Optimum parameters of preparation of Epidermal Cell Protoplasts within the Allium cepa L

Objective to optimize the isolation condition of Epidermal Cell Protoplasts within the Allium cepa L. Methods According to orthogonal experiment design L₂₅(5⁶) of enzyme, osmotic pressure stabilizer(D-Mannitol), pH, enzymolysis time, we got the optimal isolation condition by the number of protoplasts. Results The optimal isolation condition is showed in results: enzymolysis time 2h, pH 5.5, D-Mannitol 0.6 mol/L, 1.5% cellulose and 0.1% pectolyase and 0.1%BSA, temperature 28°C. It also means that optimal isolation condition is D₅B₃C₄A₂. Conclusion The optimal isolation condition is viable.