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Method for improving plant tolerance to drought by down-regulating eifiso4g1 gene and eifiso4g2 gene

A plant and gene technology, applied in the fields of botanical equipment and methods, chemical instruments and methods, biochemical equipment and methods, etc., can solve problems such as the difficulty of stress tolerance traits

Inactive Publication Date: 2021-03-23
TSINGHUA UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

It is relatively difficult to cultivate and improve stress tolerance traits by traditional breeding techniques, and it is difficult to obtain excellent drought-resistant varieties

Method used

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  • Method for improving plant tolerance to drought by down-regulating eifiso4g1 gene and eifiso4g2 gene
  • Method for improving plant tolerance to drought by down-regulating eifiso4g1 gene and eifiso4g2 gene
  • Method for improving plant tolerance to drought by down-regulating eifiso4g1 gene and eifiso4g2 gene

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0061] Embodiment 1, the acquisition and identification of mutants

[0062] 1. The acquisition of mutant ei4g1-1

[0063] Seeds of Arabidopsis thaliana with the number SALK_098730 (the background is Arabidopsis ecotype Columbia, a T-DNA insertion mutant) were purchased from the Arabidopsis Biological Research Center and grown into plants. The genomic DNA of the plants at the seedling stage was extracted, and the primer pairs composed of ei4g1-1LP and ei4g1-1RP and the primer pairs composed of LBb1.3 and ei4g1-1RP were used for PCR identification. If the amplified product is not detected by using the primer pair composed of ei4g1-1LP and ei4g1-1RP, and the amplified product can be detected by using the primer pair composed of LBb1.3 and ei4g1-1RP, it proves that the plant has the same two chromosomes homozygous mutant. Homozygous mutants were sequenced for verification.

[0064] ei4g1-1LP: 5'-TCAAGGGCAAACATATCATCC-3';

[0065] ei4g1-1RP: 5'-TTTTGACTTCACGTTTCCGTC-3';

[006...

Embodiment 2

[0106] Embodiment 2, the resistance of plant to drought stress

[0107] 1. ABA promotes stomatal closure and inhibits stomatal opening experiment

[0108] ABA is an important signaling molecule for plants to resist external adversity stress. Under drought conditions, ABA can promote stomatal closure to reduce water loss by regulating ion channels in stomatal guard cells. Calcium ions, protein kinases, and phosphatases are all involved in ABA-mediated guard cell signal transduction and the signal regulation of ABA-related stress resistance genes. Therefore, ABA-induced stomatal movement experiments were used to detect whether eIFiso4G1 and eIFiso4G2 proteins play a role in the process of ABA regulating plant drought stress response.

[0109] Strip buffer: MES-KOH (pH 6.15) containing 50 mM KCl and 10 mM.

[0110] The light intensity for culturing under a cold light source was 200 μM Photons m -2 the s -1 .

[0111] 1. ABA inhibits stomatal opening experiment

[0112] The...

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Abstract

The invention discloses a method for improving the tolerance of a plant to droughts through down-regulation of expression of an eIFiso4G1 gene and an eIFiso4G2 gene. Application of the method in inhibiting expression substances of a specific gene A and a specific gene B in the plant comprises regulation and control over drought tolerance of the plant and seed selection for plant varieties of the improved drought tolerance. The specific gene A is a gene for encoding eIFiso4G1 protein; the specific gene B is a gene for encoding eIFiso4G2 protein. The inventor discovers that through knockout of the eIFiso4G1 gene and the eIFiso4G2 gene, the tolerance of the plant to droughts can be significantly improved. The method accords with the demands of the sustainable agricultural development and hasimportant practical value and an important market prospect in the aspects of studying the drought-tolerance molecule mechanism of the plant, improving the hereditary characters of the plant, improvingthe stress resistance of the plant to natural climatic disasters and bad soil environments, cultivating new efficient drought-tolerance varieties and the like.

Description

technical field [0001] The invention belongs to the field of biotechnology, and in particular relates to a method for improving plant tolerance to drought by down-regulating eIFiso4G1 and eIFiso4G2 genes. Background technique [0002] Terrestrial plants may be affected by a variety of external adversity stresses throughout their growth and development stages. Among them, abiotic stresses such as drought, salinity, low temperature, and high temperature, and biotic stresses such as pests and diseases significantly affect plant growth and crop yield. The research on plant stress resistance has always been a hot spot in the field of botany. It is relatively difficult to cultivate and improve stress-tolerant traits by traditional breeding techniques, and it is difficult to obtain excellent drought-resistant varieties. With the development of molecular biology technology and the in-depth study of the molecular mechanism of plant stress resistance, the genetic engineering of plant...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C07K14/415C07K19/00C12N15/82C12N15/29A01H5/00A01H6/20
CPCC07K14/415C07K2319/00C12N15/8273
Inventor 张大鹏马宇毕超
Owner TSINGHUA UNIV
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