Optimization of Electrofusion for the Formation and Culture of Protoplast Heterokaryons
Electrofusion has been demonstrated with a wide variety of plant protoplasts. Nonetheless, it has not been shown that the fusion products are viable, nor has the technique been refined to make it a practical alternative to fusion by PEG. Protoplasts derived from suspension cultures of Nicotiana plumbaginifolia have been fused with N. tabacum mesophyll protoplasts. By fusing mesophyll protoplasts with suspension culture protoplasts, hybrid cells can be visually identified in both living and fixed material. This system has been used to determine the optimal conditions for electrofusion and for development of a protocol for the isolation and culture of protoplast heterokaryons.