CRISPR/Cas9 technology is a versatile tool for targeted mutagenesis in many organisms, including plants. However, this technique has not been applied to the Japanese morning glory (Ipomoea [Pharbitis] nil), a traditional garden plant chosen for the National BioResource Project in Japan. We selected dihydroflavonol-4-reductase-B (DFR-B) of I. nil, encoding an anthocyanin biosynthesis enzyme, as the target gene, and changes in the stem colour were observed during the early stages of plant tissue culture by Rhizobium [Agrobacterium]-mediated transformation. Twenty-four of the 32 (75%) transgenic plants bore anthocyanin-less white flowers with bi-allelic mutations at the Cas9 cleavage site in DFR-B, exhibiting a single base insertion or deletions of more than two bases. Thus, these results demonstrate that CRISPR/Cas9 technology enables the exploration of gene functions in this model horticultural plant. To our knowledge, this report is the first concerning flower colour changes in higher plants using CRISPR/Cas9 technology.

中文翻译：

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CRISPR / Cas9介导的日本牵牛花番薯属（Pharbitis）无二氢黄酮醇4-还原酶-B（DFR-B）基因座的诱变。

CRISPR / Cas9技术是用于多种生物（包括植物）定向诱变的多功能工具。但是，该技术尚未应用于日本牵牛花（Ipomoea [Pharbitis] nil），后者是日本国家生物资源计划中选择的传统园林植物。我们选择了编码花色苷生物合成酶的无.. I. nil的二氢黄酮醇-4-还原酶-B（DFR-B）作为靶基因，并且在植物组织培养的早期通过根瘤菌观察了茎的颜色变化。 ]介导的转化。32种转基因植物中有24种（75％）在DFR-B的Cas9切割位点上花无花青素的白花带有双等位基因突变，表现出单碱基插入或多于两个碱基的缺失。因此，这些结果表明，CRISPR / Cas9技术可以在这种园艺模型中探索基因功能。据我们所知，该报告是第一个关于使用CRISPR / Cas9技术的高等植物中花色变化的报告。