Cyclaniliprole is a novel third-generation anthranilic diamide insecticide which has broad-spectrum efficacy against various pests, including aphids. Rhopalosiphum padi and Schizaphis graminum are the two primary aphid species that infest wheat crops. This investigation evaluates cyclaniliprole’s acute toxicity and sublethal and transgenerational effects on both aphid species. The acute toxicity assessment revealed obvious insecticidal activity, with 24 h LC₅₀ values of 38.56 mg/L for R. padi and 33.71 mg/L for S. graminum. Sublethal exposure (LC₁₅ and LC₃₅ in R. padi; LC₃₅ in S. graminum) significantly reduced adult longevity and fecundity in the F₀ generation. In the F₁ generation, cyclaniliprole at LC₃₅ significantly reduced the fourth nymph stage in R. padi, whereas at LC₁₅, it shortened the third nymph stage duration in S. graminum compared to the control. Sublethal concentrations (LC₁₅ and LC₃₅) of cyclaniliprole significantly reduced age-stage-specific survival rate (s_xj), age-specific survival rate (l_x), age-specific maternity (l_xm_x), and age-stage life expectancy (e_xj) in the F₁ generation of R. padi, while only LC₃₅ decreased l_x, l_xm_x, and e_xj in S. graminum. Additionally, LC₃₅ significantly reduced the net reproductive rate (R₀) in the F₁ generation of R. padi compared to the control. These findings suggest that cyclaniliprole exhibit notable acute toxicity against both aphid species and that sublethal concentrations adversely affected the F₀ generation, with no observed hormetic effects in the F₁ generations of R. padi and S. graminum. The findings offer valuable insights for assessing the comprehensive insecticidal potential of cyclaniliprole.

Hyblaea puera is a major pest of teak and mangroves. Reliable RT-qPCR normalization requires stable reference genes, none of which have been validated in H. puera. In this study, we assessed the expression stability of ten candidate reference genes under different experimental conditions. Stability was evaluated using the ΔCt method, BestKeeper, NormFinder, and geNorm, and a comprehensive stability ranking was generated using the RefFinder online tool. Our results indicated that amplification efficiencies ranged from 91.67% to 100.82%, with R² values exceeding 0.9901. The optimal reference gene combinations varied by condition: Ribosomal Protein L27 (RPL27) and Ribosomal Protein L10 (RPL10) for temperature treatments; Actin and RPL10 for larval instars; Ribosomal protein S5 (RPS5) and Elongation factor-1α (EF-1a) for adult sexes; RPL10 and EF-1a for tall developmental stages; RPL10 and RPS5 for tissues; as well as EF-1α and Actin for all combined conditions. Finally, the expression profiles of target gene Lethal were evaluated, and the outcomes further confirm the importance of selecting fitting reference genes for normalization of qRT-PCR data. These results provide the evaluated reference gene sets for H. puera, facilitating more accurate RT-qPCR normalization in future molecular studies of host plant adaptation (teak vs. mangroves), temperature tolerance, and larval development in this pest.