Engineering of the Rose Flavonoid Biosynthetic Pathway Successfully Generated Blue-Hued Flowers Accu.pdfVIP

Engineering of the Rose Flavonoid Biosynthetic Pathway Successfully Generated Blue-Hued Flowers Accumulating Delphinidin Yukihisa Katsumoto 1, Masako Fukuchi-Mizutani 1, Yuko Fukui 1, Filippa Brugliera 2, Timothy A. Holton 2, Mirko Karan 2, Noriko Nakamura 1, Keiko Yonekura-Sakakibara 1, 3, Junichi Togami 1, Alix Pigeaire 2, Guo-Qing Tao 2, Narender S. Nehra 2, Chin-Yi Lu 2, Barry K. Dyson 2, Shinzo Tsuda 1, Toshihiko Ashikari 1, 2, Takaaki Kusumi 1, John G. Mason 2 and Yoshikazu Tanaka 1, * 1 Suntory Limited Research Center, Institute for Plant Science, 1-1-1 Wakayamadai, Shimamoto-cho, Mishima-gun, Osaka, 618-8503 Japan 2 Florigene Pty. Ltd, 1 Park Drive, Bundoora, VIC 3083, Australia 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 consti- tuents of violet and blue flowers, because roses do not possess flavonoid 30,50-hydoxylase (F3050H), 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 F3050H 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 Iris3hollandica DFR gene in addition to the viola F3050H 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