| Summary: | We performed next-generation sequencing of the 18S rDNA–ITS1–5.8S rDNA region along with traditional Sanger sequencing of <i>rbc</i>L, <i>mat</i>K, <i>ndh</i>F, and ITS1–5.8S rDNA–ITS2 to clarify the hybridization pattern in the subtribe Alopecurinae and in the genus <i>Alopecurus</i> in particular. Our data support the hybrid origin of <i>Alopecurus</i> × <i>brachystylus</i> from hybridization between <i>A. geniculatus</i> (sect. <i>Alopecurium</i>) and <i>A. pratensis</i> (sect. <i>Alopecurus</i>). Moreover, in the rDNA of hybrid <i>A.</i> × <i>brachystylus</i>, only <i>A. aequalis</i>-like ribotypes from tetraploid <i>A. geniculatus</i> participated. Surprisingly, we found the traces of introgression of <i>A. arundinaceus</i>-like ribotypes not only in hybrid <i>A.</i> × <i>marssonii</i> (<i>A. geniculatus</i> × <i>A. arundinaceus</i>) but in <i>A. aequalis</i> s. str. as well. A high-polyploid group from the section <i>Alopecurus</i>, <i>A</i>. aggr. <i>alpinus</i> has undoubted hybrid origin: e. g., <i>A. brachystachyus</i> has rDNA from the sect. <i>Alopecurium</i>. <i>Alopecurus alpinus</i>, with its allies, is clearly distinct from other members of the sect. <i>Alopecurus</i> (especially by maternal line) and thus we can re-establish a previous opinion about the separate group to which <i>A. alpinus</i> belongs. Species from the section <i>Colobachne</i> (presumably Alpine grasses from Ancient Mediterranean region) probably hybridized with the <i>A. alpinus</i> group. Even <i>A. myosuroides</i> (sect. <i>Pseudophalaris</i>) that could be referred to the separate genus has ribotypes common with the species of the section <i>Alopecurium</i> (<i>A. aequalis</i>, <i>A. geniculatus</i>) in one of the accessions. Additionally, we found that the possible polyphyletic origin of the genus <i>Limnas</i>. <i>Limnas stelleri</i> is very close to <i>Alopecurus magellanicus</i> according to NGS data, while <i>L. malyschevii</i> is more or less distinct from other studied species of the genus <i>Alopecurus</i>.
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