Utility of MtCOI polymerase chain reaction-restriction fragment length polymorphism in differentiating between Q and B whitefly Bemisia tabaci biotypes

The invasive, insecticide-resistant, Q whitefly biotype, has gradually spread to other countries including the US via human-mediated movement of plant materials. We assessed the utility of the VspI-based mtCOI (mitochondrion cytochrome oxidase I) polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) technique as a rapid, cost-effective, and reliable alternative for differentiating the Q from the dominant B biotype in Arizona. Using the standard mtCOI gene sequencing and mtCOI PCR-RFLP techniques, we biotyped eight whitefly strains of five individuals each collected from poinsettia and cotton at different locations in Arizona. Complete concordance was observed between the two methods, with three strains being identified as the Q biotype and five samples as the B biotype. We also scanned the mtCOI gene sequences for VspI polymorphisms in the B and Q biotype whiteflies currently available in the GenBank database. This global screening revealed the existence of three and four VspI polymorphic types for the Q and B biotypes, respectively. Nevertheless, all three VspI polymorphic Q biotype whiteflies shared a common and unique VspI site that can be used to differentiate Q biotype from the four VspI polymorphic B biotype whiteflies identified. These results demonstrate that the VspI-based mtCOI gene PCR-RFLP provides a reliable diagnostic tool for differentiating the Q and B biotype whiteflies in the US and elsewhere.