Single-cell RNA sequencing (scRNA-seq) has emerged as a powerful tool for studying complex cellular composition and gene expression dynamics of biological systems. In this study, we analyzed the midgut of the fall armyworm (FAW), Spodoptera frugiperda, utilizing scRNA-seq technology. scRNA-seq analysis yielded high-quality sequencing data from two replicates, showcasing robust sequencing integrity, mapping efficiency, and reproducibility. We identified twelve clusters of midgut cells, including enterocytes, enteroblasts, enteroendocrine cells, goblet cells, and stem cells, each with unique marker gene expression indicative of their specialized functions. Further analysis revealed intricate gene expression profiles and enriched biological pathways associated with each cell type, shedding light on the molecular mechanisms underlying midgut function. Additionally, lineage trajectory analysis identified the differentiation pathways of midgut cell populations, confirming canonical relationships among stem cells, enteroblasts, enterocytes, and goblet cells. Furthermore, we also studied the expression of genes coding for insecticide target sites and metabolizing enzymes in different midgut cell types. Overall, our studies provide a comprehensive understanding of midgut cellular diversity and gene expression dynamics in the FAW, offering valuable information that could be used to develop methods for managing this and other pests.

单细胞 RNA 测序 (scRNA-seq) 已成为研究生物系统复杂细胞组成和基因表达动态的强大工具。在本研究中，我们利用 scRNA-seq 技术分析了草地贪夜蛾 (FAW)、草地贪夜蛾 ( Spodoptera frugiperda)的中肠。 scRNA-seq 分析从两次重复中产生了高质量的测序数据，展示了强大的测序完整性、作图效率和重现性。我们鉴定了十二个中肠细胞簇，包括肠细胞、肠母细胞、肠内分泌细胞、杯状细胞和干细胞，每个细胞都具有表明其特殊功能的独特标记基因表达。进一步的分析揭示了与每种细胞类型相关的复杂的基因表达谱和丰富的生物途径，揭示了中肠功能的分子机制。此外，谱系轨迹分析确定了中肠细胞群的分化途径，证实了干细胞、肠母细胞、肠上皮细胞和杯状细胞之间的典型关系。此外，我们还研究了不同中肠细胞类型中编码杀虫剂靶位点和代谢酶的基因的表达。总的来说，我们的研究提供了对秋粘虫中肠细胞多样性和基因表达动态的全面了解，提供了有价值的信息，可用于开发管理这种害虫和其他害虫的方法。