Biogeography and genetic diversity of freshwater diatoms: The potential of large combined rbcL metabarcoding datasets

Aiming to gain a general picture of rbcL diversity within freshwater diatom species, this study assembles and analyzes multiple metabarcoding datasets spanning various geographical regions. From these datasets, we inferred >10,000 amplicon sequence variants (ASVs) of 263-bp length. More than half of the 1000 most abundant ASVs were recorded in both Eurasia and N America and there was only limited evidence for continent-specific lineages. The geographical range was extended for some species, illustrating the potential of metabarcoding datasets for such checks. For detailed analysis of intraspecific diversity, 73 freshwater species were selected, corresponding to 360 ASVs assigned phylogenetically. We found notable variation, some species being represented by only one or a few ASVs, while others were represented by a higher number. Furthermore, within species, ASVs exhibited different dominance and distribution patterns, in some cases with a head-tail pattern, in others a more equal spread of abundance or unresolved reticulate relationships. Except for Ulnaria ulna, no geographical structure among species' ASVs was detectable in haplotype networks using the 263-bp rbcL marker. Observed heterogeneity within species was categorized by computing several metrics of genetic variation and classified into three groups, reflecting optimal sampling strategies based on the patterns of intraspecific variation in the 73 target species There was a significant relationship between intraspecific diversity and the traditional separation between 'centric' and 'pennate' diatoms, with centric species exhibiting significantly fewer variants than pennates, possibly because of different plastid inheritance patterns.