Cloning and expression analysis of eight WRKY genes in Camellia sinensis / 中草药

ABSTRACT

Objective: To clone eight members of WRKY of transcription factor family in Camellia sinensis, and analyze their bioinformatics and expression under abiotic stress. Methods: Eight WRKY transcription factor genes were cloned from Tieguanyin cultivar by RT-PCR, and the physicochemical properties of the eight WRKY protein were analyzed by bioinformatics Methods:. At the same time, the establishment of phylogenetic tree, comparison of multiple sequences, and analysis of conserved motifs were carried out by comparing WRKY of C. sinensis with homologous genes of Arabidopsis thaliana. Quantitative real-time PCR (qRT-PCR) was used to detect the expression of eight WRKY genes under low temperature, drought, and ABA stress treatment. Results: The ORF lengths of eight WRKY genes were 1 407, 2 208, 1 302, 849, 978, 879, 1 443, and 810 bp, encoding 468, 735, 433, 282, 325, 292, 480, and 269 amino acids, respectively. GenBank accession numbers were MG298951, MG298952, MG298955, MG298956, MG298957, MG298959, MG298960, and MG298963, respectively. Phylogenetic tree and sequence alignment analysis showed that eight CsWRKYs could be divided into two groups and contained WRKYGQK conserved domain and zinc finger structures, except that CsWRKY39 lacked zinc finger structure. The expression pattern of CsWRKYs was induced under the condition of low temperature, drought, and ABA stress. The expression of CsWRKY2, CsWRKY21, CsWRKY23, CsWRKY44 and CsWRKY65 increased to more than 2 after low temperature treatment with significant response to low temperature stress. The expression of CsWRKY21, CsWRKY23, CsWRKY3,9 and CsWRKY65 was up-regulated under 12 h of drought stress and 6 h of ABA treatment. This result indicated that CsWRKYs might be closely related to stress response in C. sinensis. Conclusion: Eight CsWRKY genes from different groups were cloned, and this result indicated that CsWRKYs might be closely related to stress response in C. sinensis.