The Flower CRES-T project

Norihiro Ohtsubo. . . . . . . 111 [PDF]

Project Review

Beyond the blue rose: Modification of floral architecture with plant-specific chimeric repressors

Norihiro Ohtsubo. . . . . . . 113 [PDF]

This review describes the concepts and research strategies for the Flower CRES-T project, which has been launched in 2005 to overcome difficulties associated with the commercialization of GM flowers, through an overview of recent progress gathered in this special issue.

Original Papers

The new FioreDB database provides comprehensive information on plant transcription factors and phenotypes induced by CRES-T in ornamental and model plants

Nobutaka Mitsuda, Yuko Takiguchi, Masahito Shikata, Kimiyo Sage-Ono, Michiyuki Ono, Katsutomo Sasaki, Hiroyasu Yamaguchi, Takako Narumi, Yuri Tanaka, Masao Sugiyama, Tomomichi Yamamura, Teruhiko Terakawa, Keiko Gion, Ryota Suzuri, Yoshikazu Tanaka, Takashi Nakatsuka, Souichi Kimura, Masahiro Nishihara, Tomoyuki Sakai, Reiko Endo-Onodera, Kumi Saitoh, Kanji Isuzugawa, Yoshimi Oshima, Tomotsugu Koyama, Miho Ikeda, Megumi Narukawa, Kyoko Matsui, Masaru Nakata, Norihiro Ohtsubo, Masaru Ohme-Takagi. . . . . . . 123 [PDF] [Supplement]

he development of new FioreDB database is reported. It provides more than 300 phenotypic information of various plants, linked to more than 100 TFs and comprehensive information about Arabidopsis TFs.

Arabidopsis chimeric TCP3 repressor produces novel floral traits in Torenia fournieri and Chrysanthemum morifolium

Takako Narumi, Ryutaro Aida, Tomotsugu Koyama, Hiroyasu Yamaguchi, Katsutomo Sasaki, Masahito Shikata, Masayoshi Nakayama, Masaru Ohme-Takagi, Norihiro Ohtsubo. . . . . . . 131 [PDF] [Supplement1] [Supplement2]

Transgenic torenia and chrysanthemum expressing TCP3-SRDX exhibited fringed margins in petals and leaves as in Arabidopsis. We demonstrated that CRES-T functions beyond plant species irrespective of their ploidy, and is effective for creation of novel flower phenotypes.

Creating ruffled flower petals in Cyclamen persicum by expression of the chimeric cyclamen TCP repressor

Yuri Tanaka, Tomomichi Yamamura1, Yoshimi Oshima, Nobutaka Mitsuda, Tomotsugu Koyama, Masaru Ohme-Takagi, Teruhiko Terakawa. . . . . . . 141 [PDF] [Supplement]

We isolated a TCP homolog gene from cyclamen (CpTCP1) to modify flower shape in cyclamen. A chimeric repressor gene construct for CpTCP1 driven by the CaMV35S promoter (35S:CpTCP1SRDX) was transformed into cyclamen. The phenotypic changes resembled those of the chimeric repressor derived from AtTCP3. CpTCP1SRDX expression resulted in cyclamen with serrated wavy leaves and curled petals.

Transgenic Note

Morphological changes of Rosa x hybrida by a chimeric repressor of Arabidopsis TCP3

Keiko Gion, Ryota Suzuri, Masahito Shikata, Nobutaka Mitsuda, Yoshimi Oshima, Tomotsugu Koyama, Masaru Ohme-Takagi, Norihiro Ohtsubo, Yoshikazu Tanaka. . . . . . . 149 [PDF]

To obtain new rose cultivars that have ornamental values, we attempted to alter the morphology of Rosa x hybrida cv. Lavande with TCP3SRDX which consists of Arabidopsis TCP3 and an EAR-motif repression domain. The TCP3SRDX transgenic rose plants increased the number of leaflets and the size of leaf teeth. The petals were wavy, and the sepals were compound-leafy. We succeeded in altering rose morphology using Arabidopsis TCP3 without the sequence information of a TCP3 homologue in the target plant species.

