A microbial agent for improving rice yield and quality and preparation method thereof

Abstract

The present invention discloses a microbial agent for improving rice yield and quality and a preparation method thereof, and belongs to the technical field of microbial organic fertilizer. The present invention selects three strains of Trichoderma pseudokoningii, Pseudomonas yamana and Bacillus subtilis with high-efficiency disease resistance and growth promotion functions to form core functional microorganisms, which can produce obvious synergistic effects, show significant inhibitory effects on rice blast pathogens, and have significant disease resistance and growth promotion effects. Secondly, the present invention adds yeast extract, which can effectively promote the colonization of microorganisms and ensure that the microorganisms continue to play an effective role; dextrin, sodium oleate, maltose and nicotinamide are added as microbial promoters, and finally urea, fulvic acid and other substances are added to provide supplementary nutrients for crops, further improving rice yield and quality. Under the joint action of various raw materials of the present invention, rice blast, which is often found in rice, is effectively prevented and controlled, and rice products and quality are greatly improved.

Description

Technical Field

[0001] The invention belongs to the technical field of microbial organic fertilizers, and particularly relates to a microbial agent for improving rice yield and quality and a preparation method thereof. Background Art

[0002] Food is the basic means of survival for human beings. It is a special product related to economic development, social stability and national independence. As one of the most important food crops for human beings, rice is even more important. In the process of modern agricultural production, fertilizers and pesticides are important means of production to increase crop yields.

[0003] In the process of food production, excessive reliance on chemical fertilizers and excessive, blind and unscientific application will not only fail to achieve the goal of increasing production, but also reduce the quality of crops, causing soil fertility decline, farmland degradation, soil compaction and acidification, deterioration of soil microbial community structure, and groundwater and farmland environmental pollution, which is not conducive to the sustainable development of agriculture. Microbial fertilizer is a type of fertilizer developed in recent years. It contains a large number of beneficial active microorganisms, which can provide nutrition to plants and regulate plant growth through the specific activities of microorganisms.

[0004] The application of microbial fertilizer in rice production can not only increase rice yield, but also improve soil fertility, improve rice quality, increase the effective utilization rate of chemical fertilizers, reduce environmental pollution, etc., which is conducive to the development of green agriculture. Plant growth-promoting rhizobacteria (PGPR) are the key components of microbial fertilizers that promote the growth and development of crops, and are also an important source of bacteria. The application of PGPR is widely regarded as a feasible strategy to promote the development of green agriculture. PGPR has the ability to promote plant growth directly or indirectly. Usually, PGPR directly converts nutrients, such as nitrogen fixation, phosphorus solubilization, and potassium solubilization; or secretes metabolites, such as exopolysaccharides, organic acids, 1-aminocyclopropane-1-carboxylic acid (ACC) deaminase, indoleacetic acid, gibberellins, and siderophores for plant absorption and utilization, thereby affecting plant stress response; or indirectly promotes plant growth by helping plants resist the invasion of pathogens such as bacteria and inducing systemic resistance in plants. Therefore, screening PGPR with excellent growth-promoting potential and developing efficient PGPR agents have become one of the current research hotspots at home and abroad.

[0005] For example, Chinese patent application CN202110771488.8 discloses a composite microbial agent, which includes banana stem endophytic fungus Bacillus velezensis wxh1 and soil biocontrol bacteria Bacillus subtiliswxh2. These two strains were screened by the applicant from the rhizosphere soil of bananas. Identification showed that strain wxh1 and strain wxh2 both have the effect of inhibiting the growth of Foc1 and Foc4 Fusarium oxysporum, and can effectively prevent and control banana wilt disease and accelerate the fruiting time of bananas. The above two microbial agents are prepared into a fermentation agent and added to organic fertilizer, which can improve the disease resistance of bananas to Fusarium wilt.

[0006] For example, Chinese patent application CN202211240583.6 relates to a microbial agent, a biological organic fertilizer and a preparation method thereof, wherein a microbial agent is obtained by extracting microbial colonies different from those in CIS soil samples and / or CIP soil samples and CK soil samples, and the microbial agent includes 1 to 2 parts of Bacillus subtilis, 1 part of Bacillus amyloliquefaciens, 1 to 2 parts of Pseudomonas, 1 to 2 parts of Trichoderma harzianum and 1 part of Mortierella alpina. The microbial agent can effectively solve the problem of continuous cropping in the process of celery cultivation, without the need for intercropping or crop rotation with other crops, and avoids the decrease in celery yield due to intercropping or crop rotation.

[0007] However, most of the current microbial agents are independently isolated or used in combination with a variety of existing mature strains. They have broad-spectrum growth-promoting properties, but poor targeting, single function, and difficulty in stably colonizing in the soil root system, which leads to a decrease in application effect and an increase in cost. In actual agricultural production, it is difficult to be effectively promoted and applied. Summary of the invention

[0008] The present invention aims to solve the problems existing in the prior art and provides a growth-promoting microbial flora for rice, which has a growth-promoting function and a significant prevention and control effect on rice blast disease, and has good colonization ability, and can continuously and effectively exert the effect of disease resistance and growth promotion.

[0009] In order to achieve the above technical objectives, the technical solution adopted by the present invention is:

[0010] A microbial agent for improving rice yield and quality, comprising disease-preventing and growth-promoting microbial liquid, yeast extract, microbial promoter and organic and inorganic nutrients, wherein the mass ratio of the four is 1:(0.5-1):(0.5-0.8):(50-100).

[0011] Furthermore, the disease prevention and growth promotion microbial bacterial liquid includes Trichoderma pseudokoningii ( Trichoderma koningiopsis ), Pseudomonas yamana (Pseudomonas yamanorum ) and Bacillus subtilis ( Bacillus subtilis ), the volume ratio of the three is 1:1:1.

[0012] Furthermore, the strain number of the Trichoderma koningii is CGMCC No.5.1243, purchased from the General Microbiology Center of the China National Microbiological Culture Collection Administration; the strain number of the Pseudomonas yamana is CGMCC No.1.16088, purchased from the General Microbiology Center of the China National Microbiological Culture Collection Administration; the strain number of the Bacillus subtilis is CGMCC No.1.7740, purchased from the General Microbiology Center of the China National Microbiological Culture Collection Administration.

[0013] The strains used in the present invention can be purchased through the General Microbiological Center of the China Microbiological Culture Collection Administration, without the need for repeated biological preservation.

