The use of flow cytometry for fungal nuclear DNA quantification

Genome size information is sparse across fungi, with information being available for less than 2000 species. So far, most records have been obtained using static, microscope‐based cytometry methods or derived from genome sequencing projects. Flow cytometry is now considered the state‐of‐the‐art meth...

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Bibliographic Details
Published in:Cytometry. Part A 2021-04, Vol.99 (4), p.343-347
Main Authors: Talhinhas, Pedro, Carvalho, Rita, Loureiro, João
Format: Article
Language:English
Subjects:
Best practice
Deoxyribonucleic acid
DNA
DNA standards
Flow cytometry
Fungi
Gene sequencing
genome size
Genomes
Mutualism
mycology
nuclear extraction
Parasitism
Phylogeny
Ploidy
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Summary:Genome size information is sparse across fungi, with information being available for less than 2000 species. So far, most records have been obtained using static, microscope‐based cytometry methods or derived from genome sequencing projects. Flow cytometry is now considered the state‐of‐the‐art method for obtaining genome size measurements, and appropriate methods and DNA standards are available, enabling the analysis of most genome size ranges in a rapid, robust and inexpensive way. The average fungal genome size is 60 Mbp, but sizes vary across phylogeny, ranging from 2.2 (Encephalitozoon romaleae) to 3706 Mbp (Jafnea semitosta). In several fungal clades, genome size expansion seems to accompany evolution either to plant mutualism or to plant parasitism (particularly biotrophy), and fungi that interact with plants seem to have larger genomes than saprobes and those that interact with animals. Whereas flow cytometry for nuclear DNA quantification is routinely employed in plant sciences for genome size and ploidy studies, its use in fungal biology is still infrequent. Appropriate standards, methods and best practices are described here, with the aim of stimulating a more generalized and widespread use of flow cytometry for fungal genome size measurement.
ISSN:1552-4922
1552-4930
DOI:10.1002/cyto.a.24335