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Bacillus amyloliquefaciens with broad-spectrum pathogen inhibition function and application of bacillus amyloliquefaciens

A technology of amylolytic spores and pathogenic bacteria, applied in the direction of application, chemicals for biological control, bacteria, etc., can solve the problem that seedlings have no obvious insect resistance, and achieve the effect of good repeatability and stable antibacterial performance

Active Publication Date: 2017-11-10
ZHEJIANG SCI-TECH UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Goose view grass with a relatively high seed germ content has a high germination rate, neat emergence, fast growth, and many tillers, but the seedlings have no obvious insect resistance

Method used

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  • Bacillus amyloliquefaciens with broad-spectrum pathogen inhibition function and application of bacillus amyloliquefaciens
  • Bacillus amyloliquefaciens with broad-spectrum pathogen inhibition function and application of bacillus amyloliquefaciens
  • Bacillus amyloliquefaciens with broad-spectrum pathogen inhibition function and application of bacillus amyloliquefaciens

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0044] Embodiment 1, the isolation of Bacillus amyloliquefaciens and bacterial strain identification

[0045] 1. Plant sample collection

[0046] Suaeda glauca Bunge: collected in Honghaitan Wetland Nature Reserve, Panjin City, Liaoning Province.

[0047] 2. Isolation and screening of bacterial strains

[0048] Take fresh, pest-free Suaeda salsa tissue, soak it in 75% alcohol for 1 minute, rinse it with sterile water for 3 to 4 times, soak it with 0.1% mercury for 5 minutes, and rinse it with sterile water for 3 to 4 times. Use a sterile scalpel to cut the Suaeda salsa tissue into 5-10 mm in size, divide it into two parts, put one part on the LB solid medium, cultivate overnight to observe whether there are colonies near the tissue block (control); the other part, the tissue block After grinding, cook at 70°C for 30 minutes, cool down, dilute, inoculate on LB solid medium and culture until colonies appear. Use an inoculation needle to pick out the well-growing bacteria at t...

Embodiment 2

[0052] Embodiment 2, the standard growth curve drawing of Bacillus amyloliquefaciens of the present invention

[0053] It has been determined that the bacterium can grow at 15-45°C, and the optimum growth temperature is 37°C. Under the optimum 37°C and natural pH conditions, the content of conventional LB medium was used as the substrate, cultured on a shaker at 180 rpm, and the sampling interval was 2 to take a sample for OD600 determination. Sampling was carried out by Nanodrop2000 micro-spectrophotometer for OD value measurement, with OD600 value as the slave coordinate and culture time as the abscissa, and the bacterial growth curve was drawn as image 3 a.

Embodiment 3

[0054] Embodiment 3, PCR amplification, sequencing and dendrogram drawing of the 16srDNA of Bacillus amyloliquefaciens of the present invention

[0055] 16srDN primer sequence:

[0056] 27F5'-AGTTTGATCMTGGCTCAG-3' / 1492R5'-GGTTACCTTGTTACGACTT-3' total DNA was extracted using TIANGEN extraction kit. The extraction steps were according to the instructions of the kit. Taq uses NEB's general PCR enzyme DNA Polymerase (Catalog #M0480S) and High Fidelity Ultra-fidelity 2X Master Mix (Cat. No.: #M0491V), used for mixing to ensure the authenticity of PCR products;

[0057] Composition of PCR reaction system (100ul):

[0058] 10x amplification buffer 10uL

[0059] 4 kinds of dNTP mixture 1.5ul (each 10umol / L)

[0060] Primer P 1 (27F) 1uL (concentration 10pmol)

[0061] Primer P 2 (1492R) 1uL (concentration 10pmol)

[0062] Template DNA 2uL (about 0.1~10ng)

[0063] Taq DNA polymerase 1Ul

[0064] ddH 2 0 to 100ul;

[0065] PCR reaction conditions:

[0066] 94℃4min;

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Abstract

The invention discloses bacillus amyloliquefaciens. Bacillus amyloliquefaciens has a conservation number of CGMCC No.13492. Bacillus amyloliquefaciens is used for inhibiting bacteria, molds and phytopathogen; the bacteria are escherichia coli, staphylococcus, tetrads and bacillus subtilis; the molds are mucor, aspergillus niger and penicillium; phytopathogen is rhizoctonia cereal, lily root rot, fusarium oxysporum vasinfectum, fusarium oxyspirum F sp niveum, colletotrichum capsici, botrytis cinerea, apple ring spot and peach brown rot.

Description

technical field [0001] The invention relates to a bacillus amyloliquefaciens capable of broad-spectrum inhibition of pathogenic bacteria and an application thereof, belonging to the technical field of microbial application. Background technique [0002] As a special group of microorganisms (fungi, actinomycetes or bacteria) living in healthy plant tissues and organs during the whole life cycle or at a certain stage - plant endophyte bacteria (Endophyte) are a class of abundant secondary metabolites and broad application prospects. resources of microorganisms (Petrini et al.1992). Studies have shown that plant endophytes can produce a variety of secondary metabolites, which can not only enhance the stress resistance of plants (Skylar C 2011), but also have drug and pesticide activities. They are new lead compounds such as antibacterial and antitumor, An important resource bank of natural medicines and bioactive ingredients (Venkatachalam R2008), an important way to develop n...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/20A01N63/00A01N63/02A01P1/00A01P3/00C12R1/07
CPCA01N63/00A01N63/10C12N1/20C12N1/205C12R2001/07
Inventor 赵洪新郝雅荞吴言韦璇沈琦肖成建王立言
Owner ZHEJIANG SCI-TECH UNIV
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