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

Wang, Jingwen; Ming, Feng; Han, Yingying; Shen, Daleng

doi:10.1007/s10529-005-5718-6

Cited by 14 publications

(12 citation statements)

References 20 publications

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“…The unknown-function CYP76C1 gene was highly expressed in the flowers of A. thaliana, and at lower level in the leaves (Mizutani et al 1998). The expression pattern of CYP76AB1 we describe here in the petals of V. coerulea is similar to the expression patterns of F3'5'H in the petals of Eustoma grandiflorum (Nielsen and Podivinsky 1997), Nierembergia (Yukiko et al 2006) and Phalaenopsis (Wang et al 2006). In Vinca major, although the transcripts of the F3'5'H gene were detected in all five developmental stages of the flower, strong signals were only detected in the petals of flowers in stages 2 -4 ( Mori et al 2004).…”

supporting

confidence: 78%

Expression of a CYP76AB1 correlates with the sequential white-blue-white colour transition of Vanda coerulea petals

Ratanasut¹,

Wongkhamprai²,

Maknoi³

2011

Biol. plant.

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The blue colour of Vanda coerulea petals is slowly produced during flower development but quickly disappears when the flowers are pollinated. To investigate the molecular basis of the phenomenon, we isolated a novel cytochrome P450 gene, CYP76AB1, from this plant by polymerase chain reaction (PCR) based on the conserved regions of flavonoid 3´,5´-hydroxylase amino acid sequences. CYP76AB1 transcripts were detectable at low level in the late phase of flower development when the petals were light blue, but became abundant in the subsequent opening flower stage, as the petals turned blue. When the flowers were pollinated they turned white, and the CYP76AB1 transcripts returned to low level. This indicates that the expression of CYP76AB1 may regulate petal colour of V. coerulea, and be a target for developing permanently blue flowers.

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supporting

confidence: 78%

Expression of a CYP76AB1 correlates with the sequential white-blue-white colour transition of Vanda coerulea petals

Ratanasut¹,

Wongkhamprai²,

Maknoi³

2011

Biol. plant.

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“…The Lasergene program MegAlign (DNASTAR Inc., Madison, USA) was used to compare the CpF3'5'H deduced amino acid sequence with ten F3'5'H sequences (the CYP75A group), ten F3'H sequences (CYP75B) and two 'outlier' cytochrome P450 sequences (data not shown). Amino acid identity of CpF3'5'H to other F3'5'H sequences was in the range from 75-82%, except for the Campanula medium F3'5'H sequence (BAA03440), 68% identity, which is suggested to have a distinct F3'5'H structure [16] and the monocot Phalaenopsis hybrida F3'5'H sequence (AAZ79451, 50% identity) [17]. A phylogenetic tree was formed using the CLUSTAL W algorithm http://www-bimas.cit.nih.gov/clustalw/clustalw.html with the MegAlign data (Figure.…”

Section: Resultsmentioning

confidence: 99%

Isolation and antisense suppression of flavonoid 3', 5'-hydroxylase modifies flower pigments and colour in cyclamen

et al. 2010

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BackgroundCyclamen is a popular and economically significant pot plant crop in several countries. Molecular breeding technologies provide opportunities to metabolically engineer the well-characterized flavonoid biosynthetic pathway for altered anthocyanin profile and hence the colour of the flower. Previously we reported on a genetic transformation system for cyclamen. Our aim in this study was to change pigment profiles and flower colours in cyclamen through the suppression of flavonoid 3', 5'-hydroxylase, an enzyme in the flavonoid pathway that plays a determining role in the colour of anthocyanin pigments.ResultsA full-length cDNA putatively identified as a F3'5'H (CpF3'5'H) was isolated from cyclamen flower tissue. Amino acid and phylogeny analyses indicated the CpF3'5'H encodes a F3'5'H enzyme. Two cultivars of minicyclamen were transformed via Agrobacterium tumefaciens with an antisense CpF3'5'H construct. Flowers of the transgenic lines showed modified colour and this correlated positively with the loss of endogenous F3'5'H transcript. Changes in observed colour were confirmed by colorimeter measurements, with an overall loss in intensity of colour (C) in the transgenic lines and a shift in hue from purple to red/pink in one cultivar. HPLC analysis showed that delphinidin-derived pigment levels were reduced in transgenic lines relative to control lines while the percentage of cyanidin-derived pigments increased. Total anthocyanin concentration was reduced up to 80% in some transgenic lines and a smaller increase in flavonol concentration was recorded. Differences were also seen in the ratio of flavonol types that accumulated.ConclusionTo our knowledge this is the first report of genetic modification of the anthocyanin pathway in the commercially important species cyclamen. The effects of suppressing a key enzyme, F3'5'H, were wide ranging, extending from anthocyanins to other branches of the flavonoid pathway. The results illustrate the complexity involved in modifying a biosynthetic pathway with multiple branch points to different end products and provides important information for future flower colour modification experiments in cyclamen.

