Studies On The Genetic Relationship Of Nai (Prunus) By RAPD Analysis & Cloning F3h Gene And Dfr Gene Related To Anthocyanin Biosynthesis From Riped Fruit Of "Red Bartlett"

The method for isolating genomic DNA from nai (Primus salicina Lindl. Var.cordata J.Y.Zhang et al.) was studied in this paper. Based on the genomic DNA , we compared some essential factors that might affect the results of RAPD and established the optimal RAPD reaction system for nai. In a volume of 20 ul, amplification reactions system contained 2.5mM MgC^, 200 uMdNTP, 0.2 uM random primer, 50ng genomic DNA, 1-2U Taq DNA polymerase . Some cultivars of nai, plum and peach were examined by means of RAPDs. Cluster analysis based on similarity coefficients showed that nai was one of species in Prunus instead of the hybrid between peach and plum. We propose RAPDs as a valuable tool for the identification of specific genotypes and analysis of genetic relationships.The total RNA was extracted from riped "Red Bartllet" fruit by modified CTAB method. Two fragments of f3h gene which related to anthocyanin biosynthesis were cloned by RACE method. A 1095bp cDNA sequence was obtained by splicing the two fragments and the sequence showed that it was 97% homologous identity to that of f3h cDNA from "Fuji" apple. But dfr gene wasn't obtained from the peel of pear by RACE method.