Original Paper

Induction of double flowers in Pharbitis nil using a class-C MADS-box transcription factor with Chimeric Repressor

Kimiyo Sage-Ono, Yuko Ozeki, Satomi Hiyama, Yohei Higuchi, Hiroshi Kamada, Nobutaka Mitsuda, Masaru Ohme-Takagi, Michiyuki Ono. . . . . . . 153 [PDF] [Supplement]

We developed the GAL4SRDX/UAS system driven by inducible promoters (alcohol-inducible and heat-shock promoters) as a new gene-silencing technology for repression of overexpressing transgenes. Normal regeneration was observed by inducible repression of DPSRDX during in vitro redifferentiation, and the double-flowered Pharbitis was generated.

Gene Note

Identification and expression analysis of the Cyclamen persicum MADS-box gene family

Yuri Tanaka, Tomomichi Yamamura, Teruhiko Terakawa. . . . . . . 167 [PDF] [Supplement1] [Supplement2]

We identified and characterized 10 putative cyclamen MADS-box genes. These genes are related to the APETALA1, PISTILLATA, APETALA3, AGAMOUS, SEEDSTICK, and SEPALLATA transcription factor subfamilies. Respective genes are expressed in each whorl according to the ABCDE function, but there are slight differences in the expression of several genes in various tissues.

Original Papers

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

Takashi Nakatsuka, Misa Saito, Eri Yamada, Masahiro Nishihara. . . . . . . 173 [PDF]

A chimeric repressor of the anthocyanin biosynthetic regulator gene GtMYB3 was introduced into blue-flowered gentian. These transgenic gentians exhibited a picotee flower phenotype with a lack of pigmentation in the lower part of the petal. CRES-T system could be useful for generating novel flower colors and patterns.

Utilization of a floral organ-expressing AP1 promoter for generation of new floral traits in Torenia fournieri Lind

Katsutomo Sasaki, Hiroyasu Yamaguchi, Takako Narumi, Masahito Shikata, Yoshimi Oshima, Masaru Nakata, Nobutaka Mitsuda, Masaru Ohme-Takagi, Norihiro Ohtsubo. . . . . . . 181 [PDF] [Supplement1] [Supplement2]

A 35S:MYB24-SRDX transgenic torenia exhibited unopened flower buds and dark green leaves with curled margins. Utilization of a floral organ-specific AP1 promoter instead of the 35S promoter resulted in opening of flowers with a characteristic configuration and a normal leaf phenotype in torenia.

Efficient production of novel floral traits in torenia by collective transformation with chimeric repressors of Arabidopsis transcription factors

Masahito Shikata, Takako Narumi, Hiroyasu Yamaguchi, Katsutomo Sasaki, Ryutaro Aida, Yoshimi Oshima, Yuko Takiguchi, Masaru Ohme-Takagi, Nobutaka Mitsuda, Norihiro Ohtsubo. . . . . . . 189 [PDF]

To facilitate novel flower production in floricultural plants, we collectively introduced 42 and 50 chimeric repressors of Arabidopsis transcription factors into torenia. We efficiently obtained 241 transgenic plants with phenotypic modification in flower colors and/or shapes.

Novel vector systems to accelerate functional analysis of transcription factors using chimeric repressor gene-silencing technology (CRES-T)

Yoshimi Oshima, Nobutaka Mitsuda, Masaru Nakata, Tsuyoshi Nakagawa, Shingo Nagaya, Ko Kato, Masaru Ohme-Takagi. . . . . . . 201 [PDF] [Supplement1] [Supplement2]

New CRES-T vectors that are efficient and convenient to use were developed by employing the Gateway system, a new vector backbone and a terminator derived from the heat shock protein 18.2 (HSP) gene.

A flavonol synthase gene GtFLS defines anther-specific flavonol accumulation in gentian

Soichi Kimura, Takashi Nakatsuka, Eri Yamada, Misa Saito, Masahiro Nishihara. . . . . . . 211 [PDF] [Supplement]

The accumulation profiles of flavonoid components and expression of their biosynthetic genes in gentian anthers have been investigated. Flavonols were the flavonoid compounds accumulated in the anthers, and their accumulations were controlled by anther-specific expression of the GtFLS gene in gentian.

