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Application of mad1 protein in regulation of fungal sporulation and germination and plant linolenic acid metabolism pathway

A technology of fungi and plants, applied in the fields of application, fungi, plant genetic improvement, etc., can solve unknown problems

Active Publication Date: 2022-04-08
INST OF PLANT PROTECTION CHINESE ACAD OF AGRI SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, Mad1, as an adhesion protein, assists Metarhizium anisopliae to colonize plant roots in the interaction between Metarhizium anisopliae and plants is currently unknown.

Method used

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  • Application of mad1 protein in regulation of fungal sporulation and germination and plant linolenic acid metabolism pathway
  • Application of mad1 protein in regulation of fungal sporulation and germination and plant linolenic acid metabolism pathway
  • Application of mad1 protein in regulation of fungal sporulation and germination and plant linolenic acid metabolism pathway

Examples

Experimental program
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Effect test

Embodiment 1

[0073] Example 1, Construction of Mad1 knockout strain (Δmad1) and determination of its biological characteristics

[0074] 1. Construction of Mad1 knockout strain (Δmad1)

[0075] 1. Construction of target carrier

[0076] 1) Obtaining hyg sequence and homologous sequence S1, S2

[0077] Using the plasmid pKH-KO as a template and hyg-F / hyg-R as primers, the hygromycin hyg sequence (1406bp in size) was amplified. Genomic DNA of Metarhizium anisopliae Ma9-41 was extracted as a template, and S1-F / S1-R and S2-F / S2-R were used as primers to amplify the upstream and downstream homologous sequences of the mad1 gene with a length of 1092bp and 1014bp respectively (S1, S2). The electrophoresis detection results of hyg sequence and homologous sequence S1, S2 are as follows figure 1 shown.

[0078] 2) Obtaining S1-hyg vector

[0079] The mad1 homologous forearm S1 and the hygromycin resistance selection gene hyg were respectively connected to the cloning vector PBM23 to obtain the...

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Abstract

The invention discloses the application of Mad1 protein in regulating fungal sporulation and germination. In the present invention, using Metarhizium anisopliae as the starting bacterium, a mad1 gene knockout strain (Δmad1) is obtained through a homologous recombination method, and the biological characteristics of growth rate, sporulation and germination of the wild strain and Δmad1 are measured. The present invention also soaks the roots of peanut seedlings with wild strain and Δmad1 spore suspension to detect the transcription level of linolenic acid metabolism pathway genes in peanut roots after treatment. The experimental results showed that compared with the wild strain, the germination rate and sporulation of Δmad1 were significantly reduced, and Δmad1 significantly inhibited the transcription of linolenic acid metabolism pathway genes after acting on plants, indicating that Mad1 can promote the sporulation and germination of Metarhizium anisopliae , up-regulated gene transcription levels of linolenic acid metabolism pathway. The invention provides theoretical support for the improvement and industrial production of fungal strains, and provides basic data for Mad1 inducing plant responses to regulate the interaction between Metarhizium anisopliae and plants.

Description

technical field [0001] The invention belongs to the field of biotechnology, and specifically relates to the application of Mad1 protein in regulating fungal sporulation and germination and plant linolenic acid metabolism. Background technique [0002] As an entomopathogenic fungus, Metarhizium anisopliae has been used to control agricultural and forestry pests due to its broad insecticidal ability. Such as locusts, grubs and so on. After a long period of biological evolution, three lifestyles have been derived: saprophytic, plant symbiotic, and insect pathogenic. In recent years, Metarhizium anisopliae has been developed into various types of biological agents for the control of locust pests, and the biological control effect is remarkable. At present, the infection level, germination rate and virulence of different strains of Metarhizium anisopliae are different, which have a significant impact on the control ability of the developed biological agents. Therefore, it is n...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C07K14/37C12N15/31C12N15/80C12N1/15A01G7/06C12R1/645
CPCC07K14/37C12N15/80A01G7/06
Inventor 农向群闫多子蔡霓王广君涂雄兵张泽华
Owner INST OF PLANT PROTECTION CHINESE ACAD OF AGRI SCI
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