摘要
原生质体已被广泛应用于亚细胞定位、基因表达分析、蛋白-蛋白互作、基因编辑等方面.笔者总结了酶解法分离植物原生质体的影响因素,电穿孔和PEG介导的方法对原生质体瞬时转化效率的影响和瞬时转化的应用.植物的种类、不同的PEG浓度、转染时间、DNA浓度和原生质体数量都会影响PEG介导法对原生质体瞬时转化效率.电穿孔法必须在脉冲电压、脉冲长度、脉冲数、细胞数、DNA浓度都适宜的条件下才能达到最佳转化效率.本文为更多植物原生质体分离体系和瞬时转化体系的建立提供参考.
Abstract
The cell wall was removed from plant cells,and the cytoplasm wrapped by a single cellmembrane is called protoplast.It has been widely used in protein subcellular localization,gene expression analysis,protein-protein interaction and gene editing among others.The factors influencing plant protoplasts isolation by enzymatic method,the effects of electroporation and PEG-mediated methods on the transient transformation efficiency of protoplasts and their application in transient transformation were summarized in this review.Plant species,different PEG concentrations,transfection time,DNA concentration,and number of protoplasts were found to affect the PEG-mediated transient transformation efficiency of protoplasts.The electroporation had an optimum transformation efficiency of protoplasts only under the adequate conditions of pulse voltage,pulse duration,pulse number,cell number,and DNA concentration.This review might provide reference for establishment of more plant protoplast isolation systems and transient transformation systems for plant protoplasts.
维普
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