Cloning and expression analysis of 1-hydroxy-2-methyl-2-(E)-butenyl-4-diph- osphatereductase gene in Dendrobium officinale / 中草药

ABSTRACT

Objective: To clone the full-length cDNA encoding 1-hydroxy-2-methyl-2-(E)-butenyl-4-diphosphatereductase (HDR) gene from Dendrobium officinale (DoHDR), then to analyze the expression difference in different tissues and expression patterns of DoHDR induced by signal molecule. Methods: RT-PCR and RACE technologies were used to clone the full length cDNA of DoHDR. The analyses of homologous comparison and phylogenetic tree were performed using DNAMAN and MEGA6.0 softwares, then the expression patterns of DoHDR were studied by real-time PCR. Results: The DoHDR gene was successfully obtained (GenBank accession number KC344827), and the full-length cDNA was 1 658 bp, coding the protein containing 460 amino acids. DoHDR had high homology (≥ 80%) with HDR proteins from other plants. Tissue expression analysis showed that DoHDR had the highest expression in the leaves, followed by roots, stems, and protocorm. Quantitative PCR results showed that DoHDR could be induced by signal molecule such as abscisic acid (ABA) and salicylic acid (SA). Conclusion: The cDNA encoding DoHDR is cloned. It is helpful for the future research on the mechanism of terpenoid biosynthesis in D. officinale.