Centre for Functional Ecology, Department of Life Sciences, University of Coimbra, Coimbra, Portugal.
Department of Botany, Faculty of Science, Charles University, Prague, Czech Republic.
Methods Mol Biol. 2023;2672:25-64. doi: 10.1007/978-1-0716-3226-0_2.
Flow cytometry has emerged as a uniquely flexible, accurate, and widely applicable technology for the analysis of plant cells. One of its most important applications centers on the measurement of nuclear DNA contents. This chapter describes the essential features of this measurement, outlining the overall methods and strategies, but going on to provide a wealth of technical details to ensure the most accurate and reproducible results. The chapter is aimed to be equally accessible to experienced plant cytometrists as well as those newly entering the field. Besides providing a step-by-step guide for estimating genome sizes and DNA-ploidy levels from fresh tissues, special attention is paid to the use of seeds and desiccated tissues for such purposes. Methodological aspects regarding field sampling, transport, and storage of plant material are also given in detail. Finally, troubleshooting information for the most common problems that may arise during the application of these methods is provided.
流式细胞术已成为一种独特的灵活、准确和广泛适用的技术，可用于分析植物细胞。它最重要的应用之一集中在测量核 DNA 含量上。本章介绍了这种测量的基本特征，概述了总体方法和策略，但接着提供了大量的技术细节，以确保最准确和可重复的结果。本章的目的是让有经验的植物细胞学家和刚进入该领域的人都能轻松阅读。除了提供从新鲜组织中估计基因组大小和 DNA 倍性水平的分步指南外，还特别关注种子和干燥组织在这些目的中的应用。还详细介绍了有关植物材料的田间采样、运输和储存的方法学方面。最后，提供了在应用这些方法时可能出现的最常见问题的故障排除信息。
