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Regeneration of fertile plants from protoplasts of sunflower (Helianthus annuus L.) has met with considerable difficulties in spite of numerous efforts over several years. The isolation of protoplasts and their culture up to the callus stage does not appear to present major difficulties (Bohorova et al. 1986; Lenee and Chupeau 1986; Moyne et al. 1988; Chanabe et al. 1989; Guilley and Hahne 1989; Schmitz and Schnabl 1989; Dupuis et al. 1990). Most currently used donor materials, including hypocotyl, cotyledons, petioles, leaves, and suspension cultures, allow the isolation of protoplasts at comparatively high yields, and media have been developed for the regeneration of callus with good plating efficiencies from all of these types.