[0014] Furthermore, the preparation method of the disease prevention and growth promotion microbial bacterial liquid is as follows: single colonies of Trichoderma pseudokoningii, Pseudomonas yamana and Bacillus subtilis are taken respectively, inoculated into 100 mL of LB culture medium, and placed in a constant temperature shaking shaker at 25-30°C and 180-200r / min for 24 hours to obtain seed liquid of the three bacteria; the seed liquid of Trichoderma pseudokoningii, Pseudomonas yamana and Bacillus subtilis is transferred into 500 mL of LB culture medium at a transfer rate of 1%, and cultured at 25-30°C and 180-200r / min for 48-56 hours, and the effective viable count is tested to be (1-2)×10 8 CUF / mL, the culture was terminated to obtain the fermentation broth of the three bacteria, and the three fermentation broths were mixed in a volume ratio of 1:1:1 to obtain the disease-preventing and growth-promoting microbial liquid.

[0015] Furthermore, the method for obtaining single colonies of Trichoderma koningii, Pseudomonas yamana and Bacillus subtilis is as follows: take out the target strain stored in cryopreservation, use an inoculation loop to dip a small amount of bacterial liquid and streak it on LB solid culture medium, culture it at 25-30°C for 10-12h to complete the strain activation and obtain a single colony.

[0016] Furthermore, the preparation method of the yeast extract is as follows: using active yeast as a raw material, adding the active yeast to deionized water to prepare an active yeast solution with a mass concentration of 10-20%; then adding a zinc compound to the active yeast solution according to a solid-liquid ratio of 10g:1L, and then stirring at 50°C for 2 hours, then adjusting the pH of the solution to 6-7, and standing for 1-2 hours; then adding 0.1-0.15% of the weight of yeast wall-breaking enzyme to the mixed solution, and treating it at 45°C for 3 hours; after being treated with the yeast wall-breaking enzyme, adding 0.03% of the weight of neutral protease per liter of solution, and treating it at 45°C for 3 hours to obtain a yeast extract.

[0017] Furthermore, the enzymatic activity of the yeast cell wall-breaking enzyme is 30,000 U / g, the enzymatic activity of the neutral protease is 100,000 U / g, and the zinc compound can be a water-soluble zinc-containing compound, such as zinc nitrate, zinc chloride, zinc sulfate, and zinc acetate.

[0018] The active yeast can be commercially available active dry yeast. The embodiment of the present invention uses beer high-activity dry yeast produced by Jiangsu Yuanzheng Biotechnology Co., Ltd.

[0019] Furthermore, the microbial promoter is obtained by mixing dextrin, sodium oleate, maltose and nicotinamide in a mass ratio of 1:1:10:0.1.

[0020] Furthermore, the organic and inorganic nutrients include, by weight: 10-15 parts of urea, 5-10 parts of dipotassium hydrogen phosphate, 2-7 parts of calcium chloride, 3-5 parts of zinc sulfate, 12-16 parts of sucrose, 10-12 parts of fulvic acid, and 50-80 parts of water.

[0021] A method for preparing a microbial agent for improving rice yield and quality comprises the following preparation steps:

[0022] (1) preparing disease prevention and growth promotion microbial liquid;

[0023] (2) preparing yeast extract;

[0024] (3) The disease prevention and growth promotion microbial liquid, yeast extract, microbial promoter and organic and inorganic nutrients are fully mixed in proportion to obtain the final product microbial agent.

[0025] The method of using the microbial agent of the present invention is as follows: at the young fruit stage, dilute the microbial agent 50 times for root irrigation treatment, about 50-100 mL per plant; spray at the tillering stage and heading stage of rice, respectively, every 667m 2 Apply 5L each time and dilute 50 times before use.

[0026] Beneficial effects:

[0027] (1) The present invention screens three strains of Trichoderma pseudokoningii, Pseudomonas yamana and Bacillus subtilis with high-efficiency disease resistance and growth promotion functions to form core functional microorganisms. Among them, Trichoderma pseudokoningii can secrete a variety of biologically active substances, such as gliotoxin, etc. These substances can directly kill rice blast fungi or interfere with their normal physiological activities, thereby inhibiting the development of diseases; Pseudomonas yamana can not only directly inhibit pathogens, but also enhance the disease resistance of plants by promoting defense reactions in plants. At the same time, Pseudomonas yamana regulates plant hormone levels by producing IAA and other secondary metabolites, thereby promoting plant growth. The screened Bacillus subtilis can produce a variety of physiologically active substances and amino acids such as gibberellins, indoleacetic acid, cytokinins, etc., promote plant growth, and enhance plant disease resistance. In addition, it can also improve the effectiveness of soil nutrients through biological nitrogen fixation, phosphorus dissolution, potassium dissolution, etc., thereby indirectly enhancing the activity of other microorganisms. The simultaneous use of the three can produce obvious synergistic effects, showing a significant inhibitory effect on the rice blast pathogen, and having significant disease resistance and growth-promoting effects.

[0028] (2) Secondly, the present invention adds yeast extract. Yeast cells and their metabolites contain a variety of trace elements and bioactive substances, such as amino acids, vitamins, peptides, etc. The addition of yeast extract can effectively promote the growth of microorganisms. The added zinc compound can, on the one hand, improve the efficiency of enzymatic hydrolysis and promote the efficient dissolution of nutrients. On the other hand, when used together with microorganisms, it can effectively promote the colonization of microorganisms and ensure that the microorganisms continue to play an effective role.

[0029] (3) Add dextrin, sodium oleate, maltose and nicotinamide as microbial promoters. Dextrin, as a carbon source, can provide energy and nutrition for microorganisms. Sodium oleate helps to improve the activity and diversity of microorganisms in the soil. In addition to providing energy for microorganisms, maltose can also promote microbial colonization. Nicotinamide can promote the growth and activity of beneficial soil microorganisms, including directly increasing soil enzyme activity, affecting plant gene expression and thus changing the structure of microbial communities, and participating in important metabolic pathways, thereby providing more nutrients and energy sources for the soil ecosystem.

[0030] (4) Finally, urea, fulvic acid and other substances are added to provide supplementary nutrients for crops, further improving rice yield and quality. Under the joint action of various raw materials of the present invention, rice blast disease, which is common in rice, is effectively prevented and controlled, and rice products and quality are greatly improved. BRIEF DESCRIPTION OF THE DRAWINGS

[0031] Figure 1 The antagonistic effect experimental diagram of the three strains of Trichoderma pseudokoningii, Pseudomonas yamana and Bacillus subtilis of the present invention;

[0032] Figure 2The antibacterial effect diagram of the embodiments of the present invention and the comparative examples;

[0033] Figure 3 It is a colonization experiment result diagram of the embodiment of the present invention and the comparative example. DETAILED DESCRIPTION

[0034] The technical solution of the present invention is further described below in conjunction with specific embodiments, but is not limited thereto.

