Flower color modification of gentian plants by RNAi-mediated gene silencing

Nakatsuka, Takashi; Mishiba, Kei‐ichiro; Abe, Yoshiko; Kubota, Akiko; Kakizaki, Yuko; Yamamura, Saburo; Nishihara, Masahiro

doi:10.5511/plantbiotechnology.25.61

Cited by 56 publications

(41 citation statements)

References 28 publications

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“…Blue gentian flowers accumulate gentiodelphin, a stable blue delphinidin-based anthocyanin modified with two aromatic acyl groups. Suppression of the endogenous F3 0 5 0 H gene resulted in a decrease in delphinidin and an increase of cyanidin levels and with a colour change to magenta [79]. Additional suppression of the aromatic acyltransferase that is involved in biosynthesis of gentiodelphin resulted in lilac or pale-blue flower colour [80].…”

Section: Efforts To Redmentioning

confidence: 99%

Flower colour and cytochromes P450

Tanaka

Brugliera²

2013

Phil. Trans. R. Soc. B

268

146

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Cytochromes P450 play important roles in biosynthesis of flavonoids and their coloured class of compounds, anthocyanins, both of which are major floral pigments. The number of hydroxyl groups on the B-ring of anthocyanidins (the chromophores and precursors of anthocyanins) impact the anthocyanin colour, the more the bluer. The hydroxylation pattern is determined by two cytochromes P450, flavonoid 3′-hydroxylase (F3′H) and flavonoid 3′,5′-hydroxylase (F3′5′H) and thus they play a crucial role in the determination of flower colour. F3′H and F3′5′H mostly belong to CYP75B and CYP75A, respectively, except for the F3′5′Hs in Compositae that were derived from gene duplication of CYP75B and neofunctionalization. Roses and carnations lack blue/violet flower colours owing to the deficiency of F3′5′H and therefore lack the B-ring-trihydroxylated anthocyanins based upon delphinidin. Successful redirection of the anthocyanin biosynthesis pathway to delphinidin was achieved by expressing F3′5′H coding regions resulting in carnations and roses with novel blue hues that have been commercialized. Suppression of F3′5′H and F3′H in delphinidin-producing plants reduced the number of hydroxyl groups on the anthocyanidin B-ring resulting in the production of monohydroxylated anthocyanins based on pelargonidin with a shift in flower colour to orange/red. Pelargonidin biosynthesis is enhanced by additional expression of a dihydroflavonol 4-reductase that can use the monohydroxylated dihydrokaempferol (the pelargonidin precursor). Flavone synthase II (FNSII)-catalysing flavone biosynthesis from flavanones is also a P450 (CYP93B) and contributes to flower colour, because flavones act as co-pigments to anthocyanins and can cause blueing and darkening of colour. However, transgenic plants expression of a FNSII gene yielded paler flowers owing to a reduction of anthocyanins because flavanones are precursors of anthocyanins and flavones.

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Section: Efforts To Redmentioning

confidence: 99%

Flower colour and cytochromes P450

Tanaka

Brugliera²

2013

Phil. Trans. R. Soc. B

268

146

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“…However, the 35S promoter did not work in certain ornamental crops, and transgenes controlled by the 35S promoter were not expressed in gentian due to the gene silencing of the introduced transgene (Mishiba et al 2005, Mishiba et al 2010. Therefore, the Agrobacterium rhizogenes rolC promoter and Arabidopsis Actin2 promoter were used for the transgene expression in gentian instead of the 35S promoter (Nakatsuka et al 2008, Nakatsuka et al 2011. For utilizing chimeric repressors, the expression of introduced transgenes should be tested in target plant species.…”

Section: Modification Of Chimeric Repressor Constructsmentioning

confidence: 99%

Promotion of Efficient Molecular Breeding Using Chimeric Repressors in Ornamental Flowers

