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

  • L.C. Papayiannis (a1), J.K. Brown (a2), N.A. Seraphides (a1), M. Hadjistylli (a3), N. Ioannou (a4) and N.I. Katis (a5)...

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.

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*Author for correspondence Fax: +35722316770 E-mail: l.papayiannis@arinet.ari.gov.cy

References

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Bedford, I.D., Briddon, R.W., Brown, J.K., Rosell, R.C. & Markham, P.G. (1994) Geminivirus transmission and biological characterization of Bemisia tabaci (Gennadius) biotypes from different geographic regions. Annals of Applied Biology 125, 311325.
Berry, S., Rey, M.E.C., Rogan, D., Fondong, V.N., Fauquet, C.M. & Brown, J.K. (2004) Molecular evidence for distinct Bemisia tabaci geographicaly genotypes from cassava in Africa. Annals of the Entomological Society of America 97, 852859.
Boonham, N., González Pérez, L., Mendez, M.S., Lilia Peralta, E., Blockley, A., Walsh, K., Barker, I. & Mumford, R.A. (2004) Development of a real-time RT-PCR assay for the detection of Potato spindle tuber viroid. Journal of Virological Methods 116, 139146.
Bosco, D., Loria, A., Sartor, C. & Cenis, J.L. (2006) PCR-RFLP Identification of Bemisia tabaci Biotypes in the Mediterranean Basin. Phytoparasitica 34, 243251.
Brown, J.K. (2000) Molecular markers for the identification and global tracking of whitefly vector-Begomovirus complexes. Virus Research 71, 233260.
Brown, J.K., Frohlich, D. & Rosell, R. (1995a) The sweetpotato/silverleaf whiteflies: biotypes of Bemisia tabaci (Genn.), or a species complex? Annual Review of Entomology 40, 511534.
Brown, J.K., Coats, S.A., Bedford, I.D., Markham, P.G., Bird, J. & Frohlich, D.R. (1995b) Characterization and distribution of esterase electromorphs in the whitefly, Bemisia tabaci (Genn.) (Homoptera: Aleyrodidae). Biochemical Genetics 33, 205214.
Brown, J.K., Perring, T.M., Cooper, A.D., Bedford, I.D. & Markham, P.G. (2000) Genetic analysis of Bemisia tabaci (Hemiptera: Aleyrodidae) populations by isoelectric focusing electrophoresis. Biochemical Genetics 38, 1325.
Campbell, B.C., Steffen-Campbell, J.D. & Gill, R. (1994) Evolutionary origin of whiteflies (Hemiptera: Sternorrhyncha: Aleyrodidae) inferred from 18S rDNA sequences. Insect Molecular Biology 3, 7389.
Cenis, J.L., Perez, P. & Fereres, A. (1993) Identification of aphid (Homoptera: Aphididae) species and clones by random amplified polymorphic DNA. Annals of the Entomological Society of America 86, 545550.
Cervera, M.T., Cabezas, J.A., Simón, B., Martínez-Zapater, J.M., Beitia, F. & Cenis, J.L. (2000) Genetic relationships among biotypes of Bemisia tabaci (Hemiptera: Aleyrodidae) based on AFLP analysis. Bulletin of Entomological Research 90, 391396.
Costa, H.S. & Brown, J.K. (1991) Variation in biological characteristics and in esterase patterns among populations of Bemisia tabaci (Genn.) and the association of one population with silverleaf symptom development. Entomologia et Experimentalis Applicata 61, 211219.
Costa, H.S., Brown, J.K., Sivasupramaniam, S. & Bird, J. (1993) Regional distribution, insecticide resistance, and reciprocal crosses between the ‘A’ and ‘B’ biotypes of Bemisia tabaci. Insect Science and Application 14, 127138.
De Barro, P.J., Driver, F., Trueman, J.W.H. & Curran, J. (2000) Phylogenetic relationships of world populations of Bemisia tabaci (Gennadius) using ribosomal ITS1. Molecular Biology and Evolution 16, 2936.
De la Rúa, P., Simón, B., Cifuentes, D., Martínez-Mora, C. & Cenis, J.L. (2006) New insights into the mitochondrial phylogeny of the whitefly Bemisia tabaci (Hemiptera: Aleyrodidae) in the Mediterranean Basin. Journal Zoological Systematics and Evolution Research 44, 2533.
Frohlich, D.R., Torres-Jerez, I., Bedford, I.D., Markham, P.G. & Brown, J.K. (1999) A phylogeographical analysis of the Bemisia tabaci species complex based on mitochondrial DNA markers. Molecular Ecology 8, 16831691.
Horowitz, A.R., Denholm, I., Gorman, K., Cenis, J.L., Kontsedalov, S. & Ishaaya, I. (2003) Biotype Q of Bemisia tabaci identified in Israel. Phytoparasitica 31, 9498.
