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Report No.

Comparative analysis of flavonoid biosynthesis genes between ${it Cyclamen graecum}$ and its white-flowered mutant

Akita, Yusuke; Ishizaka, Hiroshi*; Nakayama, Masayoshi*; Shimada, Akihiko; Kitamura, Satoshi; Hase, Yoshihiro; Tanaka, Atsushi; Narumi, Issei

To reveal the relationship between floral pigmentation and flavonoid biosynthesis genes in cyclamen (${it Cyclamen}$ species), we analyzed flower pigments from wild-type cyclamen (${it C. graecum}$) and its white-flowered mutant. Analysis with high performance liquid chromatography (HPLC) showed that major anthocyanins of the wild-type petal were malvidin 3,5-diglucosides. The white-flowered mutant possessed smaller amount of anthocyanins but higher amount of flavonols than the wild-type, suggesting the change of metabolic flow by disruption of anthocyanin biosynthesis. By degenerate-PCR with total RNAs from wild-type petals, we isolated some flavonoid biosynthesis gene. RT-PCR using a specific primer set for each gene demonstrated that the expression of ${it CgraDFR2}$ decreased in white-flowered mutant compared with wild-type, whereas the expressions of other genes did not appear to differ greatly. The genomic construction of ${it CgraDFR2}$ was not different between wild-type and white-flowered mutant, inferring that reduced expression of ${it CgraDFR2}$ correlates with white-flowered mutation. These results suggest that ${it CgraDFR2}$ is involved in anthocyanin biosynthesis pathway in ${it C. graecum}$, and the white-flowered mutation could be caused by transcriptional regulation of ${it CgraDFR2}$ gene.



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