An Electrophoresis Method for Bulk Manipulation of Isolated Protoplasts from Higher Plants

Abstract

Summary An apparatus and method for the bulk electrophoresis of isolated higher plant protoplasts is described and was used in the electrophoresis of mixtures of protoplasts isolated from: (i) Pisum sativum cv. Meteor leaves and Parthenocissus tricuspidata cell suspension cultures and (ii) Zea mays cv. Campo leaves and P. tricuspidata cell suspension cultures. The P. tricuspidata protoplasts had lower mobilities than did the leaf protoplasts. Partial separation of the mixtures occurred and the majority of maize protoplasts were recovered in fractions containing 7% or fewer P. tricuspidata protoplasts. When P. tricuspidata and leaf protoplasts were fused by treatnlent with PEG and Ca2+ and the fusion inducing medium was removed by washing, hybrid protoplasts and those parent protoplasts which had not fused with the other parent, had similar mobilities (near 1 x10−8m2s−1V−1) which, for the leaf protoplasts, were lower than without the treatment with PEG and Ca2+. Surprisingly, for P. tricuspidata protoplasts this value was about the sanle as that obtained without treatment with PEG and Ca2+.