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“…The expression level of the F3 ′ 5 ′ H homolog in Torenia hybrida was higher during the early period than the late period (Ueyama et al ); however, the opposite expression pattern exists in Gentiana triflora (Nakatsuka et al ). Although F3 ′ 5 ′ H was highly expressed during the late period, its expression was not detected during the early period in Eustoma grandiflorum , Phalaenopsis and V. vinifera (Noda et al , Bogs et al , Wang et al ). Thus, the expression pattern of cineraria F3 ′ 5 ′ H during floral development is specific.…”

Section: Discussionmentioning

confidence: 99%

“…Only ligulate florets of purple and blue cineraria contain delphinidin derivatives (Sun et al ); consequently, the difference in PCFH expression during the primary period is a likely reason for the production of diverse anthocyanidins to form various colors. Except in the berry skin of V. vinifera (Bogs et al ), the transcripts of F3 ′ 5 ′ H were detected only in the floral organs of T. hybrida , E. grandiflorum , G. triflora , Phalaenopsis and Solanum lycopersicum (Ueyama et al , Noda et al , Nakatsuka et al , Wang et al , Olsen et al ). This study found for the first time that F3 ′ 5 ′ H is expressed in cineraria leaves.…”

Section: Discussionmentioning

confidence: 99%

“…Therefore, the F3 ′ 5 ′ H gene is called the blue gene (Holton and Tanaka ). Full‐length F3 ′ 5 ′ H cDNA sequences have been isolated from many plants containing delphinidin derivatives (Holton et al , Tanaka et al , Nielsen and Podivinsky , Kaltenbach et al , Suzuki et al , Okinaka et al , Mori et al , Seitz et al , Togami et al , Wang et al , Olsen et al ). According to the analysis of the amino acid sequences and biochemical characters, F3′5′Hs belong to CYP75A subfamily of the cytochrome P450‐dependent monooxygenase superfamily (P450s), a vast class of heme‐containing mixed‐function oxidases catalyzing NADPH‐ or NADH‐dependent oxygenation reactions (Tanaka et al ), while flavonoid 3′‐hydroxylases (F3′Hs), which catalyze hydroxylation of the flavonoid B‐ring at the 3′‐position, belong to the CYP75B subfamily (Brugliera et al ).…”

Section: Introductionmentioning

confidence: 99%

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Isolation and functional analysis of a homolog of flavonoid 3′,5′‐hydroxylase gene from Pericallis × hybrida

Sun

et al. 2013

Physiologia Plantarum

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As the key enzyme in the biosynthesis of blue flower color pigments, flavonoid 3',5'-hydroxylase (F3'5'H) can catalyze the conversion of its major substrates, 2-S naringenin and dihydrokaempferol, into 3',4',5'-hydroxylated pentahydroxyflavanone and dihydromyricetin, respectively. Unlike other F3'5'Hs belonging to the CYP75A subfamily, Asteraceae-specific F3'5'Hs belong to the CYP75B subfamily. Furthermore, cineraria F3'5'H expressed in yeast exhibited not only F3'H (flavonoid 3'-hydroxylase) activity but also F3'5'H activity in vitro. In this study, Southern blotting showed that there was only one copy of a homolog of the F3'5'H gene PCFH in the Pericallis × hybrida genome. This gene could be detected by Northern blot in the primary developmental stages of ligulate florets of the purple- and blue-flowered cultivars, and its transcripts also accumulated in the leaves. Heterologous expression of PCFH could produce new delphinidin derivatives in the corollas of transgenic tobacco plants, increased the content of cyanidin derivatives and lead to the blue- and red-shifting of flower color in T₀ generation plants. These results indicate that cineraria F3'5'H exhibited both F3'5'H- and F3'H-activity in vivo. The types and contents of anthocyanins and flower color phenotypes of the T₁ generation were similar to those of T₀ generation plants. PCFH exhibited stable inheritance and normal functions between generations. This study supplies new evidence to understand Asteraceae-specific F3'5'Hs and provides important references for the further study of molecular breeding of blue-flowered chrysanthemums using the PCFH gene.

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

Cited by 14 publications

References 20 publications

Expression of a CYP76AB1 correlates with the sequential white-blue-white colour transition of Vanda coerulea petals

Expression of a CYP76AB1 correlates with the sequential white-blue-white colour transition of Vanda coerulea petals

Isolation and antisense suppression of flavonoid 3', 5'-hydroxylase modifies flower pigments and colour in cyclamen

Isolation and functional analysis of a homolog of flavonoid 3′,5′‐hydroxylase gene from Pericallis × hybrida

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