Eye pigmentation genes have been utilized as visible markers for constructing genetic control prototypes in several insect vectors of human disease. Here, orthologs of two ommochrome pathway genes, kynurenine 3-hydroxylase (kmo) and cardinal, were investigated in Plutella xylostella, a globally distributed, economically important pest of Brassica crops. Both somatic mosaic and germline mutations were efficiently created using the CRISPR/Cas9 system, and null mutant strains of Pxkmo and Pxcardinal were obtained. A frame-shift mutation in Pxkmo caused yellow compound eyes at adult stage while an in-frame mutation lacking two amino acids resulted in a hypomorphic red eye phenotypes. In contrast, Pxcardinal-deficient moths with a frame-shift mutation exhibited yellow eye pigmentation in newly emerged adults which turned to red as the adults aged. Additionally, differences were observed in the coloration of larval ocelli, brains and testes in Pxkmo and Pxcardinal yellow-eye mutant lines. Our work identifies the important roles of Pxkmo and Pxcardinal in P. xylostella eye pigmentation and provides tools for future genetic manipulation of this important crop pest.

中文翻译：

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Ommochrome 途径基因犬尿氨酸 3-羟化酶和红衣主教参与小菜蛾眼部色素沉着。

眼睛色素沉着基因已被用作在人类疾病的几种昆虫载体中构建遗传控制原型的可见标记。在这里，在小菜蛾（Plutella xylostella）中研究了两个全色素途径基因犬尿氨酸 3-羟化酶 (kmo) 和红衣主教的直系同源物，小菜蛾是一种全球分布的、具有重要经济意义的芸苔属作物害虫。使用 CRISPR/Cas9 系统有效地创建了体细胞镶嵌和种系突变，并获得了 Pxkmo 和 Pxcardinal 的无效突变株。Pxkmo 中的移码突变在成年阶段导致黄色复眼，而缺少两个氨基酸的框内突变导致亚型红眼表型。相比之下，具有移码突变的 Pxcardinal 缺陷型飞蛾在新出现的成虫中表现出黄色的眼睛色素沉着，随着成虫的衰老而变成红色。此外，在 Pxkmo 和 Pxcardinal 黄眼突变体系中观察到幼虫单眼、大脑和睾丸的颜色存在差异。我们的工作确定了 Pxkmo 和 Pxcardinal 在小菜蛾眼睛色素沉着中的重要作用，并为未来对这种重要作物害虫进行遗传操作提供了工具。