감마선 처리에 의한 스프레이형 국화 화색변이체로부터 Flavonoid 3’-Hydroxylase(F3’H) 유전자의 분리 및 특성 구명

다국어 초록 (Multilingual Abstract)

The objectives of this study were to isolate and the sequence of novel F3’H 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’H 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’H 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’H 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’H gene for the color variation in chrysanthemum.

참고문헌 (Reference)

1 박인숙, "화훼류의 방사선 돌연변이 육종 현황" 한국화훼학회 13 (13): 184-200, 2005

2 박인숙, "감마선 조사와 조직배양에 의한 스프레이 국화 'Argus'의 돌연변이 육종" 한국화훼학회 15 (15): 52-57, 2007

3 Nielson, K.M, "cDNA cloning and endogenous expression of a flavonoid 3’5’-hydroxylase from petals of lisianthus (Eustoma grandiflorum)" 129 : 167-174, 1997

4 이근주, "Variation in the phenotypic features and transcripts of color mutants of chrysanthemum ( Dendranthema grandiflorum ) derived from gamma ray mutagenesis" 한국식물학회 51 (51): 418-423, 2008

5 Nakatsuka, T, "Two different mutations are involved in the formation of white-flowered gentian plants" 169 : 949-958, 2005

6 Saitou, N, "The neighbor-joining method: A new method for reconstructing phylogenetic trees" 4 : 406-425, 1987

7 Zufall, R.A, "The genetic basis of a flower color polymorphism in the common morning glory (Ipomoea purpurea)" 94 : 442-448, 2003

8 Forkmann, G, "Metabolic engineering and applications of flavonoids" 12 : 155-160, 2001

9 Tamura, K, "MEGA4: molecular evolutionary genetics analysis (MEGA) software version 4.0" 24 : 1596-1599, 2007

10 정성진, "Isolation of a Leucoanthocyanidin Dioxygenase (LDOX) Gene from a Spray-type Chrysanthemum (Dendranthema × grandiflorum) and Its Colored Mutants" 한국원예학회 28 (28): 818-827, 2010

11 Brugliera, F, "Isolation and characterization of a flavonoid 3’-hydroxylase cDNA clone corresponding to the Ht1 locus of Petunia hybrida" 19 : 441-451, 1999

12 김주현, "Inheritance and Characteristics of a New Dwarf Mutant in Lettuce" 한국원예학회 45 (45): 277-280, 2004

13 Schum, A, "Induced mutations in ornamental plants, In Somaclonal variation and induced mutations in crops improvement" Kluwer Academic Publ 363-366, 1998

14 Schoenbohm, C, "Identification of the Arabidopsis thaliana flavonoid 3’-hydroxylase gene and functional expression of the encoded P450 enzyme" 381 : 749-753, 2000

15 Holton, T.A, "Genetics and biochemistry of anthocyanin biosynthesis" 7 : 1071-1083, 1995

16 Hoshino, A, "Gene duplication and mobile genetic elements in the morning glories" 265 : 1-10, 2001

17 Dixon, R.A, "Flavonoids and isoflavonoids - A gold mine for metabolic engineering" 4 : 394-400, 1999

18 Gerats, A.G.M, "Flavonoid synthesis in Petunia hybrida: Genetics and molecular biology of flower colour, In Phenolic metabolism in plants" Plenum Press 165-199, 1992

19 Winkel-Shirley, B, "Flavonoid biosynthesis: A colorful model for genetics, biochemistry, cell biology, and biotechnology" 126 : 485-493, 2001

20 Jeong, S.T, "Expression of the flavonoid 3’-hydroxylase and flavonoid 3’,5’-hydrxylase genes and flavonoid composition in grape (Vitis vinifera)" 170 : 61-69, 2006

21 Boddu, J, "Expression of a putative flavonoid 3’-hydroxylase in sorghum mesocotyls synthesizing 3-deoxyanthocyanidin phytoalexins" 65 : 101-113, 2004

22 Seitz, C, "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" 61 : 365-381, 2006

23 Zabala, G, "Cloning of the pleiotropic T locus in soybean and two recessive alleles that differentially affect structure and expression of the encoded flavonoid 3’ hydroxylase" 163 : 295-309, 2003

24 Xu, B.B, "Cloning and molecular characterization of a functional flavonoid 3’-hydroxylase gene from Brassica napus" 164 : 350-363, 2007

25 Schlangen, K, "Chalcone 3-hydroxylation is not a general property of flavonoid 3’-hydroxylase" 177 : 97-102, 2009

26 Boss, P.K, "Analysis of the expression of anthocyanin pathway genes in developing Vitis vinifera L. cv Shiraz grape berries and the implications for pathway regulation" 111 : 1059-1066, 1996

27 Harborne, J.B, "Advances in flavonoid research since 1992" 55 : 481-504, 1992

28 Toda, K, "A single-base deletion in soybean flavonoid 3’-hydroxylase gene is associated with gray pubescence color" 50 : 187-196, 2002