Abstract
Most cells of bacteria, fungi, and green plants are surrounded by rigid cell walls. Removal of these walls makes them susceptible to various types of biochemical and genetic manipulation. This chapter presents experiments on the isolation of protoplasts from germinating sporangiospores of Phycomyces , the regeneration of isolated protoplasts, the fusion of protoplasts from different auxotrophs, and the formation of heterokaryotic regenerants. High yields of isolated protoplasts and high regeneration rates are obtained. Protoplast fusion is induced by seawater and calcium nitrate. Significantly higher fusion frequencies are achieved in solutions of polyethylene glycol (MW 6000). Up to 4% of the viable protoplasts exhibited complementation of nutritional deficiencies when calcium ions and high pH were used in addition to polyethylene glycol. Somatic hybridization in fungi through fusion of auxotrophic mutants is investigated in great detail by Ferenczy and co-workers.
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