利用原子力显微术在纳米尺度上检测植物细胞质膜完整性  

Plant plasma membrane integrity assay by atomic force microscopy at the nanoscale level

作　　者：苏晓 高永伟[3] 林金星[1,2] 李晓娟 SU Xiao;GAO Yong-wei;LIN Jin-xing;LI Xiao-juan(Key Laboratory of Genetics and Breeding in Forest Trees and Ornamental Plants,Ministry of Education,College of Biological Sciences and Technology,Beijing Forestry University,Beijing 100083;Institute of Tree Development and Genome Editing,Beijing Forestry University,Beijing 100083;Public Analysis and Test Center,Beijing Forestry University,Beijing 100083,China)

机构地区：[1]北京林业大学生物科学与技术学院,林木花卉遗传育种教育部重点实验室,北京100083 [2]北京林业大学树木发育与基因编辑研究院,北京100083 [3]北京林业大学公共测试中心,北京100083

出　　处：《电子显微学报》2023年第1期68-74,共7页Journal of Chinese Electron Microscopy Society

基　　金：国家自然科学基金资助项目(No.91954202);北京林业大学“杰出青年人才”培育计划(No.2019JQ03003)。

摘　　要：目的:探究原子力显微镜检测植物细胞质膜的损伤程度及其定量分析的方法。方法:采用酶解法制备拟南芥原生质体,原生质体经MβCD(Methyl-β-cyclodextrin)处理造成质膜损伤后,利用原子力显微镜进行成像,最后通过NanoScope Analysis软件进行成像分析,对粗糙度Ra(平均粗糙度)、Rq(均方根粗糙度)和Rmax(最高数据点和最低数据点之间的最大垂直距离)值以及从孔隙周围的长轴和短轴上测量获得孔隙的直径和深度进行了系统的测算。结果:与对照组对比,经MβCD处理的样品表面出现大量的孔隙,且粗糙度Ra、Rq和Rmax值均显著高于对照组。结论:原子力显微镜可以检测拟南芥细胞质膜的损伤,并对损伤程度进行定量分析,为探究质膜完整性提供了一个可行的新思路。Objective: To explore the feasibility of detecting plasma membrane(PM) damage in plant cells by atomic force microscopy(AFM).Methods: Protoplasts were prepared from Arabidopsis thaliana seeding. After treating with Methyl-β-cyclodextrin(MβCD), protoplasts were imaged by AFM. The average roughness(Ra), root mean square roughness(Rq), and the maximum vertical distance between the highest and lowest data points(Rmax) were measured from the images using NanoScope analysis software, the diameter and depth of each pore were statistically analyzed.Results: A large number of pores appeared on the PM of the samples treated with MβCD. Moreover, the Ra, Rq and Rmax values from the PM treated with MβCD were significantly higher(*p<0.05) than those from the control PM.Conclusions: AFM can detect the physical properties of PM in plant protoplasts. More importantly, it can be used to quantitatively analyze the degree of PM damage, providing a feasible new approach for exploring the integrity of PM.

关 键 词：原子力显微镜 植物质膜 质膜完整性 

分 类 号：Q241[生物学—细胞生物学] TH742[机械工程—光学工程] Q336[机械工程—仪器科学与技术]