Identification of Aphid (Homoptera: Aphididae) Species and Clones by Random Amplified Polymorphic DNA

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Abstract
Identification of six aphid species was possible using the random polymorphic amplified DNA (RAPD-PCR) technique. This is a variant of the polymerase chain reaction (PCR) in which a single primer of random sequence is annealed at low temperatures. A set of six primers was evaluated in the identification of a total of 19 clones of the following species: Myzus persicae (Sulzer), Rhopalosiphum padi (L.), Aphis gossypii (Glover), Aphis fabae (Scopoli), Aphis craccivora (Koch), and Acyrthosiphon pisum (Harris). Reactions made with three primers (OPA-02, OPA-07, and OPA-09) produced species-specific DNA bands. Some intraspecific polymorphism was also detected between clones of R. padi and M. persicae when reactions were primed with OPA-02 and OPA-07. Reproducibility of the RAPD-PCR reactions was tested when using two different DNA extraction procedures. The results indicated that the best reproducibility was obtained when using the isopropanol-precipitation extraction procedure. Amplification products were also obtained when working with single aphid extracts, although the percentage of successful reactions was partially reduced.

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