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Agronomically important thrips: development of species-specific primers in multiplex PCR and microarray assay using internal transcribed spacer 1 (ITS1) sequences for identification

Published online by Cambridge University Press:  22 October 2014

W.B. Yeh ,
M.J. Tseng ,
N.T. Chang ,
S.Y. Wu  and
Y.S. Tsai
W.B. Yeh*
Affiliation:
Department of Entomology, National Chung Hsing University, 250 Kuan-Kung Rd., Taichung 40227, Taiwan
M.J. Tseng
Affiliation:
Department of Entomology, National Chung Hsing University, 250 Kuan-Kung Rd., Taichung 40227, Taiwan
N.T. Chang
Affiliation:
Department of Plant Medicine, National Pingtung University of Science and Technology, 1 Shuefu Rd., Neipu, Pingtung 91201, Taiwan
S.Y. Wu
Affiliation:
Department of Entomology, National Chung Hsing University, 250 Kuan-Kung Rd., Taichung 40227, Taiwan
Y.S. Tsai
Affiliation:
Department of Entomology, National Chung Hsing University, 250 Kuan-Kung Rd., Taichung 40227, Taiwan
*
*Author for correspondence Phone: +886-4-22840799 ext. 558 Fax: +886-4-22875024 E-mail: [email protected]
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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.

Keywords

thripsITS1multiplex PCRspecies-specific probesmicroarray
Type
Research Papers
Information
Bulletin of Entomological Research , Volume 105 , Issue 1 , February 2015 , pp. 52 - 59
Copyright
Copyright © Cambridge University Press 2014 

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