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Application of Mad1 protein in regulation and control of fungal sporulation and germination and plant linolenic acid metabolic pathway

A technology of linolenic acid and fungi, which is applied in the fields of application, fungi, and plant genetic improvement, and can solve unknown problems

Active Publication Date: 2021-02-23
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 and control of fungal sporulation and germination and plant linolenic acid metabolic pathway
  • Application of Mad1 protein in regulation and control of fungal sporulation and germination and plant linolenic acid metabolic pathway
  • Application of Mad1 protein in regulation and control of fungal sporulation and germination and plant linolenic acid metabolic pathway

Examples

Experimental program
Comparison scheme
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 application of Mad1 protein in regulation and control of fungal sporulation and germination. Metarhizium anisopliae is used as a starting strain, a mad1 gene knockout strain (delta mad1) is obtained through a homologous recombination method, and the growth rate, sporulation and germination biological characteristics of a wild strain and the delta mad1 are measured. The wildstrain and delta mad1 spore suspensions are used for soaking the root of a peanut seedling to detect the transcription level of a linolenic acid metabolic pathway gene of the treated peanut root. Experimental results show that compared with the wild strain, the germination rate and sporulation quantity of the delta mad1 are remarkably reduced, the delta mad1 remarkably inhibits transcription of the linolenic acid metabolic pathway gene after acting on a plant, and thus the Mad1 can promote sporulation and germination of the Metarhizium anisopliae and up-regulate the transcription level of thelinolenic acid metabolic pathway gene. Theoretical support is provided for improvement and industrial production of fungal strains, and basic data are provided for Mad1 induced plant response to regulation and control of interaction between the Metarhizium anisopliae and the plant.

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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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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