Unexpected species richness of the lichen genus Protoblastenia (Lecanorales, Psoraceae) in Finland

193 Unexpected species richness of the lichen genus Protoblastenia (Lecanorales, Psoraceae) in Finland Juha Pykälä1, Leena Myllys2 1 Nature Solutions, Finnish Environmental Institute, Latokartanonkaari 11, FI-00790 Helsinki, Finland 2 Botany and Mycology Unit, Finnish Museum of Natural History, P. O. Box 7, FI-00014 University of Helsinki, Helsinki, Finland Corresponding author: Juha Pykälä ([email protected]) Copyright: © Juha Pykälä & Leena Myllys. This is an open access article distributed under terms of the Creative Commons Attribution License (Attribution 4.0 International – CC BY 4.0). Research Article Abstract The taxonomy of Protoblastenia in Finland was studied, based on morphology and molecular data (nuITS rDNA sequences). Twenty species were recognised, with sixteen species being newly described here: P. arupii sp. nov., P. borealis sp. nov., P. compressa sp. nov., P. dolomitica sp. nov., P. ekmanii sp. nov., P. fennoarctica sp. nov., P. minuta sp. nov., P. oulankaensis sp. nov., P. pseudocompressa sp. nov., P. pseudoterricola sp. nov., P. remota sp. nov., P. rikkinenii sp. nov., P. saanaensis sp. nov., P. timdalii sp. nov., P. violacea sp. nov. and P. westbergii sp. nov. All species are confined to calcareous rocks, except P. terricola which also grows on calcareous soil. The calcareous fells in Enontekiö in NW Finland were identified as hot spots of Protoblastenia diversity. Nine newly-described species (P. arupii, P. fennoarctica, P. minuta, P. pseudoterricola, P. rikkinenii, P. saanaensis, P. timdalii, P. violacea and P. westbergii) are restricted to this area in Finland. Several of the species are semi-cryptic. On average, they may have minor morphological differences, but many specimens cannot be identified, based on morphology only. Protoblastenia rikkinenii is reported from Norway and P. oulankaensis from Italy, based on GenBank sequences. Full descriptions and a preliminary key of Protoblastenia in Finland are provided. Key words: Calcareous rocks, DNA barcoding, Italy, ITS, lichenised fungi, new species, Norway, phylogeny Introduction Protoblastenia (Zahlbr.) J. Steiner is a lichenised genus in the family Psoraceae. The genus includes ca. 15–25 accepted, mainly calcicolous, species (Castello and Nimis 1995; Kainz 2004; Kainz and Rambold 2004). Most species occur on calcareous rocks and a few on soil over calcareous rock. Species of Protoblastenia have anthraquinones in the apothecia, causing predominantly orange-coloured discs. Other typical features are predominantly convex apothecia and pale, non-septate medium-sized spores (Kainz and Rambold 2004). Protoblastenia differs from the closely-related genus Psora Hoffm. in the absence or weakly developed upper cortex in the thallus and the absence of calcium oxalate crystals in the hypothecium (Timdal 1987; Kainz and Rambold 2004). Based on the phylogenetic studies of Kainz and Rambold (2004), Ekman and Blaalid (2011) and Miadlikowska et al. (2014), Protoblastenia forms a monophyletic group. Academic editor: Thorsten Lumbsch Received: 22 June 2025 Accepted: 3 September 2025 Published: 29 October 2025 Citation: Pykälä J, Myllys L (2025) Unexpected species richness of the lichen genus Protoblastenia (Lecanorales, Psoraceae) in Finland. MycoKeys 124: 