Live-Cell Imaging of Meiotic Spindle and Chromosome Dynamics in Maize (Zea mays)

Q1 Agricultural and Biological Sciences Current protocols in plant biology Pub Date : 2016-12-01 DOI:10.1002/cppb.20035

Natalie J. Nannas, R. Kelly Dawe

引用次数: 4

Abstract

Live-cell imaging is a powerful tool that allows investigators to directly observe the dynamics of cellular processes. Live imaging has proven particularly useful in studying mitotic and meiotic chromosome segregation, where the assembly of spindles and movement of chromosomes can be quantified in ways not possible with fixed cells. This protocol describes how to image live meiosis in the agriculturally important plant, maize. The creation of fluorescently tagged tubulin allows visualization of maize spindles, and nucleic acid dyestain chromosomes. This protocol describes all steps required for live imaging, including how to grow plants, screen for relevant genotypes, harvest meiotic cells, and collect live movies of meiosis. While this protocol was developed for imaging fluorescently tagged tubulin, it can be easily modified to observe the meiotic dynamics of any fluorescently labeled protein of interest. © 2016 by John Wiley & Sons, Inc.

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玉米减数分裂纺锤体活细胞成像及染色体动力学

活细胞成像是一种强大的工具，它允许研究人员直接观察细胞过程的动力学。实时成像已被证明在研究有丝分裂和减数分裂染色体分离方面特别有用，其中纺锤体的组装和染色体的运动可以以固定细胞无法实现的方式量化。这个程序描述了如何成像活减数分裂在农业上重要的植物，玉米。荧光标记微管蛋白的产生使玉米纺锤体可视化，核酸染色染色体。本协议描述了实时成像所需的所有步骤，包括如何种植植物，筛选相关基因型，收获减数分裂细胞，并收集减数分裂的实时影像。虽然该方案是为成像荧光标记的微管蛋白而开发的，但它可以很容易地修改，以观察任何感兴趣的荧光标记蛋白的减数分裂动力学。©2016 by John Wiley &儿子,Inc。

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Current protocols in plant biology Agricultural and Biological Sciences-Plant Science

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期刊介绍： Sound and reproducible laboratory methods are the foundation of scientific discovery. Yet nuances that are critical for an experiment''s success are not captured in the primary literature but exist only as part of a lab''s oral tradition. Current Protocols in Plant Biology provides reproducible step-by-step instructions for protocols relevant to plant research. Furthermore, Current Protocols content is thoughtfully organized by topic for optimal usage and to maximize contextual knowledge. Quarterly issues allow Current Protocols in Plant Biology to constantly evolve to keep pace with the newest discoveries and developments. Current Protocols in Plant Biology is the comprehensive source for protocols in the multidisciplinary field of plant biology, providing an extensive range of protocols from basic to cutting edge. Coverage includes: Extraction and analysis of DNA, RNA, proteins Chromosome analysis Transcriptional analysis Protein expression Metabolites Plant enzymology Epigenetics Plant genetic transformation Mutagenesis Arabidopsis, Maize, Poplar, Rice, and Soybean, and more.