Home > Browse Issues > Vol.39 No.9
Cloning and Expression Analysis of Tropinone Reductase Gene from Dendrobium huoshanense
Lin Rongyan1,2,3, Ye Xiuxian1,2,3*, Zhong Huaiqin1,2,3, Huang Minling1,2,3*
1Crop Research Institute, Fujian Academy of Agricultural Sciences, Fuzhou 350013, China; 2Flower Research Center, Fujian Academy of Agricultural Sciences, Fuzhou 350013, China; 3Fujian Engineering Research Center for Characteristic Floriculture, Fuzhou 350013, China
Abstract: In this study， the cDNA of tropinone reductase (TR) gene was successfully cloned from the protocorm of Dendrobium huoshanense by RT-PCR and RACE to explore the function of tropinone reductase， its accession number was KY912085 in GenBank. The full-length of TR-I gene was 1 043 bp， containing a 807 bp open reading frame (ORF) encoding 268 amino acids. Bioinformatics analysis showed that the protein might be a stable hydrophobic protein， with 14 functional sites and 47 potential phosphorylation sites and without signal peptide and coil helix. The secondary structure was composed of helixs， sheets and coils. The phylogenetic tree analysis showed that the protein had closest relationship with TR-I of Dendrobium catenatum and Dendrobium nobile， at the same branch. qPCR results indicated that the relative expression of TR-I gene was stems>leaves>roots in all stages， the expression levels of TR-I in stems and leaves appeared the tendency of low-high-low with the prolong of growth. Among the different varieties， the expression level of Dendrobium nobile was the highest and Dendrobium catenatum the lowest. Under the conditions in different concentrations of methyl jasmonate (MeJA)， the highest expression level appeared when the adding concentration was at 100 μmol/L. The results suggested that the gene might be involved in the synthesis of alkaloids in Dendrobium huoshanense.
CN
EN