Preparation method of microbial agent for degrading herbicide longguojing

Abstract

The invention discloses a preparation method of a microbial agent for degrading a herbicide longguojing, the microbial agent comprises bacillus subtilis, a carrier substance, a nutrient substance and a protective agent, the effective viable count of the bacillus subtilis is greater than or equal to 12 billion / g, the carrier substance is used for adsorbing and bearing bacillus subtilis powder, and the nutrient substance is used for adsorbing and bearing the bacillus subtilis powder. And the nutrient substances comprise nutrient elements of nitrogen, phosphorus and potassium, trace elements and growth factors, and are used for promoting the growth and reproduction of the bacillus subtilis. During implementation, the method has the following effects that the bacillus subtilis can generate various antibacterial substances, the substances have a remarkable inhibition effect on harmful microorganisms in soil, infection of the harmful microorganisms on cotton can be reduced, and the disease occurrence rate is reduced; the bacillus subtilis can be rapidly propagated in the soil, and forms nutrition and space site competition with the harmful bacteria, so that the growth and propagation of the harmful bacteria are effectively limited.

Description

Technical Field

[0001] The invention relates to the technical field of microbial agents, and in particular to a method for preparing a microbial agent for degrading the herbicide chlorfenapyr. Background Art

[0002] With the continuous development of agricultural production, soil health issues have become increasingly prominent. The long-term and large-scale use of chemical fertilizers and pesticides has led to soil salinization, severe compaction, and a decrease in organic matter content. The balance of soil microecology has been disrupted, and pests and diseases have occurred frequently, especially in cotton fields.

[0003] In cotton fields, the herbicide "Longcaojing" is used for weed control, and the residue of the herbicide "Longcaojing" in the field threatens the sustainable development of agricultural production. It is particularly important to find a method that can improve the soil and promote the healthy growth of crops. To this end, this application proposes a method for preparing a microbial agent for degrading the herbicide Longcaojing. Summary of the invention

[0004] To this end, the present invention provides a method for preparing a microbial agent for degrading the herbicide chlorfenapyr to solve the above-mentioned problems in the prior art.

[0005] In order to achieve the above object, the present invention provides the following technical solutions:

[0006] According to a first aspect of the present invention, a method for preparing a microbial agent for degrading the herbicide chlorfenapyr comprises Bacillus subtilis, a carrier material, a nutrient material and a protective agent.

[0007] The effective viable count of Bacillus subtilis is ≥ 120 billion / gram,

[0008] The carrier material is used to adsorb and carry Bacillus subtilis powder, and improve the stability and persistence of Bacillus subtilis in the soil.

[0009] The nutrients are nitrogen, phosphorus, potassium, trace elements and growth factors, which are used to promote the growth and reproduction of Bacillus subtilis.

[0010] The protective agent is bentonite or silica gel, which is used to protect the Bacillus subtilis from being damaged during storage and transportation, and to improve the stability of the Bacillus subtilis.

[0011] Preferably, trace elements are also included, including dipotassium hydrogen phosphate and magnesium sulfate, which are used to promote the growth and metabolism of Bacillus subtilis.

[0012] Preferably, the content of Bacillus subtilis is 1-5%, the content of the carrier substance is 85-94%, the content of the nutrient substance is 1-5%, the content of the trace element is 0.1-1%, and the content of the protective agent is 0-1%.

[0013] Preferably, the carrier substances are glucose and peptone, and the nutrients are dipotassium hydrogen phosphate, magnesium sulfate and yeast extract. 20-30 g / L of glucose, 10-15 g / L of peptone, 5-10 g / L of yeast extract, 5-10 g / L of sodium chloride, 1-2 g / L of dipotassium hydrogen phosphate, and 0.5-1 g / L of magnesium sulfate are added to distilled water to prepare a culture medium for Bacillus subtilis, and then Bacillus subtilis is inoculated onto the culture medium.

[0014] Preferably, the specific steps of preparing the Bacillus subtilis agent are:

[0015] Step 1: Add the carrier material and nutrients into distilled water in proportion, stir evenly, and adjust the pH value to between 7.0 and 7.5.

[0016] Step 2: Place the components prepared in step 1 into a sterile container and sterilize them under high temperature and high pressure.

[0017] Step 3: Inoculate the Bacillus subtilis strain into the sterilized culture medium under sterile conditions.

