Abstract
Summary Prototrophy was induced by the technique of nuclear transplantation. Nuclei were isolated from cell suspension cultures of Brassica nigra (L.) Koch. and introduced into the protoplasts of a pantothenate-requiring auxotrophic mutant of Datura innoxia P. Mill. The uptake of the nuclei was induced in the presence of 0.1 M calcium chloride, 30 % polyethylene glycol and 0.05 M glycine at pH 6.8. After nuclear uptake, the auxotrophic protoplasts were cultured in a nonselective medium for two weeks and later transferred to selective medium lacking pantothenate. The selection of prototrophic clones was carried out on feeder plates having wild-type Datura cells as the nurse tissue. Surviving prototrophic clones were capable of growth on minimal medium, regained the ability to synthesize chlorophyll and one of the clones developed nodular, hairy callus on a cytokinin-supplemented medium.
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