Cortinarius s.l. is a globally distributed agaricoid genus that has been well studied in Europe with over 1000 described species. However, the information about their taxonomy and diversity in eastern Central Europe is still limited. Only 124 species have been reported so far from Romania, based solely on morphological observations. The aim of this study was to re-examine the diversity of the genus Cortinarius s.l. in the Romanian Carpathian area, employing molecular phylogenetic and morphological methods. During intensive field work in the period 2017–2020, a total of 234 Cortinarius s.l. specimens were collected and studied with integrative taxonomic methods. For all the samples, we amplified and sequenced the nrDNA ITS region, which is the widely used official barcode marker of fungi. These sequences were compared to the data found in public databases (GenBank, UNITE, BOLD). Based on phylogenetic analyses, we identified 109 Cortinarius s.l. species, which represent 40 sections and 3 clades. Out of these species, 43 have previously been documented from Romania based on morphological identification methods, while 66 species are reported as new to the country.

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doi: bioRxiv preprint 1. Once widespread in the large European rivers, Palingenia longicauda underwent a drastic range contraction as a result of the intense pollution and hydromorphological interventions of the 19 th and 20 th centuries. For the last decades it was considered to be restricted only to the Tisa River and its tributaries, and to the Rába River, but new reports indicated its presence in the Danube River in Hungary, in the Danube Delta in Romania and Ukraine, and in the Prut River in the Republic of Moldova. 2. The objective of this study is to analyze the phylogeographic pattern between the two main eco-regions (Pannon and Pontic) of the species distribution, based on the combined mitochondrial COI (472 bp) and 16S (464 bp) sequences generated for individuals collected in Romania and Ukraine, and from publicly available ones, representing the Tisa catchment populations. 3. The presence of viable populations in the Danube Delta and on the Prut River in Romania is confirmed, and additional presence on the Mure and Bega rivers from Romania, and on the Styr and Horyn' rivers in Northern Ukraine is shown. 4. The phylogeographic results indicate that the presence of the analyzed populations are not the result of recent founding events from the Pannon region, confirming the survival and expansion of cryptic local lineages. 5. The recent recovery of the species may be related to the improvement of water quality as a result of the implementation of the EU Water Framework Directive and the EU Floods Directive after 2000.

Once widespread in the large European rivers, Palingenia longicauda underwent a drastic range contraction as a result of the intense pollution and hydromorphological interventions of the 19th and 20th centuries. For the last decades it was considered to be restricted only to the Tisa River and its tributaries, and to the Rba&aacute River, but new reports indicated its presence in the Danube River in Hungary, in the Danube Delta in Romania and Ukraine, and in the Prut River in the Republic of Moldova. The objective of this study is to analyze the phylogeographic pattern between the two main eco-regions (Pannon and Pontic) of the species distribution, based on the combined mitochondrial COI (472 bp) and 16S (464 bp) sequences generated for individuals collected in Romania and Ukraine, and from publicly available ones, representing the Tisa catchment populations. The presence of viable populations in the Danube Delta and on the Prut River in Romania is confirmed, and additional presence on the Mureș and Bega rivers from Romania, and on the Styr and Horyn rivers in Northern Ukraine is shown. The phylogeographic results indicate that the presence of the analyzed populations are not the result of recent founding events from the Pannon region, confirming the survival and expansion of cryptic local lineages. The recent recovery of the species may be related to the improvement of water quality as a result of the implementation of the EU Water Framework Directive and the EU Floods Directive after 2000.