Fermentation method of bacillus amyloliquefaciens Bam22 for preventing and treating cruciferae plasmodiophora brassicae

A technology of amylolytic spores and fermentation methods, which is applied in the field of fermentation of Bacillus amyloliquefaciens Bam22 to prevent and control cruciferous clubroot, can solve the problems of limited biocontrol methods and restrictions, and achieve excellent activity, good biological activity, and many spores Effect

Active Publication Date: 2016-02-03
INST OF PLANT PROTECTION SICHUAN ACAD OF AGRI SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, due to the fact that Brassicaceae clubroot bacteria cannot be cultured in vitro to a certain extent, the selection of biocontrol strains for this disease is limited to a certain extent, so there are on

Method used

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  • Fermentation method of bacillus amyloliquefaciens Bam22 for preventing and treating cruciferae plasmodiophora brassicae
  • Fermentation method of bacillus amyloliquefaciens Bam22 for preventing and treating cruciferae plasmodiophora brassicae
  • Fermentation method of bacillus amyloliquefaciens Bam22 for preventing and treating cruciferae plasmodiophora brassicae

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Effect test

Embodiment 1

[0019] 1) Pick a single colony of Bacillus amyloliquefaciens Bam22 that has been cultured on a solid medium, place it in a liquid medium, and culture it with shaking at 31°C for 11 hours at a rotational speed of 200rpm, as a seed solution; the liquid medium The ingredients are calculated by mass percentage: yeast extract 0.2%, beef extract 0.1%, peptone 0.7%, NaCl 0.3%, the balance is water, pH=7.0;

[0020] 2) Put the product obtained in step 1) into a fermenter for 40 hours of fermentation, the tank temperature is 30°C, the stirring speed of the fermenter is 250rpm, and the air flow rate is 0.8:1 (V / Vm); the composition of the medium in the fermenter is as follows: The mass percentage is: soybean powder 4%, peptone 0.1%, glucose 0.5%, starch 2%, manganese sulfate 0.1%, ferrous sulfate 0.06%, dipotassium hydrogen phosphate 0.2%, calcium carbonate 0.1%, defoamer 0.01% , The balance is water, pH=7.0.

Embodiment 2

[0022] 1) Pick a single colony of Bacillus amyloliquefaciens Bam22 that has been cultured on a solid medium, place it in a liquid medium, and culture it with shaking at 32°C for 10 hours at a rotation speed of 220rpm as a seed solution; the liquid medium The ingredients are calculated by mass percentage: yeast extract 0.3%, beef extract 0.2%, peptone 0.8%, NaCl 0.6%, the balance is water, pH=7.0;

[0023] 2) Put the product obtained in step 1) into a fermenter for 50 hours of fermentation, the tank temperature is 27°C, the stirring speed of the fermenter is 220rpm, and the air flow rate is 0.9:1 (V / Vm); the composition of the medium in the fermenter is as follows: The mass percentage is: soybean powder 5%, peptone 0.15%, glucose 0.7%, starch 3%, manganese sulfate 0.1%, ferrous sulfate 0.06%, dipotassium hydrogen phosphate 0.2%, calcium carbonate 0.1%, defoamer 0.01% , The balance is water, pH=7.4.

Embodiment 3

[0025] 1) Pick a single colony of Bacillus amyloliquefaciens Bam22 that has been cultured on a solid medium, place it in a liquid medium, and culture it with shaking at 30°C for 12 hours at a speed of 210rpm, as a seed solution; the liquid medium The ingredients are calculated by mass percentage: yeast extract 0.25%, beef extract 0.12%, peptone 0.75%, NaCl 0.5%, the balance is water, pH=7.0;

[0026] 2) Put the product obtained in step 1) into a fermenter for 48 hours of fermentation, the temperature of the tank is 28°C, the stirring speed of the fermenter is 220rpm, and the air flow rate is 1:1 (V / Vm); the composition of the medium in the fermenter is as follows: The mass percentage is: soybean powder 4.5%, peptone 0.12%, glucose 0.6%, starch 2.5%, manganese sulfate 0.1%, ferrous sulfate 0.06%, dipotassium hydrogen phosphate 0.2%, calcium carbonate 0.1%, defoamer 0.01% , The balance is water, pH=7.3.

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Abstract

The invention provides a fermentation method of bacillus amyloliquefaciens Bam22 for preventing and treating cruciferae plasmodiophora brassicae. The method comprises the following steps: (1) picking up a bacillus amyloliquefaciens Bam22 colony cultivated on a solid culture medium, and carrying out shake culture in a liquid culture medium at 30-32 DEG C for 10-12 hours, wherein the rotating speed is 200-220rpm; and the product is taken as a seed solution; (2) fermenting the seed solution obtained in the step (1) in a fermentation tank for 40-50 hours, wherein the access amount of the seed solution is 8%-10%; the temperature of the tank is 27-30 DEG C; the stirring speed of the fermentation tank is 220-250rpm; the air flow is 0.8:1 to 1:1 (V/V.m); the efficient bacillus amyloliquefaciens Bam22 for cruciferae plasmodiophora brassicae is preserved at the General Microbiology Center of the China Committee for Culture Collection of Microorganisms on September 6, 2015; and the preservation number is CGMCC No.11329. The fermentation method provided by the invention is very favorable to cultivation of the bacillus amyloliquefaciens Bam22; and multiple bacteria are obtained and have good activity.

Description

technical field [0001] The invention belongs to the technical field of biological pesticides, and in particular relates to a fermentation method for preventing and treating cruciferous clubroot bacillus amyloliquefaciens Bam22. Background technique [0002] In just a few decades, cruciferous clubroot has spread rapidly around the world, seriously affecting the growth of cruciferous crops and causing huge economic losses. [0003] Given that the use of chemical pesticides is harmful to the environment and poses food safety hazards, biocontrol technology is increasingly favored by researchers and agricultural practitioners. However, due to the fact that Brassicaceae clubroot bacteria cannot be cultured in vitro to a certain extent, the selection of biocontrol strains for this disease is limited to a certain extent, so there are only a few strains that have good control effects on Brassicaceae clubroot disease. species, seriously limited the further development of biocontrol m...

Claims

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Application Information

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IPC IPC(8): C12N1/20A01P3/00C12R1/07
Inventor 刘勇黄小琴张蕾伍文宪刘红雨周西全
Owner INST OF PLANT PROTECTION SICHUAN ACAD OF AGRI SCI
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