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A real-time PCR assay to differentiate the B and Q biotypes of the Bemisia tabaci complex in Cyprus

Published online by Cambridge University Press:  10 February 2009

L.C. Papayiannis ,
J.K. Brown ,
N.A. Seraphides ,
M. Hadjistylli ,
N. Ioannou  and
N.I. Katis
L.C. Papayiannis*
Affiliation:
Agricultural Research Institute, 1516 Nicosia, PO Box 22016 Cyprus
J.K. Brown
Affiliation:
Department of Plant Sciences, University of Arizona, Tucson, AZ 85721, USA
N.A. Seraphides
Affiliation:
Agricultural Research Institute, 1516 Nicosia, PO Box 22016 Cyprus
M. Hadjistylli
Affiliation:
Department of Agriculture, 1412 Nicosia, Cyprus
N. Ioannou
Affiliation:
Department of Agricultural Sciences, Biotechnology and Food Science, Cyprus University of Technology, Limassol, Cyprus
N.I. Katis
Affiliation:
Aristotle University of Thessaloniki, School of Agriculture, Plant Pathology Laboratory, 54 124, Thessaloniki, Greece
*
*Author for correspondence Fax: +35722316770 E-mail: l.papayiannis@arinet.ari.gov.cy
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Abstract

A real-time PCR assay based on TaqMan® technology was developed and evaluated for the rapid detection of the B and Q biotypes of Bemisia tabaci (Gennadius) (Hemiptera: Aleyrodidae). A survey was conducted during 2005–2007 in order to identify the distribution and prevalence of B. tabaci biotypes in Cyprus using the real-time PCR assay. More than 700 adult whiteflies collected from 35 cultivated and weed plant species were individually haplotyped using TaqMan® PCR, and the results of the assay were validated by restriction fragment length polymorphism analysis and DNA sequencing of the mitochondrial cytochrome oxidase I (mtCOI) gene. Two biotypes, B and Q, were identified in the collected plant species on the island. The real-time PCR and RFLP assay consistently yielded the same results, although the real-time assay was more sensitive and less time consuming. Phylogenetic analysis of the mtCOI DNA sequences corroborated the identity of the B and Q biotypes 100% of the time and by phylogenetic analysis the haplotypes grouped, as expected, in the major North African-Mediterranean-Middle Eastern clade of the B. tabaci complex.

Keywords

Bemisia tabaci complexreal-time PCRhaplotypeRFLP analysismitochondrial COI gene
Type
Research Paper
Information
Bulletin of Entomological Research , Volume 99 , Issue 6 , December 2009 , pp. 573 - 582
Copyright
Copyright © 2009 Cambridge University Press

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