The diversity markings and pigment patterns in insects are outcomes of adaptive evolution. The elucidation of the molecular mechanism underlying variations in pigment patterns may improve our understanding of the origin and evolution of these spectacular diverse phenotypes. Melanin, ommochrome, and pteridine are the three main types of insect pigments, and the genes that directly participate in pigment biosynthesis have been extensively studied. However, available information on gene interactions and the whole pigment regulatory network is limited. In this study, we performed integument transcriptome sequencing to analyze three larval marking allelic mutants, namely, multi lunar (L), L^C, and L^Ca, which have similar twin-spot markings on the dorsal side of multiple segments. Further analysis identified 336 differentially expressed genes (DEGs) between L and Dazao (wild type which exhibits normal markings), 68 DEGs between L^C/+ and +^LC/+^LC, and 188 DEGs between L^Ca/+ and +^LCa/+^LCa. Gene Ontology (GO) analysis indicated a significant DEG enrichment of the functional terms catalytic activity, binding, metabolic process, and cellular process. Furthermore, three mutants share six common enriched KEGG pathways. We finally identified eight common DEGs among three pairwise comparisons, including Krueppel-like factor, TATA-binding protein, protein patched, UDP-glycosyltransferase, an unknown secreted protein, and three cuticular proteins. Microarray-based gene expression analysis revealed that the eight genes are upregulated during molting, which coincides with marking formation, and are significantly differentially expressed between marking and non-marking regions. The results suggest that the eight common genes are involved in the construction of the multiple twin-spot marking patterns in the three mutants.

中文翻译：

---

珠被转录组的比较分析确定了参与家蚕三个等位基因突变体中标记模式形成的基因。

昆虫的多样性标记和色素图案是适应性进化的结果。阐明色素图案变化背后的分子机制可能会增进我们对这些引人注目的多样表型的起源和进化的理解。黑色素，全色素和蝶啶是昆虫色素的三种主要类型，并且已经直接研究了直接参与色素生物合成的基因。但是，有关基因相互作用和整个色素调控网络的可用信息有限。在这项研究中，我们进行了外皮转录组测序，分析了三个幼虫标记等位基因突变体，即多月（L），L ^C和L ^Ca，在多个节段的背侧都有类似的双点标记。进一步的分析确定了L和Dazao之间的336个差异表达基因（DEG）（野生型，表现出正常的标记），L ^C / +和+ ^LC / + ^LC之间有68个DEG ，L ^Ca / +和+ ^LCa / +之间有188 DEG。^LCa。基因本体论（GO）分析表明，催化活性，结合，代谢过程和细胞过程等功能性术语的DEG显着丰富。此外，三个突变体共有六个共同的丰富的KEGG途径。我们最终确定了三个成对比较中的八个常见DEG，包括Krueppel类因子，TATA结合蛋白，修补蛋白，UDP-糖基转移酶，未知分泌蛋白和三种表皮蛋白。基于微阵列的基因表达分析表明，在蜕皮过程中这八个基因被上调，这与标记形成相吻合，并且在标记和非标记区域之间显着差异表达。结果表明，在三个突变体中，八个共有基因参与了多个双点标记模式的构建。