PCR-based,rapid diagnosis of parasitism of Lygus spp. (Hemiptera: Miridae) by Peristenus relictus (Hymenoptera: Braconidae)

Several species of Lygus (Heteroptera: Miridae: Mirini) are serious crop pests in North America where their parasitism rate by native nymphal parasitoids is generally lower than in Europe. Peristenus relictus (Ruthe) (formerly P. stygicus Loan) (Hymenoptera: Braconidae: Euphorinae) is the predominant nymphal parasitoid of several Lygus spp. in the warm Mediterranean region and has been a candidate for introduction against Lygus hesperus Knight and L. lineolaris (Palisot de Beauvois) in the southern US. We report a rapid, sensitive, and specific PCR-based assay for diagnosis of P. relictus immature stages within Lygus nymphs that entails three steps: DNA extraction, PCR of the partial mitochondrial COI gene, and agarose gel electrophoresis. The PCR-based methodology is species-specific because the target DNA of other sympatric, congeneric species was not amplified with use of the primers developed for P. relictus diagnosis. The sensitivity of the PCR method, assessed through spike tests, was established by the detection of a ratio of 1:10,000 P. relictus DNA to Lygus DNA. Molecular diagnosis of parasitism of field collected nymphs is achievable in one day, eliminating the need to rear nymphs to obtain adult parasitoids for morphological identification.