A kind of alfalfa MsEXPA4 gene and its application in regulating low temperature resistance

By screening and verifying the MsDREB1C interacting protein MsEXPA4 in alfalfa, and constructing a recombinant vector that overexpresses the MsEXPA4 gene, the problem of insufficient low temperature resistance in alfalfa was solved, and better anti-freeze performance under low temperature stress was achieved.

CN118773203BActive Publication Date: 2025-03-21INSTITUTE OF ANIMAL SCIENCES OF CHINESE ACADEMY OF AGRICULTURAL SCIENCES
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Patent Information

Application Number
CN202410893982.5
Authority / Receiving Office
CN · China
Patent Type
Patents(China)
Current Assignee / Owner
Filing Date
2024-07-04
Publication Date
2025-03-21
Estimated Expiration
2044-07-04

AI Technical Summary

Technical Problem

The prior art studies the functions and mechanisms of DREB1/CBFs at low temperatures in alfalfa. There is almost no effective gene regulation method to improve the low temperature resistance of plants.

Method used

The MsDREB1C interacting protein MsEXPA4 was screened and verified by Y2H yeast library, and a recombinant vector overexpressing the MsEXPA4 gene was constructed, and alfalfa strain overexpressing the MsEXPA4 was obtained through genetic transformation.

Benefits of technology

Alfalfa strains overexpressing MsEXPA4 showed better resistance to frost under low temperature stress, including alleviating the damage of low temperature to cell membrane permeability, reducing leaf chlorophyll damage and enhancing antioxidant enzyme activity, which significantly improved the low temperature tolerance of plants.

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Abstract

The present invention relates to the field of biotechnology, and in particular, to a gene of alfalfa MsEXPA4 and its application in regulating low temperature resistance. The present invention uses BD-MsDREB1C as a bait protein to capture a candidate interacting protein MsEXPA4 through a Y2H yeast library, and verifies the interaction between MsDREB1C and MsEXPA4; the low temperature resistance analysis of the overexpressing MsEXPA4 strain found that whether it was a freezing treatment under plant cold acclimation or a freezing treatment under in vitro leaf cold acclimation, the overexpressing alfalfa strain showed better freezing resistance than the wild type, which was manifested in relieving the damage of low temperature to the permeability of plant cell membranes, relieving the damage caused by low temperature to chlorophyll in plant leaves, and enhancing the activity of antioxidant enzymes to alleviate the oxidative damage caused by low temperature. The present invention discloses the direct interaction between MsEXPA4 and MsDEEB1C, and shows that the physiological responses of the two to low temperature are basically the same, and the low temperature resistance response induced by cold acclimation of alfalfa is positively regulated.
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