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Application of glycosyltransferase gene UGT85A5 of Arabidopsis thaliana to improvement of salt tolerance of plants

A technology of glycosyltransferase and Arabidopsis sugar, which is applied to the application of the Arabidopsis glycosyltransferase gene UGT85A5 in improving the salt tolerance of plants, the glycosyltransferase gene and its application field, which can solve the problem of Arabidopsis thaliana. Glycosyltransferase gene has not been reported and other problems, to achieve the effect of improving salt tolerance

Inactive Publication Date: 2012-07-18
SHANDONG UNIV
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, after searching, Arabidopsis glycosyltransferase gene UGT85A5 The application in enhancing plant salt tolerance has not been reported so far

Method used

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  • Application of glycosyltransferase gene UGT85A5 of Arabidopsis thaliana to improvement of salt tolerance of plants
  • Application of glycosyltransferase gene UGT85A5 of Arabidopsis thaliana to improvement of salt tolerance of plants
  • Application of glycosyltransferase gene UGT85A5 of Arabidopsis thaliana to improvement of salt tolerance of plants

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Embodiment 1

[0016] Example 1 Arabidopsis glycosyltransferase gene UGT85A5 molecular cloning

[0017] 1. Arabidopsis glycosyltransferase gene UGT85A5 clone

[0018] Obtained through the public website http: / / www.cazy.org UGT85A5 cDNA sequence of the gene. Primers were designed according to the cDNA sequence, wherein the forward primer was 85A5-F: 5'-GCGTCTAGAATGGCGTCTCATGCTGTTAC-3'; the reverse primer was 85A5-R: 5'-GCGGAGCTCCTACTCCCCCTAAAAGAACCT-3'. Arabidopsis RNA was extracted using TRIzol kit and amplified by RT-PCR UGT85A5 The full-length cDNA sequence of the gene. The RT-PCR amplification program was as follows: 94°C for 2 min; 94°C for 40 s, 55°C for 40 s, 72°C for 1 min, a total of 36 cycles; finally, 72°C for 7 min. will get UGT85A5 The gene cDNA was cloned into the pBluescript II SK(+) vector (a universal vector) after enzyme digestion and ligation, which is called pK85A5 intermediate vector, and then the full-length PCR verification and double enzyme digestion verificatio...

Embodiment 2

[0024] Example 2 Arabidopsis glycosyltransferase gene UGT85A5 transgenic application

[0025] 1. contains UGT85A5 Construction of cDNA expression vector for coding region

[0026] use Xba I and Sac Ⅰ Obtained by double digestion of pK85A5 intermediate vector UGT85A5 full-length sequence, and then with the Xba I and Sac ⅠThe part of pBI121 vector digested with double restriction enzymes was connected to get the glycosyltransferase gene driven by CaMV 35S promoter UGT85A 5 plant overexpression vector, this vector is named pB85A5. Finally, through gene PCR verification and double enzyme digestion verification, it was identified that the construction of the plant expression vector was correct.

[0027] 2. Agrobacterium-mediated Plant Genetic Transformation

[0028] Firstly, the plant expression vector pB85A5 was transformed into Agrobacterium LBA4404, and it was verified by PCR and double enzyme digestion that the vector had been transformed into Agrobacterium.

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Abstract

The invention discloses application of a glycosyltransferase gene UGT85A5 of Arabidopsis thaliana to the improvement of salt tolerance of plants. A nucleotide sequence of the glycosyltransferase gene UGT85A5 is shown as SEQ ID No.1; and the glycosyltransferase gene UGT85A5 is cloned from the Arabidopsis thaliana by a reverse transcription-polymerase chain reaction (RT-PCR) technology. The gene UGT85A5 is utilized to construct a plant overexpression vector and perform transgenic operation of the plants to obtain transgenic plants. The detection shows that the salt tolerance of the transgenic plants is remarkably improved, so that a novel salt-tolerant plant can be created after the invention is implemented; and the gene can be used for subsequent crop variety improvement, and has a great significance for the agricultural production in China.

Description

technical field [0001] The present invention relates to a glycosyltransferase gene and its application, in particular to an Arabidopsis glycosyltransferase gene UGT85A5 The application in improving the salt tolerance of plants 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 important rol...

Claims

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Application Information

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IPC IPC(8): C12N15/54A01H5/00
Inventor 侯丙凯孙延国
Owner SHANDONG UNIV
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