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Lycogala epidendrum  is one of the most widely known myxomycete species and the first-ever discovered representative of this group. Using 687 original DNA sequences from 330 herbarium specimens from Europe, Asia, North and Central America, and Australia, we constructed the first detailed phylogenies of the genus  Lycogala , based on two independently inherited genetic markers, the ribosome small subunit 18S rRNA nuclear gene (18S rDNA) and the mitochondrial cytochrome oxidase subunit I gene (COI). In both phylogenies,  L. epidendrum  appeared to be a polyphyletic group, represented by numerous clades. The four other recognized species of the genus ( L. confusum, L. conicum, L. exiguum , and  L. flavofuscum ) are scattered between branches corresponding to  L. epidendrum . A barcode gap analysis revealed 60 18S rDNA phylogroups of  L. epidendrum , which are distant from each other not less than from other species of the genus  Lycogala . For 18 of these phylogroups with both 18S rDNA and COI sequences available, recombination patterns were analyzed to test for reproductive isolation. In contrast to the results of a simulation assuming panmixis, no crossing between ribosomal and mitochondrial phylogroups was found, thus allowing the conclusion that all tested phylogroups represent biospecies. More than one third (39.6%) of the studied specimens share a single 18S rDNA phylogroup, which we consider to be  L. epidendrum  s. str. This group displays the broadest geographic distribution and the highest intraspecific genetic variability. Nearly all (93.3%) of the remaining non-singleton 18S rDNA phylogroups are restricted to certain continents or even regions. At the same time, various reproductively isolated phylogroups occur sympatric at a given location.

Two independent genetic markers support separation of the myxomycete Lycogala epidendrum into numerous biological species