Alteration of flower colour in <Emphasis Type="Italic">Ipomoea nil</Emphasis> through CRISPR/Cas9-mediated mutagenesis of <Emphasis Type="Italic">carotenoid cleavage dioxygenase 4</Emphasis> |
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Authors: | Kenta Watanabe Chihiro Oda-Yamamizo Kimiyo Sage-Ono Akemi Ohmiya Michiyuki Ono |
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Institution: | 1.Graduate School of Life and Environmental Sciences,University of Tsukuba,Tsukuba,Japan;2.Institute of Vegetable and Floriculture Science,National Agriculture and Food Research Organization,Tsukuba,Japan;3.Faculty of Life and Environmental Sciences, Gene Research Center, Tsukuba Plant Innovation Research Center (T-PIRC),University of Tsukuba,Tsukuba,Japan;4.Biological Resources and Post-Harvest Division,Japan International Research Center for Agricultural Sciences,Tsukuba,Japan |
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Abstract: | Japanese morning glory, Ipomoea nil, exhibits a variety of flower colours, except yellow, reflecting the accumulation of only trace amounts of carotenoids in the petals. In a previous study, we attributed this effect to the low expression levels of carotenogenic genes in the petals, but there may be other contributing factors. In the present study, we investigated the possible involvement of carotenoid cleavage dioxygenase (CCD), which cleaves specific double bonds of the polyene chains of carotenoids, in the regulation of carotenoid accumulation in the petals of I. nil. Using bioinformatics analysis, seven InCCD genes were identified in the I. nil genome. Sequencing and expression analyses indicated potential involvement of InCCD4 in carotenoid degradation in the petals. Successful knockout of InCCD4 using the CRISPR/Cas9 system in the white-flowered cultivar I. nil cv. AK77 caused the white petals to turn pale yellow. The total amount of carotenoids in the petals of ccd4 plants was increased 20-fold relative to non-transgenic plants. This result indicates that in the petals of I. nil, not only low carotenogenic gene expression but also carotenoid degradation leads to extremely low levels of carotenoids. |
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