Abai, MR

6th International Congress of DipterologyRapid discrimination of species of several quarantineimportant tephritids (Diptera: Tephritidae) by using biochipChen, Y.-H. (1) & K.-H. Lu (1)(1) Department of Entomology, National Chung Hsing University, Taichung, TaiwanTraditionally, species discrimination of pests found in quarantineinspection has to rely mostly on their morphological characters. However,it is not easy to identify the species of most tephretid larvae or even eggs,which the stages are often occurring in fruits, by morphology. In order tosolve above-mentioned difficulty, application of DNA markers to assist inspecies identification has been adopted in recent years. In this study, weselected 6 fruit and melon flies, including Bactrocera dorsalis, B.Cucubitae, B. tau, Ceratitis capitata, Anastrepha suspensa and Rhagoletispomonella, which are important tephritid pests on the list of Taiwanesequarantine, to develop a biochip for rapid species discrimination. Threevariable genomic regions, i.e. cytochrome oxidase I (COI), intergenicspacer 1 (ITS1) and intergenic spacer 2 (ITS2), respectively, were selectedto develop specific probes for developing the biochip. We first obtainedthe DNA fragments from each species with PCR amplification usingspecific primer sets. After DNA sequencing and alignment, severalspecies-specific probes have been designed by following the most variableregions of species. However, the specificity of each probe is currentlyunder investigation. We expect to develop a chip with rapid, high sensitiveand accurate features and to be used to discriminate unknown species oftephritids for quarantine purpose.Key Words: tephritid fly, discrimination, biochip, cytochrome oxidase I,intergenic spacer 1 & 2∗∗∗∗∗∗∗∗∗∗43