Molecular cloning and characterisation of the flavanone-3-hydroxylase gene from Korean black raspberry
2008
Baek, M.-H. | Chung, B. Y. | Kim, J. H. | Wi, S. G. | An, B. C. | Kim, J. S. | Lee, S. S. | Lee, I.-J.
Anthocyanins are the major pigments contributing to the colouration of raspberry fruit.We isolated the gene encoding flavanone-3-hydroxylase (F3H), an enzyme which catalyses a key step in anthocyanin biosynthesis from Korean black raspberry (Rubus sp.). Quantitative real-time PCR analysis showed that expression of the Rubus F3H gene (RsF3H) occurred during all stages of fruit development. Expression was first detected in green fruit, declined in yellow fruit, then increased sharply again in red fruit at the start of anthocyanin accumulation. Expression of RsF3H was highest in black-coloured fruit. Immunoblot analysis showed a pattern of F3H protein expression similar to the pattern of RsF3H gene transcription observed by quantitative real-time PCR. Immunogold-labelling revealed that the distribution of F3H was tissue-specific. The distribution of F3H in the endocarp region was greater than in the epicarp and mesocarp regions at all stages of fruit development. Also, the anthocyanin content of black fruit increased over 300-fold when compared with green fruit. These results suggest that the RsF3H gene is specifically up-regulated when anthocyanin accumulation is most pronounced.
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