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Plant and Cell Physiology Advance Access originally published online on October 9, 2007
Plant and Cell Physiology 2007 48(11):1589-1600; doi:10.1093/pcp/pcm131
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© The Author 2007. Published by Oxford University Press on behalf of Japanese Society of Plant Physiologists. All rights reserved. For permissions, please email: journals.permissions@oxfordjournals.org

Engineering of the Rose Flavonoid Biosynthetic Pathway Successfully Generated Blue-Hued Flowers Accumulating Delphinidin

Yukihisa Katsumoto1, Masako Fukuchi-Mizutani1, Yuko Fukui1, Filippa Brugliera2, Timothy A. Holton2, Mirko Karan2, Noriko Nakamura1, Keiko Yonekura-Sakakibara1,3, Junichi Togami1, Alix Pigeaire2, Guo-Qing Tao2, Narender S. Nehra2, Chin-Yi Lu2, Barry K. Dyson2, Shinzo Tsuda1, Toshihiko Ashikari1,2, Takaaki Kusumi1, John G. Mason2 and Yoshikazu Tanaka1,*

1Suntory Limited Research Center, Institute for Plant Science, 1-1-1 Wakayamadai, Shimamoto-cho, Mishima-gun, Osaka, 618-8503 Japan
2Florigene Pty. Ltd, 1 Park Drive, Bundoora, VIC 3083, Australia

*Corresponding author. E-mail, Yoshikazu_Tanaka{at}suntory.co.jp; Fax, +81-75-962-3791.


  Abstract

Flower color is mainly determined by anthocyanins. Rosa hybrida lacks violet to blue flower varieties due to the absence of delphinidin-based anthocyanins, usually the major constituents of violet and blue flowers, because roses do not possess flavonoid 3',5'-hydoxylase (F3'5'H), a key enzyme for delphinidin biosynthesis. Other factors such as the presence of co-pigments and the vacuolar pH also affect flower color. We analyzed the flavonoid composition of hundreds of rose cultivars and measured the pH of their petal juice in order to select hosts of genetic transformation that would be suitable for the exclusive accumulation of delphinidin and the resulting color change toward blue. Expression of the viola F3'5'H gene in some of the selected cultivars resulted in the accumulation of a high percentage of delphinidin (up to 95%) and a novel bluish flower color. For more exclusive and dominant accumulation of delphinidin irrespective of the hosts, we down-regulated the endogenous dihydroflavonol 4-reductase (DFR) gene and overexpressed the Irisxhollandica DFR gene in addition to the viola F3'5'H gene in a rose cultivar. The resultant roses exclusively accumulated delphinidin in the petals, and the flowers had blue hues not achieved by hybridization breeding. Moreover, the ability for exclusive accumulation of delphinidin was inherited by the next generations.

Keywords: Anthocyanin - Delphinidin - Flavonoid - Flower color - Metabolic engineering - Rosa hybrida

Abbreviations: AT, hydroxycinnamoyl CoA:anthocyanin hydroxycinnamoyl transferase; CaMV, cauliflower mosaic virus; DFR, dihydroflavonol 4-reductase; F3'5'H, flavonoid 3',5'-hydroxylase; NMR, nuclear magnetic resonance; NPTII, neomycin phosphotransferase; RHSCC, Royal Horticultural Society Color Chart; RNAi, RNA interference; siRNA, small interfering RNA; TFA, trifluoroacetic acid; TOF-MS, time-of-flight-mass spectrometry.

3 Present address: RIKEN Plant Science Center, 1-7-22 Suehiro-cho, Tsurumi-ku, Yokohama City, Kanagawa, 230-0045 Japan.

(Received August 16, 2007; Accepted October 3, 2007)
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