Browse > Article
http://dx.doi.org/10.7235/hort.2012.11059

Isolation and Characterization of a Novel Flavonoid 3'-Hydroxylase (F3'H) Gene from a Chrysanthemum (Dendranthema grandiflorum) and Its Gamma-ray Irradiated Mutants  

Chung, Sung-Jin (Radiation Breeding and Research Team, Advanced Radiation Technology Institute, Korea Atomic Energy Research Institute)
Lee, Geung-Joo (Department of Horticulture, Chungnam National University)
Kim, Jin-Baek (Radiation Breeding and Research Team, Advanced Radiation Technology Institute, Korea Atomic Energy Research Institute)
Kim, Dong-Sub (Radiation Breeding and Research Team, Advanced Radiation Technology Institute, Korea Atomic Energy Research Institute)
Kim, Sang-Hoon (Radiation Breeding and Research Team, Advanced Radiation Technology Institute, Korea Atomic Energy Research Institute)
Kang, Si-Yong (Radiation Breeding and Research Team, Advanced Radiation Technology Institute, Korea Atomic Energy Research Institute)
Publication Information
Horticultural Science & Technology / v.30, no.2, 2012 , pp. 162-170 More about this Journal
Abstract
The objectives of this study were to isolate and the sequence of novel $F3 gene related to an anthocyanin pathway, and to confirm the expression patterns of the gene involved in the flower color variations of chrysanthemum mutants. In this study, we isolated the full-length cDNAs and the genomic DNAs of an $F3 gene from a wild type (WT) chrysanthemum (cv. Argus) and its three color mutants. The sequence analysis revealed a putative open reading frame of 1,527 bp that encodes a polypeptide of 509 amino acids. Sequence homology ranged from 97% to 99% between 'Argus' and its three color mutants. The sequence analysis from the genomic DNA revealed that the chrysanthemum $DgF3 gene consisted of three exons and two introns spanning a 3,830 bp length. The sizes of the gene for three mutants ranged from a shorter size of 3,828 bp to a longer size of 3,838 bp when compared to the size of WT. The total size of the two introns was 2,157 bp for WT, but those for three color mutants ranged from 2,154 bp to 2,159 bp. A result of an RT-PCR analysis indicated that the color variations of the mutants AM1 and AM2 can be partly explained by the structural modification derived from the sequencial changes in the gene caused by gamma ray. A Southern blot analysis revealed that the $DgF3 gene existing as multiple copies in the chrysanthemum genome. A systemic study will be further needed to provide a genetic mechanism responsible for the color mutation and to uncover any involvement of genetic elements for the expression of the $DgF3 gene for the color variation in chrysanthemum.
Keywords
color mutant; $DgF3; gene copy number; radiation mutation; RT-PCR;
Citations & Related Records
연도 인용수 순위
1 Seitz, C., C. Eder, B. Deiml, S. Kellner, S. Martens, and G. Forkmann. 2006. Cloning, functional identification and sequence analysis of flavonoid 3'-hydroxylase and flavonoid 3',5'-hydroxylase cDNAs reveals independent evolution of flavonoid 3',5'-hydroxylase in the Asteraceae family. Plant Mol. Biol. 61:365-381.   DOI
2 Tamura, K., J. Dudley, M. Nei, and S. Kumar. 2007. MEGA4: molecular evolutionary genetics analysis (MEGA) software version 4.0. Mol. Biol. Evol. 24:1596-1599.   DOI
3 Toda, K., D. Yang, N. Yamanaka, S. Watanabe, K. Harada, R. Takahashi. 2002. A single-base deletion in soybean flavonoid 3'-hydroxylase gene is associated with gray pubescence color. Plant Mol. Biol. 50:187-196.   DOI
4 Winkel-Shirley, B. 2001. Flavonoid biosynthesis: A colorful model for genetics, biochemistry, cell biology, and biotechnology. Plant Physiol. 126:485-493.   DOI
5 Xu, B.B., J.N. Li, X.K. Zhang, R. Wang, L.L. Xie, and Y.R. Chai. 2007. Cloning and molecular characterization of a functional flavonoid 3'-hydroxylase gene from Brassica napus. J. Plant Physiol. 164:350-363.   DOI
6 Zabala, G. and L. Vodkin. 2003. Cloning of the pleiotropic T locus in soybean and two recessive alleles that differentially affect structure and expression of the encoded flavonoid 3' hydroxylase. Genetics 163:295-309.
7 Zufall, R.A. and M.D. Rausher. 2003. The genetic basis of a flower color polymorphism in the common morning glory (Ipomoea purpurea). J. Hered. 94:442-448.   DOI
8 Lee, G.J., S.J. Chung, I.S. Park, J.S. Lee, J.B. Kim, D.S. Kim, and S.Y. Kang. 2008. Variation in the phenotypic features and transcripts of color mutants of chrysanthemum (Dendranthema grandiflorum) derived from gamma ray mutagenesis. J. Plant Biol. 51:418-423.   DOI
9 Nakatsuka, T., M. Nishihara, K. Mishiba, and S. Yamamura. 2005. Two different mutations are involved in the formation of white-flowered gentian plants. Plant Sci. 169:949-958.   DOI
10 Nielson, K.M. and E. Podivinsky. 1997. cDNA cloning and endogenous expression of a flavonoid 3'5'-hydroxylase from petals of lisianthus (Eustoma grandiflorum). Plant Sci. 129:167-174.   DOI
11 Park, I.S. and H.S. Song. 2005. Current trends of mutation breeding by radiation technology at domestic and foreign ornamentals. J. Kor. Flower Res. Soc. 13:184-200.
