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http://dx.doi.org/10.5010/JPB.2005.32.3.167

Molecular Characterization of Cinnamate 4-Hydroxylase gene in Red Hot Pepper (Capsicum annuum L.)  

Kim Kye-Won (National Institute of Agricultural Biotechnology, Institute of Biological Chemistry, Washington State University)
Ha Sun-Hwa (National Institute of Agricultural Biotechnology)
Cho Kang-Jin (National Institute of Agricultural Biotechnology)
Kim Eun-Ju (Department of Crop Science & Biotechnology, Jinju National University)
Lee Min-Kyung (Department of Crop Science & Biotechnology, Jinju National University)
Yu Jae-Ju (Department of Crop Science & Biotechnology, Jinju National University)
Kim Jong-Guk (Department of Microbiology, Kyungpook National University)
Lee Shin-Woo (National Institute of Agricultural Biotechnology, Department of Crop Science & Biotechnology, Jinju National University)
Publication Information
Journal of Plant Biotechnology / v.32, no.3, 2005 , pp. 167-173 More about this Journal
Abstract
Three different cDNAS for cinnamate 4-hydroxylase (C4H) which are involved in the second step of the general phenylpropanoid pathway were isolated and designated as pc4h1 (1,755 bp), pc4h2 (1,655 bp), and pc4h3 (1,316 bp), respectively. The nucleotide sequence analysis revealed that both pc4h1 and pc4h2 clones encode polypeptides of 505 amino acids frame but pc4h3 clone was truncated at the 5'-end of coding region. The alignment of the deduced amino acid sequences showed that PC4H1 and PC4H2 are highly homologous (95.8% identical) with each other and contain three conserved domains which are typical in cytochrome P450 monooxygenase: proline-rich region, threonine-containing binding pocket for the oxygen molecule, and heme binding region. In addition, result of the phylogenic tree analysis revealed that both pepper C4Hs belong to Class 1. pc4h2 transcription was strongly induced in wounded fruit (400%) and root (200%) relative to its very low basal level but not in leaf or stem tissue. In case of pc4h1, the basal level of transcription was higher than pc4h2 but induction by wounding was lower in fruit and root while leaf and stem tissues did not respond to wounding. The basal level of pc4h3 transcripts was not, if any, detectable and response to wounding was not observed.
Keywords
Cinnamate 4-hydroxylase; cytochrome P450 monooxygenase; phenylpropanoid pathway;
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1 Bennett DJ, Kirby GW (1968) Constitution and biosynthesis of capsaicin. J Chem Soc C:442-446
2 Betz C, McCollum TG, Mayer RT (2001) Differential expression of two cinnamate 4-hydroxylase genes in Valencia orange (Citrus sinensis Osbeck). Plant Mol Biol 46:741-748   DOI   ScienceOn
3 Blount JW, Korth KL, Masoud SA, Rasmussen S, Lamb C, Dixon R (2000) Altering expression of cinnamic acid 4-hydroxylase in transgenic plants provides evidence for a feedback loop at the entry point into the phenylpropanoid pathway. Plant Physiol 122: 107-116   DOI
4 Blount JW, Masoud SA, Sumner LW, Huhman D, Dixon R (2002) Over-expression of cinnamate 4-hydroxylase leads to increased accumulation of acetosyringone in elicited tobacco cell-suspension cultures. Planta 214: 902-910   DOI
5 Chapple C (1998) Molecular-genetic analysis of plant cytochrome P450-dependent monooxygenases. Annu Rev Plant Physiol Plant Mol Biol 49: 311-343   DOI
6 Hahlbrock K, Scheel D (1989) Physiology and molecular biology of phenylpropanoid metabolism. Annu Rev Plant Physiol Plant Mol Biol 40: 347-369   DOI
7 Tijet N, Helvig C, Feyereisen R (2001) The cytochrome P450 genes superfamily in Drosophila melanogaster. annotation, intron-exon organization and phylogeny. Gene 262: 189-198   DOI   ScienceOn
8 Werck-Reichhart D, Bak S, Paquette S (2002) Cytochrome P450. In the Arabidopsis Book, eds. CR Somerville, EM meyerwitz, American Society of Plant Biologists. Rockville, MD, doi/10.1199/tab.0028, http://aspb.org/ publications/arabidopsis.   DOI
9 Von Wachenfeldt C, Johnson EF (1995) Structures of eukaryotic cytochrome P450 enzymes. In : Ortiz de Montellano PR (eds), Cytochrome P450: Structure, mechanism, and Biochemistry, Vol. 2. pp 183-223
10 Werck-Reichhart D, Feyereisen R (2000) Cytochromes P450: a success story. Genome Biol 1: 3003-3009
11 Yamamura Y, Ogihara Y, Mizukami H (2001) Cinnamic acid 4-hydroxylase from Lithospermum erythrohizon: cDNA cloning and gene expression. Plant Cell Rep 20: 655-662   DOI
12 Kim KW, Varindra R, Cho KJ, Kim JG, Lee SW (2000) Comparison of accumulation of capsaicinoid contents with capsaicinoid synthetase activity at different developmental stages of Capsicum annuum L. Agric Chem Biotechnol 43: 152-155
13 Potter S, Moreland DE, Kreuz K, Ward E (1995) Induction of cytochrome P450 genes by ethanol in maize. Drug Metabol Drug Interact 12: 317-327
14 Nakamura M, Satoh T, Tanaka SI, Mochizuki N, Yokota T, Nagatani A (2005) Activation of the cytochrome P450 gene, CYP72C1, reduces the levels of active brassinostreoids in vivo. J Exp Bot 56:833-840   DOI   ScienceOn
15 Nedelkina S, Jupe SC, Blee KA, Schalk M, Werck-Reichhart D, Bolwell GP (1999) Novel characteristics and regulation of a divergent cinnamate 4-hydroxylase (CYP73A15) from french bean: engineering expression in yeast. Plant Mol Biol 39: 1079-1090   DOI   ScienceOn
16 Paquette SM, Bak S, Feyereisen R, (2000) Intron-exon organization and phylogeny in a large superfamily, the paralogous cytochrome P450 genes of Arabidopsis thaliana. DNA Cell Biol 19: 307-317   DOI   ScienceOn
17 Schuler MA (1996) Plant cytochrome P450 monooxygenases. Crit Rev Plant Sci 15: 235-284   DOI
18 Schuler MA, Werck-Reichhart D (2003) Functional genomics of P450s. Annu Rev Plant Biol 54: 629-667   DOI
19 Suzuki T, Kawada T, Iwai K (1981) Biosynthesis of acyl moieties of capsaicin and its analogues from valine and leucine in Capsicum fruits. Plant Cell Physiol 22: 23-32
20 Teutsch RG, Hasenfratz MP, Lesot A, Stoltz C, Garnier JM, Jeltsch JM, Durst F, Werck-Reichhart D (1993) Isolation and sequence of a cDNA encoding the Jerusalem artichoke cinnamate 4-hydroxylase, a major plant cytochrome P450 involved in the general phenylpropanoid pathway. Proc Natl Acad Sci USA 90: 4102-4107   DOI