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Poa is a complex genus taxonomically and genetically. As a result, relatively little information exists for this economically important genus with regards to reproductive mode and variability in chromosome number. We examined apomixis frequency and ploidy levels in 83 Poa accessions representing 33 species from the National Plant Germplasm System using flow cytometric techniques. In reproductive mode analysis, we analyzed at least three preparations of 50 seeds each from the accessions. In ploidy level analysis, at least three plants of each accession were analyzed. Sixty percent of the species had at least one apomictic or facultative apomictic accession; 40% were sexual. Thirty-three percent of the species had no sexual accessions. Autonomous apomixis was newly identified in P. bactriana and P. bulbosa in addition to it being previously reported in P. nervosa. Poa annua and P. trivialis were identified as obligate sexual, and P. nervosa as obligate apomictic. Two of four P. palustris accessions exhibited facultative apomixis. Ploidy levels among sexually reproducing Poa accessions ranged from 2n = 2x = 14 to 2n = 12x = 84, with most accessions between 2n = 3x = 21 to 2n = 8x = 56. Among apomictic accessions, ploidy levels ranged from 2n = 3x = 21 to 2n = 10x = 70, with most between 2n = 4x = 28 and 2n = 10x = 70. These results emphasize the wide variability in Poa, both among and within species.