Mitochondrial DNA Variability and Development of a PCR Diagnostic Test for Populations of the Whitefly Bemisia afer (Priesner and Hosny)

Abstract

The whitefly, Bemisia afer (Hemiptera; Aleyrodidae), is emerging as a major agricultural pest. The current identification methods based on adult and pupal morphology are laborious and unreliable. A diagnostic polymerase chain reaction (PCR) protocol was developed for the first time in this study to discriminate B. afer from other whitefly species. Primers specific to mitochondrial cytochrome oxidase I gene (mtCOI) were designed to amplify a band of approx 650 bp. The PCR products were sequenced from B. afer samples collected from Malawi, Tanzania, Uganda, Zanzibar, and the United Kingdom. Phylogenetic analyses of mtCOI sequences and those of reference B. afer sequences clustered the African B. afer separately from the UK and Chinese populations and from other whitefly species. The African cluster was divided into two clades by parsimony and neighbor-joining methods. This indicates the existence of at least two genotypic clusters of B. afer, which are diverged by 0.8 to 3.2% nucleotide (nt) identities. Analysis of molecular variance indicated that these differences were the result of within population variation but were insufficient to identify discrete populations. Among the whitefly species used in the analysis, B. afer was equally dissimilar to Bemisia tabaci and Bemisia tuberculata (21.3-26.2% nt identities). As is the case for B. tabaci, these data show that mtCOI sequences are informative also for identifying B. afer variants, which lack distinguishing morphological features.