Chiu, Y.C., Wu, W.J., & Shih, C.J. 2001 [Identification of three Aulacaspis species (Homoptera: Diaspididae) by PCR-RFLP analysis for quarantine application.]. Formosan Entomologist 21(4): 365-375.

Notes: Most scale insects are small and similar in appearance. It is difficult to identify these species especially in their immature stages, and this causes problems for quarantine officers. This study attempted to develop a molecular diagnostic technique using genomic DNA markers for identifying three species of the genus Aulacaspis murrayae, A. yabunikkei and A. yasumatsui. Amur 18S-3 and ITS6 primer sets were used to amplify ITS1 and its flanking regions of rDNA from genomic DNA as a template extracted from a single specimen. DNA extracted from these three species yielded a single fragment after PCR amplification. The DNA fragment sizes of the PCR products were approx equal to 450 bp (for A. yabunikkei) and 500 bp (for A. murrayae and A. yasumatsui). These PCR products were then cut with various restriction endonucleases to compare the results of the restriction fragment length polymorphism. Our results also reveal that it is possible to discriminate these three Aulacaspis species based on the species-specific patterns acquired from digesting the PCR products with the endonucleases, Taq I, Msp I, Cfo I and Hae III.