Preparation method of special microbial agent for preventing cotton verticillium wilt

Abstract

The invention discloses a preparation method of a special microbial agent for preventing cotton verticillium wilt. The preparation method comprises the following steps: respectively preparing seed solutions; the preparation method comprises the following steps: preparing a substrate, inoculating the substrate with a microbial seed solution, fermenting, drying and crushing a fermentation culture to obtain microbial powder, and mixing the microbial powder with a growth auxiliary material and a drying agent to obtain the microbial agent. The microbial bacterial powder is prepared from bacillus subtilis HFN04, bacillus amyloliquefaciens, bacillus laterosporus, bacillus megaterium and bacillus licheniformis. The microbial agent containing the bacillus subtilis HFN04 can effectively prevent cotton verticillium wilt, and can promote the growth and development of cotton, improve the quality and increase the yield; the preparation method of the microbial agent is simple in process and easy to operate, and the prepared microbial agent has no pollution to the environment.

Description

Technical Field

[0001] The invention relates to the technical field of cotton biological microbial agents, and in particular to a method for preparing a microbial agent specifically for preventing cotton Verticillium wilt. Background Art

[0002] At present, cotton Verticillium wilt is a soil-borne vascular disease of cotton caused by Verticillium dahliae. The pathogen usually infects the roots of cotton, causing diseases in the roots of cotton and even the whole plant, causing significant economic losses. There are no ideal prevention and control measures at home and abroad. The main reasons why cotton Verticillium wilt is difficult to prevent and control are: first, upland cotton has poor resistance to cotton Verticillium wilt, and most of the varieties promoted in production are disease-resistant varieties. Moderately diseased fields and severely diseased fields will still cause serious yield losses; second, due to cultivation modes such as drip irrigation, returning straw to the field, and dwarf dense early, a large number of pathogens accumulate in the soil, and the dormant bodies of pathogens have strong resistance to stress and can remain in the soil for a long time; third, after the pathogen infects cotton from the roots, it spreads through the vascular bundles of the plant. The chemical fungicides sprayed on the leaves are difficult to be transported to the vascular bundles, and the prevention and control effect is poor.

[0003] In addition, the pathogenicity of Verticillium wilt is obviously differentiated, and it co-evolves with the host, resulting in slow progress in cotton disease-resistant breeding research, which cannot meet production needs; crop rotation can reduce the number of soil pathogens and play a preventive role, but it has great limitations, with dense crop rotation, time-consuming, labor-intensive, high cost, and not conducive to large-scale promotion; at the same time, excessive use of chemical pesticides will be toxic to crops and the surrounding environment. As a means of biological control, biological agents are green and environmentally friendly, and can be used as an environmentally friendly and effective control method.

[0004] Related studies have shown that the use of Bacillus subtilis has a good application effect in the prevention and control of cotton wilt. However, Xinjiang, where cotton production accounts for more than 90% of the country's total cotton production, is one of the driest regions in my country. Apart from natural factors such as climate, Xinjiang irrigation areas use floods and high temperatures in summer to carry out salt suppression and water storage irrigation in autumn and winter, resulting in excessive deep infiltration, leading to large-scale secondary salinization of the soil; its soil has chloride or sulfate-chloride accumulation, poor saline-alkali soil structure, capillary action cavity, poor water permeability and other characteristics; existing cotton wilt prevention and control methods are not effective on Xinjiang cotton. Therefore, a special prevention and control method for cotton wilt is urgently needed. Summary of the invention

[0005] To this end, the present invention provides a method for preparing a microbial agent specifically for preventing cotton Verticillium wilt, so as to solve the above-mentioned problems in the prior art.

[0006] In order to achieve the above object, the present invention provides the following technical solutions:

[0007] A microbial agent specially used for preventing cotton Verticillium wilt; comprising the following raw materials in the following weight proportions: 30-60 parts of microbial powder, 20-40 parts of growth auxiliary materials and 10-20 parts of desiccant;

[0008] The microbial powder is composed of Bacillus subtilis HFN04, Bacillus amyloliquefaciens, Bacillus laterosporus, Bacillus megaterium and Bacillus licheniformis;

[0009] The ratio of the number of live bacteria of Bacillus subtilis HFN04, Bacillus amyloliquefaciens, Bacillus laterosporus, Bacillus megaterium and Bacillus licheniformis is 6:3:1:1:1;

[0010] The total viable count of the microbial agent is not less than 1×1010 CFU / g.

