Plant stress tolerance correlative protein, coding gene and application thereof

A technology that encodes genes and proteins, which is applied in the fields of plant gene improvement, application, plant peptides, etc., and can solve problems such as small improvement and inability to achieve plant stress resistance

Inactive Publication Date: 2009-12-02
INST OF CROP SCI CHINESE ACAD OF AGRI SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The tolerance of plants to adversity stresses such as drought and high salinity is a quantitative trait controlled by multiple genes, and its physiological and biochemical processes are regulated by gene interaction and co-regulation. Although the introduction of a single functional gene can improve the stress resistance of plants to a certain extent, However, the improvement range is not large, and the purpose of effectively enhancing plant stress resistance is often not achieved (Liu et al., 1998; Liu Qiang et al., 2000)

Method used

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  • Plant stress tolerance correlative protein, coding gene and application thereof
  • Plant stress tolerance correlative protein, coding gene and application thereof
  • Plant stress tolerance correlative protein, coding gene and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0047] Example 1. Screening of soybean stress tolerance-related protein GmAREB1 encoding gene and its cDNA clone

[0048] The 14-day-old soybean Tiefeng 8 seedlings (purchased from Yingjun Company) were treated with high salt (200mM NaCl) for 12 hours, and the total RNA of soybean was extracted with Trizol. 5'RACE kit (5'RACE System for Rapid Amplification of cDNA Ends Kit) (GIBCOBRL, CAT.NO.18374-058) and 3'RACE kit (3'RACE System for Rapid Amplification of cDNA Ends Kit) (GIBCOBRL, CAT.NO.18373-019) to obtain the full-length sequence of the GmAREB1 gene.

[0049] The total RNA of soybean Tiefeng 8 seedlings was extracted with Trizol (invitrogen), and cDNA was obtained by reverse transcription with superscriptII (invitrogene) reverse transcriptase. Primers P1 and P2 were designed according to the sequence of the coding region of GmAREB1 gene. Using the cDNA obtained by reverse transcription as a template, PCR amplification was performed with primers P1 and P2. The sequence...

Embodiment 2

[0054] Example 2, Expression characteristics of soybean GmAREB1 gene under adversity stress treatment

[0055] Soybean Tiefeng No. 8 seeds were planted in pots, and after 2 weeks of growth, the seedlings were taken and subjected to the following stress treatments: 1) Drought treatment: the soybean seedlings were taken out from the soil to absorb the moisture on the roots, and placed on dry filter paper; ) salt treatment: soybean seedlings were placed in 200 mM NaCl solution; 3) ABA treatment: soybean seedlings were placed in 200 μM ABA solution; 4) cold treatment: wheat seedlings were placed in a 4 ° C incubator; Samples were taken at 0, 0.5, 1, 3, 6, and 12 hours after various treatments, quickly frozen with liquid nitrogen, and stored at -80°C for later use.

[0056] Total RNA was extracted from samples taken 0, 0.5, 1, 3, 6, and 12 hours after drought treatment, salt treatment, ABA treatment, and cold treatment, and Northern analysis was performed using GmAREB1 DNA as a probe...

Embodiment 3

[0075] Embodiment 3, gel migration or electrophoretic mobility experiment (EMSA)

[0076] 1. Prokaryotic expression of GmAREB1 gene

[0077] The full-length sequence of GmAREB1 gene is inserted in the multiple cloning site of prokaryotic expression vector pGEX-4T-1, because there is GST (glutathione) gene in front of the multiple cloning site of pGEX-4T-1, GmAREB1 and GST protein like this The fusion protein was formed, and the constructed expression vector was transformed into recipient Escherichia coli BL21, and the production of the fusion protein (GmAREB1-GST) was induced by adding inducer IPTG to the medium, and the induced expression of the fusion protein was detected by SDS-PAGE electrophoresis .

[0078] Purify the GmAREB1-GST fusion protein according to the instructions of the MicroSpin GST Purification Module Kit.

[0079] 2. Labeling of ABRE probes

[0080] Synthesize the ABRE probe sequence according to the known sequence, the ABRE probe sequence is as follows: ...

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Abstract

The invention discloses a plant stress tolerance correlative protein, a coding gene and an application thereof. The plant stress tolerance correlative protein is the protein of the following (a) or (b): (a), a protein formed by an amino acid sequence shown in a sequence 1 in a sequence table; (b), a protein which is formed in a way that the amino acid sequence of the sequence 1 in the sequence table is substituted and/or lost and/or added by one or a plurality of amino acid residues, is correlative to plant stress tolerance and is derived from the sequence 1. The invention also provides the coding gene of the protein. The coding gene of the plant stress tolerance correlative protein is introduced into plant cells, and a transgenic plant variety with reinforced adverse circumstance stress tolerance to abiotic substances, such as salt, and the like can be obtained. The invention has very important theoretical and practical meanings for improving and reinforcing the stress tolerance of plants, increasing the yield, quickening the breeding process of stress tolerance molecules and effectively saving water resources.

Description

technical field [0001] The invention relates to a plant stress tolerance-related protein and its coding gene and application. Background technique [0002] Abiotic stresses such as drought and salinity are the main limiting factors affecting crop yield and quality. According to statistics, the world's arid and semi-arid areas account for 33% of the earth's land area, and saline-alkali land accounts for 7.6% of the earth's land area. There are 554 million mu of saline-alkali land to be reclaimed and utilized. [0003] The use of conventional breeding methods to improve the stress resistance of crops is restricted by the long cycle and the lack of excellent germplasm resources. With the development of modern molecular biology and genetic engineering, genetic engineering technology is used to study the relationship between plants and abiotic stress at the molecular level, reveal the molecular mechanism of plant stress signal transduction and gene expression regulation, and cul...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K14/415C12N15/29C12N15/63C12N5/10C12N1/15C12N1/19C12N1/21C12N15/82A01H1/00A01H5/00
Inventor 马有志高世庆陈明徐兆师李连城
Owner INST OF CROP SCI CHINESE ACAD OF AGRI SCI
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