Bacillus subtilis, and breeding method and application thereof

The technology of Bacillus aureus and thuringiensis spore is applied in the field of biopesticide preparation, which can solve the problems of environmental pollution, affecting the quality of life, excessive pesticide residues, etc., and achieve the effects of high genetic stability, good inhibitory effect and broad application prospects.

Inactive Publication Date: 2015-08-19
CHINA THREE GORGES UNIV
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, due to excessive application and abuse of pesticides, soil fertility decline and environmental pollution
E

Method used

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  • Bacillus subtilis, and breeding method and application thereof
  • Bacillus subtilis, and breeding method and application thereof
  • Bacillus subtilis, and breeding method and application thereof

Examples

Experimental program
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Embodiment 1

[0028] Breeding of Bacillus subtilis ML1:

[0029] The leaves of Cypress twigs were collected in Yanzhiba, Yichang, Hubei Province in the Three Gorges Basin, and bacteria were obtained from them by the dilution plate method;

[0030] Preparation of diluent: take 10g of Shuiba siberiana branches and leaves, add 100mL of sterile water, and grind the tissue to pieces. Aseptically absorb 1.0mL of crushed tissue fluid and add it into 9.0mL sterile water, dilute to 10% with sterile water -1 、10 -2 、10 -3 、10 -4 、10 -5 、10 -6 、10 -7 A total of 7 concentration gradients.

[0031] Cultivation: Take 100 μL of each concentration gradient dilution and spread them on the beef extract peptone medium plate (beef extract 10g / L, peptone 10g / L, NaCl 5g / L, agar 20g / L, the rest is water, pH7.2-7.5, Sterilize at 121°C for 20 minutes), and incubate at 30°C in an inverted incubator.

[0032] Take a single colony: after the colony grows on the plate, pick a single colony in 10mL sterilized b...

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Abstract

The invention provides Bacillus subtilis ML1 which is preserved in China Center for Type Culture Collection on May 5, 2014, with an accession number of CCTCC No: M2014181. A breeding method for the Bacillus subtilis ML1 comprises the following steps: adding sterile water into the tissue fluid of leaves of Myricaria laxiflora and carrying out dilution to obtain 7 concentration gradients; coating medium plates with the diluted tissue fluid, invertedly placing the medium plates in a thermotank and carrying out culture at 30 DEG C; after bacterium colonies have grown on the plates, carrying out shake cultivation at a constant temperature of 30 DEG C so as to obtain bacterium culture solutions, sterilizing the bacterium culture solutions, mixing the bacterium culture solutions with glycerin and preserving the obtained mixture in a refrigerator with a temperature of -86 DEG C; subjecting the preserved Bacillus subtilis strain to streak inoculation in a slant medium and carrying out culture at 30 DEG C for 12 h; and subjecting the strain to streak inoculation in a medium and carrying out culture at 30 DEG C for 18 h so as to obtain the Bacillus subtilis ML1. The Bacillus subtilis ML1 is applied to prevention and treatment of soft rot disease of konjak and provides a novel option for development of a novel biological pesticide variety and novel biological control inocula.

Description

technical field [0001] The invention relates to a bacillus thuringiensis and its breeding method, and its application in the field of preparing biological pesticides. Background technique [0002] Soft rot, commonly known as "half-sided madness", is the most serious disease in konjac production. Konjac soft rot belongs to the genus Erwinia (Erwinia), a Gram-negative bacterium, Erwinia carotovora subsp.carotovora, which can develop diseases during the cultivation and storage periods , affecting yield and quality. The konjac planting industry loses a lot due to soft rot every year. [0003] For centuries, chemical pesticides have played a huge role in the control of plant diseases and insect pests, and have made important contributions to increasing, stabilizing and improving agricultural production. However, due to excessive application and abuse of pesticides, soil fertility decline and environmental pollution. Especially in recent years, due to excessive pesticide resid...

Claims

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Application Information

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IPC IPC(8): C12N1/20A01N63/00A01P1/00C12R1/07
Inventor 刘士平秦王阁阁田伟曾文孔玉珊蒋维薛艳红
Owner CHINA THREE GORGES UNIV
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