Alteration of flower colour in Ipomoea nil through CRISPR/Cas9-mediated mutagenesis of carotenoid cleavage dioxygenase 4
Alteration of flower colour in Ipomoea nil through CRISPR/Cas9-mediated mutagenesis of carotenoid cleavage dioxygenase 4
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DOI：
10.1007/s11248-017-0051-0
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发表时间：
2017-12
影响因子：
3
通讯作者：
Kenta Watanabe;Chihiro Oda-Yamamizo;K. Sage-Ono;A. Ohmiya;M. Ono
中科院分区：
文献类型：
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作者：
Kenta Watanabe;Chihiro Oda-Yamamizo;K. Sage-Ono;A. Ohmiya;M. Ono
日本牵牛花Ipomoea Nil表现出除黄色外的各种花色，反映出花瓣中仅积累了微量的类胡萝卜素。在之前的研究中，我们将这种影响归因于花瓣中胡萝卜素生成基因的低表达水平，但可能还有其他因素。在本研究中，我们研究了类胡萝卜素裂解双加氧酶(CCD)可能参与调控花瓣中类胡萝卜素的积累。没有。通过生物信息学分析，共鉴定出7个InCCD基因。零基因组。测序和表达分析表明，InCCD4可能参与了花瓣中类胡萝卜素的降解。利用CRISPR/Cas9系统成功地敲除了白花品种I中的InCCD4。Nilcv.AK77使白色的花瓣变成了淡黄色。与非转基因植株相比，转基因植株花瓣中类胡萝卜素的总量增加了20倍。这一结果表明，在花瓣中。零，不仅低胡萝卜素基因的表达，而且类胡萝卜素的降解导致类胡萝卜素水平极低。
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 ofI. nil. Using bioinformatics analysis, sevenInCCDgenes were identified in theI. nilgenome. Sequencing and expression analyses indicated potential involvement ofInCCD4in carotenoid degradation in the petals. Successful knockout ofInCCD4using the CRISPR/Cas9 system in the white-flowered cultivarI. nilcv. AK77 caused the white petals to turn pale yellow. The total amount of carotenoids in the petals ofccd4plants was increased 20-fold relative to non-transgenic plants. This result indicates that in the petals ofI. nil, not only low carotenogenic gene expression but also carotenoid degradation leads to extremely low levels of carotenoids.