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Application of Arabidopsis thaliana glycosyltransferase gene ugt76d1 in reducing plant surface wax

A technology of UGT76D1 and glycosyltransferase, applied in the field of genetic engineering, can solve the problems that the function and application of surface wax have not been reported, and the special function of glycosyltransferase gene has not been revealed, so as to reduce surface wax and increase production , enhance the effect of absorption efficiency

Active Publication Date: 2017-10-27
SHANDONG UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Although the sequences of these glycosyltransferase genes have been published in the nucleic acid sequence database, the special functions of these glycosyltransferase genes have not been revealed. For example, the UGT76D1 gene in Arabidopsis glycosyltransferase family 1 can create The function and application of transgenic plants with reduced surface waxiness have not been reported so far

Method used

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  • Application of Arabidopsis thaliana glycosyltransferase gene ugt76d1 in reducing plant surface wax
  • Application of Arabidopsis thaliana glycosyltransferase gene ugt76d1 in reducing plant surface wax
  • Application of Arabidopsis thaliana glycosyltransferase gene ugt76d1 in reducing plant surface wax

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Embodiment 1

[0019] Example 1 Cloning of Arabidopsis Glycosyltransferase Gene UGT76D1

[0020] 1. Cloning of Arabidopsis thaliana glycosyltransferase gene UGT76D1

[0021] The cDNA sequence of the UGT76D1 gene was obtained through the public website http: / / www.cazy.org. Primers were designed according to the cDNA sequence, the forward primer was 76D1-F:5'-CGGGATCCATGGCAGAGATTCGCCAG-3', and the reverse primer was 76D1-R:5'-CGAGCTCTCATTGTTCGTCAATTTGCATC-3'. Arabidopsis RNA was extracted using TRIzol kit, and the full-length cDNA sequence of UGT76D1 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(+) vector digested by the corresponding enzymes to construct a sequencing intermediate vector called pK76D1, 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 correctness of the cl...

Embodiment 2

[0024] Example 2 Transgenic application of Arabidopsis thaliana glycosyltransferase gene UGT76D1

[0025] 1. Construction of cDNA expression vector containing UGT76D1 coding region

[0026] After the pK76D1 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, called pBI76D1.

[0027] 2. Agrobacterium-mediated plant transformation

[0028] Agrobacterium GV3101 has the ability to infect plants and transfer genes, so the constructed UGT76D1 plant expression vector (pBI76D1) 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 (a publis...

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Abstract

The invention discloses an application of Arabidopsis thaliana glycosyltransferase gene UGT76D1 in reducing plant wax, wherein the nucleotide sequence of the glycosyltransferase gene UGT76D1 is shown in SEQ ID No.1. The invention utilizes the gene UGT76D1 to construct a plant overexpression vector, performs plant transgenic operation, and obtains transgenic plants. Detection shows that the waxiness of the transgenic plant is significantly reduced, which indicates that after the implementation of the present invention, it will enhance the absorption efficiency of plant leaf surface micro-fertilizer, enhance light energy absorption and plant photosynthetic efficiency, increase yield, improve the taste of vegetables in the eating process, and increase ornamental The emerald green color of the plant and the improvement of perception will bring beneficial effects, which are of great significance for the development and production of new plant varieties.

Description

technical field [0001] The invention relates to the application of a glycosyltransferase gene, in particular to the application of an Arabidopsis thaliana glycosyltransferase gene UGT76D1 in reducing plant surface wax, and belongs to the field of genetic engineering. Background technique [0002] Wax is a hydrophobic protective layer on the surface of plants, often covering the leaves and stems of plants. Plant surface waxes are composed of cuticle exogenous waxes and cuticle inner waxes located in the plant epidermis. The composition of wax is different in different plants, mainly composed of extra-long chain fatty acids and their derivatives including alkanes, primary and secondary alcohols, aldehydes, ketones, terpenoids and trace secondary metabolites such as flavonoids and other compounds (Pollard et al. al., 2008; Samuels et al., 2008; Li-Beisson et al., 2013). [0003] Wax acts as a hydrophobic barrier on the surface of plants. Although it can reduce water loss, res...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N15/54C12N15/84A01H5/00
Inventor 侯丙凯黄戌戌李燕洁
Owner SHANDONG UNIV
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