C-type lectin 9 participates in the immune response, development and reproduction of Tribolium castaneum

C-type lectins (CTLs), as a large family of pattern recognition receptors (PRRs), have been reported to be involved in bacterial infection, but the role of CTLs in development has been poorly understood in insects. The orthologues of Tribolium castaneum CTL9 (TcCTL9) have been identified among insects, but its functions were currently unclear. Therefore, we performed functional analysis of TcCTL9 in this study. Our results indicated that TcCTL9 could bind to bacteria through lipopolysaccharide and peptidoglycan, and agglutinate Gram-positive and Gram-positive bacteria in a Ca²⁺-dependent manner. Silencing TcCTL9 reduced the immune resistance to Staphylococcus aureus and Escherichia coli, decreased the expression of antimicrobial peptides and prophenoloxidase, and inhibited the phenoloxidase activity. These data suggested that TcCTL9 functioned in the immune response via the Toll and IMD pathways and prophenoloxidase system. During development, TcCTL9 had high expression in the periods of egg to larva and pupa to adult, and knockdown of TcCTL9 suppressed the metamorphosis, egg production and hatchability, and ovary development through ecdysone and juvenile hormone pathways in T. castaneum. This study comprehensively clarified the functions of TcCTL9 orthologues in insects and provided the theoretical basis for developing novel targets of pesticides.