Molecular Characterization of Primary Juvenile Hormone Responders Methoprene-Tolerant (Met) and Kruppel Homolog 1 (Kr-h1) in Grapholita molesta (Lepidoptera: Tortricidae) with Clarification of Their Roles in Metamorphosis and Reproduction

Methoprene-tolerant (Met) is a putative JH intracellular receptor that transduces JH signal by activation of the inducible Kruppel homolog 1 (Kr-h1). We analyzed the gene sequences of Met and Kr-h1 and their patterns of expression in Grapholita molesta (Busck) immature and adult stages in order to better understand the roles of these primary JH responders in regulating the metamorphosis and reproduction of this global pest of fruit crops. The deduced amino acid sequences of both GmMet and GmKr-h1 were highly homologous to those of other Lepidoptera, especially the cotton bollworm, Helicoverpa armigera (Hubner) (Lepidoptera: Noctuidae). Peak expression of GmMet occurred during the last 3 to 5 d of the final instar, followed by that of GmKr-h1, in the last 3 d of final instar. Similar patterns of GmMet and GmKr-h1 expression were detected across various tissue types in the fifth-instar larvae, with the highest expression observed in the head, followed by the epidermis, and the fat body. When expression of GmMet and GmKr-h1 was knocked down via dsRNA injection in the fifth instar, the results were increased larval mortality, abnormal pupation, delayed pupal duration, reduced adult emergence, extended preoviposition period, and reduced fecundity. We infer that both GmMet and GmKr-h1 participated in regulation of metamorphosis and reproduction in G. molesta, the former acting upstream of the latter, and could present biorational targets for novel pest control compounds.