12 Park, I.S., G.J. Lee, D.S. Kim, S.J. Chung, J.B. Kim, H.S. Song, D.H. Goo, and S.Y. Kang. 2007. Mutation breeding of a spray chrysanthemum 'Argus' by gamma-ray irradiation and tissue culture. Flower Res. J. 15:52-57.
13 Saitou, N. and M. Nei. 1987. The neighbor-joining method: A new method for reconstructing phylogenetic trees. Mol. Biol. Evol. 4:406-425.
14 Boddu, J., C. Svabek, R.S. Sekhon, A. Gevens, R.L. Nicholson, A.D. Jones, J.F. Pedersen, D.L. Gustine, and S. Chopra. 2004. Expression of a putative flavonoid 3'-hydroxylase in sorghum mesocotyls synthesizing 3-deoxyanthocyanidin phytoalexins. Physiol. Mol. Plant Pathol. 65:101-113.   DOI
15 Schlangen, K., S. Miosic, F. Topuz, G. Muster, T. Marosits, C. Seitz, and H. Halbwirth. 2009. Chalcone 3-hydroxylation is not a general property of flavonoid 3'-hydroxylase. Plant Sci. 177:97-102.   DOI
16 Schoenbohm, C., S. Martens, C. Eder, G. Forkmann, and B. Weisshaar. 2000. Identification of the Arabidopsis thaliana flavonoid 3'-hydroxylase gene and functional expression of the encoded P450 enzyme. Biol. Chem. 381:749-753.   DOI
17 Schum, A. and W. Preil. 1998. Induced mutations in ornamental plants, p. 363-366. In: S.M. Jain, D.S. Brair, and B.S. Ahloowalia (eds.). Somaclonal variation and induced mutations in crops improvement. Kluwer Academic Publ., Dordrecht.
18 Chung, S.J., G.J. Lee, H.J. Lee, J.B. Kim, D.S. Kim, and S.Y. Kang. 2010. Isolation of a leucoanthocyanidin dioxygenase (LDOX) gene from a spray-type chrysanthemum (Dendranthema ${\times}$ Grandiflorum) and its colored mutants. Kor. J. Hort. Sci. Technol. 28:818-827.
19 Dixon, R.A. and C.L. Steele. 1999. Flavonoids and isoflavonoids - A gold mine for metabolic engineering. Trends Plant Sci. 4:394-400.   DOI
20 Forkmann, G. and S. Martens. 2001. Metabolic engineering and applications of flavonoids. Curr. Opin. Biotechnol. 12:155-160.   DOI
21 Gerats, A.G.M. and C. Martin. 1992. Flavonoid synthesis in Petunia hybrida: Genetics and molecular biology of flower colour, p. 165-199. In: H.A. Stafford and R.K. Ibrahim (eds.). Phenolic metabolism in plants. Plenum Press, New York.
22 Harborne, J.B. and C.A. Williams. 2000. Advances in flavonoid research since 1992. Phytochemistry 55:481-504.   DOI
23 Jung, Y.-H., K.H. Kim, M. Kim, S.J. Chun, and S.-C. Kim. 2004. Comparative analysis of single-and double flowered Camella japonica based on internal transcribed spacer sequences of nuclear ribosomal DNA. Hort. Environ. Biotechnol. 45:277-383.
24 Holton, T.A. and E.C. Cornish. 1995. Genetics and biochemistry of anthocyanin biosynthesis. Plant Cell 7:1071-1083.   DOI
25 Hoshino, A., Y. Johzuka-Hisatomi, and S. Iida. 2001. Gene duplication and mobile genetic elements in the morning glories. Gene 265:1-10.   DOI
26 Jeong, S.T., N. Goot-Yamamoto, K. Hashizume, and M. Esaka. 2006. Expression of the flavonoid 3'-hydroxylase and flavonoid 3',5'-hydrxylase genes and flavonoid composition in grape (Vitis vinifera). Plant Sci. 170:61-69.   DOI
27 Brugliera, F., G. Barri-Rewell, T.A. Holton, and J.G. Mason. 1999. Isolation and characterization of a flavonoid 3'-hydroxylase cDNA clone corresponding to the Ht1 locus of Petunia hybrida. Plant J. 19:441-451.   DOI
28 Boss, P.K., C. Davies, and S.P. Robinson. 1996. Analysis of the expression of anthocyanin pathway genes in developing Vitis vinifera L. cv Shiraz grape berries and the implications for pathway regulation. Plant Physiol. 111:1059-1066.   DOI