Abstract
ABSTRACTThe ability of isolated pea (Pisum sativum cv. Ran 1) mesophyll protoplasts to aggregate and fuse spontaneously was examined. Aiming to define whether Ca2+ released during enzymatic isolation of protoplasts and residual in the purified suspension is important for aggregation and fusion, different isolating media were used: Ca2+—free medium, buffered Ca2+—free medium, Ca2 +—free medium containing 2.5 mM EGTA, and medium containing 10 mM CaCl2. Light scattering was measured to assess aggregation and/or fusion and Arsenazo III was used to evaluate free Ca2+. Binding of Ca2+ to plasmalemma was characterized by 9-Aminoac ridine fluorescence. Since light scattering of purified suspensions of protoplasts isolated in the presence of 2.5 mM EGTA was lower as compared to the other variants, a Ca2+—dependent fusion during isolation was suggested. Binding of Ca2+ by 100 μM EGTA slightly reduced the light scattering of suspension of protoplasts isolated without EGTA, indicating existence of Ca2+ dependent aggr...
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