Bacillus amyloliquefaciens D2WM as well as preparation method and application of bacillus amyloliquefaciens D2WM

A technology of amylolytic spores and bacilli, which is applied in botany equipment and methods, biochemical equipment and methods, applications, etc., and can solve problems such as long time-consuming, cumbersome preparation process, and narrow biocontrol range

Active Publication Date: 2016-01-06
HUBEI ENG UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Sun Ming etc. have invented a kind of Bacillus subtilis BSn5 and antibacterial protein APn5 (application number: 200710053427.8) that have bacteriostatic activity to Erwinia carotovora soft rot, the bacteriostatic active substance of this Bacillus subtilis is protein APn5, can only Antagonism against Erwinia subspecies of carrot soft rot, its biocontrol range is relatively narrow; Huang Jing et al. disclosed that a kind of Bacillus amyloliquefaciens Lh-1 is effective in the prevention and treatment of cotton verticillium wilt, Chinese cabbage soft rot, bacterial wilt, and peach tree rot. disease, apple rot, etc. (application number: 201410735508.6), it is necessary to use Bacillus amyloliquefaciens and chitosan oligosaccharides to form a microecological bacterial agent, which is relatively effective only for the control of cotton verticillium wilt; Liu Junang et al. A Bacillus brevis Z26 that antagonizes camellia oleifera disease is disclosed, which can effectively inhibit the growth of pathogenic bacteria such as camellia oleifera anthracnose, root rot, and hemimania (application number: 201110027957.1). The rate reached 83.4%, and there is still room for further improvement; Zhou Guoyin

Method used

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  • Bacillus amyloliquefaciens D2WM as well as preparation method and application of bacillus amyloliquefaciens D2WM
  • Bacillus amyloliquefaciens D2WM as well as preparation method and application of bacillus amyloliquefaciens D2WM
  • Bacillus amyloliquefaciens D2WM as well as preparation method and application of bacillus amyloliquefaciens D2WM

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Effect test

Embodiment 1

[0044] A preparation method of Bacillusamyloliquefaciens D2WM, the steps of which are:

[0045] A. Isolation and purification of bacillus: Take 10g of healthy oil tea rhizosphere soil sample and add it to a triangular flask containing 90mL sterile water and 30-50 glass beads to make a 1:10 concentration suspension, at 28℃, After shaking in a shaker at 180 rpm for 30 minutes, place the bacterial suspension in a water bath at 80°C for 15 minutes. Transfer 1 mL of the soil suspension into a test tube containing 9 mL of sterile water to make 10 -2 Bacteria suspension, and so on, get 10 -3 , 10 -4 , 10 -5 And 10 -6 Bacteria suspension. After serial dilution, draw 100μL and the dilution factor is 10 -3 -10 -6 The bacterial suspension is evenly spread on the separation medium-beef extract peptone medium. The medium formula is as follows (1L): beef extract 3.0g, peptone 10.0g, NaCl5.0g, agar 15.0g, distilled water 1000mL, pH7. 2-7.4. Place the plate coated with the bacterial suspension...

Embodiment 2

[0056] An application of Bacillusamyloliquefaciens D2WM in the preparation of a medicine for treating or preventing vegetable soft rot, and the steps are:

[0057] A. The preparation of a biocontrol agent of Bacillusamyloliquefaciens D2WM in the treatment or prevention of vegetable soft rot. The steps are: pick a ring of D2WM colonies on the LB plate or slope and transplant it into a test tube for activation, 35 Cultivate in a shaker at ℃ for 16 hours, rotate at 150 rpm, the LB liquid medium in the test tube is: peptone 10g / L, yeast extract 5g / L, sodium chloride 5g / L, pH 7.0; the activated bacterial liquid is in accordance with 1%-3 % Of the inoculum was transplanted into a 250mL Erlenmeyer flask containing 100mL LB liquid medium, rotating at 150 rpm, and culturing on a shaker at 35°C for 10h-24h, when the bacterial content is greater than 2×10 8 Above cfu / mL, the fermentation bacteria liquid can be obtained and stored as a biocontrol agent.

[0058] B. Bacillus amyloliquefaciens D...

Embodiment 3

[0061] An application of Bacillusamyloliquefaciens D2WM in the preparation of a medicine for treating or preventing six kinds of pathogenic bacteria of Camellia oleifera, the steps are:

[0062] A. The preparation of a BacillusamyloliquefaciensD2WM biocontrol agent in the treatment or prevention of six kinds of pathogenic bacteria of Camellia oleifera, the steps are: pick a ring of D2WM colonies on the LB plate or slope and transplant it into a test tube for activation, 35°C Incubate on a shaker for 16 hours at a speed of 150 rpm. The LB liquid medium in the test tube is: peptone 10g / L, yeast extract 5g / L, sodium chloride 5g / L, pH 7.0; the activated bacterial liquid is 1%-3% Transplant the inoculum into a 250mL Erlenmeyer flask containing 100mLLB liquid medium, rotate at 150rpm, and cultivate on a shaker at 35℃ for 10-24h, when the bacterial content is greater than 2×10 8 Above cfu / mL, the fermentation bacteria liquid can be obtained and stored as a biocontrol agent.

[0063] B. In...

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Abstract

The invention discloses a bacillus amyloliquefaciens D2WM as well as a preparation method and application of bacillus amyloliquefaciens D2WM. The bacillus amyloliquefaciens D2WM comprises the steps of A, bacillus separation and purification, namely collecting after inoculating a slope; B, preliminary screening of bacillus, namely sequentially selecting strains having antagonistic effects on Erwinia and camellia oleifera diseases by using a point connection method and a plate confrontation method; and C, secondary screening of bacillus, namely culturing the preliminarily-screened strains with the antagonistic effects in an LB liquid culture medium by using a cup-plate method, and screening the strains simultaneously having the antagonistic effects on main diseases of Erwinia and camellia oleifera through an antagonistic bacterium fermentation liquid and sterile filtrate. When being applied to preparation of drugs for treating or preventing soft rot of vegetables and camellia oleifera diseases, the bacillus amyloliquefaciens has a broad-spectrum antagonistic effect, high in growth speed, capable of efficiently antagonizing anthracnose of camellia oleifera, leaf spot diseases of camellia oleifera, black spot of camellia oleifera, canker of camellia oleifera and various diseases and remarkable in antibacterial effect, the culture time for general bacteria and fungi is greatly shortened, and the production cost is reduced.

Description

Technical field [0001] The invention relates to the field of plant protection, and more specifically to a method for preparing Bacillusamyloliquefaciens D2WM which can simultaneously antagonize vegetable soft rot and main diseases of camellia oleifera, and also relates to a preparation method of Bacillusamyloliquefaciens D2WM, and also relates to a starch solution Application of Bacillus Bacillusamyloliquefaciens D2WM in preparing medicine for treating or preventing vegetable soft rot and camellia disease. Background technique [0002] Vegetable soft rot is also known as water rotten and bumps. In addition to harming cruciferous vegetables (such as radish, Chinese cabbage, etc.), it can also harm vegetables such as potatoes, tomatoes, lettuce, cucumber, celery and onions, causing varying degrees of loss. In the early stage of the onset of vegetable soft rot, the diseased part is water-soaked and translucent, and then expanded to light gray-brown wet rot, the diseased tissue is s...

Claims

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Application Information

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IPC IPC(8): C12N1/20A01N63/00A01N63/02A01P1/00A01P3/00C12R1/07
Inventor 魏蜜傅本重王立华李国元张伟鲁旭东
Owner HUBEI ENG UNIV
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