Abstract:

Washing buffer plays an important role in protoplasts purification and storing when protoplasts isolated. The study compared yiled and viability change of protoplasts in washing buffer of WI, W5 and MMg in 30 h, analyzed the influence of CaCl2 with different concentration on protoplasts in WI, W5 and MMg to get the suitable washing buffer for the highest viable protoplasts. The results indicated that WI was better at maintaining plasma membrane, while W5 was better on keeping viability, after 30 h W5 got the highest viable protoplasts population which had a good storing washing buffer for protoplasts in a short period; CaCl2 treatment showed little influence on protoplasts viability and its population improving effect depended on its concentration and the washing buffer type, there was a uptrend of viable protoplasts in MMg when treated with 0-50 mM CaCl2.