193–226. https://doi. org/10.3897/mycokeys.124.162802 MycoKeys 124: 193–226 (2025) DOI: 10.3897/mycokeys.124.162802 194 MycoKeys 124: 193–226 (2025), DOI: 10.3897/mycokeys.124.162802 Juha Pykälä & Leena Myllys: Lichen genus Protoblastenia in Finland Kainz and Rambold (2004) studied the phylogeny and taxonomy of Protoblastenia in Central Europe. They ITS-sequenced 11 species: Protoblastenia aurata Poelt & Vězda, P. calva (Dicks.) Zahbr., P. calvella Kainz & Rambold, P. cyclospora (Hepp ex Körb.) Poelt, P. incrustans (DC.) J. Steiner, P. laeta (Poelt) Kainz & Rambold, P. lilacina Poelt & Vězda, P. pseudoincrustans ined., P. rupestris (Scop.) J. Steiner, P. siebenhaariana (Körb.) J. Steiner and P. terricola (Anzi) Lynge. Three European species, Protoblastenia coniasis (A. Massal.) Poelt (Poelt 1957), P. geitleri Zahlbr. (Zahlbruckner 1936) and P. szaferi J. Nowak (Nowak 1974), were not sequenced. So far, seven species of Protoblastenia are known in Finland: P. calva, P. calvella, P. incrustans, P. lilacina, P. rupestris, P. siebenhaariana and P. terricola (Stenroos et al. 2016; Pykälä 2023). However, during the DNA barcoding of the Finnish lichen biota, it became apparent that the genus is much in need of revision. Here, we present the results of the revised taxonomy and show that the number of species in Finland is 20, almost three times more than previously reported. Material and methods Taxon sampling This study is based on material collected by the first author (JP) during the years 1990–2022 in Finland. The specimens are stored in H. Morphology Apothecia and thalli were hand-sectioned with razor blades. The sections were examined and measured in tap water. Asci and ascospores were also studied in squash preparations of apothecia mounted in water. Spores were measured using 10% potassium hydroxide (KOH). Additionally, the thickness of the hypothecium was examined by cutting the apothecia into two pieces and studying the pieces using a binocular microscope. The range of spore size is indicated as arithmetic mean and standard deviation. Minimum and maximum values are given in parentheses. The size of the apothecia (in diameter) is given in surface view. DNA extraction and sequencing Total genomic DNA was extracted from the apothecia (1–3) of 1 – 31-year-old herbarium specimens. We used two different techniques for extraction and sequencing. Most Finnish samples were sequenced as part of the “Finnish Barcode of Life” research project conducted from 2012–2023 (FinBOL, https://finbol.org/). The samples were placed on 96-well microplates and sent to the Canadian Centre for DNA Barcoding (CCDB). CCDB’s standard protocols (documentation available at http://ccdb.ca/resources) were used for extraction, PCR and sequencing. The primers ITS1 and ITS4 (White et al. 1990) were used both for PCR and sequencing of the nuclear ribosomal ITS region. The barcode sequences and their trace files along with all relevant collection data and photographs of the voucher specimens were uploaded to the Barcode of Life Data Systems (BOLD, https://www.boldsystems.org) database. 