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Title: SPECIES-SPECIFICITY AND SENSITIVITY OF A PCR-BASED ASSAY FOR PERISTENUS STYGICUS LOAN (BRACONIDAE), A PARASITOID OF LYGUS SPP. (MIRIDAE)

Author
item BON, MARIE-CLAUDE - USDA-ARS-EBCL
item RAMUALDE, NATHALIE - USDA-ARS-EBCL
item Hoelmer, Kim
item COUTINOT, DOMINIQUE - USDA-ARS-EBCL

Submitted to: Proceedings of First International Symposium on Biological Control of Arthropods
Publication Type: Proceedings
Publication Acceptance Date: 1/31/2003
Publication Date: 5/1/2003
Citation: BON, M., RAMUALDE, N., HOELMER, K.A., COUTINOT, D. SPECIES-SPECIFICITY AND SENSITIVITY OF A PCR-BASED ASSAY FOR PERISTENUS STYGICUS LOAN (BRACONIDAE), A PARASITOID OF LYGUS SPP. (MIRIDAE). PROCEEDINGS OF FIRST INTERNATIONAL SYMPOSIUM ON BIOLOGICAL CONTROL OF ARTHROPODS. 2003.

Interpretive Summary: Peristenus stygicus is a European natural enemy of several tarnished plant bugs of the genus Lygus. Plant bugs are serious crop pests in North America. The southern Mediterranean parasitoid wasp Peristenus stygicus is a promising biocontrol agent for reducing populations of southwestern Lygus species. Because natural populations of Lygus and their parasitoids are relatively uncommon in Europe and because several different Peristenus parasitoids often occur together at many locations, it has been necessary to devote considerable time and effort to field collections in order to obtain large numbers of parasitoids for research on this species and for shipment to cooperators in the United States. Traditional methods for monitoring the presence and identity of Lygus parasitoids in host populations include a combination of dissection and multi-year rearing, both difficult and time-consuming processes. Here we describe a method for efficient and rapid DNA isolation from Lygus nymphs for accurate diagnosis of parasitism. Our results indicate that the polymerase chain reaction method is species-specific and highly sensitive capable of detecting a ratio of as little as 1:10,000 parasitoid DNA to Lygus DNA. This technique can easily be expanded to other parasitoid-host systems, giving it broad applicability.

Technical Abstract: Peristenus styicus (Hymenoptera: Braconidae) is a european nymphal parasitoid of several Lygus spp. (Heteroptera: Miridae). Lygus spp. are serious crop pests in North America. The southern Mediterranean distribution of P. stygicus has increased interest in the biological control of the southwestern U.S. Lygus species. Traditional methods for monitoring the presence and identity of Lygus parasitoids in host populations include a combination of dissection and rearing, both difficult and time-consuming processes. Here we describe a method for efficient and rapid DNA isolation from Lygus nymphs that facilitates sensitive, species-specific PCR-based diagnosis of parasitism. Our results indicate that the PCR-based parasitoid species was not amplified with use of the primers developed for P. stygicus diagnosis. The sensitivity of the PCR method, as assessed through spike and recovery experiments, was established by the detection of a ratio of 1:10,000 parasitoid DNA to Lygus DNA. Although this technique was developed to detect P. stygicus within Lygus spp, it can be expanded to other parasitoid-host systems, giving it broad applicability.