[0011] Further: The deposit number of the Bacillus subtilis HFN04 is: CGMCC No.29091; the deposit unit is: General Microbiology Center of China Culture Collection Administration (CGMCC); the deposit address is: No. 3, Yard No. 1, Beichen West Road, Chaoyang District, Beijing; the deposit date is: November 21, 2023.

[0012] Furthermore: the growth auxiliary material is one of chitosan, algae, fulvic acid or humic acid, or any mixture thereof.

[0013] Further: the desiccant is one of bentonite, silica gel, silicate or white carbon black or any mixture thereof.

[0014] The present invention also discloses a method for preparing a microbial agent specifically for preventing cotton Verticillium wilt, which is characterized by comprising the following steps:

[0015] (1) preparing seed solutions of Bacillus subtilis HFN04, Bacillus amyloliquefaciens, Bacillus laterosporus, Bacillus megaterium, and Bacillus licheniformis respectively;

[0016] (2) wheat bran or corn grits or both are used as a matrix, the water content of which is controlled to be 20%-50%, and the seed solution prepared in step (1) is added, wherein the inoculation amount of the microbial seed solution is 10-100 ml of seed solution per kilogram of the matrix.

[0017] (3) fermenting the substrate to which the seed solution is added in step (2) at a temperature of 30-40° C. for 96-144 h, drying the fermented culture at 35-40° C., and crushing the culture to obtain microbial powder.

[0018] (4) The microbial powder in step (3) is mixed with growth aids and a desiccant to obtain a microbial agent.

[0019] When in use, the microbial agent of the present invention is drip-irrigated to the roots of cotton, with a dosage of 200 g / mu for the first and second irrigations, and 100 g / mu for the third to sixth irrigations.

[0020] The present invention has the following advantages: the microbial agent containing Bacillus subtilis HFN04 of the present invention can effectively prevent cotton Verticillium wilt, and can promote cotton growth and development, improve quality, and increase yield. The preparation method of the microbial agent of the present invention has a simple process and is easy to operate, and the prepared microbial agent has no pollution to the environment. DETAILED DESCRIPTION

[0021] The following is a description of the implementation of the present invention by specific specific examples. People familiar with the technology can easily understand other advantages and effects of the present invention from the contents disclosed in this specification. Obviously, the described embodiments are part of the embodiments of the present invention, but not all of them. It should be understood that these embodiments are only to further illustrate the present invention and cannot be understood as limiting the scope of protection of the present invention. Technical engineers in this field can make some non-essential improvements and adjustments to the present invention based on the contents of the above invention.

[0022] Based on the embodiments of the present invention, all other embodiments obtained by ordinary technicians in the field without making any creative work shall fall within the scope of protection of the present invention.

[0023] Example 1

[0024] A microbial agent specifically for preventing cotton Verticillium wilt according to an embodiment of the present invention comprises the following raw materials in the following weight ratios: 50 parts of microbial powder, 30 parts of growth aids and 10 parts of desiccant.

[0025] The microbial powder consists of Bacillus subtilis HFN04, Bacillus amyloliquefaciens, Bacillus laterosporus, Bacillus megaterium and Bacillus licheniformis.

[0026] The Bacillus subtilis HFN04 has a deposit number of CGMCC No. 29091; the deposit unit is China General Microbiology Center (CGMCC), China Culture Collection Administration; the deposit address is No. 3, Yard No. 1, Beichen West Road, Chaoyang District, Beijing; the deposit date is November 21, 2023.

[0027] The ratio of the number of live bacteria of Bacillus subtilis HFN04, Bacillus amyloliquefaciens, Bacillus laterosporus, Bacillus megaterium and Bacillus licheniformis is 6:3:1:1:1; the total number of live bacteria of the microbial agent is not less than 1×10 10 CFU / g.

[0028] The growth auxiliary material is one of chitosan, algae, fulvic acid or humic acid, or any mixture thereof.

[0029] The desiccant is one of bentonite, silica gel, silicate or white carbon black or any mixture thereof.

