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Application of protein OsZZW1 in regulating and controlling drought resistance of oryza sativa

A technology of protein and drought resistance, applied in the application, use of vectors to introduce foreign genetic material, angiosperms/flowering plants, etc., can solve problems such as difficult plant stress resistance

Active Publication Date: 2020-06-02
HEBEI NORMAL UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Due to the complexity of plant stress resistance traits, it is very difficult to improve the stress resistance of plants using traditional breeding methods. Segregation becomes the main factor limiting genetic engineering of plant stress resistance

Method used

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  • Application of protein OsZZW1 in regulating and controlling drought resistance of oryza sativa
  • Application of protein OsZZW1 in regulating and controlling drought resistance of oryza sativa
  • Application of protein OsZZW1 in regulating and controlling drought resistance of oryza sativa

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0068] Embodiment 1, the cloning of the coding gene (being OsZZW1 gene) of protein OsZZW1

[0069] 1. Use the Trizo1 method to extract the total RNA from the leaves of the 7-day-growing rice variety Nipponbare (hereinafter referred to as Nipponbare), then use reverse transcriptase to reverse transcribe the first-strand cDNA, and then use the SMART method to synthesize dscDNA to obtain Nipponbare ds -cDNA.

[0070] 2. Using the Nipponbare ds-cDNA obtained in step 1 as a template, PCR amplification was performed using a primer pair consisting of primer OsZZW1-F: 5'-ATGCCGAGCGCCTTCCACT-3' and primer OsZZW1-R: 5'-GAATGAGAACTTGAATCCTCCA-3', Obtain the PCR amplification product.

[0071] The PCR amplified products were subjected to 1% agarose gel electrophoresis.

[0072] 1% agarose gel electrophoresis results see figure 1 (1 is PCR amplification product).

[0073] 3. Recover a 1566bp DNA fragment from the PCR amplification product obtained in step 2.

[0074] The DNA fragments...

Embodiment 2

[0075] Embodiment 2, the acquisition of transgenic rice and the drought resistance identification

[0076] 1. Construction of recombinant plasmids

[0077] 1. Construction of recombinant plasmid pCAMBIA1300-OsZZW1

[0078] (1) Using the DNA fragment obtained in Step 3 of Example 1 as a template, using primer F: 5'-TTCTGCAGCG GGATCC ATGCCGAGCGCCTTCCAC-3' (the underline is the recognition site of restriction endonuclease BamHI) and primer R: 5'-CGTAACGCGT GGATCC A primer pair consisting of TTGAATGAGAACTTGAATCCTCC-3' (the underline is the recognition site of restriction endonuclease BamHI) was used for PCR amplification, and then DNA fragment A of 1597bp was recovered.

[0079] (2) The pCAMBIA1300 vector was digested with restriction endonuclease BamHI, and the 14.58kb vector backbone was recovered.

[0080] (3) Ligate the DNA fragment A and the vector backbone with infusion ligase to obtain the recombinant plasmid pCAMBIA1300-OsZZW1.

[0081] The recombinant plasmid pCAMBI...

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Abstract

The invention discloses application of a protein OsZZW1 in regulating and controlling drought resistance of oryza sativa. The amino acid sequence of the protein OsZZW1 is shown in SEQ ID NO:2. Experiments show that the drought resistance of oryza sativa can be improved by reducing expression of the protein OsZZW1 in Nipponbare; and improvement of the drought resistance lies in that the survival rate is increased, the content of proline is increased, and the content of malonaldehyde is reduced. The protein OsZZW1 is capable of regulating and controlling drought resistance of oryza sativa. The application has significant application value.

Description

technical field [0001] The invention belongs to the field of biotechnology, and in particular relates to the application of protein OsZZW1 in regulating the drought resistance of rice. Background technique [0002] Drought is the most important environmental factor affecting plant growth and crop yield. Drought has become a serious problem affecting agricultural production. The use of genetic engineering to improve the drought resistance of crops and improve the adaptability of crops and economic crops to adversity is a key and major issue that needs to be solved urgently in the breeding of new varieties. In recent years, people have done a lot of research on the mechanism of plant response to drought and other adversity stresses from the perspectives of physiology, biochemistry, metabolism, ecology, genetics, and evolution, and accumulated a wealth of data. Especially with the development of molecular biology, people The ability to understand the mechanism of plant resista...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K14/415C12N15/29C12N15/82A01H5/00A01H6/46
CPCC07K14/415C12N15/8218C12N15/8273
Inventor 朱正歌赵伟杰张倩姚海涛郑倩刘栋良
Owner HEBEI NORMAL UNIV
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