Key Laboratory of Insect Developmental and Evolutionary Biology, Institute of Plant Physiology and Ecology, Shanghai Institutes for Biological Sciences, Chinese Academy of Sciences, Shanghai 200032, China.
Department of Biology, University of Nevada, Reno, 1664 N Virginia St, Reno, NV 89557, USA.
Insect Biochem Mol Biol. 2018 Mar;94:1-9. doi: 10.1016/j.ibmb.2018.01.002. Epub 2018 Jan 11.
The yellow gene family has been identified in several model insects, but yellow genes were poorly identified in non-model insects and the functions of yellow genes are largely unknown. In this study, we identified seven yellow genes in an important agricultural pest Agrotis ipsilon. Each gene encodes a protein containing a major royal jelly domain. Phylogenetic analysis defined these genes as yellow-y, -b, -b2, -c, -d, -e, and -h, respectively. The A. ipsilon yellow genes yellow-b, -b2, and -c were stably expressed in all developmental stages and tissues analyzed, whereas the other four yellow genes had unique expression patterns, suggesting distinct physiological roles of each gene. Using the CRISPR/Cas9 system, we successfully disrupted yellow-y in A. ipsilon and obtained G insects with somatic mutations. Unlike the black of wild-type newly hatched larvae and of adults, the mutants were yellow, although in the pupal stage mutant coloration did not differ from wild-type coloration. This phenotype was inherited by G offspring. The G mutants did not show any growth deficiency compared with control insects; however, a dehydration-like phenotype was observed in newly hatched G larvae from sibling crossed mutants. Our results indicate that A. ipsilon yellow-y gene plays a role in body pigmentation and also might function in waterproofing.
黄色基因家族在几种模式昆虫中已被鉴定，但黄色基因在非模式昆虫中鉴定较少，其功能也知之甚少。本研究在重要农业害虫甜菜夜蛾中鉴定了 7 个黄色基因。每个基因编码的蛋白均含有主要蜂王浆结构域。系统发育分析将这些基因分别定义为 yellow-y、-b、-b2、-c、-d、-e 和 -h。在分析的所有发育阶段和组织中，A. ipsilon 的 yellow-b、-b2 和 -c 基因均稳定表达，而其他四个黄色基因具有独特的表达模式，表明每个基因具有不同的生理功能。利用 CRISPR/Cas9 系统，我们成功敲除了 A. ipsilon 的 yellow-y 基因，并获得了具有体细胞突变的 G 昆虫。与野生型刚孵化幼虫和成虫的黑色不同，突变体呈黄色，尽管在蛹期突变体的颜色与野生型颜色没有区别。这种表型可被 G 后代遗传。与对照昆虫相比，G 突变体没有表现出任何生长缺陷；然而，从突变体兄妹交配的刚孵化的 G 幼虫中观察到了类似脱水的表型。我们的结果表明，A. ipsilon yellow-y 基因在体色素沉着中起作用，并且可能在防水方面也有功能。
