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

• Published in: Bulletin of entomological research 2009-12, Vol.99 (6), p.573-582
• Main Authors: Papayiannis, L.C., Brown, J.K., Seraphides, N.A., Hadjistylli, M., Ioannou, N., Katis, N.I.
• Format: Article
• Language: English
• Subjects: Aleyrodidae; Animal populations; Animals; Bemisia argentifolii; Bemisia tabaci; Bemisia tabaci biotype Q; Bemisia tabaci complex; Biological and medical sciences; Biotypes; COI gene; Cyprus; cytochrome-c oxidase; Deoxyribonucleic acid; DNA; Electron Transport Complex IV - chemistry; Electron Transport Complex IV - genetics; Entomology; Fundamental and applied biological sciences. Psychology; Generalities; Genes, Mitochondrial; Genetic diversity; genetic variation; haplotype; Haplotypes; Hemiptera; Hemiptera - classification; Hemiptera - genetics; insect surveys; Insects; mitochondrial COI gene; mitochondrial DNA; molecular systematics; pest identification; Phylogenetics; Phylogeny; Phytopathology. Animal pests. Plant and forest protection; Plant species; polymerase chain reaction; Polymerase Chain Reaction - methods; Polymorphism; Polymorphism, Restriction Fragment Length; Protozoa. Invertebrates; rapid methods; real-time PCR; real-time polymerase chain reaction; restriction fragment length polymorphism; RFLP analysis; sequence analysis; Sequence Analysis, DNA
• ISSN: 0007-4853; 1475-2670
• DOI: 10.1017/S0007485308006603

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.