Cloning and Expression Analysis of Cinnamyl Alcohol Dehydrogenase (SmCAD) Gene in Salvia miltiorrhiza Bunge
GE Qian, KUANG Jing, WU Yu-Cui, ZHANG Yuan, XIAO Ya-Ping, WANG Zhe-Zhi
Key Laboratory of Ministry of Education for Medicinal Resources and Natural Pharmaceutical Chemistry, National Engineering Laboratory for Resource Developing of Endangered Chinese Crude Drugs in Northwest of China, College of Life Sciences, Shaanxi Normal University, Xi'an 710062, China
Abstract:Cinnamyl alcohol dehydrogenase (CAD) catalyzes the reduction reaction of cinnamic aldehyde and its derivatives depending on the NADPH.It is the key enzyme in the last step of lignin biosynthesis. A novel CAD gene designated as SmCAD (GenBank accession No.: HQ162287) was cloned from Salvia miltiorrhiza Bunge according to the transcription databases. It contained a 1083 bp open reading frame, including 3 introns and 4 exons, and encoded a protein of 360 amino acids. The predicted SmCAD protein had Zn1 and Zn2 binding sites and NADP(H) binding domain. A total of 1202 bp promoter sequences were obtained using BD walking method, and sequence analysis revealed that there were MeJA, ABA, GA3 response elements and MYB binding sites. Real-time quantitative PCR showed that SmCAD was expressed in different organs, and its expression was induced by MeJA and inhibited by GA3. These results suggested that this gene may be involved in the response of exogenous signals. This study contributes to the theoretical basis of the specific function of SmCAD.
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