Articles
Sequence comparison with the upstream region of F3’5’H gene related to pigment synthesis in wild evergreen azaleas
Article number
1434_18
Pages
129 – 134
Language
English
Abstract
Evergreen azaleas, for which many cultivars have been improved since the Edo period (1603-1867), have a variety of flower colors, and these are colored by flavonoids as the main pigments.
Our previous study has reported that the red purple petals of R. kiusianum contained both cyanidin and delphinidin series pigments, while the red petals of R. kaempferi contained only cyanidin series pigments, and the flavonoid 3’,5’-hydroxylase (F3’5′H) gene was always expressed in samples containing delphinidin series pigments such as R. kiusianum. In this study, we investigated the sequence of the upstream region of the F3’5’H gene in order to elucidate the factors responsible for the presence or absence of F3’5’H gene expression.
In the analysis of the upstream area of F3’5’H genes, a total of approximately 1.4 kb was obtained for R. kiusianum K-1180-10 and approximately 2.4 kb for R. kaempferi Ki-440-3 using the Genome Walking method.
Comparison of both samples was revealed that the MYB cis element, TATA box, and transcription start site were preserved although there were multiple base sequence substitutions, insertions or deletions in the approximately 760 bp upstream from the transcription start site.
In contrast, further upstream sequences of the 760 bp were completely different between R. kiusianum K-1180-10 and R. kaempferi Ki-440-3, suggesting that the presence or absence of F3’5’H gene expression was due to differences in the sequence of the upstream region.
Multiplex PCR amplification using specific primers to detect sequence differences in the F3’5’H genes of two wild species allowed to be roughly distinguished R. kiusianum and R. kaempferi and could be used as a DNA marker for flower color identification.
Our previous study has reported that the red purple petals of R. kiusianum contained both cyanidin and delphinidin series pigments, while the red petals of R. kaempferi contained only cyanidin series pigments, and the flavonoid 3’,5’-hydroxylase (F3’5′H) gene was always expressed in samples containing delphinidin series pigments such as R. kiusianum. In this study, we investigated the sequence of the upstream region of the F3’5’H gene in order to elucidate the factors responsible for the presence or absence of F3’5’H gene expression.
In the analysis of the upstream area of F3’5’H genes, a total of approximately 1.4 kb was obtained for R. kiusianum K-1180-10 and approximately 2.4 kb for R. kaempferi Ki-440-3 using the Genome Walking method.
Comparison of both samples was revealed that the MYB cis element, TATA box, and transcription start site were preserved although there were multiple base sequence substitutions, insertions or deletions in the approximately 760 bp upstream from the transcription start site.
In contrast, further upstream sequences of the 760 bp were completely different between R. kiusianum K-1180-10 and R. kaempferi Ki-440-3, suggesting that the presence or absence of F3’5’H gene expression was due to differences in the sequence of the upstream region.
Multiplex PCR amplification using specific primers to detect sequence differences in the F3’5’H genes of two wild species allowed to be roughly distinguished R. kiusianum and R. kaempferi and could be used as a DNA marker for flower color identification.
Authors
D. Mizuta, M. Ito, A. Nakatsuka, N. Kobayashi, S. Kubota
Keywords
anthocyanin, flavonoid 3’, 5’-hydroxylase, Rhododendron kiusianum, R. kaempferi, upstream sequence
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