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Bacillus thuringiensis with high insecticidal performance and environmental friendliness and application thereof

A technology of Bacillus thuringiensis and Bacillus aureus, applied in application, pesticides, bacteria and other directions, can solve the problem of limited research, and achieve the effect of enhancing insecticidal toxicity, improving insecticidal stability, and effectively resisting ultraviolet degradation.

Active Publication Date: 2019-05-24
HUNAN NORMAL UNIVERSITY
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  • Description
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Problems solved by technology

However, the current research on this gene is limited to the structural characteristics or enzymatic activity analysis of gene cloning and its protein expression products.

Method used

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  • Bacillus thuringiensis with high insecticidal performance and environmental friendliness and application thereof
  • Bacillus thuringiensis with high insecticidal performance and environmental friendliness and application thereof
  • Bacillus thuringiensis with high insecticidal performance and environmental friendliness and application thereof

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Embodiment

[0029] ① The Bacillus thuringiensis ΔleuB (Bacillus thuringiensis ΔleuB) of this example was preserved in the China Center for Type Culture Collection (CCTCC for short, address: Wuhan University, Wuhan, China) on February 25, 2019, and the strain preservation number is CCTCC NO: M 2019112.

[0030] ② Construction process of Bacillus thuringiensis ΔleuB in this embodiment:

[0031] Step 1: Construction of the knockout vector pRP8509: Using the genome of the wild-type Bt 4.0718 strain as a template, two pairs of primers UlueB-F / UlueB-R and DlueB-F / DlueB-R were used respectively (see Table 1 for the primer sequences, provided by Shanghai Biotechnology Co., Ltd. Industrial Biotechnology Co., Ltd.) PCR amplification to obtain the upper and lower homology arms of the leuB gene, the size of the PCR products were 690bp and 660bp ( figure 1 A-B). The upstream homology arm was double-digested with BamHI / SalI and the product was recovered, ligated and transformed with the pRP1028 vecto...

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Abstract

Disclosed are bacillus thuringiensis with high insecticidal performance and environmental friendliness and application thereof. The bacillus thuringiensis with high insecticidal performance and environmental friendliness is bacillus thuringiensis delta leuB of which the collection number is CCTCC NO: M 2019112. A gene leuB in the genome of the bacillus thuringiensis Bt 4.0718 is successfully knocked out by an I-SceI mediated traceless gene knockout system to obtain a mutant strain of the bacillus thuringiensis; the phenotypic change of the mutant strain delta leuB is obvious, no mature germs are found in an LB liquid culture medium, the synthetic amount of crystal protein is increased, metrocytes are not broken, and the crystal protein is wrapped into a microcapsule; the mutant strain hashigher insecticidal activity on plutella xylostella than wild strains, and the differences are remarkable.

Description

technical field [0001] The invention relates to a Bacillus thuringiensis strain with good insecticidal performance and environmental safety and its application, in particular to a Bacillus thuringiensis strain which does not produce spores, has increased crystal protein synthesis, stable insecticidal performance and is environmentally safe ( Bacillus thuringiensis ΔleuB) and its applications. Background technique [0002] Bacillus thuringiensis (Bt) is a Gram-positive bacterium widely distributed in nature. Its main feature is that it is accompanied by a large number of insecticidal crystal proteins (including Cry and Cyt crystal proteins) while forming spores. Produced, Bt parasporal crystal protein has a poisonous effect on various pests such as Lepidoptera, Coleoptera, and Diptera. Therefore, Bacillus thuringiensis, as a microbial insecticide, is widely used in the control of agricultural, forestry and sanitary pests. However, compared with traditional chemical pesticid...

Claims

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Application Information

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IPC IPC(8): C12N1/20A01N63/00A01P7/04C12R1/07
Inventor 丁学知全梅芳彭金莉谢俊雁夏立秋
Owner HUNAN NORMAL UNIVERSITY
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