Insecticidal engineered strain converted from furocoumarin gene and application thereof

A technology of gene and recombinant strains, applied in application, pesticides, microbial-based methods, etc., can solve the problems of low accumulation level and high accumulation of secondary metabolites, etc.

Inactive Publication Date: 2017-11-14
XUZHOU NORMAL UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, the accumulation level of furocoumarins in wild-type Neurospora crassa was low, the model fungus Aspergillus nidulans grew faster, the genetic background was clear, and the secondary metabolites accumulated more

Method used

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  • Insecticidal engineered strain converted from furocoumarin gene and application thereof
  • Insecticidal engineered strain converted from furocoumarin gene and application thereof
  • Insecticidal engineered strain converted from furocoumarin gene and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment Construction

[0015] (1) Construction of recombinant strains

[0016] Genomic DNA of Neurospora crassa was used as a template to amplify the pks-6 gene, and the pks-6 gene was connected to the vector pYH-WA-PyrG-KI using a seamless cloning kit. The method of protoplast transformation was used to transform pYH-WA-PyrG-KI-pks-6 into Aspergillus nidulans L08030, and the recombinant strain was screened by medium without PyrG, and verified by PCR.

[0017] (2) Large-scale cultivation of recombinant strains

[0018] After mixing the rice and tap water at a ratio of 1:1, put it into a sterilizing pot for high-pressure steam sterilization. After cooling, the liquid culture of the recombinant strain was added, stirred evenly with a sterile glass rod, and placed in an incubator with a temperature of 30° C. and a relative humidity of 50% for 14 days.

[0019] (3) Effect on Drosophila toxicity

[0020] Ethyl acetate was used to extract the metabolites of Aspergillus nidulans L08030 and pks-6 gene re...

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Abstract

The invention provides an aspergillus nidulans recombinant strain capable of massively synthesizing furocoumarin. A culture method of the aspergillus nidulans recombinant strain is simple and the cost is low. In addition, coarse extract of the strain has a relatively high insecticidal efficiency, and the product extraction method is simple. The aspergillus nidulans recombinant strain can be effectively used for preventing and treating insects. The aspergillus nidulans recombinant strain has wide practical value and economical value.

Description

technical field [0001] The invention relates to a recombinant strain of Aspergillus nidulans which can synthesize furanocoumarin in large quantities, and the insecticidal effect of its crude extraction product. Background technique [0002] In the process of interacting with the environment, filamentous fungi have formed a unique mechanism to cope with environmental stress. Among them, synthesizing mycotoxins to resist the ingestion of harmful organisms is one of its important survival strategies. Most filamentous fungi synthesize inducible toxins when insects ingest them. Some filamentous fungi such as Aspergillus fumigatus and Aspergillus flavus synthesize constitutive toxins under normal physiological conditions. Neurospora crassa genome contains six polyketide synthase genes (polyketide synthase genes, pks). Previous studies have found that pks-6 gene knockout strains are highly susceptible to being swallowed by Collembola. Ethyl acetate was used to extract the metab...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/15C12N15/80A01N63/04A01P7/04C12R1/66
CPCA01N63/30C12N9/1029C12N15/80
Inventor 赵艳霞郑维发黄文秀袁卫华胡萍殷澜张朋黄金金
Owner XUZHOU NORMAL UNIVERSITY
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