The Polyommatus butterflies have holocentric chromosomes, which are characterized by kinetic activity distributed along the entire chromosome length, and the highest range of haploid chromosome numbers (n) known within a single eukaryotic genus (from n = 10 to n = 226). Previous analyses have shown that these numbers most likely evolved gradually from an ancestral karyotype, in accordance with the Brownian motion model of chromosome change accumulation. Here we studied chromosome sets within a monophyletic group of previously non-karyotyped Polyommatus species. We demonstrate that these species have a limited interspecific chromosome number variation from n = 16 to n = 25, which is consistent with the Brownian motion model prediction. We also found intra- and interpopulation variation in the chromosome numbers. These findings support the model of karyotype evolution through the gradual accumulation of neutral or weakly underdominant rearrangements that can persist in the heterozygous state within a population. For Polyommatus poseidonides we report the phenomenon of flexible meiosis in which the chromosome multivalents are able to undergo either conventional or inverted meiosis within the same individual. We hypothesise that the ability to invert the order of the meiotic events may be adaptive and can facilitate proper chromosome segregation in chromosomal heterozygotes, thus promoting rapid karyotype evolution.