Fusion and transformation of plant protoplasts

Abstract

Publisher Summary Somatic hybridization of plants relies on the availability of efficient and reproducible methods for the induced fusion of protoplasts while maintaining high viability and subsequent growth of the product of fusion. The desired level of fusion depends on the mode of selection operating between the two species being hybridized. The applicable methods of fusion have to accommodate a wide range of protoplast systems, differences in the physiological tolerance of fusogens, and inheritently varying viabilities both pre- and postfusion. This chapter describes the fusion protocols that are modified and adapted to provide the effective and reproducible fusion of protoplasts from a wide range of species and source tissues. They fall into two categories: chemically induced fusion and electrofusion. Irrespective of fusion method, their application requires that a balance is established between the level of fusion and postfusion survival. Lower levels of fusion can be tolerated if a powerful method of selection is available. Progress toward the induced fusion of protoplasts can be monitored using naturally occurring visual markers, such as leaf mesophyll protoplasts fused with colorless cultured cell protoplasts, or introduced visual markers, such as fluorescent dyes.