[0018] Step 4: ferment and culture the mixture under suitable temperature and conditions to allow Bacillus subtilis to multiply in large quantities.

[0019] Step 5: During the fermentation process, stirring and ventilation are performed at regular intervals to promote bacterial growth.

[0020] Step 6: After the fermentation is completed, the bacteria in the culture solution are collected and separated from the culture medium by centrifugation and filtration.

[0021] Step seven, drying the bacterial cells to obtain the Bacillus subtilis agent.

[0022] Preferably, in step 2, the high temperature and high pressure sterilization condition is sterilization at 121 degrees Celsius for 20 minutes.

[0023] Preferably, the suitable temperature in step 4 is 28-30 degrees Celsius.

[0024] The present invention has the following advantages:

[0025] 1. Bacillus subtilis can produce a variety of antibacterial substances, which have a significant inhibitory effect on the residual herbicide "Longcaojing" and harmful microorganisms in the soil, and can reduce the infection of harmful bacteria on cotton and reduce the incidence of diseases;

[0026] 2. Bacillus subtilis can reproduce rapidly in the soil, competing with harmful bacteria for nutrients and spatial sites, thereby effectively limiting the growth and reproduction of harmful bacteria;

[0027] 3. Bacillus subtilis can form a probiotic environment in the soil, promote the formation of soil aggregate structure, improve the soil's ability to retain fertilizer and water, provide a better growth environment for cotton roots, and facilitate the development and growth of the root system;

[0028] 4. Bacillus subtilis can produce substances similar to cytokinins and plant growth hormones, and can induce cotton to produce substances such as indoleacetic acid, increase the level of growth stimulants in cotton, and thus promote the growth and development of cotton;

[0029] 5. The use of Bacillus subtilis can improve the disease resistance of cotton, making it more resistant to infection by pathogens;

[0030] 6. Improve stress resistance: In addition, Bacillus subtilis can also improve cotton's resistance to cold, drought and other stresses, so that cotton can still maintain a good growth state under adverse environmental conditions;

[0031] 7. Bacillus subtilis can decompose organic matter in the soil, increase humus, improve the soil aggregate structure, and increase the soil's fertility and ability to retain fertilizer and water;

[0032] 8. At the same time, Bacillus subtilis can also dissolve phosphorus, potassium and fix nitrogen, improve the effectiveness of nutrients in the soil, and promote the absorption and utilization of nutrients by cotton. DETAILED DESCRIPTION

[0033] The following is a description of the implementation of the present invention by specific embodiments. People familiar with the art 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, not all of the embodiments. Based on the embodiments of the present invention, all other embodiments obtained by ordinary technicians in this field without creative work are within the scope of protection of the present invention.

[0034] A method for preparing a microbial agent for degrading the herbicide chloranil, comprising Bacillus subtilis, a carrier material, a nutrient material and a protective agent.

[0035] The effective viable count of Bacillus subtilis is ≥ 120 billion / gram,

[0036] The carrier material is used to adsorb and carry Bacillus subtilis powder, and improve the stability and persistence of Bacillus subtilis in the soil.

[0037] The nutrients are nitrogen, phosphorus, potassium, trace elements and growth factors, which are used to promote the growth and reproduction of Bacillus subtilis.

[0038] The protective agent is bentonite or silica gel, which is used to protect the Bacillus subtilis from being damaged during storage and transportation, and to improve the stability of the Bacillus subtilis.

[0039] It also includes trace elements, including dipotassium hydrogen phosphate and magnesium sulfate, which are used to promote the growth and metabolism of Bacillus subtilis.

[0040] The content of the Bacillus subtilis is 1-5%, the content of the carrier material is 85-94%, the content of the nutrient material is 1-5%, the content of the trace element is 0.1-1%, and the content of the protective agent is 0-1%.

[0041] The carrier substances are glucose and peptone, and the nutrients are dipotassium hydrogen phosphate, magnesium sulfate and yeast extract. 20-30 g / L of glucose, 10-15 g / L of peptone, 5-10 g / L of yeast extract, 5-10 g / L of sodium chloride, 1-2 g / L of dipotassium hydrogen phosphate, and 0.5-1 g / L of magnesium sulfate are added into distilled water to prepare a culture medium for Bacillus subtilis, and then the Bacillus subtilis is inoculated onto the culture medium.

