CN-119799779-B - ZmHSP22 protein and application of coding gene thereof in regulation and control of low temperature stress tolerance of corn

Abstract

The invention relates to the technical field of genetic engineering, in particular to ZmHSP protein and application of a coding gene thereof in regulating and controlling low-temperature stress tolerance of plants. The nucleotide sequence of ZmHSP gene is shown as SEQ ID No.1, and the amino acid sequence of its protein is shown as SEQ ID No. 3. After ZmHSP genes are knocked out, the cold tolerance of the transgenic plants is improved compared with that of the plants of a control group under the low-temperature stress treatment condition. The invention verifies that ZmHSP protein has regulation and control function on low temperature stress tolerance of plants (especially corns), zmHSP protein and related biological materials thereof can be applied to plant cold tolerance regulation and control, provides excellent candidate gene resources for cultivating and improving new varieties of cold-tolerant plants, provides a cold-tolerant plant breeding method, has strong purposiveness, obviously shortens the cycle of cold-tolerant breeding, improves the efficiency of cold-tolerant breeding, lays a theoretical foundation for researching the mechanism of plants responding to stress signals and the molecular mechanism of adverse environment tolerance, and has important theoretical and practical significance for promoting the corn breeding process.

Inventors

• SHI YITING
• YANG SHUHUA
• Jiang Shengzhe

Assignees

• 中国农业大学

Dates

Publication Date

20260508

Application Date

20241114

Claims (5)

1. 1. Use of the maize ZMHSP protein or a gene encoding it, or a biological material containing the gene encoding it, in any of the following aspects: (1) Improving the cold resistance of corn; (2) Selecting transgenic corn with improved low temperature stress resistance; (3) Improving cold-resistant germplasm resources of corn; The cold resistance of the corn is improved by inhibiting or reducing the expression quantity of ZMHSP protein in the corn; the amino acid sequence of the corn ZMHSP protein is as follows: (A1) The amino acid sequence shown in SEQ ID No. 3.
2. 2. The use according to claim 1, wherein, The nucleotide sequence of the gene encoding the zein ZMHSP protein is any one of the following: (B1) A nucleotide sequence shown as SEQ ID No. 1; (B2) The nucleotide sequence shown as SEQ ID No. 2.
3. 3. The use according to claim 1 or 2, characterized in that, The biological material is an expression cassette, a vector and a host cell.
4. 4. A method for breeding cold-resistant corn is characterized in that, The plant with improved cold tolerance is obtained by inhibiting the expression quantity of ZMHSP protein, wherein the amino acid sequence of the ZMHSP protein is as follows: (A1) The amino acid sequence shown in SEQ ID No. 3.
5. 5. The method for breeding cold-resistant corn according to claim 4, wherein, Comprises the steps of utilizing DNA homologous recombination technology, cre/Loxp technology and Crispr/Gas9 technology to make the gene silencing expression or expression quantity of corn ZMHSP be reduced so as to obtain transgenic plant line.

Description

ZmHSP22 protein and application of coding gene thereof in regulation and control of low temperature stress tolerance of corn Technical Field The invention relates to the technical field of genetic engineering, in particular to application of HSP22 protein and a coding gene thereof in regulating and controlling low-temperature stress tolerance of plants. Background With the progressive decrease in the area of the cultivated land in the world and the increasing population in the world, there is an increasing demand for foodstuffs. Corn is one of three world food crops, and is a crop that is more sensitive to temperature. In all biological and environmental stresses, low temperature stress is one of the main limiting factors affecting crop survival and yield, and low temperature stress has an important effect on maize seedling germination and survival of seedlings and mature stage grain filling processes. Cold stress affects plant enzymes, membrane plasticity, changes physiology and metabolism, sometimes causes water shortage and dryness, causes stress conditions for plants, and adversely affects plant growth and development and yield. Hypothermia is also associated with protein dysfunction and denaturation, thereby inducing accumulation of heat shock proteins. In arabidopsis, tobacco, maize, rapeseed, chicory, poplar, wheat and barley, many heat shock proteins respond to cold stress and are up-regulated. Under low temperature stress, heat shock proteins are induced and transported into various organelles to protect cells from low temperature stress. Thus, studying the impact of corn tolerance to low temperature stress is of great importance to corn production. The difficulty of cultivating and improving the stress-resistant character by the traditional breeding technology is relatively large, and the excellent cold-resistant variety cannot be obtained quickly and efficiently. With the development of molecular biology technology, the deep research on the stress-resistant molecular mechanism of plants, and the great progress of human beings in genetic engineering research are carried out. The introduction of stress-resistant exogenous genes into plants by means of genetic engineering such as transgenes has become one of new approaches for improving stress resistance of plants. Disclosure of Invention Aiming at the defects existing in the prior art, the invention aims to provide ZmHSP protein and application of a coding gene thereof in regulating and controlling low-temperature stress tolerance of plants, and the detection shows that compared with the wild type low-temperature stress tolerance of the transgenic plants, the method can be used for quickly knocking out the target gene ZmHSP22 in the wild type plants by constructing a CRISPR/Cas9 material. In order to solve the technical problems, the technical scheme provided by the invention is as follows: use of the maize ZMHSP protein or a gene encoding it, or a biological material containing the gene encoding it, in any of the following aspects: (1) Improving the cold resistance of corn; (2) Selecting transgenic corn with improved low temperature stress resistance; (3) Improving the cold-resistant germplasm resource of the corn. Preferably, the method comprises the steps of, By inhibiting or reducing the expression quantity and/or activity of ZMHSP protein in corn, the cold resistance of corn is improved. Preferably, the method comprises the steps of, The amino acid sequence of the corn ZMHSP protein is any one of the following: (A1) An amino acid sequence shown in SEQ ID No. 3; (A2) The amino acid sequence shown in SEQ ID No.3 is obtained by replacing, deleting or inserting one or more amino acid residues to obtain the amino acid sequence of the protein with the same function. Preferably, the method comprises the steps of, The nucleotide sequence of the gene encoding the zein ZMHSP protein is any one of the following: (B1) A nucleotide sequence shown as SEQ ID No. 1; (B2) A nucleotide sequence shown as SEQ ID No. 2; (B3) The nucleotide sequence shown in SEQ ID No.2 is a nucleotide sequence which is substituted, deleted and/or added with one or more nucleotides and expresses the same functional protein; (B4) A nucleotide sequence which is fully complementary to the nucleotide sequence shown in SEQ ID No. 2. Preferably, the method comprises the steps of, The biological material is an expression cassette, a vector, a host cell or a recombinant bacterium. A breeding method of cold-resistant corn, The plant with improved cold tolerance is obtained by inhibiting the expression quantity and/or activity of ZMHSP protein, wherein the amino acid sequence of ZMHSP protein is any one of the following: (A1) An amino acid sequence shown in SEQ ID No. 3; (A2) The amino acid sequence shown in SEQ ID No.3 is obtained by replacing, deleting or inserting one or more amino acid residues to obtain the amino acid sequence of the protein with the same function. According to the bree