First detection of Bemisia tabaci (Hemiptera: Aleyrodidae) MED in Oklahoma and development of a high-resolution melting assay for MEAM1 and MED discrimination

The sweetpotato whitefly, Bemisia tabaci (Gennadius) (Hemiptera: Aleyrodidae), is a polyphagous pest recognized as composed of several cryptic or sibling species. The Middle East-Asia Minor 1 (MEAM1) and the Mediterranean (MED) putative species are invasive and destructive worldwide. The MEAM1 is established throughout the United States, while MED is documented in 27 states. This study determines the status of MED in Oklahoma and develops and validates a high-resolution melting (HRM) assay for discrimination of MEAM1 and MED. In August-October 2022, whiteflies were collected from different host plants in Stillwater, Oklahoma, and identified as species based on analysis of a diagnostic fragment of the mitochondrial cytochrome oxidase I (mtCOI) gene. MED was found in mixed infestations with MEAM1 on both sweetpotato in a greenhouse and cucumber in the field. Other cryptic species were not detected. Sequencing followed by phylogenetic analysis indicated that the MED specimens belonged to the Q2 mitotype. Additionally, the secondary endosymbionts in captured and progeny whiteflies were identified. For rapid discrimination of MEAM1 and MED species, an HRM assay using a single set of primer pairs targeting the mtCOI gene was developed. Species discrimination was tested in 2 laboratories using MEAM1 and MED Q2 mitotype genomic DNA, and a synthetic plasmid containing the MED Q1 mitotype mtCOI fragment. The HRM assay was validated to discriminate MEAM1 from MED Q1 and Q2 mitotypes. This is the first report of B. tabaci MED in Oklahoma and reinforces the need for continued monitoring of this insect species complex.