High Incidence of Related Wolbachia across Unrelated Leaf-Mining Diptera

SIMPLE SUMMARY: Polyphagous leaf-mining flies of the genus Liriomyza are pests that pose a serious threat to agricultural and horticultural industries. The endosymbiotic bacterium Wolbachia has been proposed as a useful biocontrol strategy for managing pests, but few studies have so far examined Wolbachia in leafminers. We find a high incidence of related Wolbachia in a survey of infections in 13 dipteran leafminer species collected from Australia and elsewhere which could potentially be useful for the incompatible insect technique (IIT) of pest suppression. We performed curing and crossing experiments on L. brassicae to demonstrate the presence of cytoplasmic incompatibility (CI) needed for IIT, providing a foundation for future transfection of CI Wolbachia from L. brassicae to other Liriomyza pests. Overall, these findings highlight a high incidence of Wolbachia in leaf-mining Diptera, potential horizontal transmission events and possible applications of Wolbachia-based biocontrol strategies for Liriomyza pests. ABSTRACT: The maternally inherited endosymbiont, Wolbachia pipientis, plays an important role in the ecology and evolution of many of its hosts by affecting host reproduction and fitness. Here, we investigated 13 dipteran leaf-mining species to characterize Wolbachia infections and the potential for this endosymbiont in biocontrol. Wolbachia infections were present in 12 species, including 10 species where the Wolbachia infection was at or near fixation. A comparison of Wolbachia relatedness based on the wsp/MLST gene set showed that unrelated leaf-mining species often shared similar Wolbachia, suggesting common horizontal transfer. We established a colony of Liriomyza brassicae and found adult Wolbachia density was stable; although Wolbachia density differed between the sexes, with females having a 20-fold higher density than males. Wolbachia density increased during L. brassicae development, with higher densities in pupae than larvae. We removed Wolbachia using tetracycline and performed reciprocal crosses between Wolbachia-infected and uninfected individuals. Cured females crossed with infected males failed to produce offspring, indicating that Wolbachia induced complete cytoplasmic incompatibility in L. brassicae. The results highlight the potential of Wolbachia to suppress Liriomyza pests based on approaches such as the incompatible insect technique, where infected males are released into populations lacking Wolbachia or with a different incompatible infection.