Flow Cytometry-Based Determination of Ploidy from Dried Leaf Specimens in Genomically Complex Collections of the Tropical Forage Grass Urochloa s. l.

A - Papers appearing in refereed journals

Tomaszewska, P., Pellny, T. K., Hernandez, L. M., Mitchell, R. A. C., Castiblanco, V., De Vega, J. J., Schwarzacher, T. and Heslop-Harrison, P. 2021. Flow Cytometry-Based Determination of Ploidy from Dried Leaf Specimens in Genomically Complex Collections of the Tropical Forage Grass Urochloa s. l. Genes. 12 (7), p. 957. https://doi.org/10.3390/genes12070957

AuthorsTomaszewska, P., Pellny, T. K., Hernandez, L. M., Mitchell, R. A. C., Castiblanco, V., De Vega, J. J., Schwarzacher, T. and Heslop-Harrison, P.
Abstract

Urochloa (including Brachiaria, Megathyrus and some Panicum) tropical grasses are native to Africa and are now, after selection and breeding, planted worldwide, particularly in South America, as important forages with huge potential for further sustainable improvement and conservation of grasslands. We aimed to develop an optimized approach to determine ploidy of germplasm collection of this tropical forage grass group using dried leaf material, including approaches to collect, dry and preserve plant samples for flow cytometry analysis. Our methods enable robust identification of ploidy levels (coefficient of variation of G0/G1 peaks, CV, typically <5%). Ploidy of some 348 forage grass accessions (ploidy range from 2x to 9x), from international genetic resource collections, showing variation in basic chromosome numbers and reproduction modes (apomixis and sexual), were determined using our defined standard protocol. Two major Urochloa agamic complexes are used in the current breeding programs at CIAT and EMBRAPA: the ’brizantha’ and ’humidicola’ agamic complexes are variable, with multiple ploidy levels. Some U. brizantha accessions have odd level of ploidy (5x), and the relative differences in fluorescence values of the peak positions between adjacent cytotypes is reduced, thus more precise examination of this species is required. Ploidy measurement of U. humidicola revealed aneuploidy.

KeywordsPloidy; Flow cytometry; Apomixis; Dried specimens; Brachiaria; Panicum; Tropical forage grasses
Year of Publication2021
JournalGenes
Journal citation12 (7), p. 957
Digital Object Identifier (DOI)https://doi.org/10.3390/genes12070957
Open accessPublished as ‘gold’ (paid) open access
FunderBiotechnology and Biological Sciences Research Council
Funder project or codes5633
BB/R022828/1
Publisher's version
Output statusPublished
Publication dates
Online23 Jun 2021
Publication process dates
Accepted16 Jun 2021

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