CN115058348B - A composite microbial inoculant and its preparation method and application - Google Patents

Abstract

The invention discloses a composite microbial agent which comprises the following raw materials in parts by weight: 20-50 parts of microbial powder, 50-80 parts of growth auxiliary materials and 5-10 parts of drying agent; the microbial powder comprises fusarium, paecilomyces lilacinus, bacillus amyloliquefaciens, bacillus subtilis and bacillus bailii. The invention also provides a preparation method of the composite microbial agent and application of the composite microbial agent in preventing and treating cotton verticillium wilt. The invention combines endophytic antagonistic fungi fusarium and paecilomyces lilacinus with soil beneficial bacteria bacillus amyloliquefaciens, bacillus subtilis and bacillus bailii to form a composite microorganism group, which can remarkably improve the prevention and treatment effect on cotton verticillium wilt, thereby improving the economic benefit of cotton; the composite microbial agent is a biological control preparation, has no pollution to the environment, is simple in preparation process, easy to operate and low in cost, and is suitable for large-scale application in cotton production.

Description

A kind of compound microbial inoculant and its preparation method and application

Technical field

The invention relates to the technical field of agricultural microbial inoculant production, and in particular to a composite microbial inoculant and its preparation method and application.

Background technique

Cotton Verticillium wilt is a soil-borne vascular disease of cotton caused by Verticillium dahliae. The pathogen usually infects cotton roots, causing disease in cotton roots and even the entire plant, causing significant economic losses. It is commonly known as "cancer" Disease, there is no ideal prevention and control measures at home and abroad. At present, different prevention and control measures have been adopted for Verticillium wilt. Selecting disease-resistant varieties is the most economical and effective way to prevent and control Verticillium wilt in cotton. However, as for the current conventional varieties, the disease resistance effect is not ideal, and the presence of Verticillium wilt bacteria is obvious. Differentiation of pathogenicity and co-evolution with the host have led to slow progress in cotton disease resistance breeding research, which is still unable to meet production needs. Crop rotation can reduce the base of soil pathogens and has a preventive effect, but it has great limitations and its stubble is dense. , time-consuming, labor-intensive, high-cost, and not conducive to large-scale promotion; at the same time, excessive use of chemical pesticides will cause toxicity to crops and the surrounding environment. Therefore, there is an urgent need to find an environmentally friendly and effective prevention and control method. Biological control has attracted people's attention and has been applied in practice because of its advantages of being green and environmentally friendly and having great development potential.

Contents of the invention

The purpose of the present invention is to solve the existing problems in the prior art that the control effect on cotton Verticillium wilt is not good and the excessive use of chemical pesticides harms the environment, and to provide a composite microbial inoculant and its preparation method and application.

In order to solve the above technical problems, the technical solution adopted by the present invention is as follows: a composite microbial inoculant, including the following parts by weight of raw materials: 20 to 50 parts of microbial powder, 50 to 80 parts of growth auxiliary materials, and 5 to 10 parts of desiccant. ; The microbial bacteria powder includes Fusarium, Paecilomyces lilacinus, Bacillus amyloliquefaciens, Bacillus subtilis and Bacillus bellesii.

Preferably, the ratio of the viable bacterial numbers of Fusarium, Paecilomyces lilacinus, Bacillus amyloliquefaciens, Bacillus subtilis and Bacillus veleis is 3~5:2~3:1~2:1~2 :1～2.

Preferably, the ratio of the viable bacterial numbers of Fusarium, Paecilomyces lilacinus, Bacillus amyloliquefaciens, Bacillus subtilis and Bacillus veleis is 4:2:1:1:1.

Preferably, the total viable bacterial count of the composite microbial agent is ≧5×10 ⁸ CFU/g.

Preferably, the growth auxiliary material is one or more of chitosan, seaweed or fulvic acid.

Preferably, the desiccant is one or more of bentonite, silica gel or attapulgite.

The present invention also provides a preparation method for the above-mentioned composite microbial inoculant, which includes the following steps:

S1. Prepare Fusarium solani, Paecilomyces lilacinus seed liquid, Bacillus amyloliquefaciens, Bacillus subtilis and Bacillus veleis seed liquid respectively, and mix them to obtain a microbial mixed seed liquid;

S2. Add the microbial mixed seed liquid prepared in S1 to the matrix. The inoculation amount of the microbial mixed seed liquid is 10 to 50 ml of seed liquid/kg matrix; the matrix is one or both of wheat bran and corn grits. The moisture content of the matrix is 30-50%;

S3. Ferment the substrate with the mixed seed liquid of microorganisms added to S2. The culture temperature is 25-35°C and the culture time is 5-7 days. Then the fermentation culture is dried at 35-40°C and crushed to obtain the microorganisms. bacterial powder;

S4. Mix the microbial bacteria powder in S3 with growth auxiliary materials and desiccant to obtain a composite microbial bacteria agent.