[0035] Example 1

[0036] A microbial agent for improving rice yield and quality, comprising disease-preventing and growth-promoting microbial liquid, yeast extract, microbial promoter and organic and inorganic nutrients, wherein the mass ratio of the four is 1:0.5:0.5:100.

[0037] The disease prevention and growth promotion microbial bacterial liquid includes Trichoderma pseudokoningii ( Trichoderma koningiopsis ), Pseudomonas yamana ( Pseudomonas yamanorum ) and Bacillus subtilis ( Bacillus subtilis ), the volume ratio of the three is 1:1:1.

[0038] The strain number of the Trichoderma koningii is CGMCC No.5.1243, purchased from the General Microbiology Center of the China National Microbiological Culture Collection Administration; the strain number of the Pseudomonas yamana is CGMCC No.1.16088, purchased from the General Microbiology Center of the China National Microbiological Culture Collection Administration; the strain number of the Bacillus subtilis is CGMCC No.1.7740, purchased from the General Microbiology Center of the China National Microbiological Culture Collection Administration.

[0039] The method for preparing the disease prevention and growth promotion microbial bacterial liquid is as follows: single colonies of Trichoderma pseudokoningii, Pseudomonas yamana and Bacillus subtilis are taken respectively, inoculated into 100 mL of LB culture medium respectively, and placed in a constant temperature shaking shaker at 25-30° C. and 180 r / min for culturing for 24 hours to obtain seed liquids of the three bacteria; the seed liquids of Trichoderma pseudokoningii, Pseudomonas yamana and Bacillus subtilis are transferred into 500 mL of LB culture medium at a transfer rate of 1%, and cultured at 25-30° C. and 180 r / min for 48-56 hours, and the effective viable bacteria count is tested to be (1-2)×10 8 CUF / mL, the culture was terminated to obtain the fermentation broth of the three bacteria, and the three fermentation broths were mixed in a volume ratio of 1:1:1 to obtain the disease-preventing and growth-promoting microbial liquid.

[0040] The method for obtaining single colonies of Trichoderma pseudokoningii, Pseudomonas yamana and Bacillus subtilis is as follows: take out the target strain stored in cryopreservation, use an inoculation loop to dip a small amount of bacterial liquid and streak it on LB solid culture medium, and culture it at 25-30°C for 10 hours to complete the strain activation and obtain a single colony.

[0041] The preparation method of the yeast extract is as follows: using active yeast as a raw material, adding the active yeast into deionized water to prepare an active yeast solution with a mass concentration of 10%; then adding a zinc compound to the active yeast solution according to a solid-liquid ratio of 10g:1L, and then stirring at 50°C for 2 hours, then adjusting the pH of the solution to 6-7, and standing for 1 hour; then adding 0.1% by weight of yeast wall-breaking enzyme to the mixed solution, and treating it at 45°C for 3 hours; after being treated with the yeast wall-breaking enzyme, adding 0.03% by weight of neutral protease per liter of solution, and treating it at 45°C for 3 hours to obtain the yeast extract.

[0042] The enzymatic activity of the yeast cell wall-breaking enzyme is 30,000 U / g, and the enzymatic activity of the neutral protease is 100,000 U / g.

[0043] The active yeast can be commercially available active dry yeast. In this embodiment, the active dry yeast used is beer high-activity dry yeast produced by Jiangsu Yuanzheng Biotechnology Co., Ltd.

[0044] The microbial promoter is obtained by mixing dextrin, sodium oleate, maltose and nicotinamide in a mass ratio of 1:1:10:0.1.

[0045] The organic and inorganic nutrients include, by weight: 10 parts of urea, 5 parts of dipotassium hydrogen phosphate, 2 parts of calcium chloride, 3 parts of zinc sulfate, 12 parts of sucrose, 10 parts of fulvic acid, and 80 parts of water.

[0046] A method for preparing a microbial agent for improving rice yield and quality comprises the following preparation steps:

[0047] (1) preparing disease prevention and growth promotion microbial bacterial liquid;

[0048] (2) preparing yeast extract;

[0049] (3) The disease prevention and growth promotion microbial liquid, yeast extract, microbial promoter and organic and inorganic nutrients are fully mixed in proportion to obtain the final product microbial agent.

[0050] Example 2

[0051] A microbial agent for improving rice yield and quality, the microbial agent comprising disease-preventing and growth-promoting microbial liquid, yeast extract, microbial promoter and organic and inorganic nutrients, the mass ratio of the four being 1:0.6:0.7:75.

[0052] The disease prevention and growth promotion microbial bacterial liquid includes Trichoderma pseudokoningii ( Trichoderma koningiopsis ), Pseudomonas yamana ( Pseudomonas yamanorum ) and Bacillus subtilis ( Bacillus subtilis), the volume ratio of the three is 1:1:1.

[0053] The strain number of the Trichoderma koningii is CGMCC No.5.1243, purchased from the General Microbiology Center of the China National Microbiological Culture Collection Administration; the strain number of the Pseudomonas yamana is CGMCC No.1.16088, purchased from the General Microbiology Center of the China National Microbiological Culture Collection Administration; the strain number of the Bacillus subtilis is CGMCC No.1.7740, purchased from the General Microbiology Center of the China National Microbiological Culture Collection Administration.

[0054] The method for preparing the disease prevention and growth promotion microbial bacterial liquid is as follows: single colonies of Trichoderma pseudokoningii, Pseudomonas yamana and Bacillus subtilis are taken respectively, inoculated into 100 mL of LB culture medium respectively, and placed in a constant temperature shaking shaker at 25-30°C and 200 r / min for culturing for 24 hours to obtain seed liquids of the three bacteria; the seed liquids of Trichoderma pseudokoningii, Pseudomonas yamana and Bacillus subtilis are transferred into 500 mL of LB culture medium at a transfer rate of 1%, and cultured at 25-30°C and 180 r / min for 48-56 hours, and the number of effective live bacteria tested is (1-2)×10 8 CUF / mL, the culture was terminated to obtain the fermentation broth of the three bacteria, and the three fermentation broths were mixed in a volume ratio of 1:1:1 to obtain the disease-preventing and growth-promoting microbial liquid.

[0055] The method for obtaining single colonies of Trichoderma pseudokoningii, Pseudomonas yamana and Bacillus subtilis is as follows: take out the target strain stored in cryopreservation, use an inoculation loop to dip a small amount of bacterial liquid and streak it on LB solid culture medium, culture it at 25-30°C for 11 hours to complete the strain activation and obtain a single colony.

