Photosynthetic activities of spinach leaf protoplasts

Photosynthetic activities of protoplasts isolated from spinach leaf (Spinacia oleracea L.) were investigated. The protoplasts were stable up to 9 hr, without loss of the original activity of CO(2) fixation (33-75 mumoles CO(2)/mg Chl.hr) and light-dependent O(2) evolution (33-40 mumoles O(2)/mg Chl.hr), when stored in 0.8 m mannitol-0.05 m N-tris (hydroxymethyl)-methylglycine-NaOH buffer, pH 7, at 4 C in dark. The optimum pH of 8.5 for CO(2) fixation reaction carried out in the present experimental condition employed is about the same as that reported for intact spinach chloroplasts. The CO(2) concentration for half-maximal rate of CO(2) fixation by protoplasts. "Km (CO(2))," were determined to be 19.8 mum (pH 7) and 42 mum (pH 8.5) and are similar to those observed for intact spinach chloroplasts. Protoplasts showed postillumination CO(2) fixation. Over-all results indicate that spinach protoplasts are as active as the intact plant leaf tissues in their photosynthetic activities.