High mountains of central Europe as a refuge of surprising cytotype diversity of Huperzia selago (Lycopodiaceae)

Polyploidization is pivotal in plant speciation, affecting adaptability, ecological tolerance and specific geographical distribution patterns. While cytotype diversity has been extensively studied in angiosperms and ferns, knowledge in homosporous lycophytes remains very limited. Our study addresses this gap, focusing on the homosporous lycophyte Huperzia selago in Central Europe. The genome size of 1330 individuals from 208 populations were assessed via flow cytometry, revealing five distinct cytotypes (2x, 3x, 4x, 5x, 6x). Challenging chromosome counting using gemmae roots yielded average counts of 140 (2x), 204 (3x), and 262 (4x) chromosomes. The hexaploid genome size (29 pg) approached the upper limits reported for lycophytes. Tetraploids were the most abundant (72.7%), while triploids (21.3%) were less common, while the remaining cytotypes comprised less than 5% of the dataset. Geographical separation of cytotypes was not observed. However, uncommon cytotypes were associated with the Alps and triploids also occured in the highest parts of Western Carpathians. Around 27.3% of populations exhibited multiple cytotypes. Except for atypical diploids, spore abortion differed between even (21.8%) and odd ploidies (65.2%). Microcharacter (stoma, spore) proportions did not linearly correspond with increasing ploidy levels. The high ploidy-level diversity and cytotype coexistence in Central European H. selago match the well documented patterns in ferns and angiosperms. These findings provide valuable insights into lycophyte polyploidy, underscoring the necessity for broader geographical sampling and application of molecular studies to elucidate phylogenetic relationships and taxonomic classifications within the genus Huperzia.