Preparation and application of ribonucleic acid (RNA) polymerase mutant for highly yielding antifungal substance Iturin A

A technology of RNA polymerase and mutants, applied in the field of screening of RNA polymerase mutants, can solve the problems of restricting the production and promotion of microbial pesticides, and low content of active ingredients

Inactive Publication Date: 2013-01-16
刘常宏
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Problems solved by technology

[0003] However, due to the low content of active ingredients in microbial pesticides, the production and promotion of microbial pesticides are seriously restricted.

Method used

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  • Preparation and application of ribonucleic acid (RNA) polymerase mutant for highly yielding antifungal substance Iturin A
  • Preparation and application of ribonucleic acid (RNA) polymerase mutant for highly yielding antifungal substance Iturin A
  • Preparation and application of ribonucleic acid (RNA) polymerase mutant for highly yielding antifungal substance Iturin A

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Experimental program
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Embodiment Construction

[0029] 1. Screening of RNA polymerase mutants

[0030] Activation of the strain

[0031] The wild-type Acinetobacter baumannii LCH0606 (WT) and its RNA polymerase mutant were inoculated in 4 mL / tube of LB liquid medium with a 1% inoculum amount, activated at 37°C and 120 rpm for 48 hours, and then inoculated with the same Quantity and culture conditions for secondary activation in 8 LB liquid medium, cultivated for 24h.

[0032] Preparation of rifampicin-containing plates

[0033] Add the rifampicin solution to the LB solid medium at 50-60°C to a final concentration of 50 μg / mL, which is the rifampicin-containing plate.

[0034] Will The secondary activation seed solution (8 test tubes) prepared in , was evenly spread on the plate containing rifampicin according to the amount of 200 μL / dish, cultured in the dark at 37°C for 2 days, and the colonies were observed after 2 days, which was the possible mutation of RNA polymerase A total of 78 mutants were screened.

[0...

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Abstract

The invention belongs to the technical field of biopesticides and particularly relates to a ribonucleic acid (RNA) polymerase mutant, which is screened by a rifampicin-containing (50mu g / ml) Luria-Bertani (LB) plate and used for highly yielding Iturin A, of an Acinetobacter baumannii LCH0606 strain. Mutational sites of the mutant are positioned at the 1,619th basic group, the 1,603th basic group and the 1,604th basic group of an rpoB gene for encoding a beta sub-unit of RNA polymerase. Compared with a wild type LCH0606 strain, the mutant has the advantages that the ability of producing the antifungal substance Iturin A is improved by 95 to 300 percent, and the antifungal activity is improved by 20 to 70 percent; and the mutant can be used for the control of various crop diseases and continuous cropping obstacles of medicinal plants.

Description

technical field [0001] The invention belongs to the technical field of biopesticides, and in particular relates to the screening and application of an RNA polymerase mutant of Acinetobacter baumannii LCH0606 strain with high yield of antifungal substance Iturin A. Background technique [0002] With the improvement of people's quality of life requirements and the various crises faced by the environment and ecology, the biggest challenge facing agricultural production is how to find environmentally friendly substitutes for chemical pesticides in the future to resist agricultural diseases. , biopesticides derived from beneficial microorganisms are the most promising substitutes for such products (Marc Ongena et al., Cell, 2007, 16(3): 115-125). On the one hand, microorganisms are the largest biological group on the earth. They not only have a wide range of biodiversity, but also can greatly increase the production of target substances through strain selection, metabolic regulat...

Claims

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Application Information

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IPC IPC(8): C12N1/20C12N15/01C12N9/12A01N63/00A01N63/02A01P3/00C12R1/01
Inventor 刘常宏
Owner 刘常宏
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