Plants stress tolerance correlation transcription factor GmWRKY54, encoding gene and application thereof

A coding gene and gene technology, applied in soybean stress tolerance-related proteins and their coding genes and application fields, to achieve the effects of increasing yield, improving salt tolerance, and improving drought tolerance

Inactive Publication Date: 2008-05-07
INST OF GENETICS & DEVELOPMENTAL BIOLOGY CHINESE ACAD OF SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Such transcription factors associated with abiotic stress in soybean have not yet been cloned

Method used

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  • Plants stress tolerance correlation transcription factor GmWRKY54, encoding gene and application thereof
  • Plants stress tolerance correlation transcription factor GmWRKY54, encoding gene and application thereof
  • Plants stress tolerance correlation transcription factor GmWRKY54, encoding gene and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0037] Example 1. Screening of soybean stress tolerance-related protein GmWRKY54 encoding gene and its cDNA clone

[0038] BLAST search was performed in the soybean EST database, and 64 WRKY-like gene fragments including the complete WRKY domain were clustered. 64 pairs of primers were designed according to the sequences of 64 WRKY gene fragments, and total RNA was extracted from soybean Kefeng 1 seedlings treated with 250mM NaCl, drought, -4°C and untreated for 3 weeks by conventional methods, and RT-RCR method was used to extract total RNA The relationship between the expression of 64 genes and stress treatment was determined. Screening resulted in a gene whose expression was induced by drought or salt stress and slightly induced by low temperature.

[0039] Total RNA was extracted from soybean Kefeng 1 seedlings, and cDNA was synthesized from RNA with reverse transcriptase. The primers were designed according to the gene sequence obtained from the above retrieval as follo...

Embodiment 2

[0043] Example 2, Expression characteristics of soybean GmWRKY54 gene under adversity stress treatment

[0044] Soybean Kefeng No. 1 seeds were planted in pots, and after 2 weeks of growth, the seedlings were taken and subjected to the following stress treatments: 1) salt stress treatment: the soybean seedlings were moved into 250 mM NaCl solution; 2) osmotic stress treatment: the soybean seedlings were moved into 20 %PEG solution; 3) low temperature stress treatment: the soybean seedlings were moved into 4°C aqueous solution. Illumination culture, samples were taken at 0, 0.5, 1, 3, 6, and 12 hours after the above-mentioned various treatments, and 1 g of fresh leaves were collected and ground in liquid nitrogen, suspended in 4 mol / L guanidine thiohydrogen, and the mixture was washed with acid phenol, Extract with chloroform, and add ethanol to the supernatant to precipitate to obtain total RNA. Northern analysis was performed using GmWRKY54 DNA as a probe (sequence 1 in the ...

Embodiment 3

[0046] Embodiment 3, use GmWRKY54 gene to cultivate salt-tolerant and drought-resistant plants

[0047] 1) Construction of GmWRKY54 expression vector pBin438-GmWRKY54

[0048] Using the cDNA obtained by reverse transcription of the total RNA of soybean Kefeng 1 as a template, GmWRKY54 (sequence 1 in the sequence listing) was amplified by PCR with specific primers containing BamHI and KpnI linker sequences; then BamHI and KpnI double-digested the PCR product , Recovered, GmWRKY54 was inserted forward into the plant binary expression vector pBin438 (Li Taiyuan, Tian Yingchuan, Qin Xiaofeng, etc. Research on efficient insect-resistant transgenic tobacco [J]. Chinese Science (Series B), 1994, 24 (3): 276- 282.) between the BamHI and KpnI restriction sites behind the CaMV 35S promoter, the recombinant vector pBin438-GmWRKY54 was obtained (Fig. 2).

[0049] The primer sequences are as follows:

[0050] 5’-ATC AGG ATC CAT GAC AGT AGA TCT GGT AGG-3’

[0051] 5'-GAT GGG TAC CTG ACA ...

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Abstract

The invention discloses adverse-resistant relative albumen of a plant and the code gene and the application thereof. The plant adverse-resistant relative albumen is of the protein of following (a) or (b) that: (a) is the protein formed by listing amino acid shown in the sequence 2 of a sequence table; (b) is the protein formed by listing the amino acid in the sequence 2 of the sequence table through the replacing and / or loosing and / or adding of one or a plurality of amino acid residues and derived from the sequence 2. The code gene of the plant adverse-resistant relative albumen is guided into a plant cell, and then the transgenic plant variety with increased adverse-resistant force to the abiological adversity stress of drought and salt can be obtained.

Description

technical field [0001] The invention relates to a plant stress tolerance-related protein and its coding gene and application, especially the stress tolerance-related protein derived from soybean, its coding gene and its application. Background technique [0002] Changes in physical and chemical factors in the environment, such as drought, salinity, low temperature and other stress factors, have an important impact on the growth and development of plants. In severe cases, they will cause large-scale crop yield reduction. Cultivating stress-tolerant crops is one of the main goals of planting. At present, genetic engineering breeding has become one of the important methods to enhance crop stress tolerance. Higher plant cells have multiple pathways to respond to various stresses in the environment, and transcription factors play a role in regulating the expression of stress tolerance-related effector genes. Many types of transcription factors have been found in plants related t...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K14/415C12N15/29C12N15/82
CPCC12N15/8261C07K14/415C12N15/8273Y02A40/146
Inventor 陈受宜张劲松周绮云田爱国何锶洁杜保兴
Owner INST OF GENETICS & DEVELOPMENTAL BIOLOGY CHINESE ACAD OF SCI
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