Determination Dihydroflavonol-4-Reductase’Activity,Cloing And Bioinformatics Analysis Of Black-Grained Wheat

The black–grained wheat came from hybridization between Bread Wheat and weedAgropyron glaucum.It has a stable genetic traits. Black-grained wheat keeps ahigh-profile because it has much more nutritional value and oxidation resistancefunctions than common wheat. The synthesis and metabolic pathways of anthocyaninshave been widely studied in many plants, but have not been reported in the coloredwheat. In order to explore the anthocyanins synthesis and the mechanism of metabolicpathways of black-grained wheat, our experiments chose the ‘Hashi3-37’black-grainedwheat as the material, analyzing DFR and compared DFR activity and physical andchemical properties, cloning DFR gene, and predicted the genetic structure. The mainidea was to explore the anthocyanins synthesis of the black-grained wheat synthesis andmetabolic mechanism of anthocyanins more clearly.The results show that DFR plays a key role in the formation of grain anthocyaninsand the color is affected by the synthesis and accumulation of anthocyanins inblack-grained wheat. We designed primers and successfully cloned DFR gene fromblack-grained wheat refer to the DFR gene sequences from NCBI. DFR gene contains alength of1065bp ORF, encoding345amino acids, which gDNA sequence containsthree introns, following the rule of GT–AG（NCBI. KF780538.1）. Bioinformaticsanalysis showed that DFR gene was conservative in the process of genetic evolution ofblack-grained wheat. The gene contained a conservative structure combined withNADPH and has a ‘Asn type’ specific binding site.The protein positioning incytoplasmic is acidic,stable, hydrophilic and unsecretory which does not have signalpeptide and transmembrane domain structure.The forecast of the DFR showsthree-dimensional structure in the black-grained wheat and the surface is coveredwith alpha-helix and surrounded by beta-fold zone. The alpha-helixs of the gene areconnected to the beta folding by H key constitute, the structure of the protein is stableand has a great deal of similarity measure in the grapes which have been confirmed inDFR crystal structure.