Abstract
Large-scale screenings for Wolbachia usually assess whether the endosymbiont is present in the specimens examined, but do not draw inferences on strain diversity within individuals, populations or species. However, multiple Wolbachia infections are common, and strain composition may have strong influence on the expression of Wolbachia phenotypes. Recently, high-resolution melting analysis (HRM) was introduced as a tool for the fast identification of characterized Wolbachia strains, with the major limitation of ambiguous results in multiply infected individuals. We present a different approach and utilize HRM for the fast and cost-effective de novo characterization of Wolbachia communities. Completely omitting the traditional end point PCR diagnostic step, a large sample of specimens with unknown infection status is subjected to amplification of a ca. 125 bp stretch containing the wsp HVR4 region and HRM analysis. By this, (i) a preliminary Wolbachia diagnosis for each sample and (ii) a set of melting curve profiles is retrieved. Subsequent cloning and sequencing of wsp and MLST genes can be focused on a few specimens representative for the distinct HRM profiles. We demonstrate the suitability of this approach by the de novo characterization of the Wolbachia communities of two arthropod species.