195 MycoKeys 124: 193–226 (2025), DOI: 10.3897/mycokeys.124.162802 Juha Pykälä & Leena Myllys: Lichen genus Protoblastenia in Finland DNA of the 31 specimens (Pykälä 5963, 8547, 29704, 33037, 34043, 35585, 39693, 39851, 39987, 41511b, 43105, 43194, 53252, 43340, 43524, 43605, 43624, 43855, 43910, 44115, 44130, 44177, 44275, 44314, 44431, 44438, 44876, 46413, 57501, 58392, 58457) was extracted using DNeasy® Blood & Tissue kits by Qiagen following the protocol described in Myllys et al. (2011). PCR reactions were prepared using PuReTaq Ready-To-Go PCR beads (GE Healthcare). The 25 µl reaction volume contained 19 µl of dH2O, 0.4 µM of each primer and 4 µl of extracted DNA. PCR was run under the following conditions: initial denaturation for 5 min at 95 °C followed by five cycles of 30 s at 95 °C (denaturation), 30 s at 58 °C (annealing) and 1 min at 72 °C (extension); in the remaining 35 cycles, the annealing temperature was decreased to 56 °C; the PCR schedule ended with a final extension for 7 min at 72 °C. PCR products were purified and sequenced by Macrogen Inc. (Amsterdam, the Netherlands; https:// www.macrogen.com) or alternatively cleaned with ExoSAP (Affymetrix, Santa Clara, California, USA) and sequenced by FIMM Genomics (https://www2.helsinki.fi/en/infrastructures/genome-analysis/infrastructures/fimm-genomics). The primers ITS1F (Gardes and Bruns 1993) and ITS4 (White et al. 1990) were used both for PCR amplification and sequencing of the ITS regions. Phylogenetic analyses All the ITS sequences of Protoblastenia available in GenBank (n = 55) were downloaded (1.3.2025). Psora decipiens (Hedw.) Hoffm. and P. rubiformis (Ach.) Hook. were used as outgroups, based on the phylogenies of Miadlikowska et al. (2014) and Evankow et al. (2025). A total of 141 ITS sequences were aligned (Table 1) with MUSCLE v.3.8.31 (Edgar 2004) using EMBL-EBI’s freely available web service (http://www.ebi. ac.uk/Tools/msa/muscle/). The aligned dataset was subjected to Maximum Likelihood analysis (ML). The analysis was performed with RAxML v.8.1.15 (Stamatakis 2014) on the CSC – IT Center for Science server ((http://www. csc.fi). The ITS region was divided into three partitions: ITS1, 5.8S and ITS2. These partitions were analysed under the universal GTR-GAMMA model. Node support was estimated with 1000 bootstrap replications using the rapid bootstrap algorithm. Branches with bootstrap values ≥ 70% were considered strongly supported. Results and discussion We generated 83 new ITS sequences obtained from the Finnish Protoblastenia specimens. In the ITS phylogeny (Fig. 1), the sequenced Finnish specimens were divided into 20 lineages of which 16 do not match any described species. These lineages, when represented by multiple samples, received high support values (96–100%) and are described here as new species. Typically, infraspecific differences in ITS in the Finnish Protoblastenia species are small. As expected, the phylogeny, based on only ITS sequences, remains unsupported at deeper nodes. However, six well-supported (ML bootstrap support > 70%) lineages represented by more than one species were identified: 1) P. fennoarctica sp. nov. + P. rikkinenii sp. nov. + P. saanaensis sp. nov. + P. timdalii sp. nov. + P. pseudoincrustans ined. 2) P. dolomitica sp. nov. + P. westbergii sp. nov., 196 MycoKeys 124: 193–226 (2025), DOI: 10.3897/mycokeys.124.162802 Juha Pykälä & Leena Myllys: Lichen genus Protoblastenia in Finland Table 1. Specimens used in the phylogenetic analyses. New sequences are in bold. Taxon Country Voucher specimen GenBank Accession number (ITS) Protoblastenia arupii Finland Pykälä 