Plant stress-tolerant associated protein TaDREB4B and encoding gene and application thereof

A technology of stress tolerance and genetics, applied in the direction of plant genetic improvement, botany equipment and methods, applications, etc., can solve the problems of comprehensive improvement of stress resistance of difficult plants, and achieve the effect of improving drought resistance

Active Publication Date: 2011-11-09
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

Since the stress tolerance of plants is a complex trait regulated by multiple genes, it is difficult to achieve comprehensive improvement of plant stress resistance by introducing a single functional protein gene

Method used

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  • Plant stress-tolerant associated protein TaDREB4B and encoding gene and application thereof
  • Plant stress-tolerant associated protein TaDREB4B and encoding gene and application thereof
  • Plant stress-tolerant associated protein TaDREB4B and encoding gene and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0054] Embodiment 1, the cloning of TaDREB4B

[0055] 1. Isolation of mRNA

[0056] The three-leaf stage seedlings of triticale wheat (National Germplasm Bank, No. ZM242) grown for about 10 days were subjected to drought treatment for 2 hours, quick-frozen with liquid nitrogen, and stored at -80°C for later use. Isolation of mRNA was performed using Quikprep Micro mRNA Purification Kit (Pharmacia).

[0057] 2. Construction of cDNA library and titer determination

[0058] 1. Construction of cDNA library

[0059] Using Timesaver TM cDNA Synthesis Kit (Pharmacia) synthesizes cDNA double strands from the mRNA obtained in step 1, and adds EcoRI / NotI adapter; using ZAP Express Predigested Gigapack IIIGold Cloning Kit (Stratagene) was used to construct the cDNA library, and a total of 500ul library solution was obtained.

[0060] 2. Determination of titer

[0061] (1) Dilute 1ul of library solution with SM Buffer 1000 times;

[0062] (2) Take 1ul, 10ul, and 100ul dilutions...

Embodiment 2

[0138] Example 2, real-time fluorescent quantitative PCR analysis of the expression characteristics of TaDREB4B

[0139] 1. Various coercive treatments

[0140] Seedling age is the triticale seedling of 10 days, carries out following treatment

[0141] (1) Drought treatment: Take out the hydroponic wheat seedlings to absorb the water on the roots, place them on dry filter paper, and cultivate them in a dry environment for 30 minutes, 1 hour, 2 hours, 4 hours, 8 hours, 12 hours, and 24 hours. Materials were quick-frozen in liquid nitrogen and stored at -80°C for later use.

[0142] (2) Salt treatment: the wheat seedlings were placed in 2% of NaCl and Na 2 SO 4 In the sodium salt solution (NaCl and Na 2 SO 4 The mass percentage is 3:2), light cultured for 30 minutes, 1 hour, 2 hours, 4 hours, 8 hours, 12 hours, and 24 hours, respectively, the materials were taken out, quick-frozen in liquid nitrogen, and stored at -80°C for later use.

[0143] (3) Abscisic acid treatment: ...

Embodiment 3

[0162] Embodiment 3, the activation characteristic of TaDREB4B

[0163] The main principle of using the yeast one-hybrid system to demonstrate the activation properties of transcription factors is as follows: image 3 As shown, the DRE cis-acting element and the mutant DRE cis-acting element were respectively constructed on the upstream of the basic promoter Pmin (minimal promoter) of the pHISi-1 vector and pLacZi vector, and the downstream of the Pmin promoter was connected with the reporter gene (His3, LacZ and URA3). After the expression vector YEP-GAP (without activation function) connected with the target gene encoding the transcription factor is transformed into the yeast cells connected with the DRE cis-acting element and the mutant DRE cis-acting element, if the mutant DRE The reporter gene in yeast cells with cis-acting elements cannot be expressed, but the reporter gene in yeast cells with specific DRE cis-acting elements can be expressed, indicating that the transc...

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Abstract

The invention discloses plant stress-tolerant associated protein TaDREB4B, an encoding gene and an application thereof. The protein provided by the invention is (a) or (b) as follows: (a) protein formed by the amino acid sequence as shown in sequence 1 in the sequence table; (b) protein which is obtained by substitution and / or deletion and / or addition of one or more amino acid residues for the amino acid sequence of sequence 1, is associated with plant stress tolerance, and is derived from sequence 1. The TaDREB4B of the invention is expressed by the induction of drought, high salt, high temperature, low temperature, pathogen, ABA, ethane, JA, SA, and can specifically regulate the transcriptional expression of genes containing DRE / CRT cis-element (core sequence: CCGAC); and can increase drought resistance, salt tolerance, high temperature resistance, and powdery mildew pathogen resistance of plants. The TaDREB4B of the invention provides a base for the artificial control of the expression of stress-resistant and stress-tolerant associated genes, and play an important role in cultivating plants with enhanced stress resistance and stress tolerance.

Description

technical field [0001] The invention relates to a plant stress tolerance-related protein TaDREB4B, its coding gene and application. Background technique [0002] Adversity stresses such as drought, high salinity and low temperature are obstacles to the growth and development of wheat. Therefore, understanding the response and signal transduction mechanism of wheat to stress conditions and improving the stress resistance of wheat varieties have become one of the important tasks of wheat genetic research and wheat variety improvement. Under adversity stress, plants will produce a series of responses, accompanied by many physiological, biochemical and developmental changes. Clarifying the response mechanism of plants to stress will provide scientific evidence for the research and application of stress-resistant genetic engineering. At present, the research on plant stress resistance has gradually penetrated into the cellular and molecular levels, combined with genetics and ge...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K14/415C12N15/29C12N15/63C12N5/10C12N1/15C12N1/19C12N1/21A01H5/00
CPCC12N15/8282C12N15/8271C12N15/8281C12N15/8273
Inventor 马有志徐兆师李连城陈明
Owner INST OF CROP SCI CHINESE ACAD OF AGRI SCI
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