Isolation and characterization of dihydroflavonol 4-reductase gene from Dendrobium Sonia 'earsakul'
2011
Pitakdantham, W. | Sutabutra, T., Kasetsart Univ.,Bangkok 10900 (Thailand) | Pitaksutheepong, C., National Center for Genetic Engineering and Biotechnology (BIOTEC), 113 Phaholyothin Road, Klong 1, Klong Luang, Pathumthani 12120 (Thailand) | Chiemsombat, P., Kasetsart Univ., Kamphaeng Saen Campus, Nakhom Pathom 73140 (Thailand)
This paper meant to isolate and characterize dihydroflavonol 4-reductase gene in anthocyanin biosynthetic pathway during flower development of Dendrobium Sonia 'Earsakul'. The gene was isolated from floral tissues of the orchid by reverse transcriptase polymerase chain reaction procedure. Characterization of the gene is considered to its relevancy to dihydroflavonol 4-reductase gene in other plants elucidated by construction of a neighbor-joining phylogenetic tree. Gene expression pattern related to flower development and pigmentation was investigated by relative quantification using real time polymerase chain reaction procedure. A complete coding sequence was obtained and sequence analysis revealed that the gene of Dendrobrium Sonia 'Earsakul' consisted of 1,059 base pairs. Blast analysis and multiple alignments showed that hydroflavonol 4-reductase gene of Dendrobrium Sonia 'Earsakul' shares high homology to dihydroflavonol 4-reductase gene of Dendrobium genes particularly Dendrobium hybrid genes particularly Dendrobrium hybrid 'Uniwai prince' and Dendrobium hybrid 'Geeting Fragrance'. Phylogenetic tree revealed for the first time that dihydroflavonol 4-reductase gene of Dendrobium genus are highly conserved. The dihydroflavonol 4-reductase gene of Dendrobium Sonia 'Earsakul' was highly expressed in young flower bud with no pigmentation and the expression was abundant when young flower bud started accumulation of pigments. The expression was decreased in half open flowers but slightly increased with fully opened flowers.
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