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Antioxidation-related genes of wheat as well as coding proteins thereof and application thereof

A related protein and anti-oxidation technology, which is applied in the fields of application, genetic engineering, and plant gene improvement, can solve the problems of increased degree of photoinhibition and decreased ability to resist photobleaching, and achieve the effect of increasing biomass and broad prospects

Inactive Publication Date: 2013-02-20
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

In Arabidopsis atsrx deletion mutants, the degree of photoinhibition of the chloroplast photosynthetic system is greatly increased, and the ability to resist photobleaching is also greatly reduced

Method used

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  • Antioxidation-related genes of wheat as well as coding proteins thereof and application thereof
  • Antioxidation-related genes of wheat as well as coding proteins thereof and application thereof
  • Antioxidation-related genes of wheat as well as coding proteins thereof and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0064] Example 1: Homologous cloning and gene structure analysis of TaSRX gene

[0065] All EST sequences of SRX genes were searched by NCBI-Blastn, and the full-length EST sequences of three types of SRX genes found in hexaploid wheat (subgenome group AABBDD) were obtained by alignment and splicing. Design conservative primers at ATG and TGA to clone these 3 copies at the same time, perform PCR amplification in sequence, recover the target fragment, T-A ligation, transform bacterial competence, culture at 37°C, positive clones are obtained by colony PCR identification, positive plasmid extraction, and finally carry out Three copies of the full-length genome sequence were obtained by plasmid digestion identification and sequencing. They are compared with Arabidopsis and rice SRX genes (ie AtSRX and OsSRX) as attached figure 1 shown.

[0066] The specific steps are as follows:

[0067] The primers used were 5'-ACGAATGGCGTCGACCTCGAGCTT-3' and 5'-CATCGCTAACCACCAATTCATCGCATAT-3...

Embodiment 2

[0071] Example 2: Cloning of TaSRX Gene cDNA Sequence and Protein Sequence Prediction and Alignment

[0072] The cDNA of XY54TaSRX was cloned using the same primers as in Example 1. PCR amplification reaction conditions: 95°C pre-denaturation for 5 minutes; 94°C for 40s, 68°C for 1min, 33 cycles of two-step method; 68°C for 7min. Reaction system 50 μl: 5 μl of 10×PCR reaction buffer, 0.5 μl of LA-Taq enzyme (Takara company), 100 ng of Xiaoyan 54 cDNA, 4 μl of 2.5 mmold NTP, 1 μl of 5 μmol primers, and sterilized double-distilled water to 50 μl. The PCR product was subjected to agarose gel electrophoresis, and the target TaSRX cDNA fragment was purified, and the concentration was measured by Eppendorf BioPhotometer for enzyme digestion, ligation or storage at -20°C for later use. The purified TaSRX cDNA fragment was ligated with pGEM-T Easy vector, and transformed into Escherichia coli DH10B by heat shock. Colony PCR (primers are the above-mentioned amplification primers) ide...

Embodiment 3

[0081] Example 3: Determination of TaSRX Gene Copy Number-Fluorescently Labeled DNA Fragment Analysis Technology

[0082] Using fluorescently labeled conserved primers spanning the second intron (only labeled with D4 fluorescent marker at the 5' end of the upstream primer, Takara) to amplify Xiaoyan 54 (subgenome group is AABBDD), Jing 411 (subgenome group Divided into AABBDD, the seeds come from the laboratory of Academician Li Zhensheng, Institute of Genetics and Development, Chinese Academy of Sciences 14 , DNA from leaves at the seedling stage), Langdon (the subgenome is AABB, the seeds are purchased from Graingene in the United States), Urartu wheat (the subgenome is AA, the seeds are purchased from the Institute of Crop Science, Chinese Academy of Agricultural Sciences), Goat grass (the subgenome component is DD, and the seeds are purchased from the Institute of Crop Science, Chinese Academy of Agricultural Sciences) genomic DNA (extraction method reference example 1), t...

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Abstract

The invention discloses antioxidation-related genes TaSRX-A, TaSRX-B and TaSRX-D of wheat as well as coding proteins and application thereof. The invention also discloses the application of the genes and the coding proteins thereof in regulating the antioxidation function of plants or protecting a photosynthetic system from photoinhibition and photoxidation function. The genes have the advantagesof being capable of clearing away active oxygen in a light stress condition, protecting the photosynthetic system, enhancing the capabilities of resisting photoinhibition and photoxidation of the plants and further increasing the biomass of the plants. The genes and the coding proteins thereof disclosed by the invention have an important theoretical significance and a practical significance for researching light stress resisting mechanisms of common wheat and increasing the output of the common wheat and other important crops and have wide application prospects.

Description

technical field [0001] The invention belongs to the field of plant genetic engineering. Specifically, the present invention relates to a sulfur redox factor protein related to resistance to light inhibition and photooxidation in wheat and its coding sequence, including the cloning, expression mode and application of the gene. Background technique [0002] There are a large number of reactive oxygen species in living organisms, which have two important functions. First: Low levels of reactive oxygen species can be used as signaling molecules to regulate important physiological processes such as cell division and cell differentiation 1 ; Second: a large amount of reactive oxygen species can cause damage to proteins, DNA and membrane lipids and cause cell death 2 . Organisms tend to easily produce a large amount of reactive oxygen species during the life process, so it is very important for organisms to remove excess reactive oxygen species. [0003] Peroxiredoxin (PRX, per...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N15/29C12N15/82C12N5/10C07K14/415A01H5/00C12Q1/68G01N33/53
Inventor 王道文王燃张坤普秦焕菊刘昕
Owner INST OF GENETICS & DEVELOPMENTAL BIOLOGY CHINESE ACAD OF SCI
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