Identifying genes associated with wax deposition may contribute to the genetic improvement of ornamental kale. Here, we characterized a candidate gene for wax contents, BoORP3a, encoding an oxysterol-binding protein. We sequenced the BoORP3a gene and coding sequence from the high-wax line S0835 and the low-wax line F0819, which revealed 12 single nucleotide polymorphisms between the two lines, of which six caused five amino acids substitutions. BoORP3a appeared to be relatively well conserved in Brassicaceae, as determined by a phylogenetic analysis, and localized to the endoplasmic reticulum and the nucleus. To confirm the role of BoORP3a in wax deposition, we generated three orp3a mutants in a high-wax kale background via CRISPR/Cas9-mediated genome editing. Importantly, all three mutants exhibited lower wax contents and glossy leaves. Overall, these data suggest that BoORP3a may participate in cuticular wax deposition in ornamental kale.

中文翻译：

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BoORP3a 的失活，一种氧甾醇结合蛋白，导致观赏羽衣甘蓝的低蜡表型

鉴定与蜡沉积相关的基因可能有助于观赏羽衣甘蓝的遗传改良。在这里，我们表征了蜡含量的候选基因 BoORP3a，该基因编码氧固醇结合蛋白。我们对高蜡系 S0835 和低蜡系 F0819 的 BoORP3a 基因和编码序列进行了测序，揭示了两个品系之间的 12 个单核苷酸多态性，其中 6 个导致了 5 个氨基酸取代。BoORP3a 似乎在十字花科植物中相对保守，如系统发育分析所确定的那样，并且定位于内质网和细胞核。为了确认 BoORP3a 在蜡沉积中的作用，我们通过 CRISPR/Cas9 介导的基因组编辑在高蜡羽衣甘蓝背景中生成了三个 orp3a 突变体。重要的是，所有三个突变体都表现出较低的蜡含量和光滑的叶子。全面的，