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Simple Summary: Spodoptera frugiperda of the family Noctuidae in the order Lepidoptera, is native to the Americas. It is characterized by its polyphagous nature and high reproductive capacity. S. frugiperda has caused significant socio-economic losses in many countries, primarily affecting Poaceae crops such as maize, rice, wheat, and sorghum. This study aims to analyze the members, differentiation, structure, and function of the JHBP gene family in S. frugiperda through bioinformatics approaches. Subsequently, we utilize real-time quantitative PCR (qPCR) to investigate the expression profiles of this gene family across different developmental stages and various tissues of S. frugiperda. The purpose of this analysis is to elucidate the potential roles of these genes, providing key target genes for future functional validation through gene knockdown and RNA interference (RNAi) techniques. This research offers insights that could contribute to the development of new pesticides. Juvenile hormone binding proteins (JHBPs) are carrier proteins that bind to juvenile hormone (JH) to form a complex, which then transports the JH to target organs to regulate insect growth and development. Through bioinformatics analysis, 76 genes encoding JHBP in S. frugiperda were identified from whole genome data (SfJHBP1-SfJHBP76). These genes are unevenly distributed across 8 chromosomes, with gene differentiation primarily driven by tandem duplication. Most SfJHBP proteins are acidic, and their secondary structures are mainly composed of α-helices and random coils. Gene structure and conserved motif analyses reveal significant variations in the number of coding sequences (CDS) and a high diversity in amino acid sequences. Phylogenetic analysis classified the genes into four subfamilies, with a notable presence of directly homologous genes between S. frugiperda and S. litura, suggesting a close relationship between the two species. RNA-seq data from public databases and qPCR of selected SfJHBP genes show that SfJHBP20, SfJHBP50, and SfJHBP69 are highly expressed at most developmental stages, while SfJHBP8 and SfJHBP14 exhibit specific expression during the pupal stage and in the midgut. These findings provide a theoretical basis for future studies on the biological functions of this gene family. [ABSTRACT FROM AUTHOR]

Simple Summary: The sorghum aphid, a significant pest affecting sorghum cultivation in several continents, feeds primarily on sorghum leaves and stems by sucking sap. This feeding behavior results in the depletion of nutrients, sugars, and water in the plants, leading to inhibited growth, chlorosis, wilting, and ultimately death. The main objective of this study was to assess the stability of potential reference genes in the sorghum aphid (Melanaphis sacchari). Nine candidate reference genes underwent a rigorous selection process, and their reliability was evaluated using qRT-PCR Ct values. Various experiments were conducted to determine the best reference genes for the sorghum aphid through systematic stability analyses. The research offers a dependable collection of reference genes that can improve studies on gene expression and functional genomics related to the sorghum aphid. Appropriate reference genes must be selected for accurate qRT-PCR data to conduct a thorough gene expression analysis in the sorghum aphid (Melanaphis sacchari, Hemiptera, Aphididae). This approach will establish a foundation for gene expression analysis and determines the efficacy of RNA interference in the sorghum aphid. Nine potential reference genes, including Actin, 18S, GAPDH, RPL7, EF-1α, EF-1β, 28S, HSP70, and TATA, were assessed under various experimental conditions to gauge their suitability based on qRT-PCR Ct values. The stability of these candidate reference genes in diverse experimental setups was analyzed employing several techniques, including the ΔCt comparative method, geNorm, Normfinder, BestKeeper, and RefFinder. The findings revealed that the quantity of ideal reference genes ascertained by the geNorm method for diverse experimental conditions remained consistent. For the developmental stages of the sorghum aphid, RPL7 and 18S proved to be the most dependable reference genes, whereas GAPDH and EF-1β were recommended as the most stable reference genes for different tissues. In experiments involving wing dimorphism, EF-1α and GAPDH were identified as the optimal reference gene pair. Under varying temperatures, EF-1α and EF-1β were found to be the most dependable gene pair. For studies focusing on insecticide susceptibility, 18S and TATA emerged as the most stable candidate reference genes. Across all experimental conditions, EF-1α and EF-1β was the optimal combination of reference genes in the sorghum aphid. This research has pinpointed stable reference genes that can be utilized across various treatments, thereby enhancing gene expression studies and functional genomics research on the sorghum aphid. [ABSTRACT FROM AUTHOR]