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Recent morphology-based investigations of freshwater Arctic diatoms suggest that many species remain to be discovered, reflecting a unique polar flora. During a survey of the freshwater diatom flora of northern Billefjorden, including the Petuniabukta fjord region (Spitsbergen), several morphodemes belonging to the Achnanthidium minutissimum species complex were recorded. Molecular phylogenies based on rbcL, 28S and 18S sequences, including single cells from Canada and strains from Marion Island (sub-Antarctica) and GenBank revealed the presence of 12 distinct A. minutissimum complex lineages, of which three contained strains from Spitsbergen. One Arctic lineage is described as a new species. Achnanthidium digitatum sp. nov. is morphologically characterized by narrow, linear to only slightly lanceolate valves and usually two areolae per stria. The two remaining Arctic lineages are in need of a more complete morphological and molecular comparison with other representatives of the A. minutissimum complex to clarify their taxonomic identity. It is argued that implementation of molecular data in the taxonomy of Achnanthidium will be essential to solve the taxonomic problems associated with this group, eventually resulting in a better understanding of the biogeography and niche differentiation of different species belonging to the A. minutissimum complex. Ideally, this should be based on more variable genes than the currently widely used 18S, which does not have a species level resolution in the A. minutissimum complex.