Cloning and Expression of BsDFR in Bougainvillea spectabilis
[Objective]The dihydroflavonol-4-reductase(DFR)gene in bracts of Bougainvillea spectabilis was cloned and characterized to study the role it plays in color formation.[Method]BsDFR was cloned based on the transcriptome data on the ornamental plant to study the related bioinformatics.Molecular docking technology was employed to predict the substrate specificity,and qRT-PCR applied to examine the relative transcription levels of the genes in B.spectabilis of different colors.[Result]The full-length coding sequence of BsDFR(GenBank ID:ON417750)was 987 bp encoding 328 amino acids.The protein had a calculated molecular weight of 36.49 kDa and an isoelectric point of 6.33.It had the NADPH and substrate specific binding sites unique to DFR of Asn type without a transmembrane structure or signal peptide.The subcellular localization of the protein indicated it to be cytoplasmic.Alpha helices were the most abundant secondary structure of the protein,while the tertiary structure was a dimer.A substrate docking simulation,consistent with the structural analysis,predicted BsDFR to possess a catalytic activity on dihydrokaempferol,dihydroquercetin,and dihydromyricetin.The phylogenetic tree analysis grouped it along with caryophyllales plants.High expression of the gene was found in the orange B.spectabilis by qRT-PCR.It was speculated that the main substrate to be DHK,which was catalyzed by BsDFR into leucopelargonidin,a precursor of orange-colored anthocyanidin——pelargonidin.[Conclusion]BsDFR in B.spectabilis had typical molecular characteristics of the plant dihydroflavonol-4-reductase,which is associated with the pigment synthesis in the bracts of orange B.spectabilis.
万方数据
维普
