CN-121380174-B - Application of wheat specific subfamily gene TaHsfC-4 in improvement of wheat salt tolerance

CN121380174BCN 121380174 BCN121380174 BCN 121380174BCN-121380174-B

Abstract

The invention belongs to the technical field of genetic engineering, and particularly relates to application of TaHsfC-4 genes in improvement of salt tolerance of wheat. The gene TaHsfC-4 has the registration number of OQ680124.1, has a remarkable effect in regulating and controlling the salt tolerance of wheat, knocks out the wheat gene TaHsfC-4 through a CRISPR/Cas9 gene editing technology to obtain a homozygous mutant, and after salt stress treatment, the phenotype and physiological index of the TaHsfC-4 gene knocked-out mutant are better than those of a wild type Fielder, and the salt stress treated mutant has fewer dry leaves, good growth vigor, lower relative conductivity and malondialdehyde content of the leaves, so that the leaf cell membrane oxidation resistance of the wheat TaHsfC3-4 gene mutant is improved, the cell membrane integrity is better, and the salt tolerance of the mutant is enhanced. The invention provides a new method for quickly creating new varieties of salt-tolerant wheat.

Inventors

MENG XIANGZHAO
ZHANG HUANING
MA ZHENYU
LIU ZIHUI
LI GUOLIANG
DUAN SHUONAN
GUO XIULIN

Assignees

河北省农林科学院生物技术与食品科学研究所

Dates

Publication Date: 20260508
Application Date: 20251216

Claims (4)

1. The application of knocked-out wheat specific subfamily gene TaHsfC-4 in improving wheat salt tolerance is provided, wherein the nucleotide sequence registration number of TaHsfC3-4 gene is OQ680124.1.
2. The application of biological materials related to the gene TaHsfC-4 knocked out in the cultivation of wheat varieties; the nucleotide sequence registration number of the TaHsfC-4 gene is OQ680124.1; the application is to construct biological materials related to the gene TaHsfC-4 knocked out, so as to obtain wheat varieties with improved salt tolerance; the biological material does not include propagation material; The biological material comprises nucleic acid molecules, vectors and cells; the nucleic acid molecule comprises sgRNA for inhibiting TaHsfC-4 functions, wherein the sequence of the sgRNA is shown as SEQ ID NO.1 and SEQ ID NO. 2; the vector comprises a CRISPR/Cas9 vector; the cells comprise at least one of escherichia coli and agrobacterium.
3. A method for cultivating a wheat variety with high salt tolerance comprises the step of reducing the expression quantity and/or activity of TaHsfC-4 genes in wheat; the wheat variety comprises the following characteristics that salt tolerance is improved relative to acceptor wild type wheat; the step of reducing the expression level and/or activity of TaHsfC-4 genes in wheat is to introduce the biological material related to knockout TaHsfC3-4 genes described in claim 2 into wheat tissue or wheat cells.
4. The method of claim 3, wherein the introducing comprises one of microinjection or electroporation.

Description

Application of wheat specific subfamily gene TaHsfC-4 in improvement of wheat salt tolerance Technical Field The invention belongs to the technical field of genetic engineering, and particularly relates to application of TaHsfC-4 genes in improvement of salt tolerance of wheat. Background The efficient utilization of the saline-alkali soil has great strategic significance for promoting the grain safety production and the agricultural sustainable development in China. Wheat is an important grain crop in China, the planting area and the yield are the second national, but wheat belongs to non-halophyte, and the precondition of planting in saline-alkali soil is to cultivate a variety with good salt tolerance. The wheat salt tolerance gene is excavated, the salt tolerance of the excellent wheat variety is improved by utilizing the modern biological breeding technology, and the scientific goal of 'planting and adapting to the land' is accelerated to be realized, which is very important for efficiently utilizing the saline-alkali soil, guaranteeing the grain safety and the sustainable development of future agricultural production. Disclosure of Invention The invention aims to provide an application of TaHsfC3-4 genes in improving the salt tolerance of wheat, and TaHsfC3-4 negative regulation and control of the salt tolerance in the wheat, thereby providing a new gene resource for the salt tolerance breeding of the wheat by utilizing a genetic engineering technology. The invention adopts the following technical scheme: The invention provides an application of TaHsfC-4 genes in improving salt stress tolerance of wheat, wherein the accession number of the TaHsfC-4 genes is OQ680124.1. The invention improves the salt stress resistance of wheat by inhibiting the expression of TaHsfC < 3 > -4 genes or knocking out TaHsfC < 3 > -4 genes. As a preferred embodiment, the application is to obtain wheat varieties with improved salt tolerance by constructing biological materials related to the inhibition TaHsfC-4 genes. In some embodiments of the invention, the biological material does not include propagation material. In some embodiments of the invention, the biological material comprises a nucleic acid molecule, a vector, a cell. In some embodiments of the invention, the nucleic acid molecule comprises microRNA, siRNA, shRNA, dsRNA, sgRNA and/or antisense oligonucleotides that inhibit TaHsfC' 3-4 functions. In some embodiments of the invention, the sgRNA has a sequence as shown in SEQ ID NO. 1 and SEQ ID NO. 2. In some embodiments of the invention, the sgrnas are used in combination with a CRISPR/Cas9 vector for gene knockout purposes. In some embodiments of the invention, the CRISPR/Cas9 vector further comprises an expression vector comprising the sgRNA, although Cas9 protein or an expression vector for expressing Cas9 protein may also be included. In some embodiments of the invention, the vector includes, but is not limited to, cas9 and other vectors common in the art. In some embodiments of the invention, the cells comprise at least one of E.coli, agrobacterium. Among them, E.coli is a common host cell for constructing vectors and plasmids in the art, and Agrobacterium is a common tool for delivering DNA molecules to plants in the art. The invention also provides a method for cultivating wheat to improve salt tolerance varieties, which comprises the step of reducing the expression quantity and/or activity of TaHsfC-4 genes in wheat. In some embodiments of the invention, the wheat variety comprises the feature that salt tolerance is increased relative to a reference level, the reference level being that of a wild type. In some embodiments of the invention, the step of reducing the amount and/or activity of TaHsfC-4 gene expression in wheat is introducing into a wheat tissue or wheat cell a biological material according to the invention that is associated with inhibiting TaHsfC3-4 gene. In some embodiments of the invention, the means of introducing comprises the use of at least one of a Ti plasmid, a Ri plasmid, a plant viral vector, microinjection, electroporation. In some embodiments of the invention, the step of reducing the expression and/or activity of TaHsfC-4 protein in wheat is specifically: (1) Designing a target sequence sgRNA of TaHsfC3-4 genes, and constructing a CRISPR/Cas9 vector edited by the wheat TaHsfC3-4 genes; (2) Transferring the CRISPR/Cas9 vector in the step (1) into an agrobacterium competent cell to obtain agrobacterium containing the CRISPR/Cas9 vector; (3) And (3) infecting young embryos of the Fielder with the agrobacterium infection liquid obtained in the step (2), recovering seedlings through tissue culture, and screening a stably inherited mutant strain which contains the wheat TaHsfC-4 gene mutation, does not contain the exogenous Cas9 protein and has the mutation of a target sequence. In some embodiments of the invention, the vector is pWMB110-Cas9; In some embodiments of the invention, the