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Cloning and Expression Analysis of Dihydroflavonol-4-Redutase Gene DtpsDFR in Doritaenopsis Hybrid
Zhong Huaiqin1, 2, 3, Huang Minling1, 2, 3*, Wu Jianshe1, 2, 3, Fan Ronghui1, 2, 3, Lin Bing1, 2, 3
1Institute of Crop Sciences, Fujian Academy of Agricultural Sciences, Fuzhou 350013, China; 2Flowers Research Center, Fujian Academy of Agricultural Sciences, Fuzhou 350013, China; 3Fujian Engineering Research Center of Characteristic Floriculture, Fuzhou 350013, China
Abstract: Dihydroflavonol 4-reductase (DFR) is a key enzyme in anthocyanin biosynthesis. In this study， the full-length cDNA sequence of DFR gene was obtained from bright red petals of Doritaenopsis hybrid Queen beer ‘Red Sky’ using RT-PCR and RACE techniques and designated as DtpsDFR. The cDNA sequence was 1 286 bp， encoding a deduced polypeptide of 378 amino acids. Amino acid sequence analysis indicated that DtpsDFR protein shared more than 76% homology with DFR protein from Bromheadia finlaysoniana， Oncidium， Cymbidium and Dendrobium. DtpsDFR protein contains a FR_SDR_e domain， a NADPH binding motif and a substrate specificity binding site， belonging to the NADB_Rossmann superfamily. Phylogenetic analysis clearly showed that the DtpsDFR was more related to DFR proteins in Bromheadia finlaysoniana. The result of quantitative RT-PCR analysis indicated that expression levels of DtpsDFR gene gradually reduced with the development of flower， micro-expression in faded period. The transcript level was higher in petals and sepals than in lips， micro-expression in leaves and roots.
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