Leaf mesophyll protoplast cultures from six Nicotiana species, N. debneyi, N. rustica, N. amplexicaulis, N. glauca, N. glutinosa, and N. sylvestris were carried out. When we reduced the NH_4NO_3 and Fe·EDTA concentration to 1/3(7 mM) and 1/10(10 μM)...
Leaf mesophyll protoplast cultures from six Nicotiana species, N. debneyi, N. rustica, N. amplexicaulis, N. glauca, N. glutinosa, and N. sylvestris were carried out. When we reduced the NH_4NO_3 and Fe·EDTA concentration to 1/3(7 mM) and 1/10(10 μM) from the Murashige and Skoog medium respectively, cell division of the protoplasts was efficiently induced in four Nicotiana species, N. debneyi, N. rustica, N. amplexicaulis and N. glauca. However, other two species, N. glutinosa and N. sylvestris were failed in inducing cell division at the same culture condition. The protoclone calluses derived from four Nicotiana speices were consequently regenerated on a MS basal medium supplemented with the appropriate auxin and cytokinin.