The present invention also provides the application of the above-mentioned composite microbial inoculant in preventing and treating cotton Verticillium wilt.

Preferably, the compound microbial inoculant is applied to cotton roots through drip irrigation from mid-to-late June to early July when cotton Verticillium wilt occurs, at a dosage of 0.5 to 0.75 kg/acre each time, and is applied 1 to 2 times.

The beneficial effects of the present invention are:

1) Fusarium and Paecilomyces lilacinus in the composite microbial inoculant of the present invention have antagonistic effects on Verticillium dahliae of cotton, and Bacillus amyloliquefaciens, Bacillus subtilis, and Bacillus veleis are beneficial soil bacteria, and their metabolites It contains auxins, antibiotics, cytokinins and other regulatory substances necessary for crop growth. In addition, it also has the effect of inhibiting the reproduction of Verticillium wilt; the present invention combines the above-mentioned endophytic antagonistic fungi with soil beneficial bacteria to form a composite microbial group group, releasing it into the soil, integrating growth promotion, space occupation, antagonism and inducing immunity, which can significantly improve the control effect of cotton Verticillium wilt, thus improving the economic benefits of cotton;

2) The composite microbial inoculant in the present invention is a biological control agent, which has no pollution to the environment, has a simple preparation process, is easy to operate, is low in cost, and is suitable for large-scale application in cotton production.

Detailed ways

The present invention will be further described below in conjunction with specific embodiments.

Example 1

1. Preparation method of microbial seed liquid: Fusarium (Fusarium is deposited in the General Microbiology Center of China Microbial Culture Collection Committee, the preservation number is CGMCC NO. 11314; has been disclosed in the patent with application number CN201610425079.1), light The strains of Paecilomyces violaceum were activated and cultured in liquid for 2 days respectively. The activation medium was measured in 1 L and included NaNO ₃ 2g, K ₂ HPO ₄ 1 g, KCl 0.5g, MgSO ₄ ·7H ₂ O 0.5g, and FeSO ₄ 0.01 g. , 30g of sucrose, the activation temperature is 30°C, and the seed liquid of Fusarium and Paecilomyces lilacinus is obtained. Bacillus amyloliquefaciens, Bacillus subtilis, and Bacillus veleis were activated and cultured in liquid for 1 day respectively. The activation medium was measured in 1 L and included 10 g of peptone, 5 g of yeast extract, and 10 g of NaCl. The activation temperature was 28°C to obtain the amyloliquefaciens spores. bacillus, Bacillus subtilis, and Bacillus veleis seed liquid. (In this example, the sources of Paecilomyces lilacinus, Bacillus amyloliquefaciens, Bacillus subtilis, and Bacillus veleis strains are not particularly limited and are conventional commercially available strains)

2. Preparation method of microbial fermentation matrix: use wheat bran as the matrix, control the moisture content to 35%, add Fusarium, Paecilomyces lilacinus, Bacillus amyloliquefaciens, Bacillus subtilis, Bacillus veleis microbial seed liquid, microorganism The inoculation amount of seed liquid is 10ml seed liquid/kg substrate. The matrix added with Fusarium solani, Paecilomyces lilacinus, Bacillus amyloliquefaciens, Bacillus subtilis, and Bacillus veleis microbial seed liquid is fermented and cultured. The culture temperature is 30°C; the culture time is 5 to 7 days. Continuously monitor the total viable bacterial count of the microbial composite inoculant. When the total viable bacterial count of the microbial composite inoculant is no less than 5×10 ⁸ CFU/g, dry the fermentation culture at 38°C, crush it, and pass through 200 mesh Sieve to obtain microbial powder. Among them, the ratio of viable bacterial counts of Fusarium solani, Paecilomyces lilacinus, Bacillus amyloliquefaciens, Bacillus subtilis, and Bacillus veleis is 4:2:1:1:1.

3. Prepare the compound microbial inoculant according to the following parts by weight of raw materials: 30 parts of microbial powder, 60 parts of growth auxiliary materials, and 10 parts of desiccant. In this embodiment, the growth auxiliary material is an arbitrary mixture of chitosan and seaweed, and the desiccant is bentonite. Mix the above-mentioned weight parts of microbial bacteria powder with growth auxiliary materials and desiccant to obtain a composite microbial bacteria agent.

Example 2

1. Preparation method of microbial seed liquid: the same as the preparation method of microbial seed liquid in Example 1.

