Heterologous expression of the flavonoid 3?,5?-hydroxylase gene of Vinca major alters flower color in transgenic Petunia hybrida

Mori, Satoshi; Kobayashi, Hitoshi; Hoshi, Y.; Kondo, Motoi; Nakano, Masataka

doi:10.1007/s00299-003-0709-3

Cited by 41 publications

(24 citation statements)

References 26 publications

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“…These data are consistent with the anthocyanins accumulation pattern. In Vinca major anthocyanins were only detected in petals of stage 2-5 flowers with those of stage 4 flowers accumulating the largest amount of delphinidin (Mori et al 2004). These results suggested that Phf35h expression in the petals of Phalaenopsis is developmentally regulated and that anthocyanins accumulation may be under transcriptional control.…”

Section: The Expression Profiles Of Phalaenopsis F35h Genesmentioning

confidence: 96%

Flavonoid-3′,5′-hydroxylase from Phalaenopsis: A Novel Member of Cytochrome P450s, its cDNA Cloning, Endogenous Expression and Molecular Modeling

et al. 2006

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A novel cDNA for the flavonoid-3',5'-hydroxylase (F35H) gene was cloned from petals of Phalaenopsis, and designated as Phf35h (accession number DQ148458 in GenBank/EMBL/DDBJ). The genomic clone of Phf35h was isolated by a PCR-based strategy. Nucleotide sequence analysis revealed that its genomic clone contains one intron and an open reading frame encoding a polypeptide of 507 amino acid residues. Southern hybridization analysis indicated the presence of a single gene coding for Phf35h. RT-PCR analysis showed that the Phf35h mRNA is transcribed in late phase of petal development, which is concomitant with the appearance of anthocyanins in petal tissue. The transcript is abundant in the purple petals but not in leaves or roots. The three-dimensional model of PhF3'5'H protein is classified into an alpha-domain which contains most of the alpha-helices with three small beta-sheets, a beta-domain that contains the larger beta-sheets with three small alpha-helices by homology modeling. The substrate-binding site for dihydrokaempferol on PhF3'5'H protein was determined by molecular docking algorithm. A highly conserved HPPTPLSLPH sequence was predicted to contact the aromatic ring of dihydrokaempferol.

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Section: The Expression Profiles Of Phalaenopsis F35h Genesmentioning

confidence: 96%

Flavonoid-3′,5′-hydroxylase from Phalaenopsis: A Novel Member of Cytochrome P450s, its cDNA Cloning, Endogenous Expression and Molecular Modeling

et al. 2006

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“…After the first isolation and functional characterization of the genes encoding F3#H and F3#5#H in petunia (Petunia hybrida; Holton et al, 1993;Brugliera et al, 1999), the genes were subsequently isolated from various plant species. F3#H was characterized in Arabidopsis (Arabidopsis thaliana; Schoenbohm et al, 2000), and F3#5#H cDNAs were isolated from a number of plants, including Solanum melongena (Toguri et al, 1993), Gentiana triflora (Tanaka et al, 1996), Eustoma grandiflorum (Nielsen and Podivinsky, 1997), Eustoma rusellianum (Shimada et al, 1999), Catharanthus roseus (Kaltenbach et al, 1999), Campanula medium (Okinaka et al, 2003), and Vinca major (Mori et al, 2004). Some plants, including Arabidopsis and important members of the Rosaceae such as apples (Malus spp.)…”

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confidence: 99%

Identification of the Flavonoid Hydroxylases from Grapevine and Their Regulation during Fruit Development

