《植物生理学报》 2012, 48(9): 922-927

利用流式细胞仪研究拟南芥叶发育过程中细胞周期的调控

曾民环, 许德阳, 薛景石, 袁振环, 黄海, 崔晓峰*

中国科学院上海生命科学研究院植物生理生态研究所植物分子遗传国家重点实验室, 上海 200032

通信作者：崔晓峰；E-mail: xiaofeng@sippe.ac.cn；Tel: 021-54924102

摘　要：

叶的形态建成依赖于细胞不断地分裂增殖和不同类型细胞的特化。在叶发育早期, 叶细胞主要通过旺盛的有丝分裂 来增加原基中细胞的数目。随着叶片的生长, 叶细胞自顶部向基部逐渐退出有丝分裂进入内复制来增加细胞的倍性, 同时 伴随细胞的扩展和分化。本文介绍利用流式细胞仪研究双子叶模式植物拟南芥叶发育过程中细胞周期调控的方法和具体 研究实例。我们发现至少存在3种类型的细胞周期异常的拟南芥叶发育突变体。此外, 我们还介绍利用流式细胞仪测定 DNA复制效率的方法。

关键词：拟南芥; 细胞周期调控; 流式细胞术; 叶发育

收稿：2012-05-10   修定：2012-07-25

资助：国家自然科学基金(31070163)。

Study on the Cell Cycle Regulation during Leaf Development in Arabidopsis thaliana by Flow Cytometry

ZENG Min-Huan, XU De-Yang, XUE Jing-Shi, YUAN Zhen-Huan, HUANG Hai, CUI Xiao-Feng*

National Key Laboratory of Plant Molecular Genetics, Institute of Plant Physiology and Ecology, Shanghai Institutes for Biological Sciences, Chinese Academy of Sciences, Shanghai 200032, China

Corresponding author: CUI Xiao-Feng; E-mail: xiaofeng@sippe.ac.cn; Tel: 021-54924102

Abstract:

Leaf morphogenesis relies on continuous cell proliferation and specification of distinct types of cells. During early leaf development, leaf cells mainly undergo vigorous mitotic cell divisions to boost the number of primordial cells. Along leaf growth, leaf cells gradually exit from the mitotic cell cycle from leaf tip to base and enter the endoreduplication, which is frequently associated with cell expansion and differentiation, resulting in cells with increased nuclear ploidy levels. Here we describe the methods and several examples employing flow cytometry to investigate the cell cycle regulation during leaf development in Arabidopsis thaliana, a model dicotyledonous plant. We found that there are at least three types of leaf developmental mutants with abnormal cell cycle. In addition, we introduce the assay to measure the efficiency of DNA replication in Arabidopsis.

Key words: Arabidopsis; cell cycle regulation; flow cytometry; leaf development