College of Life Sciences, East China Normal University, Shanghai, China.
Key Laboratory of Insect Developmental and Evolutionary Biology, Institute of Plant Physiology and Ecology, Shanghai Institutes for Biological Sciences, Chinese Academy of Sciences, Shanghai, China.
Insect Sci. 2019 Dec;26(6):1011-1019. doi: 10.1111/1744-7917.12663. Epub 2019 Mar 5.
Insect body pigmentation and coloration are critical to adaption to the environment. To explore the mechanisms that drive pigmentation, we used the clustered regularly interspaced short palindromic repeats (CRISPR)/CRISPR-associated protein 9 (Cas9) genome editing system to target the ebony gene in the non-model insect Spodoptera litura. Ebony is crucial to melanin synthesis in insects. By directly injecting Cas9 messenger RNA and ebony-specific guide RNAs into S. litura embryos, we successfully induced a typical ebony-deficient phenotype of deep coloration of the puparium and induction of melanin formation during the pupal stage. Polymerase chain reaction-based genotype analysis demonstrated that various mutations had occurred at the sites targeted in ebony. Our study clearly demonstrates the function of ebony in the puparium coloration and also provides a potentially useful marker gene for functional studies in S. litura as well as other lepidopteran pests.
昆虫体色素沉着和颜色对于适应环境至关重要。为了探索驱动色素沉着的机制，我们使用了成簇的规律间隔短回文重复序列（CRISPR）/CRISPR 相关蛋白 9（Cas9）基因组编辑系统，以针对非模式昆虫斜纹夜蛾中的乌木基因。乌木对于昆虫中的黑色素合成至关重要。通过直接将 Cas9 信使 RNA 和针对 ebony 的特定指导 RNA 注射到斜纹夜蛾胚胎中，我们成功地诱导了典型的 ebony 缺陷表型，即蛹的深色化以及蛹期黑色素的形成。基于聚合酶链反应的基因型分析表明， ebony 靶向部位发生了各种突变。我们的研究清楚地表明了 ebony 在蛹的颜色形成中的功能，并且还为斜纹夜蛾以及其他鳞翅目害虫的功能研究提供了一个潜在有用的标记基因。
