Genetic engineering of the anthocyanin biosynthetic pathway with flavonoid-3′,5′-hydroxylase: specific switching of the pathway in petunia

Shimada, Yukihisa; Ohbayashi, Masaya; Nakano-Shimada, Ritsuko; Okinaka, Yasushi; Kiyokawa, Shigeto; Kikuchi, Yasuhiro

doi:10.1007/s002990100319

Cited by 31 publications

(19 citation statements)

References 13 publications

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“…When the F3¢5¢H genes in a red-purple petunia that mainly accumulated delphinidin-based anthocyanins were down-regulated, pink flowers accumulating cyanidin-based anthocyanins and flavonols were obtained (Tsuda et al 2004). Flower colour changes from deep blue to pale blue/pink were also reported (Shimada et al 2001). However, the down-regulation of the F3¢5¢H gene in Nierembergia, a close relative of petunia, only resulted in a white flower containing increased flavonols, but no cyanidin, due to its low F3¢H and high FLS expression .…”

Section: Solanaceae Species-petunia Tobacco and Nierembergiamentioning

confidence: 88%

Flower colour and cytochromes P450

2006

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Flavonoids are major constituents of flower colour. Plants accumulate specific flavonoids and thus every species often exhibits a limited flower colour range. Three cytochromes P450 play critical roles in the flavonoid biosynthetic pathway. Flavonoid 3¢-hydroxylase (F3¢H, CYP75B) and flavonoid 3¢,5¢-hydroxylase (F3¢5¢H, CYP75A) catalyze the hydroxylation of the B-ring of flavonoids and are necessary to biosynthesize cyanidin-(red to magenta) and delphinidin-(violet to blue) based anthocyanins, respectively. Pelargonidin-based anthocyanins (orange to red) are synthesized in their absence. Some species such as roses, carnations and chrysanthemums do not have violet/blue flower colour due to deficiency of F3¢5¢H. Successful expression of heterologous F3¢5¢H genes in roses and carnations results in delphinidin production, causing a novel blue/violet flower colour. Down-regulation of F3¢H and F3¢5¢H genes has yielded orange petunia and pink torenia colour that accumulate pelargonidin-based anthocyanins. Flavone synthase II (CYP93B) catalyzes the synthesis of flavones that contribute to the bluing of flower colour, and modulation of FNSII gene expression in petunia and tobacco changes their flower colour. Extensive engineering of the anthocyanin pathway is therefore now possible, and can be expected to enhance the range of flower colours. Keywords Anthocyanin AE Cytochrome P450 AE Flavonoid AE Flavone AE Flower colour AE Cyanidin AE Delphinidin AE Dihydroflavonol 4-reductase AE Flavonoid 3¢-hydroxylase AE Flavonoid 3¢,5¢-hydroxylase AE Flavone synthase II AE Flavonol hydroxylase AE Monooxygenase AE Pelargonidin Abbreviations DFR dihydroflavonol 4-reductase DHK dihydrokaempferol DHQ dihydroquercetin DHM dihydromyricetin F3H flavanone 3-hydroxylase F3¢H flavonoid 3¢-hydroxylase F3¢5¢H flavonoid 3¢,5¢-hydroxylase FLS flavonol synthase FNS flavone synthase Flavonoids and anthocyaninsFlower colour is due to flavonoids, carotenoids, betalains and some other pigments (

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Section: Solanaceae Species-petunia Tobacco and Nierembergiamentioning

confidence: 88%

Flower colour and cytochromes P450

2006

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“…F3 0 5 0 H cDNAs of for example Eustoma russellianum (Shimada et al, 1999), Petunia hybrida (Shimada et al, 2001) or Campanula medium (Okinaka et al, 2003) have been extensively used for transgenic over-expression. Many efforts have been undertaken in expressing F3 0 5 0 H genes in plant species such as rose, carnation or chrysanthemum which naturally lack F3 0 5 0 H activity and therefore do not accumulate Dp-derivatives.…”

Section: Introductionmentioning

confidence: 99%

Redirection of anthocyanin synthesis in Osteospermum hybrida by a two-enzyme manipulation strategy

et al. 2007

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“…However, some flower colors are still lacking for many flower crops, since the gene pool is limited for any single species. For example, Rosa spp., Dianthus caryophyllus and Lilium spp., all of which are commercially very important flower crops, do not have blue-flowering varieties due to the lack of flavonoid 3 0 ,5 0 -hydroxylase (F3 0 5 0 H) activity in the flower petal (Shimada et al 2001). F3 0 5 0 H, a member of the cytochrome P450 family, is a key enzyme in the synthesis of the blue flower-specific anthocyanidins, 3 0 ,5 0 -hydroxylated anthocyanidins (e.g., delphinidin) (Fig.…”

Section: Introductionmentioning

confidence: 99%

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

et al. 2004

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Flavonoid 3',5'-hydroxylase (F3'5'H) is the key enzyme for the expression of blue or purple flower color. A full-length cDNA for the F3'5'H gene was cloned from petals of Vinca major, and its genomic clone, designated VmFH1 (accession number AB078781 in the GenBank/EMBL/DDBJ databases), was isolated from leaves by a PCR-based strategy. Nucleotide sequence analysis revealed that VmFH1 contains one intron and an open reading frame encoding a polypeptide of 506 amino acid residues. The deduced amino acid sequence shows between 51% and 83% identity with those of previously reported F3'5'H genes. Southern blot analysis showed that there are 3-4 copies of the F3'5'H gene in the genome of V. major. Transcripts of the F3'5'H gene were detected in young flower petals but not in leaves as revealed by RT-PCR analysis. When VmFH1 was expressed in transgenic Petunia hybrida under the control of the cauliflower mosaic virus 35S promoter, some transgenic plants showed drastic flower color alteration from red to deep red with deep purple sectors. These transgenic plants accumulated 3',5'-hydroxylated anthocyanins in their petals, which were never detected in non-transgenic plants by high-performance liquid chromatography analysis. These results indicate that VmFH1 isolated from V. major encodes F3'5'H and is active in a heterologous plant species.

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Genetic engineering of the anthocyanin biosynthetic pathway with flavonoid-3′,5′-hydroxylase: specific switching of the pathway in petunia

Cited by 31 publications

References 13 publications

Flower colour and cytochromes P450

Flower colour and cytochromes P450

Redirection of anthocyanin synthesis in Osteospermum hybrida by a two-enzyme manipulation strategy

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

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