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Application of arabidopsis glycosyl transferase gene UGT 76C2 in improving plant drought resistance

A technology of UGT76C2 and glycosyltransferase, which is applied in the field of genetic engineering, can solve problems that have not been reported and achieve the effect of improving drought tolerance

Inactive Publication Date: 2012-11-28
SHANDONG UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, after searching, the application of Arabidopsis thaliana glycosyltransferase gene UGT76C2 in enhancing plant drought tolerance has not been reported so far

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  • Application of arabidopsis glycosyl transferase gene UGT 76C2 in improving plant drought resistance
  • Application of arabidopsis glycosyl transferase gene UGT 76C2 in improving plant drought resistance
  • Application of arabidopsis glycosyl transferase gene UGT 76C2 in improving plant drought resistance

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Embodiment 1

[0016] Example 1 Cloning of Arabidopsis Glycosyltransferase Gene UGT76C2

[0017] 1. Cloning of Arabidopsis thaliana glycosyltransferase gene UGT76C2

[0018] The cDNA sequence of the UGT76C2 gene was obtained through the public website http: / / www.cazy.org. Primers were designed according to the cDNA sequence. The forward primer was 76C2-F: 5'-CGCGGATCCGCCGCCATGGAGGAGAAGAGAAATGG-3', and the reverse primer was 76C2-R: 5'-CGGGAGCTCTTACAACAATAGTATATGATTAGCT-3'. Arabidopsis RNA was extracted using TRIzol kit, and the full-length cDNA sequence of UGT76C2 gene was amplified by RT-PCR. The process of cloning the cDNA is first digested by BamHI and SacⅠ, and then connected to the pBluescript II SK(+) carrier digested by the corresponding enzymes to construct a sequencing intermediate vector called pK76C2, and then use the vector to perform full-length PCR amplification of the gene And BamHI and SacⅠ enzyme digestion verification, and finally sequence determination to verify the corr...

Embodiment 2

[0024] Example 2 Transgenic application of Arabidopsis thaliana glycosyltransferase gene UGT76C2

[0025] 1. Construction of cDNA expression vector containing UGT76C2 coding region

[0026] After the pK76C2 intermediate sequencing vector was digested with BamHI and SacⅠ, the full-length cDNA sequence with the cohesive ends was obtained. The gene fragment was connected with the pBI121 carrier part digested with corresponding enzymes to obtain a plant expression vector driven by the CaMV 35S promoter to overexpress the glycosyltransferase gene, which was called pBI76C2.

[0027] 2. Agrobacterium-mediated plant transformation

[0028] Agrobacterium GV3101 has the ability to infect plants and transfer genes, so the constructed UGT76C2 plant expression vector (pBI76C2) was transformed into Agrobacterium, and then verified by PCR and enzyme digestion. Arabidopsis flower buds were infiltrated with Agrobacterium GV3101 containing a plant expression vector using the flower dip method...

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Abstract

The invention discloses an application of an arabidopsis glycosyl transferase gene UGT 76C2 in improving plant drought resistance, wherein the nucleotide sequence of the glycosyl transferase gene UGT 76C2 is shown as SEQ ID No.1, and is cloned from arabidopsis through a reverse transcription-polymerase chain reaction (RT-PCR) technology. The application utilizes the gene UGT 76C2 to structure a plant over expression carrier, the plant transgenic operation is carried out, and a transgenic plant is obtained. Tests prove that drought resistance of the transgenic plant is obviously improved, and indicate that a novel drought-resistant plant can be created after the glycosyl transferase gene UGT 76C2 is implemented; and the glycosyl transferase gene UGT 76C2 can be used for subsequent crop variety improvement, and has an important meaning to agricultural production of China.

Description

technical field [0001] The invention relates to a glycosyltransferase gene and its application, in particular to the application of an Arabidopsis thaliana glycosyltransferase gene UGT76C2 in improving the drought tolerance of plants, which belongs to the field of genetic engineering. Background technique [0002] Glycosyltransferases are enzymes specifically responsible for catalyzing glycosylation modification reactions, which transfer active sugar groups from donors (usually UDP-glucose) to acceptor molecules. Glycosylation modification often changes the biological activity, water solubility, transport characteristics in the cell and the whole plant, subcellular localization, and mutual recognition and binding characteristics with receptors of plant molecules. In addition, it can also reduce or eliminate endogenous and exogenous Toxicity of source substances (Lim and Bowles, 2004; Bowles et al., 2006; Wang and Hou, 2009). Therefore, the glycosyltransferase gene plays an ...

Claims

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Application Information

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IPC IPC(8): C12N15/82C12N15/84A01H5/00
Inventor 侯丙凯王波
Owner SHANDONG UNIV
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