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Stress tolerance associated protein TaSAP1 of plants as well as coding gene and application thereof

A gene and plant technology, applied in the field of plant stress tolerance-related protein TaSAP1 and its coding gene and application, can solve problems such as shortage of soil salinization, limitation of crop growth and increase of yield

Inactive Publication Date: 2014-05-21
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

[0002] Increasing global water scarcity, rapid soil salinization and frequent occurrence of extreme weather severely limit crop growth and yield enhancement

Method used

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  • Stress tolerance associated protein TaSAP1 of plants as well as coding gene and application thereof
  • Stress tolerance associated protein TaSAP1 of plants as well as coding gene and application thereof
  • Stress tolerance associated protein TaSAP1 of plants as well as coding gene and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0044] Embodiment 1, the acquisition of protein TaSAP1 and its coding gene

[0045] Wheat Hanxuan No. 10 (Triticumaetivum L.) seeds of uniform size were placed in a 9cm-diameter petri dish at 22°C with a light intensity of 150μmol·m -2 ·s -1 , the photoperiod is 12 hours of light, 12 hours of darkness, and under the condition of relative humidity of 70%, add deionized water for cultivation. The leaves of Hanxuan No. 10 seedlings that had grown to one leaf and one heart were taken, quickly frozen with liquid nitrogen, and stored at -80°C for later use. Total RNA was extracted with TRIZOL, and the first-strand cDNA (Invitrogen) was synthesized with the M-MLV reverse transcription kit. Using this cDNA as a template, primers F1: 5′-TCCTCCTATCAGTTAAATCAAAGC-3′ and R1: 5′-AATGCGAAAGCCACAATACC-3′ PCR amplification was performed, and the amplified product was subjected to 1.0% agarose gel electrophoresis to recover and purify a 0.8 kb fragment for sequencing. As a result, the full-l...

Embodiment 2

[0047] Example 2, real-time fluorescent quantitative PCR analysis of the expression characteristics of the TaSAP1 gene

[0048] 1. Responses of TaSAP1 gene to different stresses

[0049] Use 75% alcohol to disinfect Wheat Hanxuan No. 10 seeds of uniform size for 1 minute, then rinse them with running tap water for several minutes, place them in a 9cm-diameter petri dish, add deionized water for cultivation, and seal with plastic wrap to avoid moisture evaporation. When the height of the seedlings reaches the plastic wrap, pick holes on the film to make the seedlings stick out along the holes. The culture conditions were controlled at 22°C, relative humidity 70%, and light intensity 150 μmol m -2 ·s -1 , the photoperiod was 12 hours of light and 12 hours of darkness. When the seedlings grow to one leaf and one heart, the following treatments are carried out respectively:

[0050] 1) Drought treatment: absorb the original water in the petri dish, and add -0.5MPa (19.2%) sta...

Embodiment 3

[0074] Embodiment 3, utilize TaSAP1 gene to improve the stress tolerance of plant

[0075] 1. Construction of recombinant expression vector

[0076] 1. Cloning of TaSAP1 gene

[0077] A pair of primers (F3 and R3) were designed according to the sequence of the TaSAP1 gene, and the 5' ends of the primers were respectively introduced into SacI and BamHI restriction enzyme recognition sites, and the cDNA of Wheat Hanxuan No. 10 obtained in Example 1 was used as a template to amplify the TaSAP1 gene by PCR. The PCR amplification product was subjected to 1% agarose gel electrophoresis, and a band of about 500 bp was recovered and purified.

[0078] F3: 5'- GAGCTC ATGGAGCACAAGGAGACCGG-3′;

[0079] R3: 5'- GGATCC GATCTTGTCGAGCTTCTCCGC-3'.

[0080] 2. Construction of recombinant expression vector

[0081] ① Use restriction endonucleases SacⅠ and BamH I to digest the purified PCR product in step 1, and recover the digested product;

[0082] ② Digest the vector pCH3-GFP with rest...

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Abstract

The invention discloses a stress tolerance associated protein TaSAP1 of plants as well as a coding gene and an application thereof. Experiments prove that the survival rate of a T3 generation TaSAP1 transformed homozygous plant obtained by using the recombinant expression vector pCH3-GFP-TaSAP1 of DNA (deoxyribonucleic acid) molecules shown by nucleotide sequences at sites from 26th to 532nd in a sequence table 2 for converting the arabidopsis in drought tolerance experiment is 54.2%-66.7%; the survival rates of a wild type plant and an empty vector-transformed plant are respectively 14.6% and 20.8%; in a salt tolerance experiment, compared with a wild type contrast plant and an empty vector-transformed plant contrast plant, the TaSAP1 transformed plant has the advantages that the reduction of chlorophyll content in the TaSAP1 transformed plant is slow in comparison than that before the salt tolerance stress. The TaSAP1 protein and the coding gene thereof provided by the invention have important significance in improvement of plant adverse resistance, provide a basis for artificial control expression of stress tolerance associated gene and play an important role in breeding plants with high stress tolerance such as strong drought tolerance and strong salt tolerance.

Description

technical field [0001] The invention relates to a plant stress tolerance-related protein TaSAP1, its coding gene and application. Background technique [0002] The increasing shortage of global water resources, the rapid increase of soil salinization and the frequent occurrence of extreme weather have severely limited crop growth and yield improvement. Among them, drought and salinization have become common problems in many parts of the world, and it is estimated that more than 50% of the available land will be seriously threatened by salinization by 2050. At the same time, the world's population is growing, and agriculture must feed a growing population while competing with urban development for arable land, increasing pressure on global food demand. As the main food crop of human beings, the production and development of wheat are seriously restricted by unfavorable environmental conditions such as drought and water shortage. Therefore, improving the drought resistance o...

Claims

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Application Information

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IPC IPC(8): C07K14/415C12N15/29C12N15/63C12N5/10C12N1/21C12N7/01C12N15/82C12N15/84A01H5/00
CPCC07K14/415C12N15/8273
Inventor 景蕊莲王彩香毛新国李昂昌小平
Owner INST OF CROP SCI CHINESE ACAD OF AGRI SCI
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