Characterizing polyploidy inArabidopsis lyratausing chromosome counts and flow cytometry

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Abstract
Protocols were developed for both chromosome counts and flow cytometry to assess ploidy level and DNA content for populations of Arabidopsis lyrata L. sampled from Europe (Arabidopsis lyrata subsp. petraea), North America (Arabidopsis lyrata subsp. lyrata), and Japan (Arabidopsis lyrata subsp. kawasakiana). Ploidy variation within this species is not clear, with previous studies having documented both diploid and tetraploid populations. Chromosome counts in this study confirmed ploidy expectations for all populations examined. Individuals from Iceland and North America were diploid (2n = 2x = 16), whereas those from Japanese and Austrian populations were tetraploid (2n = 4x = 32). Flow cytometry was also used successfully to distinguish between ploidy levels, but the need to calibrate DNA content measures with chromosome counts was demonstrated by a deviation from the expected 2:1 ratio between tetraploid and diploid values among European populations (A. lyrata subsp. petraea). This deviation might be explained by a hybrid (allopolyploid) origin or by genomic changes following polyploidization, emphasizing the dynamic nature of polyploid genomes. Variation in DNA content among families was found only for North American populations, but these individuals were sampled from a broader geographic range than those from other regions.Key words: cytogenetics, flow cytometry, polyploidy, Arabidopsis lyrata, genome size, chromosome counts.