2. Preparation method of microbial fermentation substrate: Use corn grits as the substrate, control the moisture content to 35%, add Fusarium, Paecilomyces lilacinus, Bacillus amyloliquefaciens, Bacillus subtilis, Bacillus veleis microbial seed liquid, microorganism The inoculation amount of seed liquid is 50ml seed liquid/kg substrate. The matrix added with Fusarium solani, Paecilomyces lilacinus, Bacillus amyloliquefaciens, Bacillus subtilis, and Bacillus veleis microbial seed liquids is fermented and cultured. The culture temperature is 35°C; the culture time is 5 to 7 days. Continuously monitor the total viable bacterial count of the microbial composite inoculant. When the total viable bacterial count of the microbial composite inoculant is no less than 5×10 ⁸ CFU/g, dry the fermentation culture at 40°C, crush it, and pass through 200 mesh Sieve to obtain microbial powder. Among them, the ratio of the number of viable bacteria of Fusarium, Paecilomyces lilacinus, Bacillus amyloliquefaciens, Bacillus subtilis, and Bacillus veleis is 4:3:1:1:1.

3. Prepare the compound microbial inoculant according to the following parts by weight of raw materials: 20 parts of microbial powder, 80 parts of growth auxiliary materials, and 5 parts of desiccant. In this embodiment, the growth auxiliary material is fulvic acid, and the desiccant is silica gel. Mix the above-mentioned weight parts of microbial bacteria powder with growth auxiliary materials and desiccant to obtain a composite microbial bacteria agent.

Example 3

1. Preparation method of microbial seed liquid: the same as the preparation method of microbial seed liquid in Example 1.

2. Preparation method of microbial fermentation substrate: use wheat bran as the substrate, control the moisture content to 40%, add Fusarium, Paecilomyces lilacinus, Bacillus amyloliquefaciens, Bacillus subtilis, Bacillus veleis microbial seed liquid, microorganism The inoculation amount of seed liquid is 30ml seed liquid/kg substrate. The matrix added with Fusarium solani, Paecilomyces lilacinus, Bacillus amyloliquefaciens, Bacillus subtilis, and Bacillus veleis microbial seed liquid is fermented and cultured at a culture temperature of 33°C; the culture time is 5 to 7 days. Continuously monitor the total viable bacterial count of the microbial composite inoculant. When the total viable bacterial count of the microbial composite inoculant is no less than 5×10 ⁸ CFU/g, dry the fermentation culture at 40°C, crush it, and pass through 200 mesh Sieve to obtain microbial powder. Among them, the ratio of the number of viable bacteria of Fusarium, Paecilomyces lilacinus, Bacillus amyloliquefaciens, Bacillus subtilis, and Bacillus veleis is 5:3:2:2:2.

3. Prepare the compound microbial inoculant according to the following parts by weight of raw materials: 50 parts of microbial powder, 50 parts of growth auxiliary materials, and 10 parts of desiccant. In this embodiment, the growth auxiliary material is an arbitrary mixture of seaweed and humic acid, and the desiccant is attapulgite. Mix the above-mentioned weight parts of microbial bacteria powder with growth auxiliary materials and desiccant to obtain a composite microbial bacteria agent.

Example 4 Disease prevention effect of compound microbial inoculant in field

The composite microbial inoculant prepared in Example 1, Example 2, and Example 3 was subjected to a field test to identify the disease prevention effect on cotton Verticillium wilt after application of the composite microbial inoculant. The field experiment was conducted in Jinghe County, Bozhou, Xinjiang. Plots with relatively uniform fertility, flat terrain, and a high incidence of Verticillium wilt in the past were selected as experimental fields. The experiment consists of 4 treatments, arranged in random blocks, repeated three times, and the plot area is 100m ² . The planting density is 9,500 to 11,000 plants/666.67m ² , one film and three rows planting mode, and field management is carried out according to high-quality cotton cultivation technology.

Treatment group 1: The compound microbial inoculant obtained in Example 1 was drip-irrigated to the roots of cotton at the early stage of cotton budding, at a dosage of 0.75kg/acre, once every 10 days, and applied twice in total;

Treatment group 2: The compound microbial inoculant obtained in Example 2 was drip-irrigated to the roots of cotton at the early stage of cotton budding, with a dosage of 0.75kg/mu each time, once every 10 days, and applied twice in total;

Treatment group 3: The compound microbial inoculant obtained in Example 3 was drip-irrigated to the roots of cotton at the early stage of cotton budding, at a dosage of 0.75kg/acre, once every 10 days, and applied twice in total;

Control group: Bacillus subtilis microbial agent (commercially available) was administered with the same method and dosage as the treatment group.