Sasaki

Ohtsubo

2016

JARQ

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Useful novel floral traits have recently been generated in various ornamental crops by using Chimeric REpressor gene Silencing Technology (CRES-T). CRES-T is a plant-specific gene silencing method that targets transcription factors (TFs), causing a dominant loss-of-function phenotype. In our group project, we applied CRES-T to various ornamental crops and mainly elucidated the following points: 1) CRES-T was effective for the modification of floral traits, 2) it was useful in higher polyploidy crops, and 3) it enabled the creation of commercially valuable flowers. In CRES-T, the attachment of a short repression domain derived from a transcriptional repressor to the C-terminal region of target TFs converts transcriptional activators to dominant chimeric repressors. In the target flower species, some Arabidopsis chimeric repressors were effective for the production of new floral traits without cloning the TF genes of interest. Therefore, the use of Arabidopsis chimeric repressors could be widely effective for many ornamental crops, even when the genome and/or expressed sequence tag (EST) information of target crops is unavailable or lacking. In addition, the collective transformation of chimeric repressors into ornamental crops is effective in isolating useful and/or target floral traits, such as petal colors, petal shapes, and color patterns. This technology could help to accelerate the developmental period and reduce the cost of developing new floral traits.

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“…More recently, suppression of F3Ј5ЈH and anthocyanin 5,3Ј-alomatic acyltransferase (5/3ЈAT), involved in the modification of anthocyanin gentiodelphin, resulted in novel lilac to pale blue colored flowers (Nakatsuka et al 2010). Thus, the silencing of flavonoid biosynthetic genes in gentian flowers was demonstrated to increase the diversity of flower colors by changing the composition and quantity of anthocyanin pigments in the petals (Nakatsuka et al 2008c(Nakatsuka et al , 2010Nishihara et al 2006).…”

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confidence: 99%

“…The antisense CHS-transgenic lines produced completely white flowers and inheritance of the white flower trait and herbicide resistance was observed. Subsequently, RNAi suppression of anthocyanin biosynthetic genes, including CHS, flavonoid 3Ј,5Ј-hydroxylase (F3Ј5ЈH) and anthocyanidin synthase (ANS), was achieved and resulted in various flower colors, such as white, pink and pale blue, in transgenic gentian plants (Nakatsuka et al 2008c). More recently, suppression of F3Ј5ЈH and anthocyanin 5,3Ј-alomatic acyltransferase (5/3ЈAT), involved in the modification of anthocyanin gentiodelphin, resulted in novel lilac to pale blue colored flowers (Nakatsuka et al 2010).…”

mentioning

confidence: 99%

Production of picotee-type flowers in Japanese gentian by CRES-T

Nakatsuka¹,

Saito²,

Yamada³

et al. 2011

Plant Biotechnology

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Chimeric repressor gene-silencing technology (CRES-T) is an efficient gene suppression system in a wide variety of dicots and monocots. In this study, we demonstrated that the CRES-T system functions in Japanese gentian. A chimeric repressor of the anthocyanin biosynthetic regulator gene GtMYB3, under the control of the Arabidopsis actin2 promoter, was introduced into blue-flowered gentian. Of 12 transgenic lines, 2 exhibited a picotee flower phenotype with a lack of pigmentation in the lower part of the petal. HPLC analysis showed that the petals of these lines contained less anthocyanin and more flavone than the wild-type, suggesting competitive accumulation of these two types of compounds. The expressions of 'late' flavonoid biosynthetic genes, including F3H, F3Ј5ЈH, DFR and ANS, were strongly suppressed in petals of these transgenic plants. In contrast, the 'early' flavonoid biosynthetic genes, such as CHS and FNSII, were not affected. Since FNSII is expressed more strongly in the lower part of petals than in the upper part, the absence of pigmentation in the lower parts might be induced by flavone synthesis. These results demonstrated that the suppression of anthocyanin biosynthetic genes by CRES-T was successfully applied to Japanese gentian to change petal color; therefore, this system could be useful for generating novel flower colors and patterns. Transgenic plants produced in this study might be utilized as elite materials in the breeding of Japanese gentian in the near future.

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Flower color modification of gentian plants by RNAi-mediated gene silencing

Cited by 56 publications

References 28 publications

Flower colour and cytochromes P450

Flower colour and cytochromes P450

Promotion of Efficient Molecular Breeding Using Chimeric Repressors in Ornamental Flowers

Production of picotee-type flowers in Japanese gentian by CRES-T

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