植物原生质体分离及其瞬时转化的应用  

Isolation of plant protoplast and its application in transient transformation

作　　者：马超越 彭世清[2] 郭冬[2] 朱家红[2] 王颖[2] MA Chaoyue;PENG Shiqing;GUO Dong;ZHU Jiahong;WANG Ying(College of Forestry,Hebei Agricultural University,Baoding,Hebei 071000,China;Institute of Tropical Bioscience and Biotechnology,Chinese Academy of Tropical Agricultural Sciences,Haikou,Hainan 571101,China)

机构地区：[1]河北农业大学林学院,河北保定071000 [2]中国热带农业科学院热带生物技术研究所,海口571101

出　　处：《热带生物学报》2024年第2期241-250,共10页Journal of Tropical Biology

基　　金：海南省自然科学基金(2019RC304,323RC539)资助。

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。The cell wall was removed from plant cells,and the cytoplasm wrapped by a single cell membrane 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.

关 键 词：原生质体 酶解法 瞬时转化 

分 类 号：Q813.6[生物学—生物工程]