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荧光激活细胞分选技术在植物发育和环境响应研究中的应用。

The use of fluorescence-activated cell sorting in studying plant development and environmental responses.

作者信息

Carter Anthony D, Bonyadi Roxanna, Gifford Miriam L

机构信息

School of Life Sciences, University of Warwick, Coventry, UK.

出版信息

Int J Dev Biol. 2013;57(6-8):545-52. doi: 10.1387/ijdb.130195mg.

Abstract

Fluorescence-Activated Cell Sorting (FACS) is a powerful tool that enables plant growth and development to be studied at the cellular level. Flow cytometry is used to isolate subpopulations of cells, such as those of specific cell types, or cells at particular developmental stages that have been marked with fluorescent proteins. Transgenic technology has given us the ability to generate plants that express fluorescent proteins, not just constitutively in particular cell types, but also dynamically in response to endogenous or external factors. By processing such transgenic lines with FACS, it is possible to isolate distinct populations of cells in a wide range of likely response states for further analysis. This is particularly useful for investigating biological mechanisms in plants because the control of growth and development is manifest at the cell type level. Furthermore, the specificity of the resulting data enables fine modelling of the transcriptional networks that exert systems-level control of the transcriptome; hence key regulators of responses and processes in the plant can be identified. In this review, the current state of the art for FACS methods in plants is explored by means of case studies of research in which cell sorting allowed us to make significant new discoveries.

摘要

荧光激活细胞分选(FACS)是一种强大的工具,可使植物生长和发育在细胞水平上得到研究。流式细胞术用于分离细胞亚群,例如特定细胞类型的细胞,或已用荧光蛋白标记的处于特定发育阶段的细胞。转基因技术使我们有能力培育出不仅在特定细胞类型中组成性表达荧光蛋白,而且能对内源或外部因素做出动态响应而表达荧光蛋白的植物。通过用FACS处理此类转基因株系,有可能在广泛的可能响应状态下分离出不同的细胞群体以供进一步分析。这对于研究植物的生物学机制特别有用,因为生长和发育的控制在细胞类型水平上表现出来。此外,所得数据的特异性能够对转录网络进行精细建模,这些转录网络对转录组施加系统水平的控制;因此,可以识别植物中响应和过程的关键调节因子。在本综述中,通过细胞分选使我们能够做出重大新发现的研究案例,探讨了植物FACS方法的当前技术水平。

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