DEVELOPMENT OF PLANTS FROM PROTOPLASTS OF SOLANUM (SOLANACEAE)

Experiments were performed to determine conditions critical to the isolation and culture of protoplasts from leaf mesophyll cells of a Solanum Section Tuberarium diploid clone, a S. phureja × S. chacoense F₁ hybrid. The optimum concentration of cellulase (Cellulysin) was 0.4%, while pectinase (Macerase) was inhibitory, even at a low concentration (0.075%). Maximum yields of protoplasts were achieved when the enzyme solution was not changed during incubation, and slow oscillation (60 rpm) on a shaker was used. When detached leaves were held under low light intensities or in the darkness for 3–5 days prior to protoplast production, results were more consistent than when the leaves were used directly from the greenhouse. Following dark or low light treatment the optimum osmolarity of the isolation and growth media was approximately 0.3–0.4 M. Of nine growth media tested only those of Nitsch and Ohyama and of Upadhya supported callus development, but the callus was often loose and did not differentiate roots or shoots. Callus from protoplasts cultured in the Upadhya medium modified by addition of glycine, vitamins, and casein hydrolysate, and subsequently transferred to the medium of Lam, formed roots and shoots when cultures were maintained in light. Mature plants were obtained following transfer to modified White's medium and later transplantation to soil.