To aid in identifying key predators of the glassy-winged sharpshooter (GWSS) egg stage, we developed ‘one-step ’ molecular diagnostic markers specific toward the internal transcribed spacer 2 region (ITS2) of five primary GWSS egg parasitoid species belonging to the genus Gonatocerus Nees. In addition, these markers will also serve to aid taxonomic identifications, especially before the egg parasitoids emerge from GWSS egg masses. The molecular markers were designed toward the following species: G. triguttatus (Gtri-F/R); G. walkerjonesi (wjca-F2/R); G. morrilli (gmtx-F/R); G. ashmeadi (Gash-F/R); and G. morgani (Gmorg-F/R). Cross-reactivity assays with 21 Gonatocerus species, each with specific assay conditions, demonstrated that the developed markers were highly specific toward the species that they were designed for, as crossreactivity was not observed with non-target species. One of the markers (Gtri-F/R) was tested in predator gut content assays. Positive reactions were observed in gut assays in which lacewings were allowed to feed on parasitized (G. triguttatus) GWSS eggs. In addition, positive banding was also seen with GWSS parasitized eggs alone, that is before the egg parasitoids emerged from the GWSS egg masses. The current results confirm the specificity and utility of the various one-step molecular diagnostic markers. In addition to the predation studies, these markers should be useful to the biological control program
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