Construction and application of unmarked lysobacter enzymogenes engineering strain capable of preventing plant bacteriosis

An enzyme-producing lysobacteria, plant bacteria technology, applied in the application, bacteria, plant growth regulators and other directions, can solve the problem of target gene function loss, etc., to achieve the effect of broadening the scope of biocontrol

Inactive Publication Date: 2013-02-27
NANJING AGRICULTURAL UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Because when the foreign gene is integrated into the target gene, the target gene will be inactivated by insertion mutation, resulting in the loss of the function of the target gene

Method used

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  • Construction and application of unmarked lysobacter enzymogenes engineering strain capable of preventing plant bacteriosis
  • Construction and application of unmarked lysobacter enzymogenes engineering strain capable of preventing plant bacteriosis
  • Construction and application of unmarked lysobacter enzymogenes engineering strain capable of preventing plant bacteriosis

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0022] Example 1: Construction of a marker-free engineered strain of Lysobacterium enzymogenes containing PB500-aiiA

[0023] 1 Materials and methods

[0024] 1.1 Test materials

[0025] The Escherichia coli strain DH5α used in this experiment was purchased from Dalian Bao Biological Company (TaKaRa), and the SM10λpir and DH5αλpir strains were donated by Professor Zhu Jun, School of Life Sciences, Nanjing Agricultural University. Lysobacterium enzymogenes was isolated and preserved by the inventors. The vector pEXl8GM was kindly donated by Professor Zhu Jun, School of Life Science, Nanjing Agricultural University. Plasmid pME6863 was a kind gift from Prof. Molina of the Swiss Federal Institute of Technology. The culture of Escherichia coli strains uses LB medium, the culture temperature is 37°C, and the speed of liquid culture is 225 rpm / min. The lysobacterium enzymogenes was cultured using LB medium, the culture temperature was 30° C., and the rotation speed of the liquid...

Embodiment 2

[0038] Example 2: The biocontrol application of the unmarked engineered strain of Lysobacterium lysobacterium containing PB500-aiiA to bacterial soft rot

[0039] 1 Materials and methods

[0040] 1.1 Test material

[0041] The Agrobacterium tumefaciens used in this experiment was kindly donated by Professor Zhu Jun, School of Life Science, Nanjing Agricultural University. Lysobacterium enzymogenes and soft rot pathogen of Chinese cabbage were isolated and preserved by the inventor. The vector pME6863 was kindly provided by Prof. Molina of Swiss Federal Institute of Technology. LB medium was used for bacterial culture, the culture temperature was 30° C., and the rotation speed of liquid culture was 225 rpm / min. Antibiotics gentamicin (Gm), streptomycin (Sm), spectinomycin (Spe), tetracycline (Tc), 5-bromo-4-chloro-3-indole-β-D-galactoside (X -gal), SDS, NaCO3 and other chemical reagents were purchased from Nanjing Zhuyan Biological Company.

[0042] 1.2 Extraction and dete...

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Abstract

The invention relates to a construction strategy and a construction process of an unmarked lysobacter enzymogenes engineering strain capable of preventing plant bacteriosis, and belongs to the field of microbial genetic engineering. An excellent exogenous gene aiiA is directionally integrated onto a chromosome, and any marked genes are not brought into the unmarked lysobacter enzymogenes engineering strain. A quorum-sensing system for plant pathogenic bacteria can be efficiently damaged by the engineering strain, pathogenicity of the pathogenic bacteria on a host plant (Chinese cabbage) is remarkably reduced, and biological prevention of the plant bacteriosis is realized.

Description

(1) Technical field [0001] The invention belongs to the field of microbial genetic engineering, and in particular relates to the construction and application of an enzymolytic lysobacterium unmarked engineering strain capable of preventing and treating plant bacterial diseases. (2) Background technology [0002] Lysobacter enzymogenes is the model species of the genus Lysobacter, which belongs to the family Xanthomonadaceae. The bacterial colonies are viscous, yellow, rich in G+C%, have sliding ability, and can produce protease , chitinase, β-1,3-glucanase, cellulase and other extracellular enzymes have good antagonistic activity against various plant pathogens such as fungi, oomycetes and nematodes. [0003] The OH1 1 strain was isolated from the capsicum rhizosphere soil, and it was the first reported biocontrol bacterial strain in China. The strain is effective against Rhizoctonia solani, Pythium ultimum, Phytophthora capsici, Sclerotinia sclerotiorum, Fusarum solani, So...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/21C12N15/62C12N15/63A01N63/02A01P1/00A01P3/00C12R1/01
Inventor 刘凤权钱国良
Owner NANJING AGRICULTURAL UNIVERSITY
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