Cloning and Expression Analysis of the Promoter of Glycosyltransferase Gene in Eucommia ulmoides
WEN Yong1*, LI Biao1*, ZHAO De-Gang1,2, ZHAO Yi-Chen1*
1 College of Life Sciences/Tea College/ Key Laboratory of Mountain Plant Resources Conservation and Germplasm Innovation, Ministry of Education, Guizhou University, Guiyang 550025, China; 2 Guizhou Academy of Agricultural Sciences, Guiyang 550025, China
Abstract：Plant glycosyltransferases (GTs) are a large class of enzymes that modify the secondary metabolites for glycosylation. GTs can convert photosynthesis products into disaccharides, oligosaccharides and polysaccharides, and can also catalyze some important products including cell wall polysaccharides, glycoproteins, and many different types of small molecular species. GTs can promote the glycosylation of certain hormones and signaling molecules, and thus participate in signal regulation.In order to study the regulation pattern of the Eucommia ulmoides glycosyltransferase gene (EuCGT1), this study obtained a length of 1 671 bp upstream of the 5' end of EuCGT1 gene of E. ulmoides. The Plant CARE (http://bioinformatics.psb.ugent.be/webtools/plantcare/html/) predictive analysis revealed that the fragment contained the eukaryotic promoter core elements TATA-box and CAAT-box. At the same time, the fragment also contained a plurality of induction-related cis-acting elements such as a methyl jasmonate response element TGACG-motif, a drought-related MBS element, a gibberellin response element GARE-motif, and an auxin response element TGA-element. The promoter fragment of different lengths was used to regulate the expression of GUS gene to construct plant expression vector, and the activity and function of EuCGT1 promoter in transgenic tobacco (Nicotiana tabacum 'Xanthi') were explored. It was found that the core region of the EuCGT1 promoter was mainly located in the -268~-1 bp upstream of the gene. The GUS histochemical staining of roots, stems and leaves of transgenic tobacco and the expression analysis of EuCGT1 gene in E. ulmoides showed that the promoter of this gene was more likely to be expressed in leaves. After hormone induction and environmental stress in E. ulmoides, the expression of EuCGT1 was regulated by methyl jasmonate, auxin, gibberellin and high salt. The results of this study may provide a reference for further revealing the function of the EuCGT1 promoter of E. ulmoides and the promoter-mediated regulation of gene expression.
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