Cloning and Mutant Construction of Cold-upregulated Glycosyltransferase-like Gene (OsCUGT1) from Guizhou Landrace Rice 'Pingtang Heinuo' (Oryza sativa ssp. japonica)
CAI Ming-Liang1, CHEN Rong1, HUANG Xiao-Zhen1,2,*, ZHAO De-Gang1,3,*
1 The Key Laboratory of Plant Resources Conservation and Germplasm Innovationin Mountainous Region (Ministry of Education), College of Life Sciences, Guizhou University, Guiyang 550025, China; 2 College of Tea Sciences, Guizhou University, Guiyang 550025, China; 3 Guizhou Academy of Agricultural Sciences, Guiyang Branch of DUS Center, Ministry of Agriculture and Rural Sciences, Guiyang 550006, China;
Abstract：Plant glycosyltransferases (GTs) play an important role in regulating growth and stress adaptation. To further explore and study the biological functions of new glycosyltransferases in rice (Oryza sativa), in this study, a cold-upregulated glycosyltransferase-like gene 1 (OsCUGT1) was isolated and cloned, which was based on the cold treatment transcriptome library of Guizhou landrace rice 'Pingtang Heinuo' (O. sativa ssp. japonica). The results of qRT-PCR showed that the expression of OsCUGT1 gene was significantly induced by cold treatment. The subcellular localization results showed that it was localized in the chloroplasts. Phylogenetic analysis showed that OsCUGT1 was conserved in monocot plant species. Furthermore, sequence alignment revealed that there were several SNP sites between 'Pingtang Heinuo' and O. sativa ssp. Nipponbare in the OsCUGT1 gene. Among them, the site of 511 bp was changed from C to A, which consequently caused the 170 th amino acid to change from lysine (Q) to glutamine (K). Diversity analysis of the OsCUGT1 gene sequence revealed that there were 12 SNP sites of them were found in the exon region. To further explore the biological function of OsCUGT1, the OsCUGT1 gene was edited by CRISPR/Cas9 technique. The targeted editing mutants of oscugt1 were successfully obtained by optimizing Agrobacterium-mediated transformation method, screening of hygromycin selection and sequencing analysis. The results of this study can provide research materials and theoretical basis for further clarifying the role of OsCUGT1 in rice growth development and stress resistance．
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