Ioannou, N. (1985) Yellow leaf curl and other virus diseases of tomato in Cyprus. Plant Pathology 34, 428434.
Jones, C.M., Gorman, K., Denholm, I. & Williamson, M.S. (2008) High-throughput allelic discrimination of B and Q biotypes of the whitefly, Bemisia tabaci, using TaqMan allele-selective PCR. Pest Management Science 64, 1215.
Jones, D.R. (2003) Plant viruses transmitted by whiteflies. European Journal of Plant Pathology 109, 195219.
Khasdan, V., Levin, I., Rosner, A., Morin, S., Kontsedalov, S., Maslenin, L. & Horowitz, A.R. (2005) DNA markers for identifying biotypes B and Q of Bemisia tabaci (Hemiptera: Aleyrodidae) and studying population dynamics. Bulletin of Entomological Research 95, 605613.
Kumar, S., Tamura, K. & Nei, M. (2004) MEGA3: Integrated software for molecular evolutionary genetics analysis and sequence alignment. Briefings in Bioinformatics 5, 150163.
Martin, J.H., Mifsud, D. & Rapisarda, C. (2000) The whiteflies (Hemiptera: Aleyrodidae) of Europe and the Mediterranean Basin. Bulletin of Entomological Research 90, 407448.
Mound, L.A. & Halsey, S.H. (1978) Whiteflies of the World: A Systematic Catalogue of the Aleyrodidae (Homoptera) with Host Plant and Natural Enemy Data. Chichester, UK, British Museum (Natural History) & John Wiley & Sons.
Papayiannis, L.C., Ioannou, N., Boubourakas, I.N., Dovas, C.I., Katis, N.I. & Falk, B.W. (2005) Incidence of viruses infecting cucurbits in Cyprus. Journal of Phytopathology 153, 530535.
Papayiannis, L.C., Ioannou, N., Dovas, C.I., Maliogka, V.I. & Katis, N.I. (2006) First report of Tomato chlorosis virus in Cyprus. Plant Pathology 55, 567.
Pascual, S. & Callejas, C. (2004) Intra- and interspecific competition between biotypes B and Q of Bemisia tabaci (Hemiptera: Aleyrodidae) from Spain. Bulletin of Entomological Research 94, 369375.
Perring, T.M. (2001) The Bemisia tabaci species complex. Crop Protection 20, 725737.
Qiu, B.L., Coats, S.A., Ren, S.X., Idris, A.M., Caixia, X.U. & Brown, J.K. (2007) Phylogenetic relationships of native and introduced Bemisia tabaci (Homoptera: Aleyrodidae) from China and India based on mtCOI sequencing and host plant comparisons. Progress in Natural Sciences 17, 645654.
Schuster, D.J., Kring, J.B. & Price, J.F. (1991) Association of the Sweetpotato Whitefly with a Silverleaf Disorder of Squash. HortScience 26, 155156.
Sseruwagi, P., Legg, J.P., Maruthi, M.N., Colvin, J., Rey, M.E.C. & Brown, J.K. (2005) Genetic diversity of Bemisia tabaci (Gennadius) (Hemiptera: Aleyrodidae) populations and presence of the B and a non-B biotype that is capable of inducing silverleaf symptoms in squash, in Uganda. Annals of Applied Biology 147, 253265.
Thompson, J.D., Gibson, T.J., Plewniak, F., Jeanmougin, F. & Higgins, D.G. (1997) The ClustalX windows interface: flexible strategies for multiple sequence alignment aided by quality analysis tools. Nucleic Acids Research 24, 48764882.
Tsangarakou, A., Tsigenopoulos, C.S., Gorman, K., Lagnel, J. & Bedford, I.D. (2006) Biotype status and genetic polymorphism of the whitefly Bemisia tabaci (Hemiptera: Aleyrodidae) in Greece: mitochondrial DNA and microsatellites. Bulletin of Entomological Research 97, 2940.
Viscarret, M.M., Torres-Jerez, I., Agostini de Manero, E., López, S.N., Botto, E.E. & Brown, J.K. (2003) Mitochondrial DNA evidence for a distinct clade of New World Bemisia tabaci (Genn.) (Hemiptera:Aleyrodidae) from Argentina and Bolivia, and presence of the Old World B biotype in Argentina. Annals of the Entomological Society of America 96, 572.
Vos, P., Hogers, R., Bleeker, M., Reijians, M., Lee, T., Hornes, M., Frijters, A., Pot, J., Peleman, J., Kuiper, M. & Zabeau, M. (1995) AFLP, a new technique for DNA fingerprinting. Nucleic Acids Research 23, 44074414.
Yokomi, R.K., Hoelmer, K.A. & Osborne, L.S. (1990) Relationship between the sweetpotato whitefly and the squash silverleaf disorder. Phytopathology 80, 895900.

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

  • L.C. Papayiannis (a1), J.K. Brown (a2), N.A. Seraphides (a1), M. Hadjistylli (a3), N. Ioannou (a4) and N.I. Katis (a5)...

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