Parasitism by Diachasmimorpha longicaudata Resolved by Molecular Markers

NATALIA PETIT-MARTY; DIEGO F. SEGURA; MARIANA M. VISCARRET; LEONELA Z. CARABAJAL PALADINO; MARÍA E. UTGÉS; SILVIA LANZAVECCHIA; RUBEN ZANDOMENI; JORGE L. CLADERA

Florida, EEUU

Congreso; 5th Meeting of the Working Group on Fruit Flies of the Western Hemisphere; 2004

Fruit flies control programs are increasing the use of the biological control with parasitoids, alone or in combination with SIT. Thereby, parasitism information is important and needed when monitoring the establishment and effectiveness of parasitoids released in control programs. Currently, parasitism diagnosis involves identification at larvae stage, which requires training and specialized knowledge or waiting for adults to emerge which is to time consuming. Simple and accurate methods for parasitism rates determination may be resolved using molecular markers. Diachasmimorpha longicaudata (Himenoptera: Braconidae) is a parasitoid of several fruit flies larval stages, including Ceratitis spp., Anastrepha spp. and Bactrocera spp. (Dipera: Tephritidae). In this work we use ITS1 DNA marker to differentiate pupae of Ceratitis capitata and Anastrepha fraterculus infested by Diachasmimorpha longicaudata from those not infested. We did this study using PCR with specific primers flanking the ITS1 (internal transcribed spacer 1) of ribosomal nuclear genes and amplifying fragment sequences. In D. longicaudata: C. capitata parasitism assay, ITS1 DNA fragment obtained by PCR allowed us to distinguish easily C. capitata pupae from D. longicaudata infested C capitata pupae. In order to distinguish A. fraterculus from D. longicaudata, ITS1 fragment restriction maps were made based on DNA sequences obtained. Two differentiating endonulceases enzymes were found. Parasitism rates obtained with this molecular marker were compared with emerged adults counts. The molecular marker analyzed here proved successful.