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.

Centre for Functional Ecology Department of Life Sciences University of Coimbra Coimbra Portugal

Czech Academy of Sciences Institute of Botany Průhonice Czech Republic

Department of Botany Faculty of Science Charles University Prague Czech Republic

Department of Botany Faculty of Science University of South Bohemia České Budějovice Czech Republic

Henan University School of Life Sciences State Key Laboratory of Crop Stress Adaptation and Improvement State Key Laboratory of Cotton Biology Key Laboratory of Plant Stress Biology Kaifeng China

Institut Botànic de Barcelona Barcelona Catalonia Spain

Institute of Experimental Botany of the Czech Academy of Sciences Centre of Plant Structural and Functional Genomics Olomouc Czech Republic

Laboratory of Molecular Biology and Cytometry Department of Agricultural Biotechnology Bydgoszcz University of Science and Technology Bydgoszcz Poland

School of Plant Sciences BIO5 Institute Arizona Cancer Center Department of Biomedical Engineering University of Arizona Tucson AZ USA

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