Bacillus subtilis bacterial strain and application thereof

A technology of Bacillus subtilis and strains, applied in the field of microorganisms, can solve the problems of chemical prevention and control of environmental pollution and insecurity, and achieves the effects of high control effect, huge application potential, and stable control effect.

Active Publication Date: 2017-05-31
INST OF VEGETABLE & FLOWERS CHINESE ACAD OF AGRI SCI +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] The purpose of the present invention is to provide a kind of bacillus subtilis bacterial strain and its application

Method used

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  • Bacillus subtilis bacterial strain and application thereof
  • Bacillus subtilis bacterial strain and application thereof
  • Bacillus subtilis bacterial strain and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0023] Isolation, purification and identification of Bacillus subtilis:

[0024] Take soil samples evenly from the tomato planting fields in Shunyi, mix 10g of soil with 100mL of sterilized water to make a suspension, and then carry out gradient dilution in turn, take 10000 times diluted soil mixture 100μL and spread it on the LB medium plate, The formula of LB medium is: yeast powder 5g / L, peptone 10g / L, NaCl 10g / L, agar powder 1.3-1.5g / 100mL, dilute to 1L and seal. The medium was cultured at 37°C. After the tissue pieces grew colonies, a single colony was picked and purified on LB medium for classification and identification. The strains were morphologically determined according to the Bergey's Manual of Bacterial Identification.

[0025] Pick a single colony with a sterilized toothpick, put it into an EP tube filled with 300 μL of bacterial lysate SDS, lyse at 65°C for 2 hours, and then extract bacterial DNA with a bacterial genomic DNA extraction kit (Tiangen Biotechnolog...

Embodiment 2

[0030] 1, the preparation method of Bs-1 bacterial strain fermented liquid, comprises the following steps:

[0031] (1) Streak-inoculate Bacillus subtilis Bs-1 on LB medium, culture it on a plate in a constant temperature incubator for 60 hours, and set the culture temperature at 25°C to obtain a single colony;

[0032] (2) Inoculate a single colony of Bacillus subtilis Bs-1 in a seed medium (LB medium), and cultivate it in a constant temperature shaking table with a rotation speed of 160 rpm and a temperature of 25° C. for 24 hours; liquid seed medium formula For LB medium, namely yeast powder 5g / L, peptone 10g / L, NaCl 10g / L, dilute to 1L;

[0033] (3) Inoculate the seed liquid into the sterilized fermentation medium to carry out fermentation culture, the inoculum size is 3%, the temperature is set at 25°C, and the obtained concentration is 10 4 cfu / mL fermentation liquid (stock solution); liquid fermentation medium formula is LB medium, namely yeast powder 5g / L, peptone 10g...

Embodiment 3

[0045] 1, the preparation method of Bs-1 bacterial strain fermented liquid, comprises the following steps:

[0046] (1) Streak inoculation of Bacillus subtilis Bs-1 on the LB medium, and plate culture in a constant temperature incubator for 60 hours at a culture temperature of 25°C to obtain a single colony;

[0047] (2) Inoculate a single colony of Bacillus subtilis Bs-1 in the seed medium, and cultivate it in a constant temperature shaker with a rotation speed of 160 rpm and a temperature of 37° C. for 24 hours; the formula of the liquid seed medium is LB medium, That is, yeast powder 5g / L, peptone 10g / L, NaCl 10g / L, dilute to 1L;

[0048] (3) Inoculate the seed liquid into the sterilized fermentation medium to carry out fermentation culture, the inoculum size is 3%, the temperature is set at 25°C, and the obtained concentration is 10 4 cfu / mL fermentation broth; liquid fermentation medium formula is LB medium, that is, yeast powder 5g / L, peptone 10g / L, NaCl 10g / L, and the ...

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Abstract

The invention discloses a bacillus subtilis bacterial strain and application thereof. The strain is bacillus subtilis Bs-1, is preserved in China Microbiological Collection Management Committee General Microbiology Center, and has a preservation number of CGMCC NO.12364. The invention further discloses application of a biological agent prepared by the bacillus subtilis Bs-1 to preparation and treatment of root-knot nematode. In the invention, bacillus subtilis is separated from soil, has an excellent control effect on root-knot nematode, is easy for colonization in the soil, and is beneficial to early prevention and control of the root-knot nematode hazard; the bacterial strain has a huge application potential for solving the problem of prevention and treatment of the root-knot nematode, and is helpful for implementing high yield, safety and pollution-free production of vegetables; fermentation liquor of the bacillus subtilis Bs-1 bacterial strain reaches a fatality rate of 100% on root-knot nematode, reaches a prevention and treatment effect of 76.9% in a greenhouse, has the good and stable prevention and treatment effect, and is suitable for agricultural production requirements.

Description

technical field [0001] The invention belongs to the technical field of microorganisms, and in particular relates to a bacillus subtilis strain and an application thereof. Background technique [0002] Root-knot nematode (Meloidogyne) is a highly specialized omnivorous plant pathogenic nematode. Nematodes known to harm vegetables mainly include root-knot nematode peanut, root-knot nematode northern, root-knot nematode southern, root-knot nematode java, and root-knot nematode beet. Nematodes have a wide range of hosts, often harming more than 30 kinds of vegetables such as melons, solanaceous fruits, beans, radishes, carrots, lettuce, and cabbage, and can also spread some fungal and bacterial diseases. Root-knot nematode is an important crop pathogen, causing losses of about billions of dollars every year. Root-knot nematodes mainly infect the roots of crops. After infection, the roots swell to form huge root knots, which can be connected to form beads. Because the roots are...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/20A01N63/00A01N63/02A01P5/00C12R1/125
CPCA01N63/00A01N63/10C12N1/205C12R2001/125
Inventor 茆振川杨宇红谢丙炎凌键张晓平
Owner INST OF VEGETABLE & FLOWERS CHINESE ACAD OF AGRI SCI
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