[0056] The preparation method of the yeast extract is as follows: using active yeast as a raw material, adding the active yeast into deionized water to prepare an active yeast solution with a mass concentration of 15%; then adding a zinc compound to the active yeast solution according to a solid-liquid ratio of 10g:1L, and then stirring at 50°C for 2 hours, then adjusting the pH of the solution to 6-7, and standing for 2 hours; then adding 0.1% by weight of yeast wall-breaking enzyme to the mixed solution, and treating it at 45°C for 3 hours; after being treated with the yeast wall-breaking enzyme, adding 0.03% by weight of neutral protease per liter of solution, and treating it at 45°C for 3 hours to obtain the yeast extract.

[0057] The enzymatic activity of the yeast cell wall-breaking enzyme is 30,000 U / g, and the enzymatic activity of the neutral protease is 100,000 U / g.

[0058] The active yeast can be commercially available active dry yeast. In this embodiment, the active dry yeast used is beer high-activity dry yeast produced by Jiangsu Yuanzheng Biotechnology Co., Ltd.

[0059] The microbial promoter is obtained by mixing dextrin, sodium oleate, maltose and nicotinamide in a mass ratio of 1:1:10:0.1.

[0060] The organic and inorganic nutrients include, by weight: 11 parts of urea, 8 parts of dipotassium hydrogen phosphate, 5 parts of calcium chloride, 4 parts of zinc sulfate, 14 parts of sucrose, 11 parts of fulvic acid, and 60 parts of water.

[0061] A method for preparing a microbial agent for improving rice yield and quality comprises the following preparation steps:

[0062] (1) preparing disease prevention and growth promotion microbial liquid;

[0063] (2) preparing yeast extract;

[0064] (3) The disease prevention and growth promotion microbial liquid, yeast extract, microbial promoter and organic and inorganic nutrients are fully mixed in proportion to obtain the final product microbial agent.

[0065] Example 3

[0066] A microbial agent for improving rice yield and quality, the microbial agent comprising disease-preventing and growth-promoting microbial liquid, yeast extract, microbial promoter and organic and inorganic nutrients, the mass ratio of the four being 1:1:0.8:50.

[0067] The disease prevention and growth promotion microbial bacterial liquid includes Trichoderma pseudokoningii ( Trichoderma koningiopsis ), Pseudomonas yamana ( Pseudomonas yamanorum ) and Bacillus subtilis ( Bacillus subtilis ), the volume ratio of the three is 1:1:1.

[0068] The strain number of the Trichoderma koningii is CGMCC No.5.1243, purchased from the General Microbiology Center of the China National Microbiological Culture Collection Administration; the strain number of the Pseudomonas yamana is CGMCC No.1.16088, purchased from the General Microbiology Center of the China National Microbiological Culture Collection Administration; the strain number of the Bacillus subtilis is CGMCC No.1.7740, purchased from the General Microbiology Center of the China National Microbiological Culture Collection Administration.

[0069] The method for preparing the disease prevention and growth promotion microbial bacterial liquid is as follows: taking single colonies of Trichoderma pseudokoningii, Pseudomonas yamana and Bacillus subtilis respectively, inoculating them into 100 mL of LB culture medium respectively, placing them in a constant temperature shaking shaker at 25-30°C and 200 r / min for 24 hours, and obtaining seed liquids of the three bacteria; transferring the seed liquids of Trichoderma pseudokoningii, Pseudomonas yamana and Bacillus subtilis into 500 mL of LB culture medium at a transfer rate of 1%, culturing them at 25-30°C and 200 r / min for 48-56 hours, and testing the effective viable count to be (1-2)×10 8 CUF / mL, the culture was terminated to obtain the fermentation broth of the three bacteria, and the three fermentation broths were mixed in a volume ratio of 1:1:1 to obtain the disease-preventing and growth-promoting microbial liquid.

[0070] The method for obtaining single colonies of Trichoderma pseudokoningii, Pseudomonas yamana and Bacillus subtilis is as follows: take out the target strain stored in cryopreservation, use an inoculation loop to dip a small amount of bacterial liquid and streak it on LB solid culture medium, and culture it at 25-30°C for 12 hours to complete the strain activation and obtain a single colony.

[0071] The preparation method of the yeast extract is as follows: using active yeast as a raw material, adding the active yeast into deionized water to prepare an active yeast solution with a mass concentration of 20%; then adding a zinc compound to the active yeast solution according to a solid-liquid ratio of 10g:1L, and then stirring at 50°C for 2 hours, then adjusting the pH of the solution to 6-7, and standing for 2 hours; then adding 0.15% of the weight of yeast wall-breaking enzyme to the mixed solution, and treating it at 45°C for 3 hours; after being treated with the yeast wall-breaking enzyme, adding 0.03% of the weight of neutral protease per liter of solution, and treating it at 45°C for 3 hours to obtain the yeast extract.

[0072] The enzymatic activity of the yeast cell wall-breaking enzyme is 30,000 U / g, and the enzymatic activity of the neutral protease is 100,000 U / g.

[0073] The active yeast can be commercially available active dry yeast. The embodiment of the present invention uses beer high-activity dry yeast produced by Jiangsu Yuanzheng Biotechnology Co., Ltd.

[0074] The microbial promoter is obtained by mixing dextrin, sodium oleate, maltose and nicotinamide in a mass ratio of 1:1:10:0.1.

[0075] The organic and inorganic nutrients include, by weight: 15 parts of urea, 10 parts of dipotassium hydrogen phosphate, 7 parts of calcium chloride, 5 parts of zinc sulfate, 16 parts of sucrose, 12 parts of fulvic acid, and 50 parts of water.

[0076] A method for preparing a microbial agent for improving rice yield and quality comprises the following preparation steps:

[0077] (1) preparing disease prevention and growth promotion microbial liquid;

[0078] (2) preparing yeast extract;

[0079] (3) The disease prevention and growth promotion microbial liquid, yeast extract, microbial promoter and organic and inorganic nutrients are fully mixed in proportion to obtain the final product microbial agent.

[0080] Comparative Example 1

[0081] A microbial agent for improving rice yield and quality, comprising disease-preventing and growth-promoting microbial liquid, yeast extract, microbial promoter and organic and inorganic nutrients, wherein the mass ratio of the four is 1:0.5:0.5:100.

[0082] The method for preparing the disease prevention and growth promotion microbial bacterial liquid is as follows: a single colony of Trichoderma pseudokoningii is inoculated into 100 mL of LB culture medium, and the culture is placed in a constant temperature shaking shaker at 25-30° C. and 180 r / min for 24 hours to obtain a seed liquid; the seed liquid of Trichoderma pseudokoningii is transferred into 500 mL of LB culture medium at a transplanting amount of 1%, and the culture is carried out at 25-30° C. and 180 r / min for 48-56 hours, and the effective viable bacterial count is tested to be (1-2)×10 8 CUF / mL, the culture is terminated to obtain the fermentation liquid, which is the disease prevention and growth promotion microbial liquid. The strain number of the Trichoderma pseudokoningii is CGMCC No.5.1243, which was purchased from the General Microbiological Center of China National Microbiological Culture Collection Committee.

