Flow sorting and culture of plant protoplasts

Conditions have been established for the rapid flow analysis of leaf protoplasts of Nicotiana tabacum L. cv. Xanthi using a flow cytometer-cell sorter. A procedure based upon chlorophyll autofluorescence was devised to permit the systematic evaluation of flow conditions in order to identify those under which protoplast damage was minimized. These conditions were employed for the flow sorting of protoplasts, following which it was possible to regenerate the sorted protoplasts into complete plants. The application of flow sorting is discussed for the rapid identification and selection of somatic hybrids produced by protoplast fusion.