Integrated Transcriptomic and Small RNA Sequencing Reveal miR-11–3p/ UGT383A3 Axis Mediates Thiamethoxam Susceptibility in Diaphorina Citri

Diaphorina citri is the primary vector of Huanglongbing (HLB), posing a severe threat to global citrus production. Prolonged thiamethoxam application has led to declining susceptibility in field populations, necessitating elucidation of the underlying molecular mechanisms. Here, we performed integrated transcriptomic and small RNA sequencing on D. citri adults exposed to sublethal thiamethoxam concentrations and identified a regulatory axis comprising miR-11–3p and UGT383A3 . RNA interference and miRNA mimic/inhibitor assays demonstrated that both miR-11–3p and UGT383A3 significantly modulate thiamethoxam susceptibility. Dual-luciferase reporter assays confirmed that miR-11–3p directly targets the 3′ UTR of UGT383A3 to negatively regulate its expression. Validation across three field populations (GDGZ, GDQY, and GDJM) showed that thiamethoxam exposure downregulated miR-11–3p and upregulated UGT383A3, and UGT383A3 knockdown increased thiamethoxam susceptibility. These findings reveal a novel miR-11–3p/ UGT383A3 regulatory axis governing thiamethoxam susceptibility in D. citri, providing a potential target for RNAi-based pest management.