The oriental armyworm, Mythimna separata (Walker) is a serious pest of agriculture that does particular damage to Gramineae crops in Asia, Europe, and Oceania. Metamorphosis is a key developmental stage in insects, although the genes underlying the metamorphic transition in M. separata remain largely unknown. Here, we sequenced the transcriptomes of five stages; mature larvae (ML), wandering (W), and pupation (1, 5, and 10 days after pupation, designated P1, P5, and P10) to identify transition-associated genes. Four libraries were generated, with 22,884, 23,534, 26,643, and 33,238 differentially expressed genes (DEGs) for the ML-vs-W, W-vs-P1, P1-vs-P5, and P5-vs-P10, respectively. Gene ontology enrichment analysis of DEGs showed that genes regulating the biosynthesis of the membrane and integral components of the membrane, which includes the cuticular protein (CP), 20-hydroxyecdysone (20E), and juvenile hormone (JH) biosynthesis, were enriched. Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analysis indicated that DEGs were enriched in the metabolic pathways. Of these DEGs, thirty CP, seventeen 20E, and seven JH genes were differentially expressed across the developmental stages. For transcriptome validation, ten CP, 20E, and JH-related genes were selected and verified by real-time PCR quantitative. Collectively, our results provided a basis for further studies of the molecular mechanism of metamorphosis in M. separata.

中文翻译：

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东方粘虫 Mythimna separata (Walker) (鳞翅目: 夜蛾科) 前蛹-蛹转变过程中差异表达基因的转录鉴定

东方粘虫，蜉蝣(Walker) 是一种严重的农业害虫，对亚洲、欧洲和大洋洲的禾本科作物造成特别损害。变态是昆虫的一个关键发育阶段，尽管在昆虫的变态转变背后的基因M. separata仍然很大程度上未知。在这里，我们对五个阶段的转录组进行了测序；成熟幼虫 (ML)、游走 (W) 和化蛹（化蛹后 1、5 和 10 天，指定为 P1、P5 和 P10）以识别过渡相关基因。生成了四个文库，分别具有 22,884、23,534、26,643 和 33,238 个差异表达基因 (DEG)，分别用于 ML-vs-W、W-vs-P1、P1-vs-P5 和 P5-vs-P10。DEGs的基因本体富集分析表明，调节膜生物合成的基因和膜的组成部分，包括表皮蛋白（CP）、20-羟基蜕皮激素（20E）和保幼激素（JH）生物合成，被富集。京都基因和基因组百科全书（KEGG）通路分析表明，DEGs 在代谢通路中富集。在这些 DEG 中，30 个 CP，17 个 20E，和七个 JH 基因在发育阶段差异表达。对于转录组验证，选择了 10 个 CP、20E 和 JH 相关基因，并通过实时 PCR 定量验证。总的来说，我们的研究结果为进一步研究变态的分子机制提供了基础。M. separata.