44130 (H) PV766636 P. aurata Austria Kainz 950 (M) AY425658 P. aurata Austria Wittmann (M) AY425659 P. borealis Finland Pykälä 16567 (H) PV766637 P. borealis Finland Pykälä 23710 (H) PV766638 P. borealis Finland Pykälä 25292 (H) PV766639 P. borealis Finland Pykälä 26097 (H) PV766640 P. borealis Finland Pykälä 29704 (H) PV766641 P. borealis Finland Pykälä 32757 (H) PV766642 P. borealis Finland Pykälä 33037 (H) PV766643 P. borealis Finland Pykälä 33101 (H) PV766644 P. borealis Finland Pykälä 38784 (H) PV766645 P. borealis Finland Pykälä 39151 (H) PV766646 P. borealis Finland Pykälä 39942 (H) PV766647 P. borealis Finland Pykälä 40695 (H) PV766648 P. borealis Finland Pykälä 41355 (H) PV766649 P. borealis Finland Pykälä 42802 (H) PV766650 P. borealis Finland Pykälä 44641 (H) PV766651 P. borealis Finland Pykälä 44906 (H) PV766652 P. borealis Finland Pykälä 45759 (H) PV766653 P. borealis Finland Pykälä 47217 (H) PV766654 P. borealis Finland Pykälä 57501 (H) PV766655 P. borealis Finland Pykälä 58196 (H) PV766656 P. borealis Finland Pykälä 58392 (H) PV766657 P. borealis Finland Pykälä 60511 & Kuusisto (H) PV766658 P. calva Ireland Sipman 30678 (B) AY425628 P. calva Austria Kainz 981 (M) AY425642 P. calva Germany Kainz 501 (M) AY425643 P. calva France Rambold 6242 (M) AY425644 P. calva Germany Kainz 852 (M) AY425645 P. calva Norway Edvardsen & Ekman NO6 (BG) EF524319 “P. calva” Croatia Gueidan CG659 (DUKE) HQ650618 P. calvella Germany Kainz 764 (M) AY425646 P. calvella Germany Kainz 765 (M) AY425647 P. calvella Austria Kainz 967 (M) AY425648 P. calvella Germany Kainz 805 (M) AY425649 P. calvella Finland Pykälä 44807 (H) OQ604661 P. calvella Finland Pykälä 43503 (H) OQ604662 P. calvella Sweden Ekman & Eide Ekman 5821 (UPS) OR773065 P. compressa Finland Pykälä 35585 (H) PV766659 P. compressa Finland Pykälä 36521 (H) PV766660 P. compressa Finland Pykälä 38349 (H) PV766661 P. compressa Finland Pykälä 38646 (H) PV766662 P. compressa Finland Pykälä 38965 (H) PV766663 P. compressa Finland Pykälä 41376 (H) PV766664 197 MycoKeys 124: 193–226 (2025), DOI: 10.3897/mycokeys.124.162802 Juha Pykälä & Leena Myllys: Lichen genus Protoblastenia in Finland Taxon Country Voucher specimen GenBank Accession number (ITS) P. compressa Finland Pykälä 41511b (H) PV766665 P. compressa Finland Pykälä 42124 (H) PV766666 P. cyclospora Italy Tretiach (M) AY425667 P. cyclospora Greece Kainz 913 (M) AY425668 P. dolomitica Finland Pykälä 36072 (H) PV766667 P. dolomitica Finland Pykälä 39217 (H) PV766668 P. dolomitica Finland Pykälä 39851 (H) PV766669 P. dolomitica Finland Pykälä 43340 (H) PV766670 P. dolomitica Finland Pykälä 43466 (H) PV766671 P. dolomitica Finland Pykälä 43855 (H) PV766672 P. dolomitica Finland Pykälä 44115 (H) PV766673 P. ekmanii Finland Pykälä 37508 (H) PV766674 P. fennoarctica Finland Pykälä 43105 (H) PV766675 P. fennoarctica Finland Pykälä 43194 (H) PV766676 P. fennoarctica Finland Pykälä 43656 (H) PV766677 P. fennoarctica Finland Pykälä 44116 (H) PV766678 P. incrustans Svalbard Zhang KP314370 P. incrustans Svalbard Zhang ZT2013200 KP314440 P. incrustans Germany Kainz 498 (M) AY425630 P. incrustans Germany Kainz 855 (M) AY425631 P. incrustans Germany Kainz 842 (M) AY425632 P. incrustans Germany Kainz 228 (M) AY425669 P. incrustans Germany Kainz 220 (M) AY425670 P. laeta Germany Kainz 856 (M) AY425650 P. laeta Germany Kainz 737 (M) AY425651 P. laeta Germany Kainz 741 (M) AY425652 P. laeta France Rambold 6253 (M) AY425653 “P. lilacina” Italy Kainz 161 (M) AY425629 P. lilacina France Sipman 22890 (B) AY425660 P. lilacina Germany Kainz 809 (M) AY425661 P. lilacina Greece Kainz 900 (M) AY425662 P. lilacina Germany Kainz 788 (M) AY425663 P. lilacina Finland Pykälä 27009 (H) PV766679 P. lilacina Finland Pykälä 27629 (H) PV766680 P. lilacina Finland Pykälä 37012 (H) PV766681 P. lilacina Finland Pykälä 57976 (H) PV766682 P. minuta Finland Pykälä 44275 (H) PV766683 P. oulankaensis Finland Pykälä 39629 (H) PV766684 P. oulankaensis Finland Pykälä 39810 (H) PV766685 P. oulankaensis Finland Pykälä 40201 (H) PV766686 P. oulankaensis Finland Pykälä 44928 (H) PV766687 P. oulankaensis Finland Pykälä 44949 (H) PV766688 P. oulankaensis Finland Pykälä 44953 (H) PV766689 P. oulankaensis Finland Pykälä 55995 (H) PV766690 P. pseudocompressa Finland Pykälä 35901 (H) PV766691 P. pseudocompressa Finland Pykälä 36325 (H) PV766692 P. pseudocompressa Finland Pykälä 39367 (H) PV766693 198 MycoKeys 124: 193–226 (2025), DOI: 10.3897/mycokeys.124.162802 Juha Pykälä & Leena Myllys: Lichen genus Protoblastenia in Finland Taxon Country Voucher specimen GenBank Accession number (ITS) P. pseudocompressa Finland Pykälä 39693 (H) PV766694 P. pseudocompressa Finland Pykälä 39987 (H) PV766695 P. pseudocompressa Finland Pykälä 58457 (H) PV766696 P. pseudoincrustans ined. Austria Kainz 949 (M) AY425664 P. pseudoincrustans ined. Germany Kainz 488 (M) AY425665 P. pseudoincrustans ined. Germany Kainz 489 (M) AY425666 P. pseudoterricola Finland Pykälä 44157 (H) PV766697 P. remota Finland Pykälä 8547 (H) PV766698 P. remota Finland Pykälä 39455 (H) PV766699 P. rikkinenii Finland Pykälä 43252 (H) PV766700 P. rikkinenii Finland Pykälä 43910 (H) PV766701 P. rikkinenii Finland Pykälä 44387 (H) PV766702 P. rupestris Germany Kainz 714 (M) AY425654 P. rupestris Germany Kainz 791 (M) AY425655 P. rupestris Germany Kainz 773 (M) AY425656 P. rupestris France Rambold 6252 (M) AY425657 P. rupestris Norway Edvardsen & Ekman NO8 (BG) EF524318 P. rupestris France Gaya 415 & Guiedan (BCN) EU639582 “P. rupestris” Norway Millanes (O) KY266981 P. rupestris Canada McMullin (OAC) KT695366 P. rupestris Norway Johnsen (BG) MG926010 “P. rupestris” UK Cannon (K) MZ159571 P. rupestris Spain Garrido-Benavent IGB1018 (VAL) OQ550133 P. rupestris USA Lendemer 49650 (NY) MK092337 P. rupestris Finland Pykälä 5963 (H) PV766703 P. rupestris Finland Pykälä 27569 (H) PV766704 P. rupestris Finland Pykälä 34043 (H) PV766705 P. rupestris Finland Pykälä 46413 (H) PV766706 “P. aff. rupestris” USA Leavitt SL19096 MZ922194 “P. aff. rupestris” USA Leavitt SL19099 MZ922195 P. saanaensis Finland Pykälä 44431 (H) PV766707 P. saanaensis Finland Pykälä 44438 (H) PV766708 P. siebenhaariana Germany Kainz 188 (M) AY425627 P. terricola Austria Hafellner 42151 (GZU) AY425639 P. terricola Austria Kainz 977 (M) AY425640 P. terricola Switzerland Sippman 30291 (B) AY425641 P. terricola Finland Pykälä 44876 (H) PV766709 P. timdalii Finland Pykälä 43624 (H) PV766710 P. timdalii Finland Pykälä 44177 (H) PV766711 P. violacea Finland Pykälä 43015 (H) PV766712 P. violacea Finland Pykälä 43524 (H) PV766713 P. violacea Finland Pykälä 43605 (H) PV766714 P. westbergii Finland Pykälä 43196 (H) PV766715 P. westbergii Finland Pykälä 43279 (H) PV766716 