title: Agronomically important thrips: development of species-specific primers in multiplex PCR and microarray assay using internal transcribed spacer 1 (ITS1) sequences for identification.
abstract: Thrips, the sole vector of plant Tospovirus, are major pests of many agricultural crops throughout the world. Molecular approaches have been applied in recent decades to identify these minute and morphologically difficult to distinguish insects. In this study, sequences of internal transcribed spacer 1 (ITS1) region of 15 agronomically important thrips, including several virus transmission species, have been analyzed in order to design species-specific primers for multiplex PCR and probes for microarray assay. That the ITS1 sequence distances within species were smaller than those among species suggests that the ITS1 fragment can be used for thrips species identification. The specificity and stability of these primers, combined with universal paired primers, were tested and verified in multiplex PCR. Using these specific primers as probes, microarray assay showed that PCR products of all thrips species hybridized consistently to their corresponding probes, though some signals were weak. We have demonstrated that multiplex PCR using specific primers based on ITS1 sequences is a simple, reliable, and cost-effective diagnostic tool for thrips species identification. Moreover, the DNA microarray assay is expected to extend into a reliable high-throughput screening tool for the vast numbers of thrips.
authors.affiliation: Department of Entomology,National Chung Hsing University,250 Kuan-Kung Rd., Taichung 40227,Taiwan.
authors.forename: W B
authors.initials: WB
authors.lastname: Yeh
authors.affiliation: Department of Entomology,National Chung Hsing University,250 Kuan-Kung Rd., Taichung 40227,Taiwan.
authors.forename: M J
authors.initials: MJ
authors.lastname: Tseng
authors.affiliation: Department of Plant Medicine,National Pingtung University of Science and Technology,1 Shuefu Rd., Neipu, Pingtung 91201,Taiwan.
authors.forename: N T
authors.initials: NT
authors.lastname: Chang
authors.affiliation: Department of Entomology,National Chung Hsing University,250 Kuan-Kung Rd., Taichung 40227,Taiwan.
authors.forename: S Y
authors.initials: SY
authors.lastname: Wu
authors.affiliation: Department of Entomology,National Chung Hsing University,250 Kuan-Kung Rd., Taichung 40227,Taiwan.
authors.forename: Y S
authors.initials: YS
authors.lastname: Tsai
mesh_terms: D000818:Animals; D017931:DNA Primers / Q000235:genetics; D021903:DNA, Ribosomal Spacer; D007300:Insect Control / Q000379:methods*; D008969:Molecular Sequence Data; D060885:Multiplex Polymerase Chain Reaction; D020411:Oligonucleotide Array Sequence Analysis; D017422:Sequence Analysis, DNA; D013045:Species Specificity; D013624:Taiwan; D061149:Thysanoptera / Q000145:classification / Q000235:genetics*
chemical_list: D017931:DNA Primers; D021903:DNA, Ribosomal Spacer
keywords: ITS1; microarray; multiplex PCR; species-specific probes; thrips