title: Development of Species-Specific Primers for Agronomical Thrips and Multiplex Assay for Quarantine Identification of Western Flower Thrips.
abstract: While morphological identification of thrips species has been difficult because of their minute size and a lack of easily recognizable characteristics, molecular identification based on the development of specific molecular markers can be easily and reliably carried out. Among the known molecular markers, the nuclear internal transcribed spacer (ITS) exhibits distinguishable variations among thrips species. In this study, sequences of ITS2 region of 10 agriculturally important thrips were established to design species-specific primers for polymerase chain reaction (PCR). ITS2 sequence variations within these species were far less than those among species, indicating the suitability of this marker for species-specific primers design. These primers, though with one or two sporadically variable positions, showed a good efficacy within species. The specificity of these primers, examined on thrips species belonging to 15 genera, proved satisfactory. Furthermore, a multiplex PCR was used successfully for identifying Frankliniella occidentalis (Pergande), an insect pest monitored for quarantine purpose, and three additional thrips also commonly found in imported agricultural products and field samples, i.e., Thrips tabaci Lindeman, Thrips hawaiiensis (Morgan), and Frankliniella intonsa (Trybom). This study has demonstrated that specific primers and multiplex PCR based on ITS2 are reliable, convenient, and diagnostic tool to discriminate thrips species of quarantine and agricultural importance.
authors: Department of Entomology, National Chung Hsing University; 250 Kuan-Kung Rd., Taichung, Taiwan 40227. wbyeh@nchu.edu.tw.
authors: W B
authors: WB
authors: Yeh
authors: Department of Entomology, National Chung Hsing University; 250 Kuan-Kung Rd., Taichung, Taiwan 40227.
authors: M J
authors: MJ
authors: Tseng
authors: Department of Plant Medicine, National Pingtung University of Science and Technology; 1 Shuefu Road, Neipu, Pingtung, Taiwan 91201.
authors: N T
authors: NT
authors: Chang
authors: Department of Entomology, National Chung Hsing University; 250 Kuan-Kung Rd., Taichung, Taiwan 40227.
authors: S Y
authors: SY
authors: Wu
authors: Department of Entomology, National Chung Hsing University; 250 Kuan-Kung Rd., Taichung, Taiwan 40227.
authors: Y S
authors: YS
authors: Tsai
mesh_terms: D000818:Animals; D017931:DNA Primers / Q000032:analysis; D021901:DNA, Intergenic / Q000235:genetics*; D060885:Multiplex Polymerase Chain Reaction; D010802:Phylogeny; D011790:Quarantine; D017422:Sequence Analysis, DNA; D013045:Species Specificity; D061149:Thysanoptera / Q000145:classification* / Q000235:genetics*
chemical_list: D017931:DNA Primers; D021901:DNA, Intergenic
keywords: ITS2; multiplex PCR; specific primer; thrip