[0083] In this comparative example, except that only Trichoderma pseudokoningii was used in the disease prevention and growth promotion microbial bacterial liquid, the other raw materials and preparation methods were the same as those in Example 1.

[0084] Comparative Example 2

[0085] A microbial agent for improving rice yield and quality, comprising disease-preventing and growth-promoting microbial liquid, yeast extract, microbial promoter and organic and inorganic nutrients, wherein the mass ratio of the four is 1:0.5:0.5:100.

[0086] The preparation method of the disease prevention and growth promotion microbial bacterial liquid is as follows: a single colony of Pseudomonas yamana is inoculated into 100 mL of LB culture medium, and the culture is placed in a constant temperature shaking shaker at 25-30° C. and 180 r / min for 24 hours to obtain a seed liquid; the seed liquid of Pseudomonas yamana is transferred into 500 mL of LB culture medium at a transfer rate of 1%, and the culture is carried out at 25-30° C. and 180 r / min for 48-56 hours, and the effective viable count is tested to be (1-2)×10 8CUF / mL, the culture is terminated to obtain the fermentation liquid, which is the disease prevention and growth promotion microbial liquid. The strain number of the Pseudomonas yamana is CGMCC No.1.16088, which was purchased from the General Microbiology Center of China Microbiological Culture Collection Administration.

[0087] In this comparative example, except that only Pseudomonas yamanensis was used in the disease prevention and growth promotion microbial liquid, the other raw materials and preparation method were the same as those in Example 1.

[0088] Comparative Example 3

[0089] A microbial agent for improving rice yield and quality, comprising disease-preventing and growth-promoting microbial liquid, yeast extract, microbial promoter and organic and inorganic nutrients, wherein the mass ratio of the four is 1:0.5:0.5:100.

[0090] The preparation method of the disease prevention and growth promotion microbial bacterial liquid is as follows: a single colony of Bacillus subtilis is taken, inoculated into 100 mL of LB culture medium, placed in a constant temperature shaking shaker at 25-30° C. and 180 r / min for culturing for 24 hours to obtain a seed liquid; the seed liquid of Bacillus subtilis is transferred into 500 mL of LB culture medium at a transfer rate of 1%, and cultured at 25-30° C. and 180 r / min for 48-56 hours, and the effective viable bacterial count is tested to be (1-2)×10 8 CUF / mL, the culture was terminated to obtain the fermentation liquid, which was the disease prevention and growth promotion microbial liquid. The strain number of the Bacillus subtilis was CGMCC No.1.7740, purchased from the General Microbiological Center of the China Microbiological Culture Collection Administration

[0091] In this comparative example, except that only Bacillus subtilis was used in the disease prevention and growth promotion microbial liquid, the other raw materials and preparation method were the same as those in Example 1.

[0092] Comparative Example 4

[0093] A microbial agent for improving rice yield and quality, comprising disease-preventing and growth-promoting microbial liquid, yeast extract, microbial promoter and organic and inorganic nutrients, wherein the mass ratio of the four is 1:0.5:0.5:100.

[0094] The disease-preventing and growth-promoting microbial bacterial liquid includes Trichoderma koningiopsis and Pseudomonas yamanorum in a volume ratio of 1:1.

[0095] The strain number of the Trichoderma pseudokoningii is CGMCC No.5.1243, which was purchased from the General Microbiology Center of China National Microbiological Culture Collection Administration; the strain number of the Pseudomonas yamana is CGMCC No.1.16088, which was purchased from the General Microbiology Center of China National Microbiological Culture Collection Administration.

[0096] The method for preparing the disease prevention and growth promotion microbial bacterial liquid is as follows: single colonies of Trichoderma pseudokoningii and Pseudomonas yamana are taken respectively, inoculated into 100 mL of LB culture medium respectively, and placed in a constant temperature shaking shaker at 25-30° C. and 180 r / min for 24 hours to obtain seed liquid of the two bacteria; Trichoderma pseudokoningii and Pseudomonas yamana are transferred into 500 mL of LB culture medium at a 1% transfer rate, and cultured at 25-30° C. and 180 r / min for 48-56 hours, and the effective viable bacteria count is tested to be (1-2)×10 8 CUF / mL, the culture was terminated to obtain the fermentation broth of the two bacteria, and the two fermentation broths were mixed in a volume ratio of 1:1 to obtain the disease-preventing and growth-promoting microbial bacterial liquid.

[0097] In this comparative example, except that only Trichoderma pseudokoningii and Pseudomonas yamana were used in the disease prevention and growth promotion microbial bacterial liquid, the other raw materials and preparation methods were the same as those in Example 1.

[0098] Comparative Example 5

[0099] A microbial agent for improving rice yield and quality, comprising disease-preventing and growth-promoting microbial liquid, yeast extract, microbial promoter and organic and inorganic nutrients, wherein the mass ratio of the four is 1:0.5:0.5:100.

[0100] The disease prevention and growth promotion microbial bacterial liquid includes Trichoderma pseudokoningii ( Trichoderma koningiopsis ) and Bacillus subtilis ( Bacillus subtilis ), the volume ratio of the two is 1:1.

[0101] The strain number of the Trichoderma pseudokoningii is CGMCC No.5.1243, which was purchased from the General Microbiology Center of China National Microbiological Culture Collection Administration; the strain number of the Bacillus subtilis is CGMCC No.1.7740, which was purchased from the General Microbiology Center of China National Microbiological Culture Collection Administration.

[0102] The method for preparing the disease prevention and growth promotion microbial bacterial liquid is as follows: single colonies of Trichoderma pseudokoningii and Bacillus subtilis are taken respectively, inoculated into 100 mL of LB culture medium respectively, and placed in a constant temperature shaking shaker at 25-30°C and 180 r / min for 24 hours to obtain seed liquids of the two bacteria; the seed liquids of Trichoderma pseudokoningii and Bacillus subtilis are transferred into 500 mL of LB culture medium at a transfer rate of 1%, and cultured at 25-30°C and 180 r / min for 48-56 hours, and the effective viable bacterial count is tested to be (1-2)×10 8 CUF / mL, the culture was terminated to obtain the fermentation broth of the two bacteria, and the two fermentation broths were mixed in a volume ratio of 1:1 to obtain the disease-preventing and growth-promoting microbial bacterial liquid.

[0103] In this comparative example, except that only Trichoderma pseudokoningii and Bacillus subtilis were used in the disease prevention and growth promotion microbial bacterial liquid, the other raw materials and preparation methods were the same as those in Example 1.

