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The effects of rapid desiccation on estimates of plant genome size

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Abstract

Flow cytometry has become the dominant method for estimating nuclear DNA content in plants, either for ploidy determination or quantification of absolute genome size. Current best practices for flow cytometry involve the analysis of fresh tissue, however, this imposes significant limitations on the geographic scope and taxonomic diversity of plants that can be included in large-scale genome size studies. Dried tissue has been used increasingly in recent years, but largely in the context of ploidy analysis. Here we test rapid tissue drying with silica gel as a method for use in genome size studies, potentially enabling broader geographic sampling of plants when fresh tissue collection is not feasible. Our results indicate that rapid drying introduces comparatively minor error (<10%), which is similar to the error introduced by other common methodological variations such as instrument. Additionally, the relative effect of drying on genome size and data quality varied between species and buffers. Tissue desiccation provides a promising approach for expanding our knowledge of plant genome size diversity.

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Abbreviations

CV:

Coefficient of variation

PI:

Propidium iodide

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Acknowledgments

This study represented a collaborative project involving three research groups and was supported by Natural Sciences and Engineering Research Council of Canada (NSERC) grants to BCH, TRG, and SGN and used equipment provided through the Canada Foundation for Innovation (CFI) and the Ontario Research Fund (ORF). Additional support was provided to BCH through a Canada Research Chair and to JDB by an NSERC postgraduate scholarship. We sincerely thank P. Šmarda and J. Suda for providing constructive feedback on the manuscript. We wish to thank several other lab members for assisting with flow cytometric analysis, including Anastasia Richardson, Kelly Hadfield, Nicholas Jeffery, João Lima, Paula Nathwani, and Paola Pierossi. We also thank Hana Serajaddini, Lindsay Wilson, Benjamin Yim, and Thomas Henry for lab and field assistance.

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Authors and Affiliations

  1. Department of Integrative Biology, University of Guelph, Guelph, ON, Canada, N1G 2W1

    Jillian D. Bainard, Brian C. Husband, Sarah J. Baldwin, Aron J. Fazekas, T. Ryan Gregory, Steven G. Newmaster & Paul Kron

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  1. Jillian D. Bainard
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  2. Brian C. Husband
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Correspondence to Paul Kron.

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Responsible Editors: T. Ryan Gregory and Jillian D. Bainard.

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Bainard, J.D., Husband, B.C., Baldwin, S.J. et al. The effects of rapid desiccation on estimates of plant genome size. Chromosome Res 19, 825–842 (2011). https://doi.org/10.1007/s10577-011-9232-5

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