拟南芥子叶发育过程中叶绿体特征的量化及分布分析  

Quantification and distribution analysis of cotyledonal chloroplasts during seeding development in Arabidopsis

作　　者：马灵玉 王广超 张曦 胡子建 沈微微 林金星 MA Ling-yu;WANG Guang-chao;ZHANG Xi;HU Zi-jian;SHEN Wei-wei;LIN Jin-xing(Advanced Innovation Center for Forest Molecular Design and Breeding,Beijing Forestry University,Beijing 100083;National Engineering Laboratory of Forest Breeding,College of Biological Science and Technology,Beijing Forestry University,Beijing 100083,China)

机构地区：[1]北京林业大学林木分子设计育种高精尖创新中心,北京100083 [2]北京林业大学生物科学与技术学院林木育种国家工程实验室,北京100083

出　　处：《电子显微学报》2021年第3期270-278,共9页Journal of Chinese Electron Microscopy Society

基　　金：国家自然科学基金资助项目(No.31530084);教育部学科创新引智计划(No.B13007);教育部创新团队计划(No.IRT13047);中国博士后科学基金(No.2019M660494);青年科学基金项目(No.32000558)。

摘　　要：拟南芥子叶发育过程中叶绿体的体积、面积和形状及分布等特征对于植株生长具有重要影响,但如何对萌发过程中细胞内叶绿体的体积、面积和形状等特征进行定量和分布的相关性分析仍然存在一定空白。本研究首先使用荧光显微镜获取不同萌发时期的子叶叶绿体的二维层扫图集,然后对图集进行优化及三维重构和叶绿体分割,并对叶绿体的体积、面积、及形状等特征进行量化分析,不仅获得不同尺寸、形状的叶绿体分布比例,而且将特征和分布进行四维可视化展示。此外,研究中还使用泰森多边形算法对不同属性的叶绿体进行聚类分析,进而得到不同类别的叶绿体在组织中的连接和分布信息。另外,本文通过电子显微镜成像方法,进一步确认了随着拟南芥子叶的生长,叶绿体的超微结构发生了显著变化,逐渐变大。该图像处理方法不仅能够快速识别细胞中叶绿体的不同特征及参数,还能够根据细胞中叶绿体的密度、面积、距离等特征参数对叶绿体的连接度和分布进行特异性分析,是一种功能强大且高效灵活的多细胞分析方法。The size,shape and distribution of cotyledonal chloroplasts influence seedling growth greatly during plant development in Arabidopsis thaliana.However,there are some gaps in the distribution correlated quantitative analysis of the volume,area and shape of intracellular chloroplasts during germination.In this paper,a new procedure was introduced to perform 4-dimensional visualization of distribution correlated quantitative analysis of cotyledonal chloroplasts during seeding development.Here,2-dimensional images atlas of cotyledonal chloroplasts in different germination stages were acquired with a fluorescence microscope at first.Secondly,3-demensinal reconstruction was performed after optimize the images quality and quantitative data of volume,area,prolate ellipticity and sphericity was recorded after segmentation of single chloroplast.The 4 D visualization of chloroplasts distribution with respect to size and shape was processed and shown.The Voronoi diagram-based automatic segmentation was used to establish chloroplasts cluster to quantify the connectivity and distribution of chloroplasts in the multicellular tissue.In addition,we found that with the growth of cotyledons of Arabidopsis thaliana,the chloroplasts in cotyledons became larger and the number decreased gradually.This image processing procedure can not only quickly identify the different characteristics and parameters of chloroplasts in cells,but also perform specific analysis on the connectivity and distribution of chloroplasts based on the density,area,distance of chloroplasts.It was proved to be an efficient and flexible image process procedure for complex multicellularity analysis in plant.

关 键 词：叶绿体成像 细胞参数 四维可视化 泰森多边形 图像处理 

分 类 号：Q334[生物学—遗传学] Q336