聚乙二醇法诱导烟草原生质体融合的条件优化

摘要

采用聚乙二醇(PEG)法诱导烟草原生质体融合,对融合时PEG的浓度、用量、融合时间及融合温度进行优化,以期得到一种融合率较高的方法.结果表明:以40％的PEG融合的二元融合率最高；PEG与原生质体悬浮液体积比为3∶5,在0.5～0.6mol/L甘露醇为渗透压时,25 ～ 30℃下融合20 ～ 25 min的融合效果最佳,融合率始终保持在20％左右,甚至可达25％.%In order to improve the fusion rate of tobacco protoplast, the different factors affecting fusion rate of protoplast including the amount and concentration of PEG, fusion temperature, fusion time and mannitol concentration etc were investigated in the present work. The results showed that 40% PEG gave the best fusion rate, and could produce the most numerous heterokaryon. The optimized conditions also included 3: 5 volume proportion of PEG and protoplast, 0. 5～0.6 mol/L mannitol and continuous fusion for 20 ～ 15 min in 25 ～ 30℃ , by which 20% -25% fusion rate might be obtained.