[0030] A method for preparing a microbial agent specifically for preventing cotton Verticillium wilt comprises the following steps:

[0031] (1) preparing seed solutions of Bacillus subtilis HFN04, Bacillus amyloliquefaciens, Bacillus laterosporus, Bacillus megaterium, and Bacillus licheniformis respectively;

[0032] (2) wheat bran is used as a substrate, the water content of which is controlled to be 50%, and the seed solution prepared in step (1) is added, wherein the inoculation amount of the microbial seed solution is 100 ml of seed solution per kilogram of substrate.

[0033] (3) fermenting the substrate to which the seed solution was added in step (2) at a temperature of 35° C. for 144 h, drying the fermented culture at 40° C., and crushing the culture to obtain microbial powder.

[0034] (4) The microbial powder in step (3) is mixed with growth aids and a desiccant to obtain a microbial agent.

[0035] Example 2

[0036] A microbial agent specially used for preventing cotton Verticillium wilt comprises the following raw materials in the following weight proportions: 30 parts of microbial powder, 20 parts of growth auxiliary materials and 10 parts of desiccant.

[0037] The microbial powder consists of Bacillus subtilis HFN04, Bacillus amyloliquefaciens, Bacillus laterosporus, Bacillus megaterium and Bacillus licheniformis.

[0038] The Bacillus subtilis HFN04 has a deposit number of CGMCC No. 29091; the deposit unit is China General Microbiology Center (CGMCC), China Culture Collection Administration; the deposit address is No. 3, Yard No. 1, Beichen West Road, Chaoyang District, Beijing; the deposit date is November 21, 2023.

[0039] The ratio of the number of live bacteria of Bacillus subtilis HFN04, Bacillus amyloliquefaciens, Bacillus laterosporus, Bacillus megaterium and Bacillus licheniformis is 6:3:1:1:1; the total number of live bacteria of the microbial agent is not less than 1×10 10 CFU / g.

[0040] The growth auxiliary material is one of chitosan, algae, fulvic acid or humic acid, or any mixture thereof.

[0041] The desiccant is one of bentonite, silica gel, silicate or white carbon black or any mixture thereof.

[0042] A method for preparing a microbial agent specifically for preventing cotton Verticillium wilt comprises the following steps:

[0043] (1) preparing seed solutions of Bacillus subtilis HFN04, Bacillus amyloliquefaciens, Bacillus laterosporus, Bacillus megaterium, and Bacillus licheniformis respectively;

[0044] (2) wheat bran is used as a substrate, the water content of which is controlled to be 50%, and the seed solution prepared in step (1) is added, wherein the inoculation amount of the microbial seed solution is 100 ml of seed solution per kilogram of substrate.

[0045] (3) fermenting the substrate to which the seed solution was added in step (2) at a temperature of 35° C. for 144 h, drying the fermented culture at 40° C., and crushing the culture to obtain microbial powder.

[0046] (4) The microbial powder in step (3) is mixed with growth aids and a desiccant to obtain a microbial agent.

[0047] Test description:

[0048] The test base was set up in the Eighth Division of the Corps. The soil fertility was medium, the incidence of Fusarium wilt was medium, and the cotton fields of the previous cotton crop were selected. The demonstration area was 2,000 mu (control area was 2,000 mu), including 800 mu of the 7th Company of the 149th Regiment, 600 mu of the 11th Company of the 134th Regiment, and 600 mu of the 13th Company of the 141st Regiment. The test soil was gray desert soil with low organic matter content and clay texture. The specific indicators are shown in Table 1.

[0049] Table 1 Physical and chemical indicators of tested soil

[0050]

[0051] In the experimental base, cotton planting adopts machine-picked cotton, 1 film, 6 rows, 3 tubes, 66+10cm plant row configuration, with a theoretical plant number of 16,800 plants / mu (including 18,680 plants / 666.7m2 in the demonstration site of the 13th Company of the 141st Regiment), sowing from April 11th to April 18th (drip water for seedling emergence), topping from July 1st to July 5th, harvesting on October 5th, and irrigation 9 times during the whole growing period.

