Development of a continuous whitefly cell line [Homoptera : Aleyrodidae : Bemisia tabaci (Gennadius)] for the study of begomovirus

The whitefly Bemisia tabaci (Gennadius) is a widely distributed pest of man y important food and fiber crops, This whitefly is also a vector of more th an 70 plant-infecting viruses. A cell line was established in vitro using e mbryonic tissues from the eggs of Bemisia tabaci (Gennadius), B biotype (ps eudonym B, argentifolii Bellows & Perring), and referred to as 'Btb(Ba)97, Hunter-Polston', Cell cultures were successfully inoculated with begomoviru s (BGMV and ToMoV)-infected tomato plant sap. Embryonic tissues were seeded into Kimura's modified medium and kept at a temperature of 24 degreesC. Co ntinuous cell cultures were established and have since undergone 92 passage s in 25-cm(2) flasks. Cell doubling time is approximately 3 days and the ce lls have been successfully revived after 1 year after storage at -80 degree sC, The cell population is monolayers of predominately fibroblast with some epithelial cells. Begomoviruses (bean golden mosaic begomovirus, BGMV, and tomato mottle begomovirus, ToMoV) were inoculated to the cell cultures ind ependently and detected by labeling by an indirect immunofluorescence techn ique. The viruses were detected bound to the cell membranes and within the cell cytoplasm, This is the first report of a continuous cell line establis hed from a species of whitefly and its inoculation with two begomoviruses. The successful inoculation of whitefly cell cultures with begomoviruses sho wn in our results represents great promise for the development of systems t hat allow researchers to achieve a better understanding of the complex rela tionship between begomoviruses and their whitefly vectors. (C) 2001 Academi c Press.