Preparation method of citrus dominant function mutant

Abstract

The invention discloses a method for preparing a citrus dominant functional mutant. The method comprises the following steps: 1, taking a vector pCAMBIA1300 as a template by utilizing a first primer pair, amplifying to obtain a vector framework containing a transfer DNA structural domain, connecting a first strong promoter gene, a reporter gene and a second strong promoter gene into the left and right boundaries of the transfer DNA structural domain of the carrier framework, and constructing to obtain a destination vector of which the transfer DNA structural domain contains a target sequence;2, integrating the target sequence onto citrus seedling genome by agrobacterium tumefaciens mediated transformation, and screening to obtain a positive transformant; 3, culturing the positive transformant into a plant, planting in soil or grafting onto stock for preservation, thereby obtaining the citrus dominant functional mutant. According to the method disclosed by the invention, available citrus genetic materials at modern times can be created, and the problem that directly available citrus mutants cannot be rapidly created by utilizing a T-DNA insertion method at present can be solved.

Description

technical field

[0001] The invention belongs to the technical field of biotechnology research and development, and relates to a method for preparing a citrus dominant function mutant. Background technique

[0002] After the T-DNA sequence is inserted into the plant genome, it can destroy the transcription of the gene at the insertion site. The plants with the T-DNA sequence inserted can obtain the homozygous line of the T-DNA insertion of the gene in their progeny through methods such as self-crossing and backcrossing, that is, a mutant with the loss of the gene function can be obtained. Using the obtained T-DNA insertion homozygous mutants and wild-type plant materials to perform phenotypic detection under different growth conditions has become an important means to discover and identify functional genes in Arabidopsis, rice and other plants. However, citrus has a long childhood period, and it generally takes at least 5 years to grow from seeds to seed plants and bear frui...

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