[0042] The specific steps of making Bacillus subtilis inoculant are:

[0043] Step 1: Add the carrier material and nutrients into distilled water in proportion, stir evenly, and adjust the pH value to between 7.0 and 7.5.

[0044] Step 2: Place the components prepared in step 1 into a sterile container and sterilize them under high temperature and high pressure.

[0045] Step 3: Inoculate the Bacillus subtilis strain into the sterilized culture medium under sterile conditions.

[0046] Step 4: ferment and culture the mixture under suitable temperature and conditions to allow Bacillus subtilis to multiply in large quantities.

[0047] Step 5: During the fermentation process, stirring and ventilation are performed at regular intervals to promote bacterial growth.

[0048] Step 6: After the fermentation is completed, the bacteria in the culture solution are collected and separated from the culture medium by centrifugation and filtration.

[0049] Step 7: Dry the bacteria to obtain the Bacillus subtilis inoculum. In step 2, the high temperature and high pressure sterilization condition is sterilization at 121 degrees Celsius for 20 minutes. The appropriate temperature in step 4 is 28-30 degrees Celsius.

[0050] Embodiment 1

[0051] In this embodiment, the content of Bacillus subtilis is 1%, the content of the carrier substance is 90%, the content of the nutrient substance is 2%, the content of the trace element is 0.1%, and the content of the protective agent is 0.1%.

[0052] The carrier substances are glucose and peptone, and the nutrients are dipotassium hydrogen phosphate, magnesium sulfate and yeast extract. 20 g / L of glucose, 15 g / L of peptone, 10 g / L of yeast extract, 5 g / L of sodium chloride, 1 g / L of dipotassium hydrogen phosphate and 0.5 g / L of magnesium sulfate are added into distilled water to prepare a culture medium for Bacillus subtilis, and then the Bacillus subtilis is inoculated into the culture medium.

[0053] The specific steps of making Bacillus subtilis inoculant are:

[0054] Step 1: Add the carrier material and nutrients into distilled water in proportion, stir evenly, and adjust the pH value to between 7.2.

[0055] Step 2: Place the components prepared in step 1 into a sterile container and sterilize them under high temperature and high pressure.

[0056] Step 3: Inoculate the Bacillus subtilis strain into the sterilized culture medium under sterile conditions.

[0057] Step 4: ferment and culture at 28 degrees Celsius to allow Bacillus subtilis to multiply in large quantities.

[0058] Step 5: During the fermentation process, stirring and ventilation are performed at regular intervals to promote bacterial growth.

[0059] Step 6: After the fermentation is completed, the bacteria in the culture solution are collected and separated from the culture medium by centrifugation and filtration.

[0060] Step 7: Dry the bacterial cells to obtain the Bacillus subtilis inoculum. In step 2, the high temperature and high pressure sterilization condition is sterilization at 121 degrees Celsius for 20 minutes.

[0061] Embodiment 2

[0062] In this embodiment, the content of Bacillus subtilis is 5%, the content of the carrier substance is 94%, the content of the nutrient substance is 5%, the content of the trace element is 1%, and the content of the protective agent is 1%.

[0063] The carrier substances are glucose and peptone, and the nutrients are dipotassium hydrogen phosphate, magnesium sulfate and yeast extract. 30 g / L of glucose, 10 g / L of peptone, 5 g / L of yeast extract, 10 g / L of sodium chloride, 2 g / L of dipotassium hydrogen phosphate and 1 g / L of magnesium sulfate are added into distilled water to prepare a culture medium for Bacillus subtilis, and then the Bacillus subtilis is inoculated into the culture medium.

[0064] The specific steps of making Bacillus subtilis inoculant are:

[0065] Step 1: Add the carrier material and nutrients into distilled water in proportion, stir evenly, and adjust the pH value to between 7.4.

[0066] Step 2: Place the components prepared in step 1 into a sterile container and sterilize them under high temperature and high pressure.

[0067] Step 3: Inoculate the Bacillus subtilis strain into the sterilized culture medium under sterile conditions.

[0068] Step 4: ferment and culture at 30 degrees Celsius to allow Bacillus subtilis to multiply in large quantities.

[0069] Step 5: During the fermentation process, stirring and ventilation are performed at regular intervals to promote bacterial growth.

