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Bacillus amyloliquefaciense BA-KA3 and application thereof

A technology for dissolving starch spores and bacilli, applied in the field of microbial technology and biological control, can solve problems such as easy killing of beneficial microorganisms, drug resistance of pathogenic bacteria, and increase in the cost of tomato gray mold prevention and control, so as to promote crop growth and broad-spectrum inhibition. The effect of bacterial activity

Active Publication Date: 2015-07-08
BIOLOGY INST OF HEBEI ACAD OF SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, chemical pesticides are still the main measures to prevent and control the disease, but long-term use not only induces the resistance of pathogenic bacteria, but also increases the cost of tomato gray mold control and easily kills beneficial microorganisms in the environment

Method used

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  • Bacillus amyloliquefaciense BA-KA3 and application thereof
  • Bacillus amyloliquefaciense BA-KA3 and application thereof
  • Bacillus amyloliquefaciense BA-KA3 and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0023] Example 1 Isolation of Bacillus amyloliquefaciens BA-KA3

[0024] Bacillus amyloliquefaciens BA-KA3 was isolated from the rhizosphere soil of tomatoes in Liulintun Township, Luancheng County, Shijiazhuang City, Hebei Province.

[0025] The specific method is as follows: the rhizosphere soil of tomatoes was collected in the protected area of ​​Liulintun Township, Luancheng County, Shijiazhuang City, Hebei Province, and stored at 4°C for the isolation of antagonistic bacteria. Dilute 5g of tomato rhizosphere soil with 0.9% saline to 10 -4 、10 -5 、10 -6 . Pipette 100 μL each into the PB medium and spread evenly with a glass spreader. After culturing in an incubator at 28°C for 3 days, bacteria with different colony shapes were picked for isolation and purification. Using Botrytis cinerea as an indicator bacterium, the antagonistic bacteria of Botrytis cinerea were screened by plate confrontation method. After culturing in the dark at 22°C for 5 days, the strains wit...

Embodiment 2

[0027] Example 2 Identification of Bacillus amyloliquefaciens BA-KA3

[0028] The strain BA-KA3 does not produce pigment when grown on LB medium, and the colonies are flat or round, milky white and opaque, and the edges of the colonies are irregular. The surface is wrinkled. Gram-positive, rod-shaped, spore-forming, spore-elliptic or cylindrical. Refer to the "Common Bacteria Identification Manual" for physiological and biochemical identification of strains: V-P test positive, methyl red test negative, indole test positive, gelatin liquefaction test positive, citrate utilization test positive, starch hydrolysis test positive, lipase negative, h 2 S production test positive, nitrate reduction test positive, malonate utilization negative, tyrosine hydrolysis negative, cellulolysis positive.

[0029] 16S rDNA sequence analysis of bacterial strains: the molecular identification of strains is carried out by amplifying 16S rDNA of antagonistic bacteria.

[0030] Forward primer...

Embodiment 3

[0036] Example 3 Indoor control effect test of Bacillus amyloliquefaciens BA-KA3 on tomato

[0037] Using sterile water as a negative control and azoxystrobin as a commonly used chemical control, the control effects of Bacillus amyloliquefaciens BA-KA3 sterile fermentation broth and bacterial suspension on tomato detached leaves, fruits and potted tomatoes were determined.

[0038] The tomato fruit and detached leaves were subjected to surface disinfection treatment, soaked in 75% alcohol for 30 seconds, and washed three times with sterile water. The cell concentration was 1×10 9 CFU / mL of Bacillus amyloliquefaciens BA-KA3, centrifuge at 8000rpm for 20min at 4°C, take the supernatant, remove the bacteria with a 0.22μm microporous membrane, and obtain a sterile fermentation broth; resuspend the precipitate with sterile water to obtain a bacterial Suspension.

[0039] Determination of the control effect of BA-KA3 on tomato botrytis cinerea in vitro: sterilized pipette tip to p...

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Abstract

The invention relates to a Bacillus amyloliquefaciense BA-KA3 and application thereof. The Bacillus amyloliquefaciense BA-KA3 is preserved in China General Microbiological Culture Collection Center on October 23, 2013, and has preservation No. CGMCC8342; its on tomato gray mold germs has obviously of antagonistic anti-role, on wheat scab bacteria. The Bacillus amyloliquefaciense BA-KA3 has obvious antagonistic effect on Fusarium graminearum, Rhizoctonia cerealis, Alternaria alternata, Phytophthora capsici, Colletotrichum capsici, Cercospora personata, Alternaria alternate and Garlic sclerotinia disease, and can significantly improve plant growth indexes such as germination rate, and budding rate and root length of seeds,and especially applies to preparation of agricultural preparations for control of tomato diseases due to Botrytiscinerea invasion and has certain growth promotion effect.

Description

technical field [0001] The present invention relates to a kind of Bacillus amyloliquefaciens ( Bacillus amyloliquefaciense ) BA-KA3 and its application belong to the field of microbial technology and biological control. Background technique [0002] Botrytis cinerea is a fungal disease of tomato caused by Botrytis cinerea, which can damage tomato stems, leaves, flowers and fruits. After the onset spreads rapidly, it poses a great threat to tomato production, especially in winter and spring seasons in the north. The low temperature and high humidity climate conditions in the shed provide a favorable environment for the infection and spread of Botrytis cinerea. Cause tomato production to reduce 20%~40%, can reach more than 60% when serious. At present, chemical pesticides are still the main measures to prevent and control the disease, but long-term use not only induces the resistance of pathogenic bacteria, but also increases the cost of the control of Botrytis cinerea and e...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/20A01N63/00A01N63/02A01P3/00A01P21/00C12R1/07
CPCA01N63/00A01N63/10C12N1/205C12R2001/07
Inventor 刘昆昂黄亚丽姜倍倍马宏贾振华郝楠张晶晶宋聪赵芊黄媛媛刘方张立文宋水山
Owner BIOLOGY INST OF HEBEI ACAD OF SCI
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