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CN-122012599-A - Wheat TaRVE-4A gene and application thereof

CN122012599ACN 122012599 ACN122012599 ACN 122012599ACN-122012599-A

Abstract

The invention discloses a wheat TaRVE-4A gene and application thereof in regulating tolerance of wheat to salt stress and/or high temperature stress, and belongs to the technical field of genetic engineering. The CDS region nucleotide sequence of TaRVE-4A gene is shown in SEQ ID NO. 1. According to the invention, through cloning and analyzing the wheat TaRVE8-4A gene and combining with BSMV-VIGS technology to silence the wheat TaRVE-4A gene and carrying out functional verification on the gene by using a wheat transgene over-expression technology, taRVE-4A gene expression is found to be closely related to salt tolerance and high temperature tolerance of wheat. The invention provides theoretical basis and related genes for salt-tolerant and high-temperature-tolerant breeding and production of wheat.

Inventors

  • CAO FANGBIN
  • XU KEWEN
  • LU YIFAN

Assignees

  • 浙江大学

Dates

Publication Date
20260512
Application Date
20260325

Claims (8)

  1. The application of the TaRVE8-4A gene in regulating and controlling the tolerance of wheat to salt stress and/or high temperature stress is characterized in that the nucleotide sequence of the CDS region of the TaRVE-4A gene is shown as SEQ ID NO. 1.
  2. 2. The use according to claim 1, wherein the TaRVE-4A gene encodes a protein having the amino acid sequence shown in SEQ ID No. 2.
  3. 3. The use according to claim 1 or 2, wherein the TaRVE-4A gene negatively regulates the tolerance of wheat to salt stress and/or high temperature stress.
  4. 4. The use of claim 3, wherein wheat has significantly increased tolerance to salt stress and/or high temperature stress following TaRVE-4A gene silencing.
  5. 5. The use of claim 4, wherein sodium ion uptake is reduced following TaRVE-4A gene silencing in wheat plants.
  6. 6. The use according to claim 1, wherein the use comprises down-regulating expression or loss of function of TaRVE-4A gene in wheat plants by biotechnology means to increase the tolerance of the plants to salt stress and/or high temperature stress.
  7. 7. The use according to claim 6, wherein the TaRVE-4A gene function is deleted by gene mutation, gene knockout or gene silencing techniques to obtain a mutant plant with increased tolerance to salt stress and/or high temperature stress.
  8. 8. A breeding method for improving tolerance of wheat to salt stress and/or high temperature stress, comprising the steps of: (1) Inserting TaRVE-4A gene fragment with a nucleotide sequence shown as SEQ ID NO.3 between NheI sites of a BSMV gamma vector to construct a recombinant vector RNA gamma TaRVE-4A; (2) The vectors RNA alpha, RNA beta and RNA gamma TaRVE-4A are linearized and then transcribed in vitro, and wheat seedlings are infected, so that mutant plants with enhanced tolerance to salt stress and/or high temperature stress are obtained.

Description

Wheat TaRVE-4A gene and application thereof Technical Field The invention relates to the technical field of genetic engineering, in particular to a wheat TaRVE-4A gene and application thereof in regulating and controlling tolerance of wheat to salt stress and/or high temperature stress. Background Soil salinization, an increasingly serious global agricultural problem, has become one of the key factors limiting global crop yield and quality In recent years, global warming has led to the frequency of extremely high temperature events, causing significant adverse effects on crop production, thereby threatening grain safety and agricultural sustainable development (Challinor et al, 2014). Studies have shown that per 1 ℃ rise in global air temperature, the average yield per crop of four major crops, wheat, rice, corn and soybean, will drop by about 6.0%, 3.2%, 7.4% and 3.1%, respectively (Zhao et al, 2017). In this context, it is particularly urgent to improve the heat resistance and climate adaptation of crops. The salt resistance and the high temperature resistance of plants are complex quantitative characters, and relate to a series of physiological and biochemical reactions. To cope with high salt and high temperature stress, plants have evolved including active oxygen scavenging mechanisms, regulation of stomatal behavior, sodium ion efflux mechanisms, and the like. In actual production, high temperature and salt stress often occur simultaneously, so that cultivation of new varieties of crops with salt resistance and high temperature resistance has important significance. Wheat (Triticum aestivum l.) is one of the most important food crops worldwide, and after thousands of years of crossing and domestication, common wheat widely planted today is heterohexaploid (AABBDD), and its complex genome structure, while conferring excellent agronomic traits such as high yield, high quality, etc., may also lead to impaired stress resistance traits present in some wild closely related species including salt tolerance and heat tolerance. At present, a plurality of salt-tolerant or high-temperature-tolerant related genes are identified in wheat by adopting a forward genetics method and a reverse genetics method. However, compared with main grains such as rice, corn and the like, the number of salt tolerance and heat tolerance key genes of wheat which have completed in-situ functional verification is still limited. Therefore, the novel mechanisms of digging the genetic locus/gene of wheat for salt tolerance and high Wen Guanjian tolerance and analyzing salt tolerance are the basis for cultivating excellent materials of strong salt tolerance and Gao Wenxiao tolerance. The RVE (REVEILLE) family is an important group of MYB-related (1R-MYB) subfamilies in the MYB superfamily, including CCA1 (CIRCADIAN CLOCK ASSOCIATED 1), LHY (LATE ELONGATED HYPOCOTYL 1), RVE1-RVE8 and RVE7-like, which all contain the conserved sequences SHAQK (Y/F) F, with the exception of RVE5 and RVE6, whose expression in seedlings is biologically clock regulated (Liu et al 2023). Few studies have shown that RVE is involved in abiotic stress responses, e.g. the scan et al (2021) study found that soybean gene GmMYB133 is homologous to the arabidopsis RVE8 branching gene, and that overexpression of GmMYB133 in arabidopsis under salt stress conditions promotes seed germination and plant growth, and increases chlorophyll content and decreases Malondialdehyde (MDA) content. In agreement with this, gmMYB133 overexpression significantly upregulated the expression of 4 salt tolerant positive regulatory factors, suggesting that GmMYB133 may enhance salt stress tolerance of the plant. Bao et al (2024) constructed the Arabidopsis transgenic line overexpressing GmRVE a. As a result, it was found that transgenic seedlings grew better than wild-type under salt and drought stress, and Malondialdehyde (MDA) content in transgenic lines was significantly lower than wild-type, indicating GmRVE a may be a positive regulator of salt and drought stress responses. At present, research reports on gene functions of RVE family in wheat are not seen. Disclosure of Invention The invention aims to provide a gene which is cloned from wheat (Triticum aestivum L.) and participates in response of wheat salt stress and/or high temperature stress, and the gene is applied to breeding and production of salt-tolerant Gao Wenxiao-tolerant wheat germplasm. In order to achieve the above purpose, the invention adopts the following technical scheme: The invention identifies a gene involved in wheat salt tolerance response based on bioinformatics analysis and gene expression analysis, takes salt tolerance germplasm X118 identified in the earlier stage of a subject group as a material, clones the full-length CDS sequence of the gene, and the corresponding nucleotide sequence is shown as SEQ ID NO.1 and is named TaRVE-4A. The results of the gene sequence analysis showed that TaRVE-4A contained 8 exon