IDENTIFICATION OF APHID (HOMOPTERA, APHIDIDAE) SPECIES AND CLONES BY RANDOM AMPLIFIED POLYMORPHIC DNA

Identification of six aphid species was possible using the random poly morphic amplified DNA (RAPD-PCR) technique. This is a variant of the p olymerase chain reaction (PCR) in which a single primer of random sequ ence is annealed at low temperatures. A set of six primers was evaluat ed in the identification of a total of 19 clones of the following spec ies: Myzus persicae (Sulzer), Rhopalosiphum padi (L.), Aphis gossypii (Glover), Aphis fabae (Scopoli), Aphis craccivora (Koch), and Acyrthos iphon pisum (Harris). Reactions made with three primers (OPA-02, OPA-0 7, and OPA-09) produced species-specific DNA bands. Some intraspecific polymorphism was also detected between clones of R. padi and M. persi cae when reactions were primed with OPA-02 and OPA-07. Reproducibility of the RAPD-PCR reactions was tested when using two different DNA ext raction procedures. The results indicated that the best reproducibilit y was obtained when using the isopropanol-precipitation extraction pro cedure. Amplification products were also obtained when working with si ngle aphid extracts, although the percentage of successful reactions w as partially reduced.