[0104] Comparative Example 6

[0105] A microbial agent for improving rice yield and quality, comprising disease-preventing and growth-promoting microbial liquid, yeast extract, microbial promoter and organic and inorganic nutrients, wherein the mass ratio of the four is 1:0.5:0.5:100.

[0106] The disease-preventing and growth-promoting microbial bacterial liquid includes Pseudomonas yamana ( Pseudomonas yamanorum ) and Bacillus subtilis ( Bacillus subtilis ), the volume ratio of the two is 1:1.

[0107] The strain number of the Pseudomonas yamana is CGMCC No.1.16088, which was purchased from the General Microbiology Center of the China National Microbiological Culture Collection Administration; the strain number of the Bacillus subtilis is CGMCC No.1.7740, which was purchased from the General Microbiology Center of the China National Microbiological Culture Collection Administration.

[0108] The preparation method of the disease prevention and growth promotion microbial bacterial liquid is as follows: single colonies of Pseudomonas yamana and Bacillus subtilis are taken respectively, inoculated into 100 mL of LB culture medium respectively, and placed in a constant temperature shaking shaker at 25-30°C and 180 r / min for 24 hours to obtain seed liquid of the two bacteria; the seed liquid of Pseudomonas yamana and Bacillus subtilis are transferred into 500 mL of LB culture medium at a transfer rate of 1%, and cultured at 25-30°C and 180 r / min for 48-56 hours, and the effective viable bacteria count is tested to be (1-2)×10 8 CUF / mL, the culture was terminated to obtain the fermentation broth of the two bacteria, and the two fermentation broths were mixed in a volume ratio of 1:1 to obtain the disease-preventing and growth-promoting microbial bacterial liquid.

[0109] In this comparative example, except that only Pseudomonas yamanensis and Bacillus subtilis were used in the disease prevention and growth promotion microbial liquid, the other raw materials and preparation methods were the same as those in Example 1.

[0110] Comparative Example 7

[0111] A microbial agent for improving rice yield and quality, comprising disease-preventing and growth-promoting microbial liquid, yeast extract, microbial promoter and organic and inorganic nutrients, wherein the mass ratio of the four is 1:0.5:0.5:100.

[0112] The disease prevention and growth promotion microbial bacterial liquid includes Trichoderma pseudokoningii ( Trichoderma koningiopsis ), Pseudomonas yamana ( Pseudomonas yamanorum ) and Bacillus subtilis ( Bacillus subtilis ), the volume ratio of the three is 2:1:1.

[0113] The strain number of the Trichoderma koningii is CGMCC No.5.1243, purchased from the General Microbiology Center of the China National Microbiological Culture Collection Administration; the strain number of the Pseudomonas yamana is CGMCC No.1.16088, purchased from the General Microbiology Center of the China National Microbiological Culture Collection Administration; the strain number of the Bacillus subtilis is CGMCC No.1.7740, purchased from the General Microbiology Center of the China National Microbiological Culture Collection Administration.

[0114] The method for preparing the disease prevention and growth promotion microbial bacterial liquid is as follows: single colonies of Trichoderma pseudokoningii, Pseudomonas yamana and Bacillus subtilis are taken respectively, inoculated into 100 mL of LB culture medium respectively, and placed in a constant temperature shaking shaker at 25-30° C. and 180 r / min for culturing for 24 hours to obtain seed liquids of the three bacteria; the seed liquids of Trichoderma pseudokoningii, Pseudomonas yamana and Bacillus subtilis are transferred into 500 mL of LB culture medium at a transfer rate of 1%, and cultured at 25-30° C. and 180 r / min for 48-56 hours, and the effective viable bacteria count is tested to be (1-2)×10 8 CUF / mL, the culture was terminated to obtain the fermentation broth of the three bacteria, and the three fermentation broths were mixed in a volume ratio of 2:1:1 to obtain the disease-preventing and growth-promoting microbial liquid.

[0115] In this comparative example, except that the volume ratio of Trichoderma pseudokoningii, Pseudomonas yamana and Bacillus subtilis in the disease prevention and growth promotion microbial liquid is changed to 2:1:1, the other raw materials and preparation method are the same as those in Example 1.

[0116] Comparative Example 8

[0117] A microbial agent for improving rice yield and quality, comprising disease-preventing and growth-promoting microbial liquid, yeast extract, microbial promoter and organic and inorganic nutrients, wherein the mass ratio of the four is 1:0.5:0.5:100.

[0118] The disease prevention and growth promotion microbial bacterial liquid includes Trichoderma pseudokoningii ( Trichoderma koningiopsis ), Pseudomonas yamana ( Pseudomonas yamanorum ) and Bacillus subtilis ( Bacillus subtilis ), the volume ratio of the three is 1:2:1.

[0119] The strain number of the Trichoderma koningii is CGMCC No.5.1243, purchased from the General Microbiology Center of the China National Microbiological Culture Collection Administration; the strain number of the Pseudomonas yamana is CGMCC No.1.16088, purchased from the General Microbiology Center of the China National Microbiological Culture Collection Administration; the strain number of the Bacillus subtilis is CGMCC No.1.7740, purchased from the General Microbiology Center of the China National Microbiological Culture Collection Administration.

[0120] The method for preparing the disease prevention and growth promotion microbial bacterial liquid is as follows: single colonies of Trichoderma pseudokoningii, Pseudomonas yamana and Bacillus subtilis are taken respectively, inoculated into 100 mL of LB culture medium respectively, and placed in a constant temperature shaking shaker at 25-30° C. and 180 r / min for culturing for 24 hours to obtain seed liquids of the three bacteria; the seed liquids of Trichoderma pseudokoningii, Pseudomonas yamana and Bacillus subtilis are transferred into 500 mL of LB culture medium at a transfer rate of 1%, and cultured at 25-30° C. and 180 r / min for 48-56 hours, and the effective viable bacteria count is tested to be (1-2)×10 8 CUF / mL, the culture was terminated to obtain the fermentation broth of the three bacteria, and the three fermentation broths were mixed in a volume ratio of 1:2:1 to obtain the disease-preventing and growth-promoting microbial liquid.

[0121] In this comparative example, except that the volume ratio of Trichoderma pseudokoningii, Pseudomonas yamana and Bacillus subtilis in the disease prevention and growth promotion microbial bacterial liquid is changed to 1:2:1, the other raw materials and preparation method are the same as those in Example 1.

[0122] Comparative Example 9

[0123] A microbial agent for improving rice yield and quality, comprising disease-preventing and growth-promoting microbial liquid, yeast extract, microbial promoter and organic and inorganic nutrients, wherein the mass ratio of the four is 1:0.5:0.5:100.

