Bacillus subtilis protein elicitor AMEP412 and function thereof

A Bacillus subtilis, elicitor technology, applied in biocides, plant growth regulators, animal husbandry and other directions, can solve the problems of less identification, unfavorable plant protection and biological control research, etc., to promote plant growth and improve plant disease resistance Effect

Active Publication Date: 2018-12-28
黑龙江权晟生物科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] The first purpose of the present invention is to provide a Bacillus subtilis protein elicitor AMEP412, which overcomes the current probl

Method used

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  • Bacillus subtilis protein elicitor AMEP412 and function thereof
  • Bacillus subtilis protein elicitor AMEP412 and function thereof
  • Bacillus subtilis protein elicitor AMEP412 and function thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0033] Purification and mass spectrometry identification of Bacillus subtilis AMEP412 protein.

[0034] Bacillus subtilis BU412 was inoculated into YME liquid medium (maltose 10g / L, yeast extract powder 4g / L, glucose 4g / L, pH 7.5) for overnight culture, and the supernatant was collected by centrifugation at 12000g, filtered through a 0.22μm filter membrane, Anion-exchange chromatography was performed using an AKTA protein purifier, and the purification column used was Q hp anion-exchange column (column volume: 5 mL) of GE Company. Use 20mM Tris-HCl (pH 7.5) buffer to load the sample and balance, carry out 0-100% linear gradient elution with 0.5M NaCl, 20mM Tris-HCl buffer, collect protein components S1-S4 according to the absorption peak ( figure 1 ). The protein fraction S2 was taken, concentrated using an ultrafiltration tube with a cut-off pore size of 3 kDa, and purified by molecular sieves through an AKTA protein purifier. The purification column used was a superdex-75 m...

Embodiment 2

[0037] Prokaryotic expression and purification of protein elicitor AMEP412.

[0038] The gene sequence of AMEP412 (GenBank accession number is WP_017418614.1) was submitted to Beijing Huada Gene Bio Co., Ltd. for whole gene synthesis, and subcloned into pET32a vector, transformed into BL21 (DE3) competent cells, and obtained positive clones. Prokaryotic recombinant expression was induced by IPTG, the bacteria were collected, and the supernatant after sonication was subjected to affinity purification using a Ni-NTA purification column, and the eluted samples were subjected to SDS-PAGE to detect the recombinant target protein ( Figure 6 ). The results showed that the prokaryotic recombinant protein of the protein elicitor AMEP412 was successfully obtained.

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Abstract

The invention relates to a bacillus subtilis protein elicitor AMEP412. The amino acid sequence of the bacillus subtilis protein elicitor is shown as SEQ ID NO.1. The invention also relates to the newfunction of the bacillus subtilis protein elicitor AMEP412. The invention discloses protein eliciting functions of a protein AMEP412 with an unknown function, including plant growth promotion, improvement of disease, salt and alkali, drought and low temperature resistance ability of plants, and new materials are accumulated for biological prevention and control work.

Description

technical field [0001] The invention belongs to the field of plant protection and biological control, and relates to a subtilis protein elicitor AMEP412 and its function. Background technique [0002] Plants are closely related to the surrounding environment during the growth process, and plants will respond accordingly to environmental impacts to avoid hazards. The study of elicitors developed from this on plant disease control and stress resistance has become a new research direction. Elicitors are a class of compounds derived from the pathogen itself that can act on plants and induce plant defense responses. The elicitor is to induce the function of the plant's own immune defense system, comprehensively improve the plant's own metabolic level, enhance the ability to defend against diseases and natural stress, and then improve crop yield and quality. The elicitor does not act on the pathogen, so it will not cause the pathogen to develop drug resistance, and will not have...

Claims

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Application Information

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IPC IPC(8): C07K14/32A01N43/50A01P1/00A01P21/00
CPCA01N43/50C07K14/32
Inventor 刘权殷奎德申永瑞向君亮刘爽李章雷
Owner 黑龙江权晟生物科技有限公司
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