CN-122012578-A - Perforin CaMACPF and application thereof in construction of new germplasm of flood-fighting vegetable

Abstract

The invention discloses perforin CaMACPF and application thereof in construction of new germplasm of flood-fighting vegetables. The invention constructs an arabidopsis plant over-expressing perforin CaMACPF gene by using an agrobacterium-mediated inflorescence dip-dyeing method, and cultures the obtained transgenic plant under the condition of hypoxia stress, so that compared with a wild type control, the capacity of the transgenic plant for resisting hypoxia stress can be obviously improved by over-expressing the perforin CaMACPF gene in the plant. Based on this, perforin CaMACPF can be used to construct low-oxygen stress tolerant crop varieties, which can tolerate waterlogging and low-oxygen stress caused by other conditions, and reduce adverse effects of low-oxygen stress on crop growth. The invention is beneficial to the construction of low-oxygen stress resistant germplasm, thereby reducing the loss of crops caused by flood disasters and the like.

Inventors

• YU LUJUN
• GUO YIXIAN
• XIAO SHI

Assignees

• 中山大学

Dates

Publication Date

20260512

Application Date

20260202

Claims (10)

1. 1. Use of perforin CaMACPF6 for increasing tolerance of plants to hypoxia stress, characterized in that the amino acid sequence of perforin CaMACPF6 is shown in SEQ ID No. 2.
2. 2. Use of a gene encoding perforin CaMACPF in increasing the tolerance of a plant to hypoxia stress, characterized in that the amino acid sequence of perforin CaMACPF6 is shown in SEQ ID No. 2.
3. 3. Use of an expression enhancer of perforin CaMACPF6 for increasing tolerance of a plant to hypoxia stress, characterized in that the amino acid sequence of perforin CaMACPF6 is shown in SEQ ID No. 2.
4. 4. The application of perforin CaMACPF6 in constructing waterlogging tolerant plant line is characterized in that the amino acid sequence of perforin CaMACPF6 is shown in SEQ ID No. 2.
5. 5. The application of a gene encoding perforin CaMACPF in constructing waterlogging tolerant plant lines is characterized in that the amino acid sequence of perforin CaMACPF6 is shown as SEQ ID NO. 2.
6. 6. The application of the expression promoter of perforin CaMACPF in constructing waterlogging tolerant plant line is characterized in that the amino acid sequence of perforin CaMACPF is shown as SEQ ID NO. 2.
7. 7. The use according to any one of claims 1 to 6, wherein the plant is arabidopsis thaliana or capsicum.
8. 8. The use according to claim 2 or 5, wherein the nucleotide sequence of the gene is shown in SEQ ID No. 1.
9. 9. A method for constructing a plant line resistant to hypoxia stress is characterized in that the expression level of perforin CaMACPF6 in a plant is improved by transgenesis or by using an expression promoter, and the amino acid sequence of perforin CaMACPF6 is shown as SEQ ID NO. 2.
10. 10. The method of claim 9, wherein the plant is arabidopsis thaliana or capsicum.

Description

Perforin CaMACPF and application thereof in construction of new germplasm of flood-fighting vegetable Technical Field The invention belongs to the technical field of genetic engineering, and particularly relates to perforin CaMACPF and application thereof in construction of new germplasm of flood-resistant vegetables. Background The flood disasters are affected by global climate change, the occurrence frequency is increased, and the range is gradually expanded, so that the economic crops are seriously affected by the disasters. According to statistics of an international disaster database (The international disaster database, EM-DAT), 3,518 serious flood disasters occur in the whole world in the last twenty years, and 176 serious flood disasters occur in average each year, so that the agricultural production area of nearly 16% of the world is affected, and the grain safety is seriously threatened. In China, the overall flood risk is also in an ascending trend. Flooding can result in reduced oxygen levels around plant cells and closed pores, resulting in inhibition of the respiratory chain in mitochondria, and inhibition of aerobic respiration, causing hypoxia stress to the plant. The environmental pressure of crops in the flood-facing period can cause limitation of acquisition of O 2 and CO 2, accumulation of secondary metabolites, enrichment of toxic substances in soil and the like, so that the yield of the crops is reduced, and especially the crops of Solanaceae such as peppers and the like can be seriously damaged or even die. Therefore, the method has great significance in researching the waterlogging resistance mechanism of crops, cultivating new germplasm of the waterlogging resistance crops, being suitable for hydroponic vegetable varieties and the like by excavating related genes affecting the hypoxia tolerance of the plants. At present, the report of genes related to the hypoxia tolerance of animals is more, and the number of plants is less. There is no report on the correlation of MACPF (Membrane Attack Complex and Perforin) protein with hypoxia stress tolerance. Disclosure of Invention The invention provides perforin CaMACPF and application thereof in constructing new germplasm of flood-fighting vegetables, aiming at the defects existing in the prior art. A first object of the present invention is to provide the use of perforin CaMACPF in increasing the tolerance of plants to hypoxia stress. A second object of the present invention is to provide the use of a gene encoding perforin CaMACPF in improving the tolerance of plants to hypoxia stress. It is a third object of the present invention to provide the use of an expression enhancer of perforin CaMACPF in improving the tolerance of plants to hypoxia stress. A fourth object of the invention is to provide the use of perforin CaMACPF in the construction of waterlogging tolerant plant lines. A fifth object of the present invention is to provide the use of the gene encoding perforin CaMACPF6 in the construction of waterlogging tolerant plant lines. A sixth object of the present invention is to provide the use of an expression enhancer for perforin CaMACPF in the construction of waterlogging tolerant plant lines. It is a seventh object of the present invention to provide a method of constructing a plant line resistant to hypoxia stress. The above object of the present invention is achieved by the following technical scheme: The invention constructs an arabidopsis plant over-expressing perforin CaMACPF gene (shown as SEQ ID NO. 1) by using an agrobacterium-mediated inflorescence dip-dyeing method, and the obtained transgenic arabidopsis plant is cultured under the condition of hypoxia stress, so that compared with a wild type control, the capability of the plant for hypoxia stress can be obviously improved by over-expressing the perforin CaMACPF gene in the plant. Thus, the invention claims the use of perforin CaMACPF6 to increase the tolerance of plants to hypoxia stress. Specifically, the amino acid sequence of the perforin CaMACPF is shown as SEQ ID NO. 2. The invention also claims the use of the gene encoding said perforin CaMACPF in increasing the tolerance of plants to hypoxia stress. Alternatively, the nucleotide sequence of the gene is shown as SEQ ID NO. 1. In particular, the invention is directed to improving the hypoxia stress tolerance of plants by overexpressing the gene encoding the perforin CaMACPF in the plants. Thus, the invention also claims the use of an expression promoter of perforin CaMACPF in increasing the tolerance of plants to hypoxia stress. Specifically, the expression promoter of perforin CaMACPF is a recombinant overexpression vector containing a gene encoding perforin CaMACPF 6. The application of the perforin CaMACPF6 in the construction of plant lines resistant to hypoxia stress is also within the scope of the present invention. Similarly, the use of the gene encoding the perforin CaMACPF of the present invention, and th