et al. 2005

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(J.B., D.M., S.P.R.)Flavonoids are important secondary metabolites in many fruits, and their hydroxylation pattern determines their color, stability, and antioxidant capacity. Hydroxylation of the B-ring of flavonoids is catalyzed by flavonoid 3#-hydroxylase (F3#H) and flavonoid 3#,5#-hydroxylase (F3#5#H), and may also require cytochrome b 5 . We report the identification of genes encoding F3#H, F3#5#H, and a putative cytochrome b 5 from grapevine (Vitis vinifera L. cv Shiraz) and their transcriptional regulation in fruit. Functionality of the genes VvF3#H and VvF3#5#H1 was demonstrated by ectopic expression in petunia (Petunia hybrida), which altered flower color and flavonoid composition as expected. VvF3#H was expressed in grapes before flowering, when 3#-hydroxylated flavonols are made, and all three genes were expressed after flowering, when proanthocyanidins (PAs) are synthesized. In berry skin, expression of all three genes was low at the onset of ripening (véraison) but increased after véraison concomitant with the accumulation of 3#-and 3#,5#-hydroxylated anthocyanins. VvF3#H and VvCytoB5 were expressed in seeds but not VvF3#5#H1, consistent with the accumulation of 3#-hydroxylated PAs in this tissue. VvCytoB5 expression was correlated with expression of both VvF3#H and VvF3#5#H1 in the different grape tissues. In contrast to red grapes, where VvF3#H, VvF3#5#H1, and VvCytoB5 were highly expressed during ripening, the expression of VvF3#5#H1 and VvCytoB5 in white grapes during ripening was extremely low, suggesting a difference in transcriptional regulation. Our results show that temporal and tissue-specific expression of VvF3#H, VvF3#5#H1, and VvCytoB5 in grapes is coordinated with the accumulation of the respective hydroxylated flavonols and PAs, as well as anthocyanins. Understanding the regulation of flavonoid hydroxylases could be used to modify flavonoid composition of fruits.

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“…Genes encoding F3#H and F3#5#H have been isolated in myriad plant species, including petunia (Holton et al, 1993;Menting et al, 1994;Brugliera et al, 1999), Arabidopsis (Schoenbohm et al, 2000), lisianthus (Eustoma grandiflorum; Noda et al, 2004), and grape (Vitis vinifera; Bogs et al, 2006), among others. Manipulation of F3#H and F3#5#H genes has been effective in genetic engineering of floral crops to develop new genotypes with novel flower colors for ornamental purposes (Shimada et al, 1999;Mori et al, 2004;Nakatsuka et al, 2007).…”

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confidence: 99%

Ectopic Expression of Apple F3′H Genes Contributes to Anthocyanin Accumulation in the Arabidopsis tt7 Mutant Grown Under Nitrogen Stress

et al. 2010

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Three genes encoding flavonoid 3'-hydroxylase (F3'H) in apple (Malus x domestica), designated MdF3'HI, MdF3'HIIa, and MdF3'HIIb, have been identified. MdF3'HIIa and MdF3'HIIb are almost identical in amino acid sequences, and they are allelic, whereas MdF3'HI has 91% nucleotide sequence identity in the coding region to both MdF3'HIIa and MdF3'HIIb. MdF3'HI and MdF3'HII genes are mapped onto linkage groups 14 and 6, respectively, of the apple genome. Throughout the development of apple fruit, transcriptional levels of MdF3'H genes along with other anthocyanin biosynthesis genes are higher in the red-skinned cv Red Delicious than that in the yellow-skinned cv Golden Delicious. Moreover, patterns of MdF3'H gene expression correspond to accumulation patterns of flavonoids in apple fruit. These findings suggest that MdF3'H genes are coordinately expressed with other genes in the anthocyanin biosynthetic pathway in apple. The functionality of these apple F3'H genes has been demonstrated via their ectopic expression in both the Arabidopsis (Arabidopsis thaliana) transparent testa7-1 (tt7) mutant and tobacco (Nicotiana tabacum). When grown under nitrogen-deficient conditions, transgenic Arabidopsis tt7 seedlings expressing apple F3'H regained red color pigmentation and significantly accumulated both 4'-hydrylated pelargonidin and 3',4'-hydrylated cyanidin. When compared with wild-type plants, flowers of transgenic tobacco lines overexpressing apple F3'H genes exhibited enhanced red color pigmentation. This suggests that the F3'H enzyme may coordinately interact with other flavonoid enzymes in the anthocyanin biosynthesis pathway.

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Heterologous expression of the flavonoid 3?,5?-hydroxylase gene of Vinca major alters flower color in transgenic Petunia hybrida

Cited by 41 publications

References 26 publications

Flavonoid-3′,5′-hydroxylase from Phalaenopsis: A Novel Member of Cytochrome P450s, its cDNA Cloning, Endogenous Expression and Molecular Modeling

Flavonoid-3′,5′-hydroxylase from Phalaenopsis: A Novel Member of Cytochrome P450s, its cDNA Cloning, Endogenous Expression and Molecular Modeling

Identification of the Flavonoid Hydroxylases from Grapevine and Their Regulation during Fruit Development

Ectopic Expression of Apple F3′H Genes Contributes to Anthocyanin Accumulation in the Arabidopsis tt7 Mutant Grown Under Nitrogen Stress

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