[0052] The experimental group used microbial agent + conventional drip irrigation fertilization; the microbial agent of the present invention was drip-irrigated to the roots of cotton, with a dosage of 200 g / mu for the first and second irrigations, and 100 g / mu for the third to sixth irrigations;

[0053] The control group (CK) was subjected to local conventional drip fertilization.

[0054] Effects of microbial agent treatment on cotton growth and development at the seedling stage:

[0055] From May 2 to May 8, sampling surveys were conducted on the number of cotton seedlings and the development of cotton root systems at three test sites (see Table 2 for details). Compared with the control, the number of cotton seedlings per mu treated with microbial agents increased by 101-2033, the main root length increased by 0.38-1.3 cm, and the number of lateral roots increased by 0.4-4.1.

[0056] Table 2 Survey table of growth indexes of cotton seedlings under different treatments

[0057]

[0058] Effects of microbial agent treatment on cotton growth and development at bud stage:

[0059] The results of the bud stage survey (see Table 3 for details) showed that the use of microbial agents can promote the growth and development of cotton buds. The plant height, number of fruit branches and number of buds increased by 5.7-14.0 cm, 0.2-1.0 branches and 0.6-4.1 branches respectively compared with the control treatment.

[0060] Table 3 Survey of cotton bud growth and development at different treatments

[0061]

[0062] Effects of microbial agent treatment on cotton growth and development during flowering and boll stage:

[0063] As can be seen from Table 4, the use of microbial agents can significantly promote the development of cotton fruit branches, early flowering and early boll setting, and the number of fruit branches, flowers and bolls increased by 0.3-1.0, 0.1-3.0 and 1.0-3.0 respectively compared with the control.

[0064] Table 4 Survey table of growth indexes of cotton in flowering and boll stage under different treatments

[0065]

[0066] During the flowering and boll stage, the whole cotton plant was dried and weighed (see Table 5 for details). The dry weight of each plant treated with microbial agents increased by 16.6-37.3 g compared with the control, and the dry weight growth rate was 24.4%-75.2%. The above shows that the use of microbial agents has a significant promoting effect on the accumulation of dry matter in cotton plants.

[0067] Table 5 Survey table of dry matter weight of cotton plants at flowering and boll stage under different treatments

[0068]

[0069]

[0070] Effects of microbial agent treatment on cotton growth process:

[0071] From the survey data on cotton growth process in Table 6, it can be seen that after emergence, budding, flowering and boll opening of cotton treated with microbial agents can be respectively 3d, 2d and 2d earlier than those of the control, and the growth period can be shortened by 1-2d.

[0072] Table 6 Questionnaire on growth process of cotton under different treatments Date / Month

[0073]

[0074] Effects of microbial agent treatment on cotton yield and quality:

[0075] The use of microbial agents can improve the soil nutrient environment and promote the growth of cotton plants. As shown in Table 7, the number of bolls per plant increased by 0.2, the number of bolls per mu increased by 3495-12291, and the weight of single boll increased by 0.1-0.4g compared with the control group. The seed cotton yield of microbial agent treatment increased by 44.4-76.0kg per mu compared with the control group, an increase of 10.2-16.8%, and an average increase of 13.5%. The machine-picked yield increased by 48-57kg per mu of seed cotton, an increase of 11.0-13.9%, and an average increase of 12.4%.

[0076] Table 7 Questionnaire on cotton yield composition under different treatments

[0077]

[0078] Table 8 shows the indoor quality test of samples taken from the middle boll of cotton plants at the harvest time. The seed cotton lint score and fiber uniformity of the three test points treated with microbial agents were better than those of the control, which were 0.3-2.2 percentage points and 0.2-1.3 percentage points higher than those of the control, respectively. The microbial agents had no significant effect on quality indicators such as cotton lint length, micronaire value, and breaking strength.

[0079] Table 8 Summary of cotton quality indicators under different treatments

[0080]

[0081] Effects of microbial agent treatment on soil physical and chemical properties:

[0082] From the survey data of soil physical and chemical indicators before and after the application of microbial agents in Tables 9 and 10, it can be seen that the change trends of soil organic matter, hydrolyzed nitrogen, available potassium, water-soluble salt, pH and other indicators in the three test points were not significant; the soil available phosphorus content (except for the 141th group test point) in the microbial agent treatment and the control treatment showed an overall increasing trend after harvest compared with before sowing, with an increase of 15.9-69.6% and 14.9-43.3% respectively. The increase in the microbial agent treatment was 1.0-26.3 percentage points higher than that in the control treatment, indicating that the use of microbial agents has a significant effect on activating soil phosphorus, reducing phosphate fertilizer fixation, and increasing soil available phosphorus.

