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KMID : 0903620040450010049
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Journal of the Korean Society for Horticultural Science
2004 Volume.45 No. 1 p.49 ~ p.54
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Cloning of cDNA Coding for Dihydroflavonol-4-reductase (DFR) and Characterization of dfr Expression in Carnation (Dianthus caryophyllus) Petals
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Lee, Hee Jung
Nahm. Seok Hyeon/Ahn. Byung Joon/Joung, Hyang Young/Min, Byung Whan
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Abstract
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A heterologous cDNA probe from Petunia hybrida was used to isolate dihydroflavonol-4-reductase (DFR)-encoding cDNA clone from Dianthus caryophyllus. Comparison of the coding region of this DFR cDNA sequence including the sequences of Zea mays, Anthirrinum majus. Petunia hybrida. Matthiola incana, Callistephus chinensis, and Rosa hybrida reveals a similarity higher than 75% at the amono acid level. The DFR transcript is G/C rich in monocotyledonous plants show a strong codon bias preferring codons with a G or C in the third position. The function of this nucleotide sequences were verified by comparison with amino acid sequences of the amino-terminus and tryptic peptides from purfied plant enzyme, by Northern blotting with mRNA from wild type and mutant plants and by in vitro expression yealding an enzymatically active reductase. Northern blot analysis of the DFR wild-type and mutant lines showed that the lack of DFR activity in the stable acyanic mutants in clearly caused by a transcriptional block of the DFR gene.
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