Bacillus amyloliquefaciens ba168 and its fermentation culture method and application

A technology for desolvating amyloliquefaciens and fermenting spores, which is applied to Bacillus amyloliquefaciens Ba168 and its fermentative cultivation and application fields, can solve problems such as crop diseases, industrial losses of traditional Chinese medicinal materials, and impact on crop yields, and achieves good environmental compatibility. Control effect, high biological activity effect

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A technology for desolvating amyloliquefaciens and fermenting spores, which is applied to Bacillus amyloliquefaciens Ba168 and its fermentative cultivation and application fields, can solve problems such as crop diseases, industrial losses of traditional Chinese medicinal materials, and impact on crop yields, and achieves good environmental compatibility. Control effect, high biological activity effect

CN102839142BActive Publication Date: 2017-02-08杨凌绿都生物科技有限公司

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Embodiment 1

[0037] Example 1:Isolation and purification of Bacillus amyloliquefaciens Ba168:

[0038] (1), soil sample collection:

[0039] Select multiple locations from the original forest soil in the deep mountains of the Qinling Mountains, take 3 to 15cm of soil from each location, mix it through a coarse sieve, and take 25g for separation;

[0040] (2), medium plate preparation:

[0041] Heat and melt NA medium and LB medium and pour it into a petri dish to cool to make NA medium and LB medium plates. The NA medium formula is: glucose 10g, agar 15g, beef extract 3g, yeast extract 1g, peptone 5g, water 1L, pH 6.8~7.0; LB liquid medium formula is: peptone 10g, NaCl 10g, water 1L , yeast extract 5g, pH 7.0;

[0042] (3), the separation of bacteria:

[0043] Plate inoculation was inoculated by smear method and mixed bacteria method.

[0044] Smearing method: Take 0.05ml of the above-mentioned 5 concentrations of soil dilutions, add them to the prepared NA and LB plates respectively...

Embodiment 2

[0048] Example 2: Identification of Bacillus amyloliquefaciens Ba168:

[0049] The identification process to the bacillus amyloliquefaciens Ba168 CGMCC No.6462 of embodiment 1 gained is:

[0050] (1) Morphological characteristics: the colonies on the LB medium are round, with obvious gloss on the surface, not smooth; the edges are irregular, diffuse, and the surface is wrinkled; the colonies are milky white, opaque, turbid; they have certain adhesion; The formula of the LB medium is: 10 g of peptone, 5 g of yeast extract, 10 g of NaCl, 18 g of agar, 1000 ml of water, and the pH is 7.0.

[0051] In the early stage of culture (5~12h), the colonies are colorless and transparent droplets, which are shiny; in the later stage (after 24h), they are white and opaque, relatively dim.

[0052] The thallus is rod-shaped, about 2 μm long, solitary, with peritoneal flagella, and produces spores. Gram stain was positive.

[0053] (2), physiological and biochemical experiment: by the ph...

Embodiment 3

[0061] Example 3: The cultivation method of bacillus amyloliquefaciens (Bacillus amyloliquefaciens) Ba168 of the present invention is:

[0062] (1), strain activation:

[0063] Medium: LB solid medium, the formula of the LB solid medium is: 5g of yeast extract, 10g of peptone, 5g of NaCl, 15-20g of agar, 1000ml of water, pH 7.4-7.6;

[0064] Culture conditions: 30°C, 48h;

[0065] Continuous transfer for 3 generations to ensure the purity of the strain, and then inoculated into the fermentation broth for use.

[0066] (2), preparation of seed solution (shake flask fermentation or eggplant bottle fermentation):

[0067] Culture medium: LB liquid medium, the formula of described LB liquid medium is: yeast extract 5g, peptone 10g, NaCl 10g, water 1000ml, pH 7.0 (flask volume 250ml, filling capacity 50ml; eggplant bottle volume 1000ml, filling Liquid volume 200-300ml);

[0068] Culture conditions: 30°C, 50h-60h;

[0069] Inoculation amount: 6~8%.

[0070] (3), seed tank fe...

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Abstract

The invention discloses bacillus amyloliquefaciens (Ba 168) and a preparation method and an application of the bacillus amyloliquefaciens, and aims at providing a biological control method of phytopathogens. The bacterial strain is preserved in the China General Microbiological Culture Collection Center (CGMCC), and the preservation number is CGMCC No.6462 on August 21, 2012. The bacterial strain is separated and obtained from soil in the Qinling Mountain virgin forest, and subjected to bacteria activation, shake-flask fermentation, seed liquid fermentation and large tank fermentation to obtain fermentation liquid; and the fermentation liquid is subjected to filtration and concentration, ultrafiltration membrane separation, and spray drying and packaging to be prepared into bactericide. The bacillus amyloliquefaciens has the advantages that the bacillus amyloliquefaciens can well prevent and cure bighead atractylodes rhizome root rot, American ginseng root rot, pseudo-ginseng root rot and other root rot caused by fusarium oxysporum.

Description

technical field [0001] The invention belongs to the field of biological control of plant diseases by using beneficial microorganisms, and in particular relates to Bacillus amyloliquefaciens (Bacillus amyloquefaciens) Ba168 strain, and the fermentation production of the strain and the effect of the fermentation product on preventing and controlling plant pathogenic bacteria. Background technique [0002] The prevalence of plant diseases not only causes crop yield reduction, but also causes great economic losses to human beings, and even brings catastrophic consequences. The harm of plant diseases can also lead to poor quality of agricultural products, unsuitable for use or processing, and even lead to poisoning of humans and animals. In order to prevent and control plant diseases, chemical pesticides have once become people's first choice because of their advantages of low cost, quick effect, time-saving and labor-saving, etc. However, chemical pesticides are generally non-de...

Claims

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Application Information

Patent Timeline

08 Feb 2017

Publication

CN102839142B

IPC: C12N1/20; A01N63/02; A01P3/00; C12R1/07

Inventors: 杨轩; 翟枫