Bacillus subtilis and culture method thereof

A technology of Bacillus subtilis and Bacillus subtilis is applied in the field of biological pesticides and fertilizers, which can solve the problems of no breakthrough, and achieve the effects of improving production efficiency, reducing production costs, and having strong reproductive ability.

Active Publication Date: 2018-05-18
WUHAN KERNEL BIO-TECH CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] In addition, the existing Bacillus subtilis fermentation level has been in the bottleneck period, and there is no major breakthrough

Method used

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  • Bacillus subtilis and culture method thereof
  • Bacillus subtilis and culture method thereof
  • Bacillus subtilis and culture method thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0024] Example 1 Screening and Identification of Bacillus subtilis KN-104

[0025] 1. Heat treatment screening of strains

[0026] Weigh 1.0 g of soil sample (crop rhizosphere soil that has been seriously infested with rice blast disease) and put it into a sterilized Erlenmeyer flask, add 90 ml of sterilized culture solution (starch 5-20 g / L, corn steep liquor 10-20 g / L, yeast extract 1-10g / L, corn flour 2-15g / L, peptone 5-10g / L, dipotassium hydrogen phosphate 0.5-5g / L, pH7.2-7.5), at a temperature of 30°C After culturing for 3 days under the condition of rotating speed of 120-180rpm, bathe in a constant temperature water bath at 80°C for 30min. After fully shaking, take the fermentation broth for concentration gradient dilution, draw 100ul dilution and spread it evenly on the NA medium (peptone 10g / L, beef extract powder 3g / L, sodium chloride 5g / L, agar powder 15g / L, pH value 7.2), after cultivating at 30°C for 3-5 days, pick a single colony for microscopic examination, an...

Embodiment 2

[0041] The cultivation of embodiment 2 bacillus subtilis KN-104 bacterial strain

[0042] (1) Strain activation:

[0043] Pick a ring of Bacillus subtilis KN-104 strain, inoculate it on the slant medium, and cultivate it at 30°C for 60h,

[0044] Incline medium: peptone 10g / L, yeast powder 5g / L, sodium chloride 10g / L, agar powder 15g / L, pH 7.0;

[0045] (2) Seed cultivation:

[0046] The Bacillus subtilis KN-104 activated in step (1) was inoculated in the seed medium, and cultivated at 30°C and 150rpm for 12h,

[0047]Seed medium: starch 15g / L, corn steep liquor 15g / L, yeast extract 3g / L, corn flour 8g / L, peptone 6g / L, dipotassium hydrogen phosphate 0.5g / L, pH7.2;

[0048] (3) Fermentation culture:

[0049] Put the cultured seed liquid into the fermentation medium, and cultivate it under the conditions of 32°C and 250rpm for 40h to obtain the Bacillus subtilis KN-104 fermentation liquid, the number of viable spores in the fermentation liquid reaches 35 billion CFU / ml,

[...

Embodiment 3

[0051] Embodiment 3 Bacillus subtilis KN-104 is to the crop disease-resistant specificity test

[0052] 1) Test agent: Bacillus subtilis KN-104 fermentation broth supernatant

[0053] 2) Targets for testing: common diseases of crops, the present invention only lists rice blast, leaf blast, sheath blight, cabbage black spot, tomato gray mold, watermelon wilt, apple anthracnose, soybean root rot, tomato Leaf mold.

[0054] 3) Determination by plate culture method: get 6ml of Bacillus subtilis KN-104 fermentation liquid supernatant and quantitative 54ml sterilized PDA culture medium and mix evenly, pour into 4 sterilized culture mediums evenly respectively, after cooling, make containing Medicine tablet. Set no drug blank control.

[0055] 4) The cultivated pathogenic bacteria (rice blast, leaf blast, sheath blight, cabbage black spot, tomato gray mold, watermelon wilt, apple anthracnose, soybean root rot, tomato leaf mold), in the absence of Under bacterial conditions, use a...

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Abstract

The invention discloses a bacillus subtilis KN-104 strain. The strain is deposited in China Center for Type Culture Collection with the deposit number of CCTCC NO: M2017697. By directional target screening, the obtained bacillus subtilis is high in specificity of disease resistance, has a remarkable prevention effect on rice blast, leaf blast, rice sheath blight and alternaria brassicae, but doesnot have prevention effect on diseases such as botrytis cinerea, watermelon wilt disease, apple anthracnose, soybean root rot and tomato leaf mould. Meanwhile, the strain is high in fertility and doesnot have hemolysis.

Description

technical field [0001] The invention relates to the technical field of biological pesticides and fertilizers, in particular to the screening and application of Bacillus subtilis. Background technique [0002] At present, the main means of preventing and controlling crop diseases are planting disease-resistant varieties, applying chemical pesticides and using biological pesticides. The main chemical pesticides such as tricyclazole, rice blastin EC, carbendazim, thiophanate-methyl, chlorothalonil, etc. are easy to make bacteria resistant to drugs, and the control effect is continuously reduced. Improper use will also cause damage to the ecological environment and human Health brings serious hazards that cannot be ignored, so biological control has gradually become the main means of targeting crop diseases. [0003] Bacillus subtilis is a kind of Bacillus genus. Because Bacillus subtilis has the advantages of fast reproduction, strong colonization ability, harmless to humans a...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/20A01P3/00C12R1/125
CPCC12N1/20C12N1/205C12R2001/125
Inventor 周莉潘渝刘守德刘华梅程治国邹维万俊王猛孙刚忠王晓辉
Owner WUHAN KERNEL BIO-TECH CO LTD
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