ANNOTATION OF WHITEFLY EXPRESSED SEQUENCE TAGS AND VALIDATION OF GENES WITH POTENTIAL SIGNIFICANCE TO BEGOMOVIRUS TRANSMISSION

Abstract

The whitefly Bemisia tabaci (Gennadius) (Hemiptera) complex is the sole arthropod vector of the genus, Begomovirus (family, Geminiviridae), which causes debilitating diseases of plants, worldwide. Virus-vector specificity is conferred through co-evolved, whitefly vector-viral capsid protein-protein interactions. Membrane-bound receptors are thought to facilitate virion passage across the gut-hemolymph and hemolymph-salivary gland interfaces, and virion circulation is expected to elicit innate defense and stress-related proteins. Our goal was to select and validate genes involved in whitefly-mediated transmission. Whitefly expressed sequence tags (ESTs) from a previous study were re-annotated, taking advantage of newly available insect EST, UniGene, and Protein sequences. Six whitefly genes and transcripts, actin, cyclophilin, GBLP, GAPDH 3, knottin, and whitefly endosymbiont HSP60, representing three gene ontology (GO) categories, were analyzed using PCR or RT- PCR, respectively, followed by cloning and DNA sequencing. Analysis confirmed the presence of all six whitefly genes and five transcripts, with the knottin transcript being undetectable.