TRV2 viral vector and application thereof in mutant gene library

Abstract

The invention provides a TRV2 viral vector and application thereof in a mutant gene library, and relates to the field of plant protein ad plant herbicide resistance. The mutant gene library is constructed by using the TRV2 viral vector, and meanwhile, the efficiency of a mutant is higher in combination with two efficient manners including herbicide screening and virus infection. The mutant libraryhas higher flexibility and target, and mutation sites of the mutant are easier to identify. Mutant plants are screened by using the characteristic that ALS mutant protein is easy to screen by a herbicide, the herbicide is spayed on harvested seeds after germination acceleration and sowing, seedlings with gene mutation in a CCA-CT form have herbicide resistance and can survive, and target genes ofa library gRNA of the seedlings are also mutated at the same probability; herbicide-resistant seeds of the herbicide are harvested and subjected to selfing for genotype separation, and a large quantity of tomato homozygous mutants can be obtained.

Description

technical field [0001] The invention belongs to the fields of plant protein and plant herbicide resistance, and in particular relates to a TRV2 virus vector and its application in a mutant gene library. Background technique [0002] Traditional methods for obtaining mutant libraries, such as EMS mutagenesis, radiation mutagenesis, etc., have great uncertainty in the mutation sites on the genome, and the mutation efficiency is not high, and there are too few beneficial mutations, making mutant screening inefficient 1. Efficient cloning of functional genes is difficult, and it is impossible to realize directed mutagenesis breeding in the true sense; modern methods, such as genetic transformation of sgRNA libraries, require a large amount of genetic transformation work to obtain a large number of mutants, which is time-consuming and labor-intensive, and applicable The scope is very limited, and it can only be carried out in species that have established a complete genetic trans...

AI Technical Summary

Problems solved by technology

[0002] Traditional methods for obtaining mutant libraries, such as EMS mutagenesis, radiation mutagenesis, etc., have great uncertainty in the mutation sites on the genome, and the mutation efficiency is not high, and there are too few beneficial mutations, making mutant screening inefficient 1. Efficient cloning of functional genes is difficult, and it is impossible to realize directed mutagenesis breeding in the true sense; modern methods, such as genetic transformation of sgRNA libraries, require a large amount of genetic transformation work to obtain a large number of mutants, which is time-consuming and labor-intensive, and applicable The scope is very limited, and it can only be carried out in species that have established a complete genetic transformation system, such as rice, etc.

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