Corn 3-phosphate dehydrogenase ZmGPDH4 and application of coding gene of corn 3-phosphate dehydrogenase ZMGPDH4 in regulating plant stress tolerance

A technology of plant stress tolerance and glycerol phosphate, which can be used in applications, plant products, genetic engineering, etc., and can solve the problems of few research reports

Inactive Publication Date: 2020-01-14
HEILONGJIANG BAYI AGRICULTURAL UNIVERSITY
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

Except for the model plant Arabidopsis thaliana and some algae, the GPDH f

Method used

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  • Corn 3-phosphate dehydrogenase ZmGPDH4 and application of coding gene of corn 3-phosphate dehydrogenase ZMGPDH4 in regulating plant stress tolerance
  • Corn 3-phosphate dehydrogenase ZmGPDH4 and application of coding gene of corn 3-phosphate dehydrogenase ZMGPDH4 in regulating plant stress tolerance
  • Corn 3-phosphate dehydrogenase ZmGPDH4 and application of coding gene of corn 3-phosphate dehydrogenase ZMGPDH4 in regulating plant stress tolerance

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0073] Example 1. Expression of ZmGPDH4 gene in maize under pathogen stress The data of ZmGPDH4 expression in maize under stress was downloaded from NCBI (SRP039594).

[0074] The result is as figure 1 shown. The results showed that the expression of ZmGPDH4 gene was significantly up-regulated in different maize varieties (R: Stress-tolerant variety S: Sensitive variety) after pathogen stress treatment. The results showed that ZmGPDH4 could be induced and expressed by pathogens in different degrees in maize, indicating that ZmGPDH4 is an important regulator in the process of maize adapting to biological stress.

Embodiment 2

[0075] Example 2. Acquisition and Stress Tolerance Analysis of ZmGPDH4 Transgenic Arabidopsis 1. Acquisition and Identification of ZmGPDH4 Transgenic Arabidopsis

[0076] 1. Taking maize root cDNA as a template, using ZmGPDH4-GFP-F and ZmGPDH4-GFP-R primers for PCR amplification to obtain PCR amplification products. The primer sequences are as follows (the underlined sequence is the enzyme recognition site):

[0077] ZmGPDH4-GFP-F: 5'-GC TCTAGA ATGGAGATGGAGAACGGGCAC-3';

[0078] ZmGPDH4-GFP-R:5'-ACGC GTC GAC GTAGAATGGAGTATGACCACCG-3'.

[0079] 2. Carry out double digestion and ligation of the pBI121-GFP vector and the PCR amplification product obtained in step 1 with restriction endonucleases XbaI and SalI to obtain the recombinant plasmid 35S:ZmGPDH4-GFP. and validated by sequencing.

[0080] Sequencing results show that the recombinant vector 35S:ZmGPDH4-GFP replaces the DNA fragment between the XbaI and SalI restriction sites of the pBI121-GFP vector with the ZmGPDH4...

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Abstract

The invention discloses corn 3-phosphate dehydrogenase ZmGPDH4 and application of a coding gene of the corn 3-phosphate dehydrogenase ZmGPDH4 in regulating plant stress tolerance. For the corn 3-phosphate dehydrogenase ZmGPDH4 and the application disclosed by the invention, by taking a corn GPDH gene family member ZmGPDH4 as a research object, and the corn GPDH gene family member ZmGPDH4 is transferred to an arabidopsis mutant to obtain T3 generation homozygous transformants; two transformants of COM-1 and COM-2 in the T3 generation homozygous transformants are selected to perform disease-resistant function identification; and by taking the arabidopsis mutant as a control, changes in contents of 3-glycerophosphate and glycerinum and physiological phenotype of the ZmGPDH4 transgenic arabidopsis under the germ stress are studied. A result shows that under the germ Pst DC3000 stress treatment condition, the contents of 3-glycerophosphate and glycerinum in the ZmGPDH4 transgenic arabidopsis are significantly higher than those in the arabidopsis mutant, and the disease infection condition of the the ZmGPDH4 transgenic arabidopsis is significantly better than that of the control. The result shows that the ZmGPDH4 can significantly improve the disease resistance of transgenic plants and can be applied to cultivation of a corn anti-reverse variety as an inverse gene.

Description

technical field [0001] The invention belongs to the field of biotechnology, and specifically relates to the application of corn 3-phosphate glycerol dehydrogenase ZmGPDH4 and its coding gene in regulating plant stress tolerance. Background technique [0002] Improving the stress tolerance of crops has become a hot and difficult point in the current technical research in the field of agriculture and animal husbandry, and it is also a major problem that needs to be solved urgently. In recent years, with the development of functional genomics and molecular biology, excavating key genes of stress tolerance and using genetic engineering technology to breed new crop varieties with good stress tolerance have become one of the means to effectively improve crop stress tolerance. [0003] Glycerol-3-phosphate (G-3-P) is an important intermediate product in the process of lipid metabolism and participates in various physiological and biochemical processes in plants. Glycerol 3-phospha...

Claims

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Application Information

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IPC IPC(8): C12N9/04C12N15/53C12N15/82A01H5/00A01H6/20
CPCC12N9/0006C12Y101/01008C12N15/8281C12N15/8243
Inventor 徐晶宇刘梦李佐同赵莹贺琳魏金鹏赵长江
Owner HEILONGJIANG BAYI AGRICULTURAL UNIVERSITY
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