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

Abstract

The whitefly Bemisia tabaci (Gennadius) is a widely distributed pest of many important food and fiber crops. This whitefly is also a vector of more than 70 plant-infecting viruses. A cell line was established in vitro using embryonic tissues from the eggs of Bemisia tabaci (Gennadius), B biotype (pseudonym B. argentifolii Bellows & Perring), and referred to as 'Btb(Ba)97, Hunter-Polston'. Cell cultures were successfully inoculated with begomovirus (BGMV and ToMoV)-infected tomato plant sap. Embryonic tissues were seeded into Kimura's modified medium and kept at a temperature of 24 degrees C. Continuous cell cultures were established and have since undergone 92 passages in 25-cm(2) flasks. Cell doubling time is approximately 3 days and the cells have been successfully revived after 1 year after storage at -80 degrees C. 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 independently and detected by labeling by an indirect immunofluorescence technique. The viruses were detected bound to the cell membranes and within the cell cytoplasm. This is the first report of a continuous cell line established from a species of whitefly and its inoculation with two begomoviruses. The successful inoculation of whitefly cell cultures with begomoviruses shown in our results represents great promise for the development of systems that allow researchers to achieve a better understanding of the complex relationship between begomoviruses and their whitefly vectors.