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Molecules 2018, 23(4), 861;

Molecular Cloning and Functional Characterization of a Dihydroflavonol 4-Reductase from Vitis bellula

1,* , 1
Hunan Provincial Key Laboratory of Plant Resources Conservation and Utilization, College of Biology and Environmental Sciences, Jishou University, No. 120 Ren Min Nan Lu, Jishou City 416000, China
Department of Plant Biology, North Carolina State University, 100 Derieux Place, Raleigh, NC 27695, USA
Authors to whom correspondence should be addressed.
Received: 14 March 2018 / Revised: 30 March 2018 / Accepted: 2 April 2018 / Published: 10 April 2018
(This article belongs to the Section Metabolites)
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Vitis bellula is a new grape crop in southern China. Berries of this species are rich in antioxidative anthocyanins and proanthocyanidins. This study reports cloning and functional characterization of a cDNA encoding a V. bellula dihydroflavonol reductase (VbDFR) involved in the biosynthesis of anthocyanins and proanthocyanidins. A cDNA including 1014 bp was cloned from young leaves and its open reading frame (ORF) was deduced encoding 337 amino acids, highly similar to V. vinifera DFR (VvDFR). Green florescence protein fusion and confocal microscopy analysis determined the cytosolic localization of VbDFR in plant cells. A soluble recombinant VbDFR was induced and purified from E. coli for enzyme assay. In the presence of NADPH, the recombinant enzyme catalyzed dihydrokaempferol (DHK) and dihydroquercetin (DHQ) to their corresponding leucoanthocyanidins. The VbDFR cDNA was introduced into tobacco plants via Agrobacterium-mediated transformation. The overexpression of VbDFR increased anthocyanin production in flowers. Anthocyanin hydrolysis and chromatographic analysis revealed that transgenic flowers produced pelargonidin and delphinidin, which were not detected in control flowers. These data demonstrated that the overexpression of VbDFR produced new tobacco anthocyanidins. In summary, all data demonstrate that VbDFR is a useful gene to provide three types of substrates for metabolic engineering of anthocyanins and proanthocyanidins in grape crops and other crops. View Full-Text
Keywords: Vitis bellula; dihydroflavonol reductase; dihydroflavonol; leucoanthocyanidin; anthocyanin; proanthocyanidin Vitis bellula; dihydroflavonol reductase; dihydroflavonol; leucoanthocyanidin; anthocyanin; proanthocyanidin

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This is an open access article distributed under the Creative Commons Attribution License which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited (CC BY 4.0).

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Zhu, Y.; Peng, Q.; Li, K.; Xie, D.-Y. Molecular Cloning and Functional Characterization of a Dihydroflavonol 4-Reductase from Vitis bellula. Molecules 2018, 23, 861.

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