烟草脱外壁花粉与完整花粉电激导入效果的比较研究

利用膜不通透的荧光染料钙黄素作为指示剂，比较了烟草脱外壁花粉与完整的未萌发花粉和萌发花粉电激导入的效果．研究了电激过程中的电场强度、脉冲持续时间、电激液的介质成分和浓度对三者的导入率及电激后花粉萌发率的影响．结果表明，从未萌发花粉、萌发花粉到脱外壁花粉，导入率逐渐升高，证实了脱外壁花粉由于摆脱了外壁的阻碍而有利于外源物质的电激导入．但电激后花粉萌发率的变化规律相反．荧光指示剂的应用便于优化选择适宜电激参数，为外源基因的导入实验提供了参考依据．

COMPARATIVE INVESTIGATIONS ON ELECTROPORATION EFFECTS OF DE EXINED POLLEN AND INTACT POLLEN IN NICOTIANA TABACUM

Using calcein as an indicator, the effects of electroporation on de exined pollen, ungerminated and germinated intact pollen in Nicotiana tabacum were compared. Main factors affecting introduction efficiency such as field strength, time constant of pulse, constituents and concentration of the electroporation buffer were investigated as well as the germination rate after electroporation. The results showed that under optimal electroporation conditions, the introduction efficiency of the de exined pollen was the highest, the germinated pollen was lower, and the ungerminated pollen was the lowest. This confirmed that the pollen free of exine barrier was beneficial for introducing exogenous molecules. However, the germination rate of de exined pollen was the lowest. The use of fluorescent indicator favored choice of appropriate electroporation parameters, that might be helpful for experiments on foreign gene introduction by electroporation.