RNAi-mediated knockdown of HcCAT2 depresses the adaptive capacity of Hyphantria cunea larvae to cytisine and coumarin

The diversity of host plants is an important reason for the global spread of Hyphantria cunea. However, no studies have explored the role of the antioxidant defense system with catalase (CAT) as the core at the molecular level in the adaptation of the H. cunea to host plant secondary metabolites. Herein, the purpose is to explore how HcCAT2, highly expressed in cytisine- or coumarin-treated H. cunea larvae, mediates the adaptation of H. cunea to cytisine and coumarin, and to develop nucleic acid pesticides targeting HcCAT2. Findings revealed that H. cunea larvae treated with dsHcCAT2 alongside cytisine or coumarin exhibited significantly reduced body weight, survival rate, and expression levels of growth-related genes, energy metabolism genes, and oxidative damage regulatory genes compared to treated with cytisine or coumarin alone. HcCAT2 overexpression enhanced cell viability, lowered apoptosis rates, Ca²⁺ concentrations, ROS levels, and MPTP opening, and increased mitochondrial membrane potential in cytisine or coumarin-treated SF9 cells. Encapsulation of dsHcCAT2 in chitosan (CS) improved stability and gene silencing efficacy. CS-dsHcCAT2 treatment did not significantly affect SF9 cell, Lymantria dispar larvae, and Arma chinensis nymphs. However, H. cunea larvae treated with CS-dsHcCAT2 combined with cytisine or coumarin showed significantly reduced body weight and survival compared to those receiving secondary metabolites alone. Therefore, HcCAT2 is a critical antioxidant defense gene for adaptation of H. cunea larvae to cytisine and coumarin stress, with the ability of maintaining energy metabolism homeostasis and antioxidant defense level. The constructed CS-dsHcCAT2 can be developed as a synergistic agent for plant-derived pesticides.