Nitarska Daria, Boehm Robert, Debener Thomas, Lucaciu Rares Calin, Halbwirth Heidi
Institute of Chemical, Environmental and Bioscience Engineering, Technische Universität Wien, 1060 Vienna, Austria.
Klemm+Sohn GmbH & Co, 70379 Stuttgart, Germany.
Plant Cell Tissue Organ Cult. 2021;147(1):49-60. doi: 10.1007/s11240-021-02103-5. Epub 2021 May 26.
The CRISPR/Cas9 system is a remarkably promising tool for targeted gene mutagenesis, and becoming ever more popular for modification of ornamental plants. In this study we performed the knockout of ' (') with application of CRISPR/Cas9 in the red flowering poinsettia ( cultivar 'Christmas Eve', in order to obtain plants with orange bract colour, which accumulate prevalently pelargonidin. F3'H is an enzyme that is necessary for formation of cyanidin type anthocyanins, which are responsible for the red colour of poinsettia bracts. Even though F3'H was not completely inactivated, the bract colour of transgenic plants changed from vivid red (RHS 45B) to vivid reddish orange (RHS 33A), and cyanidin levels decreased significantly compared with the wild type. In the genetically modified plants, an increased ratio of pelargonidin to cyanidin was observed. By cloning and expression of mutated proteins, the lack of F3'H activity was confirmed. This confirms that a loss of function mutation in the poinsettia gene is sufficient for obtaining poinsettia with orange bract colour. This is the first report of successful use of CRISPR/Cas9 for genome editing in poinsettia.
The online version contains supplementary material available at 10.1007/s11240-021-02103-5.
CRISPR/Cas9系统是一种极具前景的靶向基因诱变工具，在观赏植物改良中越来越受欢迎。在本研究中，我们应用CRISPR/Cas9对红色一品红（品种‘平安夜’）中的 基因进行敲除，以获得苞片颜色为橙色的植株，这些植株主要积累天竺葵色素。F3'H是矢车菊素型花青素形成所必需的一种酶，矢车菊素型花青素是一品红苞片红色的原因。尽管F3'H没有完全失活，但转基因植株的苞片颜色从鲜艳的红色（RHS 45B）变为鲜艳的红橙色（RHS 33A），与野生型相比，矢车菊素水平显著降低。在转基因植株中，观察到天竺葵色素与矢车菊素的比例增加。通过克隆和表达突变蛋白，证实了F3'H活性的缺失。这证实了一品红 基因的功能缺失突变足以获得苞片颜色为橙色的一品红。这是关于成功使用CRISPR/Cas9对一品红进行基因组编辑的首次报道。
在线版本包含可在10.1007/s11240-021-02103-5获取的补充材料。
