 700 times; treatment F was PHA + ectoine (8:2) diluted 700 times; treatment G was PHA + ectoine (9:1) diluted 700 times; treatment CK1 was the non-stress control; treatment CK2 was the salt-alkali stress control.

[0068] The test results show that after the roots of rice were irrigated with treatments C and D, the root length of the rice was significantly higher than that of other treatments, indicating that the finished product powder prepared in Example 3 of the method of the present invention diluted 50,000 times and 200,000 times can both enable rice to resist salt-alkali stress and enhance the ability of rice roots to root in adverse conditions.

[0069] The above are only the preferred embodiments of the present invention and do not limit the present invention. Any modifications, equivalent replacements, improvements, etc. made within the spirit and principle of the present invention shall be included within the protection scope of the present invention.

Claims

1. A method for preparing Trichoderma harzianum and its metabolites, characterized in that: The method comprises the following preparation steps: S1. Inoculate Trichoderma harzianum into PDB medium and culture it to obtain Trichoderma harzianum culture solution; S2, inoculating the culture fluid of Trichoderma harzianum into a solid culture medium containing rice, catecholamines and trimethoprim for fermentation, and centrifuging to obtain the supernatant to obtain a fermentation fluid; S3, countercurrently eluting the fermented bacterial liquid to obtain a purified bacterial liquid; S4, filtering the purified bacterial liquid, distilling under reduced pressure, concentrating, drying, and powdering to obtain finished powder.

2. The method for preparing Trichoderma harzianum and its metabolites according to claim 1, characterized in that: Step S2, the mass ratio of rice, catecholamines and trimethoprim is 1:0.05-0.08:0.02-0.04, and the spore production of the solid culture medium after fermentation is greater than 1×10 8 Pieces / mL.

3. The method for preparing Trichoderma harzianum and its metabolites according to claim 1, characterized in that: Step S2, the culture temperature is 24°C-26°C, the humidity is 45%-75%, the pH value is 5.5-7.0, and the culture time is 6-8 hours; the centrifugal speed is 3500-4000r / min, and the centrifugal time is 15-20min.

4. The method for preparing Trichoderma harzianum and its metabolites according to claim 1, characterized in that: Step S3, the inorganic salt used in the countercurrent elution is one of magnesium chloride, sodium chloride, magnesium sulfate, and sodium sulfate; the polar solvent is one of methanol, ethanol, and acetonitrile; the mass percentage of the inorganic salt in the eluent is less than 3%, the volume percentage of the polar solvent is 20%-75%, and the remainder is water; the elution volume ratio of the fermentation liquid to the eluent is 1:1-2.

5. The method for preparing Trichoderma harzianum and its metabolites according to claim 3, characterized in that: Step S3, the elution temperature is 20-25°C, the flow rate of the fermentation liquid is 5-7ml / min, and the flow rate of the eluent is 20-25ml / min. Step S4, the pressure of the reduced pressure distillation is 0.08-1.0MPa, and the temperature is 40°C-45°C.

6. The method for preparing Trichoderma harzianum and its metabolites according to claim 1, characterized in that: Step S4, the ingredients of the finished powder include 6-phosphate-glucose, acetylglucosamine, chitosan, free α-amino acids, β-amino acids, fatty acids, hydroxycinnamic acid, 6-pentyl-α-pyrone, 5,6-dihydro-6-pentyl-2H-pyran-2-one, isonitrile, khatzian lactone, collomycin, trichomycin, khatzianin, and gibberellic acid.

7. Use of Trichoderma harzianum and its metabolites obtained by the preparation method according to any one of claims 1 to 6 in resisting Pythium ulminans, Pythium ultimum, Phytophthora capsici, Gibberella tritici, and Fusarium oxysporum, wherein the dilution multiple of the finished product powder is 30 times.

8. Application of the Trichoderma harzianum and its metabolites prepared by the preparation method according to any one of claims 1 to 6 to promote rooting and seedling growth after coating corn roots, wherein the amount of the finished powder as a coating agent is 0.1g / 100Kg-10g / 100Kg; application of the Trichoderma harzianum metabolites to promote rice germination rate after coating wheat and rice roots, wherein the amount of the finished powder as a coating agent is 0.2g / 100Kg-5g / 100Kg.

9. Use of Trichoderma harzianum and its metabolites obtained by the preparation method according to any one of claims 1 to 6 for rice resistance to salt-alkali stress and adverse rooting, wherein the dilution amount of the finished powder and water is 20,000-200,000 times.

10. Application of Trichoderma harzianum and its metabolites obtained by the preparation method according to any one of claims 1 to 6 in promoting growth and preventing diseases in pepper, bitter melon and pea, wherein the dilution amount of the finished powder and water in pepper planting is 20,000 times, the dilution amount of the finished powder and water in bitter melon planting is 30,000 times, and the dilution amount of the finished powder and water in pea planting is 10,000 times, and the application method is root irrigation.

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