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Bacillus amyloliquefaciens for inhibiting bird rape plasmodiophora brassicae and separation and application of active substances thereof

A technology for dissolving amyloid spores, Pseudomonas brassicae, applied in applications, chemicals for biological control, bacteria, etc.

Active Publication Date: 2012-03-21
HUBEI BIOPESTICIDE ENG RES CENT
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

At present, there are few types of commercialized Bacillus amyloliquefaciens preparations, such as Bacillus subtilis var. amyloliquefaciens FZB24 which has obtained a limited commercial production application license

Method used

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  • Bacillus amyloliquefaciens for inhibiting bird rape plasmodiophora brassicae and separation and application of active substances thereof
  • Bacillus amyloliquefaciens for inhibiting bird rape plasmodiophora brassicae and separation and application of active substances thereof
  • Bacillus amyloliquefaciens for inhibiting bird rape plasmodiophora brassicae and separation and application of active substances thereof

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Embodiment Construction

[0040] The Bacillus amyloliquefaciens (Bacillus amyloliquefaciens) HB-26 of the present invention was preserved by the China Center for Type Culture Collection on July 18, 2011, with the number CCTCC NO.M2011259. The cell size of the bacterial strain is (0.4-1.0)umx(1.0-1.5)um, the cells are short rod-shaped, the spores grow in the middle, the Gram staining is positive, and it can move. The colonies formed on the LB agarose plate were milky white, oval, with raised and wrinkled surfaces. The optimum growth temperature is 30°C, pH6-8 all grow well, and the optimum growth pH is 7.0-7.2.

[0041] The present invention is described in detail below:

[0042] One, the culture method of bacillus amyloliquefaciens, concrete steps are as follows:

[0043] 1. Isolation and preservation of bacteria

[0044] Soil samples and liquid samples collected from all over the country (such as biogas slurry collected from Jingzhou, Hubei) were separated, cultured and preserved.

[0045] The spe...

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Abstract

The invention relates to a bacillus amyloliquefaciens for inhibiting bird rape plasmodiophora brassicae and separation, application and purposes of active substances thereof. Bacillus amyloliquefaciens HB-26 is preserved in China Center for Type Culture Collection (CCTCC), the preserving number is CCTCCNO: M2011259. A culture method is as follows: inoculating biogas slurry separated bacterial strain in an LB liquid nutrient medium, preserving the biogas slurry separated bacterial strain at 30 DEG C for one night, pouring fermentation liquor at the root of Chinese cabbage and finding the bacterial strains with bacteriostatic activity on plasmodiophora brassicae. The separated active substance is characterized in that the HB-26 is inoculated in the LB liquid nutrient medium, the fermentation liquor is dried by freezing, the frozen strain powder is added with ethyl acetate for extraction, is vibrated, rotated and evaporated, is dissolved by methyl alcohol, and then subjected to HPLC (High Performance Liquid Chromatrography) separation, the components are collected, the mass spectrum identification is conducted, and the component of the active substance can be judged according to the condition of the generation of plasmodiophora brassicae disease of the Chinese cabbage. The molecular weight of the bacteriostatic activity substance BA20 is 1494.0, and the nuclear magnetic resonance hydrogen spectrum shows that the chemical nature of the bacteriostatic activity substance is carbohydrate bacteriocin. The active substance is used for preparing a microbial agent for preventing the plasmodiophora brassicae disease of the Chinese cabbage.

Description

technical field [0001] The invention relates to a strain of Bacillus amyloliquefaciens, a method for cultivating the Bacillus amyloliquefaciens, a method for isolating an active substance having an antibacterial effect on Plasmodium brassicae, and the application of the active substance are related to the technical field of microbial pesticides. Background technique [0002] Bacillus is a non-pathogenic bacterium that widely exists in nature and can produce a variety of antibacterial substances, most of which are polypeptides, mainly inhibiting Gram-positive bacteria; some can also inhibit Gram-negative bacteria, molds and yeasts (Liu Jing et al., Research on the properties of antibacterial substances of Bacillus subtilis JA and the separation and purification of antimicrobial peptides, Acta Microbiology, 2004, 44(4): 511-513). They are easy to reproduce, have strong ability to adapt to the environment, and have great development and utilization value. Bacillus amyloliquefa...

Claims

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Application Information

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IPC IPC(8): C12N1/20C12P19/00A01N63/02A01P3/00C12R1/07
Inventor 刘晓艳杨自文王开梅万中义闵勇江爱兵曹春霞张志刚周荣华
Owner HUBEI BIOPESTICIDE ENG RES CENT
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