Objective:To find out the best isolation conditions of protoplasts of Lonicera japonica. Method:Some influence factors on isolation of protoplasts

such as composition of enzyme solution

methods

hours and temperature of enzyme dissolving

osmotic pressure

and so on

were compared and analyzed to find out the best conditions of protoplasts isolation of L. japonica. Result:The callus flourishing well of L. japonica was used as materials for protoplasts isolation. 0.7 mol·L-1 mannitol+1.5% cellulose Onozuka R-10+0.25% pectolase Yakult Y-23 was the best composition of enzyme solution

which was dissolved in cell protoplast wash medium(CPW) solution. 12-hour enzyme dissolving at 25℃ was the best methods of protoplasts isolation. And standing was better than shaking. Conclusion:Abundant and superior protoplasts of L. japonica can be obtained under the best isolation conditions that this study has found out which will give the necessary help for protoplasts culture

cell fusion and genetic transformation of L. japonica.