De novo DNA methylation of the 35S enhancer revealed by high-resolution methylation analysis of an entire T-DNA segment in transgenic gentian

Satoshi Yamasaki, Masayuki Oda, Nozomu Koizumi, Kazuhiko Mitsukuri, Masafumi Johkan, Takashi Nakatsuka, Masahiro Nishihara, Kei-ichiro Mishiba. . . . . . . 223 [PDF] [Supplement1] [Supplement2]

Detailed DNA methylation profile of the entire T-DNA sequence integrated into transgenic gentians showed that highly methylated CpHpH sequences were found within the 35S enhancer region (-148 to -85 and -298 to -241). EMSA showed that gentian nuclear extracts could bind to the corresponding probes (-149 to -124 and -275 to -250), and that the probes could compete with one another. Thus, a nuclear factor might be involved in the de novo methylation of the two regions.

Isolation of cDNAs encoding tetrahydroxychalcone 2′-glucosyltransferase activity from carnation, cyclamen, and catharanthus

Junichi Togami, Hiroaki Okuhara, Noriko Nakamura, Kanako Ishiguro, Chika Hirose, Misa Ochiai, Yuko Fukui, Masa-atsu Yamaguchi, Yoshikazu Tanaka. . . . . . . 231 [PDF]

4,2′,4′,6′-Tetrahydroxychalcone (THC) 2′- glucoside confers yellow color. Five, one and one types of glucosyltransferase (GT) cDNAs encoding THC 2′- GT activity were isolated from carnation, cyclamen, and catharanthus. Expression of three of the carnation cDNAs in petunia resulted in small amount of THC 2′- glucoside.

Gene Note

Biochemical and molecular characterization of anthocyanidin/flavonol 3-glucosylation pathways in Rosa × hybrida

Masako Fukuchi-Mizutani, Misako Akagi, Kanako Ishiguro, Yukihisa Katsumoto, Yuko Fukui, Junichi Togami, Noriko Nakamura, Yoshikazu Tanaka. . . . . . . 239 [PDF]

We isolated a rose flavonoid 3-glucosyltransferase homolog that is specific to anthocyanidin and expresses in mature petals synthesizing anthocyanins. We also found that another homolog which preferably catalyzes glucosylation of flavonols to anthocyanidins and expresses in young petals biosynthesizes anthocyanidin 3-glucosides in most rose petals.

Original Papers

Molecular based evidence for a lack of gene-flow between Rosa × hybrida and wild Rosa species in Japan

Noriko Nakamura, Ursula Tems, Masako Fukuchi-Mizutani, Steve Chandler, Yoshiie Matsuda, Shunsuke Takeuchi, Shogo Matsumoto, Yoshikazu Tanaka. . . . . . . 245 [PDF]

Gene flow from cultivated roses to wild rose species was measured with using the KSN locus that is specific to the cultivated roses as the molecular marker. No gene flow to from the cultivated rose was detected even in wild Rosa species grown in close proximity to the cultivated roses, which indicates the flow is very low if any.

Environmental risk assessment and field performance of rose (Rosa x hybrida) genetically modified for delphinidin production

Noriko Nakamura, Masako Fukuchi-Mizutani, Yukihisa Katsumoto, Junichi Togami, Mick Senior, Yoshie Matsuda, Keiji Furuichi, Mie Yoshimoto, Akihiro Matsunaga, Kanako Ishiguro, Mitsuhiro Aida, Masao Tasaka, Hirokazu Fukui, Shinzo Tsuda, Steve Chandler, Yoshikazu Tanaka. . . . . . . 251 [PDF] [Supplement]

Risk assessment of two genetically modified blue rose lines on the basis of the Cartagena law revealed that their release is most unlikely affect Japanese biodiversity. Furthermore, the roses contained the transgenes only in L1 cells. General release permission was granted for the rose.

Technical Note

Trehalose drastically extends the in vitro vegetative culture period and facilitates maintenance of Torenia fournieri plants

Hiroyasu Yamaguchi, Katsutomo Sasaki, Masahito Shikata, Ryutaro Aida, Norihiro Ohtsubo. . . . . . . 263 [PDF] [Supplement1] [Supplement2]

In in vitro culture of torenia, substituting trehalose for sucrose drastically extended more than twice the culture period (for 2.5 months). Furthermore, the use of the low price commercial food additive, Okome-ni-TREHA® rather than reagent-grade trehalose enabled us to reduce the costs and labor.