On June 20 and June 30, drip irrigation compound microbial inoculant and Bacillus subtilis microbial inoculant were carried out respectively. On July 28 and August 20, the incidence of cotton Verticillium wilt was investigated. On September 25, field investigation and Seed test (2 points are randomly selected from each plot, and 20 plants are continuously selected from each point for field investigation and seed test. The number of bolls and shedding numbers are investigated, and 30 bolls are picked and weighed indoors to weigh the single boll). Cotton picking started on October 3, and all picking was completed on October 25. During the harvest period, the single receipt of each community is called the cumulative production. The results are shown in Table 1 and Table 2. Treating cotton with composite microbial inoculants can significantly reduce the damage of cotton Verticillium wilt. The cotton Verticillium wilt disease index on July 28 was 15.66-23.83, and the disease prevention effect was 47.15%-65.27%. , the disease index on August 20 was 20.66~28.34, and the disease prevention effect was 55.08%~67.25%. The compound microbial inoculant also has a significant effect on increasing cotton yield. The number of bolls per mu of cotton treated with the compound microbial inoculant was 76841 to 79667, an increase of 12.59% to 16.73% compared with the control group. The cotton yield per mu of cotton treated with the compound microbial inoculant was 423.67 to 435.49. kg, an increase of 10.09% to 13.16% compared with the control group.

Table 1 The control effect of compound microbial agent treatment on cotton Verticillium wilt in the field

Note: Different lowercase letters in the same column indicate significant differences (P<0.05).

Table 2 Determination results of cotton yield caused by compound microbial agent treatment in field

Note: Different lowercase letters in the same column indicate significant differences (P<0.05).

In summary, the composite microbial agent in the present invention has good control effect on cotton Verticillium wilt, with the best control effect being more than 60%; it can significantly increase cotton yield, with an increase rate of more than 10%; and at the same time, it can also improve fiber quality, thereby Improve the overall economic benefits of cotton. Moreover, the composite microbial inoculant in the present invention is a biological control agent, which has no pollution to the environment, has a simple preparation process, is easy to operate, and has low cost, and is very suitable for large-scale application in cotton production.

The description of the present invention is considered to be illustrative rather than restrictive. Based on the present invention, those skilled in the art can make some substitutions and modifications to some of the technical features without creative efforts based on the disclosed technical content. All deformations are within the protection scope of the present invention.

Claims (7)

1. A composite microbial inoculant, characterized in that it includes the following raw materials in parts by weight: 20 to 50 parts of microbial powder, 50 to 80 parts of growth auxiliary materials, and 5 to 10 parts of desiccant; the microbial powder is made from Fusarium Bacillus, Paecilomyces lilacinus, Bacillus amyloliquefaciens, Bacillus subtilis and Bacillus veleis; The ratio of the number of viable bacteria of Fusarium solani, Paecilomyces lilacinus, Bacillus amyloliquefaciens, Bacillus subtilis, and Bacillus veleis is 5:3:2:2:2; The deposit number of the Fusarium species is CGMCC NO. 11314. 2. The composite microbial inoculant according to claim 1, characterized in that the total viable bacterial count of the composite microbial inoculant is ≧5×10 ⁸ CFU/g. 3. The composite microbial inoculant according to claim 1, characterized in that the growth auxiliary material is one or more of chitosan, seaweed or fulvic acid. 4. The composite microbial inoculant according to claim 1, characterized in that the desiccant is one or more of bentonite, silica gel or attapulgite. 5. The preparation method of composite microbial inoculant as claimed in claim 1, characterized in that it includes the following steps: S1. Prepare Fusarium solani, Paecilomyces lilacinus seed liquid, Bacillus amyloliquefaciens, Bacillus subtilis and Bacillus veleis seed liquid respectively, and mix them to obtain a microbial mixed seed liquid; S2. Add the microbial mixed seed liquid prepared in S1 to the matrix. The inoculation amount of the microbial mixed seed liquid is 10-50 ml seed liquid/kg matrix; the matrix is one or both of wheat bran and corn grits. , the moisture content of the matrix is 30 to 50%; S3. Ferment and culture the substrate with the mixed microbial seed liquid added to S2. The culture temperature is 25-35°C and the culture time is 5-7 d. Then the fermentation culture is dried at 35-40°C and pulverized to obtain Microbial powder; S4. Mix the microbial bacteria powder in S3 with growth auxiliary materials and desiccant to obtain a composite microbial bacteria agent. 6. Application of the composite microbial inoculant according to any one of claims 1 to 4 in preventing and treating cotton Verticillium wilt. 7. The application according to claim 6, characterized in that the compound microbial inoculant is applied to cotton roots through drip irrigation from mid-to-late June to early July at the initial stage of cotton Verticillium wilt disease, with a dosage of 0.5 to 0.75 kg/mu each time. , apply 1 to 2 times.