Testing the usage of ITS2 and 28S-D2 markers in identifying some species of Ammophila, Prionyx, and Sphex genera (Hymenoptera: Sphecidae)

In addition to traditional methods, investigation of rDNA gene regions with molecular techniques is widely used to identify insect taxa. In this study, the usability of the primers (TrITS2, 28S-D2) belonging to second expansion segment (D2) of the 28S subunit (28S-D2) and second internal transcribed spacer (ITS2) rDNA gene regions were tested to identify and differentiate six species of Sphecidae family namely Ammophila sabulosa, Ammophila heydeni, Prionyx kirbyi, Prionyx nudatus, Sphex flavipennis, Sphex funerarius for the first time. DNA isolated from samples were amplified by Polymerase Chain Reaction (PCR) and digested by restriction enzymes. PCR products and restriction enzyme analysis of ITS2 obtained from Prionyx species displayed species-specific DNA bands indicating that these species could be identified by using TrITS2 primers and restriction enzymes. However, neither DNA bands nor the sequence analysis of 28S-D2rDNA primers provided species-specific data. Consequently, ITS2 marker was suitable for only one of the genera (Prionyx), whereas 28S-D2 marker was suitable for none of the taxa examined.