Flow cytometry (FCM) using DNA-selective fluorochromes is now the prevailing method for the measurement of nuclear DNA content in plants. Ease of sample preparation and high sample throughput make it generally better suited than other methods such as Feulgen densitometry to estimate genome size, level of generative polyploidy, nuclear replication state and endopolyploidy (polysomaty). Here we present four protocols for sample preparation (suspensions of intact cell nuclei) and describe the analysis of nuclear DNA amounts using FCM. We consider the chemicals and equipment necessary, the measurement process, data analysis, and describe the most frequent problems encountered with plant material such as the interference of secondary metabolites. The purpose and requirement of internal and external standardization are discussed. The importance of using a correct terminology for DNA amounts and genome size is underlined, and its basic principles are explained.

Laboratory of Molecular Cytogenetics and Cytometry Institute of Experimental Botany Sokolovská 6 Olomouc Czech Republic

References provided by Crossref.org

New estimates and synthesis of chromosome numbers, ploidy levels and genome size variation in Allium sect. Codonoprasum: advancing our understanding of the unresolved diversification and evolution of this section

Genome assembly of a diversity panel of Chenopodium quinoa

Effect of nanoparticles on the ex-vitro performance of cryopreservation-derived plant material

SOS1 tonoplast neo-localization and the RGG protein SALTY are important in the extreme salinity tolerance of Salicornia bigelovii

Diploid and tetraploid cytotypes of the flagship Cape species Dicerothamnus rhinocerotis (Asteraceae): variation in distribution, ecological niche, morphology and genetics

Cytomolecular trends in Chamaecrista Moench (Caesalpinioideae, Leguminosae) diversification

Cryptic invasion suggested by a cytogeographic analysis of the halophytic Puccinellia distans complex (Poaceae) in Central Europe

Incidence and evolutionary relevance of autotriploid cytotypes in a relict member of the genus Daphne (Thymelaeaceae)

The Use of Flow Cytometry for Estimating Genome Sizes and DNA Ploidy Levels in Plants

SWEET11b transports both sugar and cytokinin in developing barley grains

The giant diploid faba genome unlocks variation in a global protein crop

Genomics and biochemical analyses reveal a metabolon key to β-L-ODAP biosynthesis in Lathyrus sativus

Morphological and environmental differentiation as prezygotic reproductive barriers between parapatric and allopatric Campanula rotundifolia agg. cytotypes

The relationship between transposable elements and ecological niches in the Greater Cape Floristic Region: A study on the genus Pteronia (Asteraceae)

Stoichiometry versus ecology: the relationships between genome size and guanine-cytosine content, and tissue nitrogen and phosphorus in grassland herbs

Evolutionary importance of the relationship between cytogeography and climate: New insights on creosote bushes from North and South America

Reference standards for flow cytometric estimation of absolute nuclear DNA content in plants

Morphological, ecological and geographic differences between diploids and tetraploids of Symphytum officinale (Boraginaceae) justify both cytotypes as separate species

Genome size of alpine plants does not predict temperature resistance

Pre- and postzygotic mechanisms preventing hybridization in co-occurring species of the Impatiens purpureoviolacea complex