[0124] The disease prevention and growth promotion microbial bacterial liquid includes Trichoderma pseudokoningii ( Trichoderma koningiopsis ), Pseudomonas yamana ( Pseudomonas yamanorum ) and Bacillus subtilis ( Bacillus subtilis), the volume ratio of the three is 1:1:2.

[0125] The strain number of the Trichoderma koningii is CGMCC No.5.1243, purchased from the General Microbiology Center of the China National Microbiological Culture Collection Administration; the strain number of the Pseudomonas yamana is CGMCC No.1.16088, purchased from the General Microbiology Center of the China National Microbiological Culture Collection Administration; the strain number of the Bacillus subtilis is CGMCC No.1.7740, purchased from the General Microbiology Center of the China National Microbiological Culture Collection Administration.

[0126] The method for preparing the disease prevention and growth promotion microbial bacterial liquid is as follows: single colonies of Trichoderma pseudokoningii, Pseudomonas yamana and Bacillus subtilis are taken respectively, inoculated into 100 mL of LB culture medium respectively, and placed in a constant temperature shaking shaker at 25-30° C. and 180 r / min for culturing for 24 hours to obtain seed liquids of the three bacteria; the seed liquids of Trichoderma pseudokoningii, Pseudomonas yamana and Bacillus subtilis are transferred into 500 mL of LB culture medium at a transfer rate of 1%, and cultured at 25-30° C. and 180 r / min for 48-56 hours, and the effective viable bacteria count is tested to be (1-2)×10 8 CUF / mL, the culture was terminated to obtain the fermentation broth of the three bacteria, and the three fermentation broths were mixed in a volume ratio of 1:1:2 to obtain the disease-preventing and growth-promoting microbial liquid.

[0127] In this comparative example, except that the volume ratio of Trichoderma pseudokoningii, Pseudomonas yamana and Bacillus subtilis in the disease prevention and growth promotion microbial liquid was changed to 1:1:2, the other raw materials and preparation method were the same as those in Example 1.

[0128] Comparative Example 10

[0129] A microbial agent for improving rice yield and quality, comprising disease-preventing and growth-promoting microbial liquid, yeast extract, microbial promoter and organic and inorganic nutrients, wherein the mass ratio of the four is 1:0.5:0.5:100.

[0130] In this comparative example, commercial yeast extract was used. Except for this, the other raw materials and preparation methods were the same as those in Example 1. The commercial yeast extract was purchased from Wuhan Kangqiong Biopharmaceutical Technology Co., Ltd. and was a brewer's yeast extract.

[0131] Performance Testing

[0132] Verification of strain antagonism:

[0133] After activating the frozen Trichoderma koningii, Pseudomonas yamana and Bacillus subtilis, they were streaked on LB medium plates and cultured at 25-30℃ for 3-5 days. The results showed that the strains were not antagonistic to each other ( Figure 1). This indicates that there is no obvious inhibitory activity between the three strains, and they can be combined into a composite microbial agent.

[0134] Identification of the growth-promoting ability of strains:

[0135] The molybdenum antimony colorimetric method was used to determine the ability of the strain to solubilize phosphorus; the flame atomic spectrophotometer was used to determine the ability of the strain to solubilize potassium; the carbon nitrogen analyzer was used to determine the ability of the strain to fix nitrogen; the concentrated sulfuric acid reaction method was used to determine the ability of the strain to secrete gibberellins; the Salkowski colorimetric solution method was used to determine the ability of the strain to secrete indoleacetic acid; and the CAS detection solution method was used to determine the ability of the strain to synthesize iron carriers. Each group was repeated 3 times and the results were averaged.

[0136] Table 1 Identification of growth-promoting ability of strains

[0137]

[0138] Verification of the antibacterial effect of different disease prevention and growth promotion microbial liquids:

[0139] The disease prevention and growth promotion microbial bacterial liquid prepared in Examples 1-3 and Comparative Examples 1-9 was added to the melted and cooled LB solid culture medium at a volume ratio of 5%, mixed thoroughly and poured into a culture dish to prepare a culture medium, with the addition of clean water as the control group (CK). A 5mm diameter puncher was used to take freshly activated 5d rice blast pathogen cakes and inoculated them into the center of the above culture medium, cultured at 25-30°C, and the colony diameters were measured by the cross method after 5d, and the average value was taken. Each test group was repeated 3 times, and the test was repeated 3 times. Calculate the inhibition rate of rice blast mycelium growth.

[0140] Growth inhibition rate = [(control colony diameter - bacterial cake diameter) - (treated colony diameter - bacterial cake diameter)] / [(control colony diameter - bacterial cake diameter)], where the bacterial cake diameter is 0.5 cm.

[0141] The inhibitory effects of the embodiments and comparative examples on rice blast pathogens are shown in Table 2:

[0142] Table 2 Inhibition effect test results

[0143]

[0144] As can be seen from Table 2, the strain combination of the present invention has a good inhibitory effect on the rice blast pathogen, while in Comparative Examples 1-9 where the bacterial solution composition is changed, the synergistic effect between the three strains is weakened or disappears, and the antibacterial effect decreases. Figure 2 shown.

[0145] Planting experiment:

[0146] The planting experiment was carried out at our company's experimental base, which is a mountainous area with little sunshine, large temperature difference between day and night, and high field humidity, which is suitable for the occurrence of rice blast.

[0147] The experiment set up 14 treatments, each treatment was repeated 4 times, and all experimental results were averaged.

[0148] The treatment groups were:

[0149] Example 1-3, Comparative Example 1-10: Apply at the tillering stage and heading stage of rice, dilute the test agent 100 times and spray every 667m 2 Apply 5L each time.

[0150] Set up a clean water control ck: Compare the embodiment and the comparative example, spray the same amount of clean water.

[0151] The incidence of rice blast diseased ears was investigated six months after sowing, and the severity of the diseased ears was graded, with 3 points in each plot and 100 ears in each point.

[0152] Rice blast classification standards:

[0153] Level 1: Individual branches and stalks are infected, with a loss of less than 5% per ear;

[0154] Level 2: 1 / 3 of the branches are infected, and each ear loses about 20%;

[0155] Level 3: The ear neck or main axis is diseased, the grains are half full, and each ear loses about 50%;

[0156] Level 4: The ear neck is diseased, with many shrunken grains and a loss of about 70% per ear;

[0157] Level 5: The ear neck is infected and turns into white ears, with a loss of about 90% per ear.

[0158] Diseased ear (plant) prevention effect (%) = (disease-caused ear (plant) rate in the blank control area - disease-caused ear (plant) rate in the treatment area) / disease-caused ear (plant) rate in the blank control area × 100%;

[0159] Disease index = Σ (number of diseased ears (plants) at each level × representative value at each level) / (total number of diseased ears (plants) surveyed × highest representative value) × 100;

[0160] Disease index control effect (%) = (disease index of blank control area - disease index of treated area) / disease index of blank control area × 100%.