[0070] Step 6: After the fermentation is completed, the bacteria in the culture solution are collected and separated from the culture medium by centrifugation and filtration.

[0071] Step 7: Dry the bacterial cells to obtain the Bacillus subtilis inoculum. In step 2, the high temperature and high pressure sterilization condition is sterilization at 121 degrees Celsius for 20 minutes.

[0072] The following data were obtained when the bacterial agent obtained in the above examples was used in cotton fields and cotton cultivation:

[0073] Soil type: sandy loam with slight stickiness. Land preparation was done on April 6, film was laid and sown on April 8, and seedlings emerged on April 25.

[0074] Field management: Base fertilizer input is 50kg / mu (30kg compound fertilizer + 20kg urea). Drip water 8 times in total during the growth period, a total of 270 cubic meters per mu, 32kg / mu of urea, 23kg / mu of diammonium phosphate, and 20kg / mu of potassium fertilizer.

[0075] Chemical adjustment and control: Chemical control 8 times, total dosage 21 grams.

[0076] Main pests and diseases and their control: Mainly control tiger pests, thrips, cotton aphids and cotton bollworms.

[0077] Topping period: July 5. Flower harvest period: October 10.

[0078] Effects of the microbial agent of the present application on cotton field soil and cotton growth and development

[0079] The test results show that the bacterial agent has a certain improvement and promoting effect on soil water-soluble salts, organic matter, total nitrogen, hydrolyzable nitrogen, available phosphorus and quick-acting potassium. At the same time, it inhibits harmful pathogens, and the damage caused by cotton seedling diseases is significantly reduced.

[0080] Among them: soil pH value decreased by 0.31, water-soluble salt decreased by 0.53g / kg, a decrease of 12.96%; organic matter increased by 0.12g / kg, total nitrogen increased by 9.80g / kg, hydrolyzable nitrogen increased by 16.77mg / kg, available phosphorus increased by 38.60mg / kg, and available potassium increased by 27.67mg / kg.

[0081] Impact on cotton field soil and cotton growth and development: The test results show that the yield of square-seed cotton in 1,000 mu treated with the bacterial agent increased by 83kg / mu, a 16.81% increase over the control; the yield of square-seed cotton in 100 mu increased by 115.5kg / mu, a 18.27% increase over the control.

[0082] It can be seen that the bacterial agent of the present application can effectively improve the performance of the soil when implemented, which is beneficial to the growth of cotton and significantly increases the yield.

[0083] Embodiment 3

[0084] In this embodiment, the content of Bacillus subtilis is 5%, the content of the carrier substance is 94%, the content of the nutrient substance is 5%, the content of the trace element is 1%, and the content of the protective agent is 1%.

[0085] The carrier substances are glucose and peptone, and the nutrients are dipotassium hydrogen phosphate, magnesium sulfate and yeast extract. 30 g / L of glucose, 10 g / L of peptone, 5 g / L of yeast extract, 10 g / L of sodium chloride, 2 g / L of dipotassium hydrogen phosphate and 1 g / L of magnesium sulfate are added into distilled water to prepare a culture medium for Bacillus subtilis, and then the Bacillus subtilis is inoculated into the culture medium.

[0086] The specific steps of making Bacillus subtilis inoculant are:

[0087] Step 1: Add the carrier material and nutrients into distilled water in proportion, stir evenly, and adjust the pH value to between 7.4.

[0088] Step 2: Place the components prepared in step 1 into a sterile container and sterilize them under high temperature and high pressure.

[0089] Step 3: Inoculate the Bacillus subtilis strain into the sterilized culture medium under sterile conditions.

[0090] Step 4: ferment and culture at 30 degrees Celsius to allow Bacillus subtilis to multiply in large quantities.

[0091] Step 5: During the fermentation process, stirring and ventilation are performed at regular intervals to promote bacterial growth.

[0092] Step 6: After the fermentation is completed, the bacteria in the culture solution are collected and separated from the culture medium by centrifugation and filtration.

[0093] Step 7: Dry the bacterial cells to obtain the Bacillus subtilis inoculum. In step 2, the high temperature and high pressure sterilization condition is sterilization at 121 degrees Celsius for 20 minutes.

[0094] The following data were obtained when the bacterial agent obtained in the above examples was used in cotton fields and cotton cultivation:

[0095] Soil type: sandy loam with slight stickiness. Land preparation was done on April 6, film was laid and sown on April 8, and seedlings emerged on April 25.

