Plant stress tolerance related protein TaTPRPK1, encoding gene thereof, and application thereof

A technology of plant stress tolerance and stress tolerance, 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 promoting development

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...

Method used

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  • Plant stress tolerance related protein TaTPRPK1, encoding gene thereof, and application thereof
  • Plant stress tolerance related protein TaTPRPK1, encoding gene thereof, and application thereof
  • Plant stress tolerance related protein TaTPRPK1, encoding gene thereof, and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0050] Embodiment 1, the cloning of TaTPRPK1

[0051] 1. Isolation of mRNA

[0052] The three-leaf stage seedlings of triticale triticifolia (National Germplasm Bank, No. ZM242) grown in hydroponics for about 10 days were treated with NaCl for 2 hours, quick-frozen in liquid nitrogen, and stored at -80°C for later use.

[0053] Total RNA was extracted from wheat leaves by Trizol method (TianGen), and reverse transcriptase XL (AMV) was used for first-strand cDNA synthesis. The ds cDNA was synthesized by SMART method, and the PCR product was detected by 1.0% agarose gel electrophoresis.

[0054] 2. Acquisition of the full-length sequence of the TaTPRPK1 gene

[0055] The full-length sequence of wheat dual-specificity protein kinase gene was obtained by 5'RACE and 3'RACE methods.

[0056] The protein shown in Sequence 1 of the sequence listing is named TaTPRPK1 protein, which consists of 494 amino acid residues and has a conserved dual-specificity protein kinase activation reg...

Embodiment 2

[0057] Example 2, real-time fluorescent quantitative PCR analysis of the expression characteristics of TaTPRPK1

[0058] 1. Coercion treatment

[0059] Seedling age is the triticale seedling of 10 days, carries out following treatment:

[0060] (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.

[0061] (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.

[0062] (3) Abscisic acid treatment: put the ...

Embodiment 3

[0081] Example 3, TaTPRPK1 improves the salt tolerance of Arabidopsis

[0082] 1. Construction of recombinant expression vector

[0083] 1. Cloning of TaTPRPK1 gene

[0084] A pair of primers (TaTPRPK1-121F and TaTPRPK1-121R) were designed according to the sequence of the TaTPRPK1 gene, and BamHI and XhoI restriction sites were introduced at the ends of the primers respectively, and TaTPRPK1 was amplified by PCR using the cDNA of wheat wheat as a template.

[0085] TaTPRPK1-121F: 5'-TCCCCCGGGATGGGCGCCAGGATGTC-3';

[0086] TaTPRPK1-121R: 5'-GGACTAGTTCAGTTCATATTCAGCGTCCG-3.

[0087] The PCR amplified product was subjected to 1.2% agarose gel electrophoresis, and a band of about 1.5 Kb was recovered and purified using Agarose Gel DNA Purification Kit Ver.2.0 (TaKaRa Company, Code No.: DV807A).

[0088] 2. Construction of recombinant expression vector

[0089] ① Use restriction endonucleases BamHI and XhoI to digest step 1 to recover the purified PCR product, and recover the d...

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Abstract

The invention discloses a plant stress tolerance related protein TaTPRPK1, an encoding gene thereof, and an application thereof. The protein provided by the invention is (a) or (b): (a) the protein is composed of an amino acid sequence represented by a sequence 1 in a sequence listing; and (b) the protein which is related to the plant stress tolerance, derived from the sequence 1, and obtained through substituting and/or deleting and/or adding to the amino acid sequence of the sequence 1 by one or some amino acid residues. The invention also discloses the encoding gene (TaTPRPK1) of the protein, and the application of the protein in breeding stress tolerance plants. The TaTPRPK1 can enhance the resistance of plants to NaCl, and the growth of roots. The TaTPRPK1 of the present invention lays the foundation for artificially controlling the expression of the anti-stress and stress resistance related gene, and plays an important role in breeding plant varieties with strengthened anti-stress and stress resistance.

Description

technical field [0001] The invention relates to a plant stress tolerance-related protein TaTPRPK1, 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. [0003] 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 geneti...

Claims

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Application Information

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IPC IPC(8): C07K14/415C12N15/29C12N15/63C12N5/10C12N1/15C12N1/19C12N1/21C12N15/11A01H5/00
Inventor 徐兆师马有志李连城陈明
Owner INST OF CROP SCI CHINESE ACAD OF AGRI SCI
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