Isolation and fusion studies on protoplasts from pollen tetrads

Different enzymes were tested for isolation of intact protoplasts from pollen tetrads. About 80% isolation was achieved from pollen tetrads of Cajanus cajan and Zea mays and about 60% from Luffa cylindrica and Lycopersicon esculentum after 4 h of treatment with 5% cellulase. When these mononucleate protoplasts were incubated in presence of 0.05 M CaCl2 in 0.3 M glucose at pH 10.5, 70-80% fusion was achieved. Fusion was rare in sodium nitrate solutions.