[0161] Rice yield and quality survey:

[0162] After harvest, the yield was counted and the starch and protein contents were determined. The soluble protein content was determined by Coomassie Brilliant Blue G-250 staining, and the starch content was determined by anthrone colorimetry.

[0163] Table 3 Rice planting results

[0164]

[0165] Table 4 Rice yield and quality

[0166]

[0167] From the data in Tables 3-4, we can see that the application of the bacterial agent of the embodiment of the present invention can effectively improve the yield and quality of rice, and has a good prevention and control effect on rice blast. This is because the microbial bacteria screened by the present invention, after the three strains are mixed in equal proportions, have good disease resistance and growth promotion effects. In Comparative Example 1-9, which changed the microbial composition, the synergistic balance between the strains was broken, and the disease resistance and growth promotion effects were weakened. Comparative Example 10, which used ordinary yeast extract, provided limited nutrients and weakened the colonization effect of microorganisms, which also led to a weakening of the planting effect. We further verified the colonization effects of the embodiments of the present invention and Comparative Example 10:

[0168] After the rice seeds are disinfected, soak them at 28°C for 2 days, and the water is changed every 12 hours. Wrap the seeds with gauze and germinate at 28°C for 24 hours, rinsing them with sterile water once in the middle. Pick out seeds with similar germination degree and transfer them aseptically to sterilized large test tubes containing quartz sand and 20mL 1 / 4MS nutrient solution. Transfer 3 seedlings to each tube. After culturing in a constant temperature incubator at 28°C for 7 days, add 5mL of the microbial solution obtained in Examples 1-3 and Comparative Example 10, and continue to culture under the same conditions for 7 days. During this period, water them regularly with an equal amount of 1 / 4MS nutrient solution. After 7 days, take out the rice seedlings, shake off the quartz sand, weigh 0.1g of rice roots, add them to a conical flask containing 10 ml sterile water, vortex and oscillate at 140 r / min for 30 minutes, then spread them on LB plates with gradient dilutions, and count them after culturing at 25-30°C for 2 days. Repeat 3 times for each group and take the average of the results. The experimental results are as follows Figure 3 As shown, the microbial agent of the embodiment of the present invention has good colonization ability in the rhizosphere.

[0169] It should be noted that the above embodiments are only partial embodiments of the preferred methods of implementing the present invention, rather than all embodiments. Obviously, based on the above embodiments of the present invention, all other embodiments obtained by ordinary technicians in this field without creative work should fall within the scope of protection of the present invention.

Claims

1. A microbial agent for improving rice yield and quality, characterized in that: The microbial agent comprises a disease prevention and growth promotion microbial liquid, a yeast extract, a microbial promoter and organic and inorganic nutrients, the mass ratio of the four being 1: (0.5-1): (0.5-0.8): (50-100); the disease prevention and growth promotion microbial liquid comprises Trichoderma pseudokoningii ( Trichoderma koningiopsis ), Pseudomonas yamana ( Pseudomonas yamanorum ) and Bacillus subtilis ( Bacillus subtilis ), the volume ratio of the three is 1:1:1; the strain number of the Trichoderma koningii is CGMCC No.5.1243, purchased from the General Microbiology Center of China National Microbiological Culture Collection Administration; the strain number of the Pseudomonas yamana is CGMCC No. 1.16088, purchased from the General Microbiology Center of China National Microbiological Culture Collection Administration; the strain number of the Bacillus subtilis is CGMCC No.1.7740, purchased from the General Microbiology Center of China National Microbiological Culture Collection Administration.

2. The microbial agent for improving rice yield and quality according to claim 1, characterized in that: The method for preparing the disease prevention and growth promotion microbial bacterial liquid is as follows: single colonies of Trichoderma pseudokoningii, Pseudomonas yamana and Bacillus subtilis are taken respectively, inoculated into 100 mL of LB culture medium respectively, and placed in a constant temperature shaking shaker at 25-30°C and 180-200 r / min for 24 hours to obtain seed liquids of the three bacteria; the seed liquids of Trichoderma pseudokoningii, Pseudomonas yamana and Bacillus subtilis are transferred into 500 mL of LB culture medium at a transfer rate of 1%, and cultured at 25-30°C and 180-200 r / min for 48-56 hours, and the effective viable bacteria count is tested to be (1-2)×10 8 CUF / mL, the culture was terminated to obtain the fermentation broth of the three bacteria, and the three fermentation broths were mixed in a volume ratio of 1:1:1 to obtain the disease-preventing and growth-promoting microbial liquid.

3. The microbial agent for improving rice yield and quality according to claim 1, characterized in that: The preparation method of the yeast extract comprises the following steps: using active yeast as a raw material, adding the active yeast into deionized water to prepare an active yeast solution with a mass concentration of 10-20%; then adding a zinc compound into the active yeast solution at a solid-liquid ratio of 10 g:1 L, stirring at 50° C. for 2 hours, then adjusting the pH of the solution to 6-7, and standing for 1-2 hours; then adding 0.1-0.15% by mass of yeast wall-breaking enzyme into the mixed solution, and treating at 45° C. for 3 hours; after the yeast wall-breaking enzyme treatment, adding 0.03% by mass of neutral protease per liter of solution, and treating at 45° C. for 3 hours to obtain the yeast extract.

4. The microbial agent for improving rice yield and quality according to claim 3, characterized in that: The enzymatic activity of the yeast cell wall-breaking enzyme is 30,000 U / g, and the enzymatic activity of the neutral protease is 100,000 U / g.

5. The microbial agent for improving rice yield and quality according to claim 1, characterized in that: The microbial promoter is obtained by mixing dextrin, sodium oleate, maltose and nicotinamide in a mass ratio of 1:1:10:0.

1.

6. The microbial agent for improving rice yield and quality according to claim 1, characterized in that: The organic and inorganic nutrients include, by weight: 10-15 parts of urea, 5-10 parts of dipotassium hydrogen phosphate, 2-7 parts of calcium chloride, 3-5 parts of zinc sulfate, 12-16 parts of sucrose, 10-12 parts of fulvic acid, and 50-80 parts of water.

7. A method for preparing the microbial agent for improving rice yield and quality according to any one of claims 1 to 6, characterized in that: The method comprises the following preparation steps: (1) preparing disease prevention and growth promotion microbial liquid; (2) preparing yeast extract; (3) The disease prevention and growth promotion microbial liquid, yeast extract, microbial promoter and organic and inorganic nutrients are fully mixed in proportion to obtain the final product microbial agent.

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