Isolation of protoplasts from the placental cells of Lycopersicum pimpinellifolium Mill

Living protoplasts were isolated from the interplacental regions of Lycopersicum pimpinellifolium berries by the removal of the walls from cells in tissue slices treated for 1–2 h with 10 % pectinase in 0.5 M sucrose solution. Protoplasts thus isolated, then washed and transferred to microculture chambers for observation were invariably spherical. Each protoplast contains a nucleus, a number of chloroplasts of variable shape and a vacuole with smaller vacuoles contained therein. Phase contrast optics reveal cytoplasmic granules, the size of mitochondria, which serve to indicate such dynamic processes as cyclosis. Treatment with ox bile salt and sodium lauryl sulphate cause rapid disruption of the protoplasts producing subprotoplasts and isolated tonoplasts and serves to confirm the absence of rigid cell walls.