Protein CkWRKY33 and coding gene and application thereof

A protein and coding technology, applied in the fields of application, genetic engineering, plant gene improvement, etc., can solve the problem of little research on stress-related genes

Active Publication Date: 2020-02-18
THE INST OF BIOTECHNOLOGY OF THE CHINESE ACAD OF AGRI SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] So far, studies on Caragana caragana have mainly focused on biological characteristics, physiologic...

Method used

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  • Protein CkWRKY33 and coding gene and application thereof
  • Protein CkWRKY33 and coding gene and application thereof
  • Protein CkWRKY33 and coding gene and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0063] Example 1. Cloning of the coding gene of protein CkWRKY33 (i.e. CkWRKY33 gene)

[0064] 1. Sow the seeds of Caragana korshinskii on sandy soil and manage the normal growth to obtain a Caragana korshinskii plant that has grown for one month; then stop watering for 20 days to obtain a drought-treated Caragana korshinskii plant.

[0065] The drought-treated Caragana korshinskii plant has a well-developed root system, white leaves and thick epidermal hairs, which have strong drought resistance.

[0066] 2. Take the leaves of the drought-treated Caragana korshinskii plant obtained in step 1, extract total RNA first, and then reverse transcription to obtain the cDNA of Caragana korshinskii.

[0067] 3. After completing step 2, use the cDNA of Caragana korshinskii as a template, and use primer F: 5'-ATGACTATGGATGATCATAACTG-3' and primer R: 5'-TTAGAAGTCCTTTGACATAAAT-3' to perform PCR amplification, and then recover About 1614bp PCR amplification product.

[0068] 4. Sequencing the PCR a...

Embodiment 2

[0071] Example 2. Obtainment of transgenic CkWRKY33 gene Arabidopsis and identification of drought resistance

[0072] 1. Obtaining the recombinant plasmid pBinGlyRed3-CkWRKY33

[0073] 1. Cut pBinGlyRed3 vector with restriction enzymes EcoRI and XmaI (Zhang C, Iskandarov U, Klotz ET, et al. Athraustochytrid diacylglycerol acyltransferase 2 with broadsubstrate specificity strongly increases oleic acid content in engineered Arabidopsis thaliana seeds. Journal of Experimental Botany, 2013 , 64(11):3189-3200.), and recover a vector backbone of about 9kb.

[0074] 2. Using the cDNA of Caragana korshinskii obtained in step 2 in step 1 as a template, using primer FF: 5’-ACGGGGGACT GA ATTC ATGACTATGGATGATCATAACTG-3' (underlined is the recognition site of restriction enzyme EcoRI) and primer FR: 5'-CCGCCTCGAG CCCGGG A primer pair consisting of TTAGAAGTCCTTTGACATAAAT-3' (underlined is the recognition site of restriction enzyme XmaI) was subjected to PCR amplification, and a PCR amplificat...

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Abstract

The invention discloses protein CkWRKY33 and a coding gene and application thereof. The amino acid sequence of the protein CkWRKY33 is shown as SEQID NO:2. Experiments prove that the CkWRKY33 gene istransmitted into arabidopsis thaliana Columbia-0 subtype, so that genetically modified Arabidopsis thaliana is obtained. Compared with the arabidopsis thaliana Columbia-0 subtype, the drought resistance of the genetically modified Arabidopsis thaliana is improved, improvement of the drought resistance reflects that the propanedial content is reduced, the soluble sugar content is increased, the proline content is increased, the peroxidase activity is improved, the rate of water loss is reduced, and the motility rate is increased. Therefore, the protein CkWRKY33 can improve the plant stress resistance. The protein CkWRKY33 has important application value.

Description

Technical field [0001] The invention belongs to the field of biotechnology, and specifically relates to the protein CkWRKY33 and its coding gene and application. Background technique [0002] Caragana korshinskii is a kind of leguminous shrub that mainly grows on sandy land in desert or semi-desert areas. It has a highly developed root system, strong drought tolerance and cold tolerance. Caragana korshinskii is not only an important forage and industrial raw material, but also can maintain water and soil and prevent soil desertification. It has important economic and ecological value (Fang XW, Li JH, Xiong YC, Xu DH, Fan XW, Li F M. Responses of Caraganakorshinskii Kom.to shoot removal: mechanisms underlying regrowth[J].Ecological Research, 2007, 23(5):863-871.). [0003] Up to now, the research on Caragana korshinskii has mainly focused on the biological characteristics, physiological changes and anatomical structure, but there are few studies on its resistance-related genes. Su...

Claims

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Application Information

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IPC IPC(8): C07K14/415C12N15/29C07K19/00C12N15/62C12N15/82A01H5/00A01H6/20
CPCC07K14/415C07K2319/20C07K2319/21C07K2319/22C07K2319/41C12N15/8273
Inventor 龙艳裴新梧梁凤萍
Owner THE INST OF BIOTECHNOLOGY OF THE CHINESE ACAD OF AGRI SCI
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