Isolation, Culture and Plant Regeneration From Protoplasts of Nicotiana debneyi

Abstract

Mesophyll protoplasts of two genetically distinct genotypes of N. debneyi were cultured with sustained division following a plating efficiency in excess of 50%. Fully fertile mature plants were regenerated from callus cultures derived from protoplasts. Shoots were induced in medium containing 1 mg/l 6-benzylaminopurine and 0.5 mg/I indole acetic acid. The repeatably high efficiency of protoplast culture was used to evaluate the quantitative effects of two drugs, kanamycin and trimethoprim, which effectively inhibited colony formation at concentrations of 100 and 50 µg/ml, respectively. An enhancer of DNA uptake, poly-L-ornithine, had virtually no effect on sustained protoplast division at a concentration of 7.5 µg/ml or less.