Regeneration of Plants from Embryogenic Suspension Culture Protoplasts of Wheat (Triticum aestivum L.)

Abstract

We describe the regeneration of plants from protoplasts of wheat isolated from a regenerable embryogenic suspension culture that was initiated from immature embryo-derived ‘aged’ compact callus. Proto-colonies were also obtained from a non-regenerable cell line established from friable callus that arose spontaneously on the aged callus, but did not give rise to any plants. Plating efficiencies well in excess of twenty percent were routinely obtained from protoplasts isolated from the regenerable as well as the non-regenerable cell lines in Kao and Michyaluk, as well as Murashige and Skoog based, nutrient media. Four to five week old protocolonies, derived from regenerable suspension culture protoplasts, formed distinct somatic embryos and shoots upon transfer to various regeneration media. Shoots were rooted on transfer to rooting media. Successive transfer of the plantlets to MS or half-strength MS medium, with or without activated charcoal, produced healthy plants that were successfully transferred to soil and grown in the greenhouse.