Trehalose is the principal sugar circulating in the hemolymph of insects, and trehalose synthesis is catalyzed by trehalose-6-phosphate synthase (TPS) and trehalose-6-phosphate phosphatase (TPP). Insect TPS is a fused enzyme containing both TPS domain and TPP domain. Thus, many insects do not possess TPP genes as TPSs have replaced the function of TPPs. However, TPPs are widely distributed across the dipteran insects, while the roles they play remain largely unknown. In this study, 3 TPP genes from notorious dipteran pest Bactrocera minax (BmiTPPB, BmiTPPC1, and BmiTPPC2) were identified and characterized. The different temporal-spatial expression patterns of 3 BmiTPPs implied that they exert different functions in B. minax. Recombinant BmiTPPs were heterologously expressed in yeast cells, and all purified proteins exhibited enzymatic activities, despite the remarkable disparity in performance between BmiTPPB and BmiTPPCs. RNA interference revealed that all BmiTPPs were successfully downregulated after double-stranded RNA injection, leading to decreased trehalose content and increased glucose content. Also, suppression of BmiTPPs significantly affected expression of downstream genes and increased the mortality and malformation rate. Collectively, these results indicated that all 3 BmiTPPs in B. minax are involved in trehalose synthesis and metamorphosis. Thus, these genes could be evaluated as insecticidal targets for managing B. minax, and even for other dipteran pests.
Keywords: Bactrocera minax; RNA interference; heterologous expression; metamorphosis; trehalose; trehalose-6-phosphate phosphatase.
© 2022 Institute of Zoology, Chinese Academy of Sciences.