Isolation and regeneration of rose bud callus protoplasts (Protoplasts sp. cv. soraya)

Rosa spp. (Roses): In Vitro Culture, Micropropagation, and the Production of Secondary Products

“…A key factor in the in vitro culture ofthese protoplasts was the rapid serial pre-culture of rose cell suspensions prior to protoplast isolation. The culture of rose protoplasts, with a plating efficiency of the order of 20%, which developed to the callus stage, has been reported by Krishnamurthy et al (1979). There have been no reports of the regeneration of shoot primordia from in vitro cultured rose protoplasts.…”

A protoplast to plant system in roses

“…One ml aliquots of the protoplast/agarose mixture were then added to 1 ml volumes of double strength callus regeneration medium in 30mm diameter plastic petri dishes, mixed well and allowed to set before sealing with Nescofilm. The various callus regeneration media tested in this study included DPD (Binding 1974), MMS (Krishnamurthy et al 1979), SH (Schenk & Hildebrant 1972), MS (Murashige & Skoog 1962), MS-NH4 (MS medium minus ammonium) (Ochatt & Power 1988c), MXG (Strauss & Potrykus 1980), B5 (Gamborg et al 1968) and KM8p (Kao & Michayluk 1975). Except for the KM8p medium, all these media were supplemented, irrespective of the recommended concentrations, with final concentrations of 30 g 1 l sucrose and 3 mg 1-1 2,4-D as the only sugar and growth regulator, respectively, and 90g 1-1 mannitol.…”

“…Pearce & Cocking (1973) obtained only cell wall formation, budding and abnormal cytokinesis with protoplasts obtained from cell suspension cells of Rosa × hybrida'Paul's Scarlet'. Krishnamurthy et al (1979) and Strauss and Potrykus (1980) used protoplasts from floral bud callus o f R x hybrida 'Soraya' and cell suspensions of 'Paul's Scarlet' rose, respectively. In both cases, callus colonies were obtained but no attempts were made to regenerate plantlets from them.…”