Standardised molecular diagnostic tool for the identification of cryptic species within the Bemisia tabaci complex

BACKGROUND The whitefly Bemisia tabaci complex harbours over 40 cryptic species that have been placed in 11 phylogenetically distinct clades based on the molecular characterisation of partial mitochondrial DNA COI (mtCOI) gene region. Four cryptic species are currently within the invasive clade i.e. ‘MED’, ‘MEAM1’, ‘MEAM2’ and IO’. Correct identification of these species is a critical step towards implementing reliable measures for plant biosecurity and border protection, however no standardised B. tabaci-specific primers are currently available which has caused inconsistencies in the species identification processes. RESULTS We report three sets of PCR primers developed to amplify the mtCOI region which can be used for genotyping ‘MED’, ‘MEAM1’, and ‘IO’ species and tested these primers on 91 MED, 35 MEAM1 and five IO individuals. PCR and sequencing of amplicons identified a total of 21, six and one haplotypes in MED, MEAM1 and IO respectively, of which six haplotypes were new to the B. tabaci database. CONCLUSION These primer pairs enabled standardisation and robust molecular species identification via mtCOI screening of the targeted invasive cryptic species and will improve quarantine decisions. The use of this diagnostic tool could be extended to other species within the complex.