A kind of Bacillus subtilis inoculum and its preparation method and application

A technology of Bacillus subtilis and inoculum, applied in the field of Bacillus subtilis inoculum and its preparation, can solve the problems of difficulty in forming large-scale application, unstable use effect, poor stability of inoculum, etc., so as to reduce the amount of fine powder produced , The effect of easy operation and short fermentation period

Active Publication Date: 2021-09-24
北京新正和农业科技有限公司
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

CN103146617A (a strain of Bacillus subtilis and its cultivation method and application) is cultivated at high temperature of 35-38°C, and the energy consumption of long-term fermentation for 48-72 hours is relatively large; it is only effective for the prevention and treatment of cucumber and watermelon Verticillium wilt (fungal disease) effect
CN111778173A (Bacillus subtilis Pro1A2, its bacterial agent and its preparation method and its application in melon cultivation) take 28~35℃, secondary fermentation, first stage: 12~16 hours; second stage: 36~48 hours, apply The dosage form is fermented liquid (with many impurities, this dosage form is extremely unstable, and it is difficult to form a large-scale application)
[0004] The existing method for preparing Bacillus subtilis usually adopts liquid two-stage fermentation to obtain the fermentation liquid, which is concentrated in a frame or disc centrifuge after pretreatment, and then spray-dried to obtain the original powder; the fermentation liquid is passed through a frame or disc centrifuge. Centrifuge separation, solid-liquid separation, fast separation speed but low yield
In addition, the use of multi-stage fermentation will also cause problems such as rapid strain mutation (mutation), premature aging, and high rate of miscellaneous bacteria, which makes the prepared microbial products poor in stability and unstable in use

Method used

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  • A kind of Bacillus subtilis inoculum and its preparation method and application
  • A kind of Bacillus subtilis inoculum and its preparation method and application
  • A kind of Bacillus subtilis inoculum and its preparation method and application

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0026] Embodiment 1, the activation of bacterial classification

[0027] A. According to the composition ratio of spore medium (solid medium) in Table 1, prepare strain isolation medium and test tube (bacterial strain propagation) medium;

[0028] B. Subject the prepared separation medium to 110°C at 1.5 kgf / cm 2 After sterilization, incubate at a constant temperature of 34°C for 24 hours before use, and pass the prepared test tube culture medium at 110°C at 1.5 kgf / cm 2 After sterilization, incubate at a constant temperature of 34°C for 24 hours for use;

[0029] C. Prepare the original seed spore suspension, multiply it, and cultivate it at a constant temperature of 34°C ± 2°C for 168h; refrigerate it at 0-5°C for later use;

[0030] D. Preparation of test tube slant: pick out the single colony strains in the separation medium, transfer them evenly to the test tube medium for 168 hours at a constant temperature of 34°C ± 2°C, and set aside;

[0031] E. Preparation of bact...

Embodiment 2

[0032] Embodiment 2, primary fermentation prepares Bacillus subtilis inoculant

[0033] A. According to the formula of the liquid fermentation medium in Table 1, pour all kinds of raw materials into the preparation tank and stir to constant volume. The feed liquid is then filtered through a 10-mesh filter, and the filtrate is moved to the fermentation tank. / cm 2 Under the condition of sterilization, the temperature of the feed liquid was lowered to 30° C., and the bacterial suspension prepared in Example 1 was introduced into the tank by differential pressure method for cultivation (inoculum size: 30%);

[0034] B. Culture conditions: First-level aerobic submerged liquid fermentation culture. If the pH (pH) is abnormally high, if no bacteria is confirmed, an appropriate amount of 10% glucose can be added and the air flow can be reduced to normal.

[0035] Speed: 135~160 rpm / min

[0036] Airflow: 0 -10h 320m 3 / h

[0037] 10-24h 380m 3 / h

[0038] 24-Tank 450m 3 / h

[...

Embodiment 3

[0047] Embodiment 3, preventive effect test

[0048] Test design

[0049] 1, purpose: by test, determine the control effect of the bacillus subtilis agent that embodiment 2 prepares to main eubacterial disease;

[0050] 2. Control agent: the product using the biopesticide registration certificate obtained by Deqiang Biological Co., Ltd., namely: 100 billion Bacillus subtilis (model BS-10) for the prevention and treatment of fungal diseases; 10 billion Bacillus subtilis for the prevention and treatment of bacterial diseases (Model M22);

[0051] 3. Region: Simultaneously carry out control experiments on the above-mentioned diseases in four places in Sichuan Province, Guizhou Province, Henan Province and Heilongjiang Province;

[0052] 4. The application method is shown in Table 2 below

[0053] Table 2. Field application methods

[0054]

[0055] 5. Investigation and data analysis: 30 plants in each of the control and test tissues, and the number of diseased plants in th...

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Abstract

The invention discloses a bacillus subtilis bacterial agent and its preparation method and application. The bacterial agent is obtained by activating bacillus subtilis strains, liquid fermentation culture, ceramic membrane filtration and spray drying, and can be used to prevent and control fungi of various crops and bacterial pathogen infection, the preparation method is simple, and has the advantages of short fermentation period, high number of live spores in a tank, high ceramic membrane filtration yield, good control effect and strong stability.

Description

technical field [0001] The present application relates to the technical field of microbial biology, in particular to a Bacillus subtilis bacterial agent and its preparation method and application. Background technique [0002] Fungal and bacterial diseases are two major diseases that endanger the growth of crops, and have seriously endangered agricultural production. At present, the commonly used agents for preventing and controlling true and bacterial diseases are mainly chemical pesticides. Chemical agents usually have mediocre effects on some diseases, but have a great impact on the ecological environment and the quality and safety of agricultural products. [0003] Most of the existing Bacillus subtilis preparations have a single function, either can only prevent fungal diseases, or can only prevent bacterial diseases, and the activity attenuation rate of the existing bacterial agents is ≥ 60%, the genetic variation rate is ≥ 70%, and it is resistant to environmental int...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N1/20A01N63/22A01P1/00A01P3/00C12R1/125
CPCA01N63/22C12N1/20
Inventor 易虹
Owner 北京新正和农业科技有限公司
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