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Bacillus subtilis strain A33, microbial inoculum and preparation method and application thereof

A technology of Bacillus subtilis and A33, applied in the field of agricultural microorganisms, can solve problems such as crop and human phytotoxicity, destruction of soil ecological diversity, environmental pollution, etc., and achieve remarkable field control effects, obvious disease resistance and growth promotion, and environmental friendliness Effect

Active Publication Date: 2014-07-30
MICROBIOLOGY INST OF SHAANXI
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  • Abstract
  • Description
  • Claims
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Problems solved by technology

However, the abuse of chemical pesticides has also led to serious consequences, such as environmental pollution, destruction of soil ecological diversity, and phytotoxicity to crops and humans, etc.

Method used

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  • Bacillus subtilis strain A33, microbial inoculum and preparation method and application thereof
  • Bacillus subtilis strain A33, microbial inoculum and preparation method and application thereof
  • Bacillus subtilis strain A33, microbial inoculum and preparation method and application thereof

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preparation example Construction

[0032] The preparation method of above-mentioned microbial bacterial agent, concrete steps are as follows:

[0033] (1) Strain activation: inoculate the cryopreserved A33 strain on LB plate medium (broth medium) and culture it at 28-37°C for 18-24 hours, then pick a single colony and inoculate it on LB slant medium and cultured at 28-37°C for 18-24 hours, then washed the surface of the culture medium with sterile water, and used the eluate as the inoculum;

[0034] (2) Preparation of seed solution: Add the inoculation solution prepared in step (1) to the LB liquid medium according to the ratio of 0.05-0.5% (volume percentage), and shake and cultivate at 28-37°C for 12-18 hours, as the seed liquid;

[0035] (3) Fermentation culture: put the seed liquid obtained in step (2) into the fermentation medium according to the proportion of 1-10% (volume percentage), and then cultivate it under the condition of 28-37°C and the rotating speed of the shaker at 150-23Orpm for 32 -44 hour...

Embodiment 1

[0064] The preparation of A33 microbial inoculum is carried out according to the following steps:

[0065] (1) Strain activation: Inoculate the A33 strain stored at -80°C on the LB plate medium, and cultivate it at 33°C for 18 hours, then pick a single colony and inoculate it on the LB slant medium, and inoculate it at 33°C Cultivate for 18 hours, then wash the surface of the medium with sterile water, and use the eluate as the inoculum;

[0066] (2) Preparation of seed solution: Prepare LB liquid medium according to the conventional method, fill 100mL of LB culture solution into a 250mL Erlenmeyer flask, and after autoclaving, insert the above-mentioned activated inoculation loop in step (1) into each bottle. The A33 strain was cultured with shaking on a constant temperature shaker at a rotation speed of 210 rpm, and cultured at 33°C for 24 hours, and the obtained seed liquid was obtained;

[0067] (3) Preparation of fermentation medium: according to 3% sucrose, 4% corn star...

Embodiment 2

[0071] This experiment was carried out in the greenhouse of the Institute of Plant Protection, Hebei Academy of Agriculture and Forestry Sciences, and cucumber powdery mildew was selected (provided by the Laboratory of Plant Disease Biological Control, Institute of Plant Protection, Hebei Academy of Agriculture and Forestry Sciences). Using the cotyledon spray method, the fermentation liquid of the biocontrol strain was evenly sprayed on the cucumber leaf surface with a throat sprayer, and the inoculation concentration was 5×10 after 24 hours. 5 Spores / mL Cucumber powdery mildew spore suspension, set aseptic water control. The experiment was carried out in a greenhouse with a relative humidity of 30%-70% and a temperature of 22°C. After the blank control was fully onset, the disease grade was investigated, and the disease index and control effect were calculated. Cucumber powdery mildew grade classification standards and control efficacy calculation methods refer to "GB / T1798...

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Abstract

The invention discloses a Bacillus subtilis strain A33, which is preserved in China General Microbiological Culture Collection Center and has a preservation number of CGMCC No.7468. The invention also discloses a microbial inoculum prepared from the A33 strain, and a preparation method and application thereof. The A33 strain provided by the invention can be used for the control of common foliage diseases such as powdery mildew, downy mildew, gray mold and wilt of facility crops, and has the characteristics of high efficiency, wide antimicrobial spectrum, simple usage, low cost and no pollution, etc.

Description

technical field [0001] The invention belongs to the technical field of agricultural microorganisms, and in particular relates to a bacillus subtilis strain A33, a bacterial agent, a preparation method and an application. Background technique [0002] Facility agricultural production is a semi-closed ecosystem under human control. In addition to summer and autumn sun-shade net-covered cultivation, compared with open-field cultivation ecosystems, it has the characteristics of high temperature, large temperature difference, low light, high humidity and slow airflow, which is conducive to occurrence of plant diseases. The resulting crop loss is close to about one-third of the total production, causing huge economic losses. For a long time, the control of plant diseases has mainly relied on the use of chemical pesticides. However, the abuse of chemical pesticides has also led to serious consequences, such as environmental pollution, destruction of soil ecological diversity, and...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/20A01N63/00A01N63/02A01P3/00C12R1/125
Inventor 马瑜柯杨李勃
Owner MICROBIOLOGY INST OF SHAANXI
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