[0096] Field management: Base fertilizer input is 50kg / mu (30kg compound fertilizer + 20kg urea). Drip water 8 times in total during the growth period, a total of 270 cubic meters per mu, 32kg / mu of urea, 23kg / mu of diammonium phosphate, and 20kg / mu of potassium fertilizer.

[0097] Chemical adjustment and control: Chemical control 8 times, total dosage 21 grams.

[0098] Main pests and diseases and their control: Mainly control tiger pests, thrips, cotton aphids and cotton bollworms.

[0099] Topping period: July 5. Flower harvest period: October 10.

[0100] The effect of the bacterial agent of the present application on the degradation of grass roots in cotton fields.

[0101] The test results show that the bacterial agent can effectively degrade the grass roots in the soil.

Claims

1. A method for preparing a microbial agent for degrading the herbicide chlorfenapyr, characterized in that: It includes Bacillus subtilis, a carrier substance, a nutrient substance and a protective agent; The effective viable count of the Bacillus subtilis is ≥ 120 billion / g; The carrier material is used to adsorb and carry Bacillus subtilis powder, and improve the stability and persistence of Bacillus subtilis in the soil; The nutrients are nitrogen, phosphorus, potassium, trace elements and growth factors, which are used to promote the growth and reproduction of Bacillus subtilis; The protective agent is bentonite or silica gel, which is used to improve the stability of Bacillus subtilis.

2. The method for preparing a microbial agent for degrading the herbicide chlorfenapyr according to claim 1, characterized in that: It also includes trace elements, including dipotassium hydrogen phosphate and magnesium sulfate, which are used to promote the growth and metabolism of Bacillus subtilis.

3. The method for preparing a microbial agent for degrading the herbicide chlorfenapyr according to claim 2, characterized in that: The content of the Bacillus subtilis is 1-5%, the content of the carrier material is 85-94%, the content of the nutrient material is 1-5%, the content of the trace element is 0.1-1%, and the content of the protective agent is 0-1%.

4. The method for preparing a microbial agent for degrading the herbicide chlorfenapyr according to claim 3, characterized in that: The carrier substances are glucose and peptone, and the nutrients are dipotassium hydrogen phosphate, magnesium sulfate and yeast extract. 20-30 g / L of glucose, 10-15 g / L of peptone, 5-10 g / L of yeast extract, 5-10 g / L of sodium chloride, 1-2 g / L of dipotassium hydrogen phosphate, and 0.5-1 g / L of magnesium sulfate are added into distilled water to prepare a culture medium for Bacillus subtilis, and then the Bacillus subtilis is inoculated onto the culture medium.

5. The method for preparing a microbial agent for degrading the herbicide chlorfenapyr according to claim 4, characterized in that: The specific steps of making Bacillus subtilis inoculant are as follows: Step 1, adding the carrier substance and the nutrient substance into distilled water in proportion, stirring evenly, and adjusting the pH value so that the pH value is between 7.0 and 7.5; Step 2, placing the components prepared in step 1 into a sterile container and sterilizing them under high temperature and high pressure; Step 3, inoculating Bacillus subtilis into a sterilized culture medium under sterile conditions; Step 4, placing the mixture under suitable temperature and conditions for fermentation culture to allow Bacillus subtilis to multiply in large quantities; Step 5: During the fermentation process, stirring and aeration are performed at regular intervals to promote bacterial growth; Step 6: After the fermentation is completed, the bacterial cells in the culture solution are collected and separated from the culture medium by centrifugation and filtration; Step seven, drying the bacterial cells to obtain the Bacillus subtilis agent.

6. The method for preparing a microbial agent for degrading the herbicide chlorfenapyr according to claim 5, characterized in that: In step 1, when adjusting the pH value, sodium hydroxide or hydrochloric acid is used for adjustment.

7. The method for preparing a microbial agent for degrading the herbicide chlorfenapyr according to claim 5, characterized in that: In step 2, the high temperature and high pressure sterilization condition is sterilization at 121 degrees Celsius for 20 minutes.

8. The method for preparing a microbial agent for degrading the herbicide chlorfenapyr according to claim 5, characterized in that: The suitable temperature in the step 4 is 28-30 degrees Celsius.