拟南芥不同组织细胞对DAPI染色的差异分析  

Analysis of DAPI Staining in Different Tissues of Arabidopsis

作　　者：于静芳 尹润竹 周俊 YU Jingfang;YIN Runzhu;ZHOU Jun(MOE Key Laboratory of Laser Life Science&Institute of Laser Life Science,College of Biophotonics,South China Normal University,Guangzhou 510631,China)

机构地区：[1]华南师范大学生物光子学研究院激光生命科学研究所暨激光生命科学教育部重点实验室,广州510631

出　　处：《激光生物学报》2020年第2期148-152,共5页Acta Laser Biology Sinica

基　　金：国家自然科学基金项目(31600288);广东省科技计划项目(2016A020210127);华南师范大学青年教师培育项目(671075)。

摘　　要：4’,6-二脒基-2-苯基吲哚(DAPI)作为一种DNA特异性荧光染料,在荧光显微观察各种生物细胞中得到了广泛应用。然而,植物不同组织细胞响应DAPI染色目前仍缺乏比较系统的表征。本文利用模式植物拟南芥,探究了其各个组织细胞对DAPI染色的差异。首先我们构建了一个细胞核定位的稳转基因植物UBQ10∶BAG5-eYFP。接下来的试验发现在非固定的BAG5-eYFP植物活细胞中,只有表皮细胞和叶肉细胞能被DAPI染色。相比于叶绿体,线粒体中的DNA对DAPI染色较为敏感,表现出DAPI荧光和线粒体marker CoxIV-RFP很好的共定位。多聚甲醛固定之后,根部分生区、气孔保卫细胞能被DAPI染色。这些结果表明,拟南芥不同组织细胞对DAPI染色呈现出很大的差异。叶片表皮细胞和叶肉细胞对于探究植物活体细胞核的研究将会是一个很好的模型。4’,6-diamidino-2-phenylindole(DAPI),as one of specific DNA fluorescent dyes,is widely used in fluorescence microscopic observation of various biological cells.However,there is still lack of a systematic characterization of different tissues response to DAPI staining in plant.In this study,using the model plant Arabidopsis thaliana,we explored the differential responses to DAPI staining in various tissues.Firstly,we generated a stable nucleus-targeted transgenic plant UBQ10∶BAG5-eYFP.The results showed that only the epidermal cells and mesophyll cells can be stained by DAPI in living plant.Compared with chloroplast,the DNA in mitochondria is very sensitive to DAPI staining,displaying a good co-localization between the fluorescence of DAPI and mitochondrial marker CoxIV-RFP.After fixed with paraformaldehyde,the root meristem zone and stomatal guard cells can be stained by DAPI.These results indicated that different tissues have great differences when being stained with DAPI in Arabidopsis.The leaf epidermal cells and mesophyll cells would be good models for studying the function of nucleus in living plant.

关 键 词：细胞核 BAG5 DAPI 拟南芥 

分 类 号：Q942.7[生物学—植物学]