Plating of isolated tobacco mesophyll protoplasts on agar medium

The cultivation of single cells of Nicotiana tabacum L. var. "Samsun" and Phaseolus vulgaris L. var. "Early Golden Cluster" on a thin agar layer in Petri dishes is described. Under these conditions about 20 per cent of the cells divided repeatedly and established tissue clones which could be isolated and maintained as growing tissue cultures. It was possible also to follow the successive divisions of isolated cells and to observe their behavior during cytogenesis under the microscope. Show more

A method for the selection of auxotrophic mutants from somatic cell cultures of Nicotiana tabacum depends upon the incorporation of 5-bromodeoxyuridine into the DNA of wild-type cells and upon its lack of incorporation into the DNA of auxotrophic cells. There may be more than one functional copy of essential genes in the haploid genome of Nicotiana tabacum. Show more