[0083] Table 9 Survey table of soil physical and chemical indicators before application of microbial agents (before sowing)

[0084]

[0085] Table 10 Survey table of soil physical and chemical indicators after application of microbial agents (after harvest)

[0086]

[0087] In summary, the use of the microbial agent of the present invention on the basis of conventional drip irrigation fertilization can promote cotton growth and development, improve quality, and increase yield. Cotton boll opening can be 2 days earlier than the control; the number of bolls per plant increases by 0.2, and the weight of a single boll increases by 0.1-0.4g; the lint percentage and fiber uniformity increase by 0.3-2.2 percentage points and 0.2-1.3 percentage points respectively; the seed cotton yield per mu can be increased by 48-57kg, an increase of 11.0-13.9%, and an average increase of 12.4%; at the same time, the use of microbial agents can reduce soil bulk density and improve soil structure.

[0088] The above description is only a preferred embodiment of the present invention and is not intended to limit the present invention. Any modifications, equivalent substitutions and improvements made within the spirit and principles of the present invention should be included in the protection scope of the present invention.

Claims

1. A microbial agent specifically for preventing cotton Verticillium wilt, characterized in that: The raw materials include the following weight ratios: 30-60 parts of microbial powder, 20-40 parts of growth auxiliary materials and 10-20 parts of desiccant; The microbial powder is composed of Bacillus subtilis HFN04, Bacillus amyloliquefaciens, Bacillus laterosporus, Bacillus megaterium and Bacillus licheniformis; The ratio of the number of live bacteria of Bacillus subtilis HFN04, Bacillus amyloliquefaciens, Bacillus laterosporus, Bacillus megaterium and Bacillus licheniformis is 6:3:1:1:1; The total viable count of the microbial agent is not less than 1×10 10 CFU / g.

2. A microbial agent for preventing cotton Verticillium wilt according to claim 1, characterized in that: The deposit number of the Bacillus subtilis HFN04 is: CGMCC No.29091; the deposit unit is: China Culture Collection Center of General Microbiology CGMCC; the deposit address is: No. 3, Yard No. 1, Beichen West Road, Chaoyang District, Beijing; the deposit date is: November 21, 2023.

3. A microbial agent for preventing cotton Verticillium wilt according to claim 1, characterized in that: The growth auxiliary material is one of chitosan, algae, fulvic acid or humic acid, or any mixture thereof.

4. A microbial agent for preventing cotton Verticillium wilt according to claim 1, characterized in that: The desiccant is one of bentonite, silica gel, silicate or white carbon black or any mixture thereof.

5. A method for preparing a microbial agent specifically for preventing cotton Verticillium wilt, characterized in that: The following steps are involved: (1) preparing seed solutions of Bacillus subtilis HFN04, Bacillus amyloliquefaciens, Bacillus laterosporus, Bacillus megaterium, and Bacillus licheniformis respectively; (2) using one or both of wheat bran and corn grits as a substrate, controlling the water content to be 20%-50%, adding the seed solution prepared in step (1), wherein the inoculation amount of the microbial seed solution is 10-100 ml of seed solution per kilogram of substrate; (3) fermenting the substrate to which the seed solution was added in step (2) at a temperature of 30-40° C. for 96-144 h, drying the fermented culture at 35-40° C., and crushing the culture to obtain microbial powder; (4) The microbial powder in step (3) is mixed with growth aids and a desiccant to obtain a microbial agent.

6. Use of a microbial agent specifically for preventing cotton Verticillium wilt as claimed in any one of claims 1 to 4 in preventing cotton Verticillium wilt.

7. The use according to claim 6, characterized in that: Apply the microbial agent to the roots of cotton by drip irrigation, with a dosage of 200g / mu for the first and second irrigations, and 100g / mu for the third to sixth irrigations.

Images