Enabling biocontained plant virus transmission studies through establishment of an axenic whitefly (Bemisia tabaci) colony on plant tissue culture

Abstract Whiteflies (Bemisia tabaci) and the diseases they transmit are active enemies of global crop yield and therefore multidisciplinary studies involving plant-insect-virus interactions are needed. Presented here is the establishment of a colony of axenic (culturable microorganism free) whiteflies on tissue-cultured plants. Colony establishment was achieved with a surface sterilization of whitefly eggs laid on axenic phototrophically tissue-cultured plants. The recovery and transfer of emerging whiteflies through coupled tissue culture vessels to new axenic plants allows robust subculturing and produces hundreds of whitefly adults per month in a 1.1 L tissue culture vessel. Whitefly proliferation on more than a dozen plant species is shown as well as in vitro testing of whitefly preference for different plants. This novel axenic system provides the high-level of biocontainment needed to conduct virus transmission experiments. Axenic whitefly adults were able to acquire and transmit a begomovirus from tissue-cultured plants, indicating the irrelevance of culturable gut microorganisms in virus transmission. The methodology described here could be used to address a wide range of questions regarding whiteflies and plant virus transmission without the expense, facilities, and contamination risks associated with standard approaches.