Methodology significantly affects genome size estimates: quantitative evidence using bryophytes

Cytometry. Part a : the Journal of the International Society for Analytical Cytology
Jillian D BainardSteven G Newmaster

Abstract

Flow cytometry (FCM) is commonly used to determine plant genome size estimates. Methodology has improved and changed during the past three decades, and researchers are encouraged to optimize protocols for their specific application. However, this step is typically omitted or undescribed in the current plant genome size literature, and this omission could have serious consequences for the genome size estimates obtained. Using four bryophyte species (Brachythecium velutinum, Fissidens taxifolius, Hedwigia ciliata, and Thuidium minutulum), three methodological approaches to the use of FCM in plant genome size estimation were tested. These included nine different buffers (Baranyi's, de Laat's, Galbraith's, General Purpose, LB01, MgSO(4), Otto's, Tris.MgCl(2), and Woody Plant), seven propidium iodide (PI) staining periods (5, 10, 15, 20, 45, 60, and 120 min), and six PI concentrations (10, 25, 50, 100, 150, and 200 microg ml(-1)). Buffer, staining period and staining concentration all had a statistically significant effect (P = 0.05) on the genome size estimates obtained for all four species. Buffer choice and PI concentration had the greatest effect, altering the 1C-values by as much as 8% and 14%, respectively. As well, the qualit...Continue Reading

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Citations

Aug 19, 2011·Chromosome Research : an International Journal on the Molecular, Supramolecular and Evolutionary Aspects of Chromosome Biology·Jillian D BainardSteven G Newmaster
Aug 27, 2011·Chromosome Research : an International Journal on the Molecular, Supramolecular and Evolutionary Aspects of Chromosome Biology·Jillian D BainardPaul Kron
Oct 31, 2013·Genome Génome / Conseil National De Recherches Canada·Jillian D Bainard, Juan Carlos Villarreal
Jul 29, 2011·Cytometry. Part a : the Journal of the International Society for Analytical Cytology·Man C CheungDaniel J Ehrlich
Jul 24, 2010·Cytometry. Part a : the Journal of the International Society for Analytical Cytology·J Suda, Ilia J Leitch
Nov 22, 2012·Cytometry. Part a : the Journal of the International Society for Analytical Cytology·Attila Tárnok
Jun 24, 2014·Cytometry. Part a : the Journal of the International Society for Analytical Cytology·Tereza VeselskáMiroslav Kolařík
Apr 30, 2013·Molecular Phylogenetics and Evolution·Jillian D BainardSteven G Newmaster
Nov 30, 2019·Annals of Botany·Jillian D BainardJessica M Budke
Mar 23, 2021·Cytometry. Part a : the Journal of the International Society for Analytical Cytology·João LoureiroSílvia Castro

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