P. westbergii Finland Pykälä 44314 (H) PV766717 P. westbergii Finland Pykälä 44316 (H) PV766718 Psora rubiformis Norway Timdal (O) KY266957 P. decipiens Greenland Timdal 10078 (O) EF524326 199 MycoKeys 124: 193–226 (2025), DOI: 10.3897/mycokeys.124.162802 Juha Pykälä & Leena Myllys: Lichen genus Protoblastenia in Finland 3) P. compressa sp. nov. + P. pseudocompressa sp. nov., 4) P. lilacina + P. oulankaensis sp. nov., 5) P. borealis sp. nov. + P. ekmanii sp. nov. + P. rupestris, 6) P. calva + P. calvella + P. pseudoterricola sp. nov. + P. terricola + P. violacea sp. nov. All the new Finnish species with more than one sequence show a barcode gap, i.e. their genetic distance to the nearest neigbour exceeds their maximum intraspecific distance (Table 2). Note that the gap is slightly smaller if GenBank sequences here identified as P. oulankaensis and P. remota are included. The maximum genetic distance within the species including the Italian sequence of P. oulankaensis (AY425629) is 1.7%. If the two North American sequences (MZ922194, MZ922195) are included in P. remota, the maximum genetic distance of P. remota is 1.6%. The specimens previously treated as P. siebenhaariana by Stenroos et al. (2016) belong to six different species, all characterised by a violet hypothecium: Protoblastenia arupii sp. nov., P. dolomitica sp. nov., P. pseudoterricola sp. nov., P. terricola, P. violacea sp. nov. and P. westbergii sp. nov. However, all these species have more densely occurring apothecia than P. siebenhaariana. Furthermore, the specimens previously identified as P. incrustans belong to P. compressa sp. nov. and P. pseudocompressa sp. nov. Specimens treated as Protoblastenia rupestris belong in nine species: P. rupestris, P. calvella, P. borealis sp. nov., P. fennoarctica sp. nov., P. minuta sp. nov., P. r e - mota sp. nov., P. rikkinenii sp. nov., P. saanaensis sp. nov. and P. timdalii sp. nov. The sequenced specimens previously identified as P. calva belong to P. lilacina, P. oulankaensis sp. nov. and P. pseudocompressa. Previously only Finnish specimens on soil have been identified as P. terricola, but the species is also epilithic in Finland. Unfortunately, sequences of P. coniasis, P. geitleri and P. szaferi were not available in GenBank. However, all three species differ morphologically from our new species: Protoblastenia coniasis resembles P. incrustans, but has a yellow thallus that gradually turns brownish-rose in the herbarium (Poelt 1957). Table 2. Genetic distances within and between the new species of Protoblastenia, based on the Finnish and GenBank sequences. Maximum intraspecific distance (%) Minimum distance to nearest neighbour (%) Barcodegap (%) Nearest neighbour P. arupii (n = 1) 9.9 P. dolomitica P. borealis (n = 22) 0.6 2.5 1.9 P. ekmanii P. compressa (n = 8) 0.8 2.0 1.2 P. pseudocompressa P. dolomitica (n = 7) 1.1 3.9 2.8 P. westbergii P. ekmanii (n = 1) 2.5 P. borealis P. fennoarctica (n = 4) 0.8 4.2 3.4 P. rikkinenii P. minuta (n = 1) 9.2 P. timdalii P. oulankaensis (n = 7) 1.2 9.6 8.4 P. lilacina P. pseudocompressa (n = 6) 0.6 2.0 1.4 P. compressa P. pseudoterricola (n = 1) 4.9 P. terricola P. remota (n = 2) 0.4 14.0 13.6 P. incrustans P. rikkinenii (n = 3) 0.6 4.4 3.8 P. fennoarctica P. saanaensis (n = 2) 0.2 5.2 5.0 P. timdalii P. timdalii (n = 2) 0.7 2.8 2.1 P. pseudoincrustans ined. P. violacea (n = 3) 0.2 6.9 6.7 P. terricola P. westbergii (n = 4) 2.2 4.6 2.4 P. dolomitica 200 MycoKeys 124: 193–226 (2025), DOI: 10.3897/mycokeys.124.162802 Juha Pykälä & Leena Myllys: Lichen genus Protoblastenia in Finland Protoblastenia siebenbaariana AY425627 P. aurata AY425659 P. aurata AY425658 P. rikkinenii 43252 P. rikkinenii 43910 P. rikkinenii 44387 ”P. rupestris” KY266981 P. fennoarctica 44116 P. fennoarctica 43194 P. fennoarctica 43105 P. fennoarctica 43656 P. pseudoincrustans ined. AY425564 P. pseudoincrustans ined. AY425566 P. pseudoincrustans ined. AY425565 P. timdalii 43624 P. timdalii 44177 P. saanaensis 44431 P. saanaensis 44438 P. minuta 44275 P. westbergii 43196 P. westbergii 44314 P. westbergii 44316 P. westbergii 43279 P. dolomitica 44115 P. dolomitica 36072 P. dolomitica 39851 P. dolomitica 43855 P. dolomitica 43340 P. dolomitica 39217 P. dolomitica 43466 P. arupii 44130 P. incrustans AY425630 P. incrustans AY425669 P. incrustans AY425670 ”P. rupestris ” MZ159571 P. incrustans KP314370 P. incrustans AY425632 P. incrustans AY425631 P. incrustans KP314440 P. pseudocompressa 39367 P. pseudocompressa 36325 P. pseudocompressa 35901 P. pseudocompressa 58457 P. pseudocompressa 39693 P. pseudocompressa 39987 P. compressa 36521 P. compressa 35585 P. compressa 42124 P. compressa 38349 P. compressa 38965 P. compressa 38646 P. compressa 41376 P. compressa 41511b P. oulankaensis 44949 P. oulankaensis 39810 P. oulankaensis 40201 P. oulankaensis 39629 P oulankaensis 55995. P. oulankaensis 44953 P. oulankaensis 44928 ”P. lilacina” AY425629 P. lilacina 27009 P. lilacina 27629 P. lilacina AY425660 P. lilacina 37012 P. lilacina AY425661 P. lilacina AY425663 P. lilacina 57976 P. lilacina AY425662 P. rupestris 46413 P. rupestris 27569 P. rupestris AY425656 P. rupestris AY425657 P. rupestris MG926010 P. rupestris AY425655 P. rupestris EF524318 P. rupestris MK092337 P. rupestris OQ550133 P. rupestris KT695366 P. rupestris AY425654 P. rupestris 34043 P. rupestris EU639582 P. rupestris 5963 P. ekmanii 37508 P. borealis 44906 P. borealis 57501 P. borealis 29704 P. borealis 16567 P. borealis 40695 P. borealis 47217 P. borealis 33101 P. borealis 44641 P. borealis 60511 P. borealis 41355 P. borealis 33037 P. borealis 23710 P. borealis 42802 P. borealis 45759 P. borealis 26097 P. borealis 38784 P. borealis 39151 P. borealis 32757 P. borealis 39942 P. borealis 58196 P. borealis 25292 P. borealis 58392 P. remota 39455 P. remota 8547 P. aff. rupestris MZ922194 P. aff. rupestris MZ922195 P. cyclospora AY425668 P. cyclospora AY425667 P. laeta AY425652 P. laeta AY425650 P. laeta AY425651 P. laeta AY425653 P. calva AY425642 P. calva AY425644 P. calva AY425645 P. calva AY425643 P. calva EF524319 P. calva AY425628 P. terricola AY425639 P. terricola AY425641 P. terricola AY425640 P. terricola 44876 P. violacea 43015 P. violacea 43605 P. violacea 43524 P. pseudoterricola 44157 P. calvella OR773065 P. calvella OQ604662 P. calvella AY425648 P. calvella AY425649 ”P. calva” HQ650618 P. calvella OQ604661 P. calvella AY425646 P. calvella AY425647 0.07 Psora rubiformis KY266957 Psora decipiens EF524326 92 79 100 70 100 94 87 99 99 99 96 100 96 100 100 71 100 99 100 100 98 88 97 97 100 95 97 100 74 100 71 73 99 98 100 91 100 100 90 67 100 100 58 100 95 75 100 95 99 99 95 91 83 87 201 MycoKeys 124: 193–226 (2025), DOI: 10.3897/mycokeys.124.162802 Juha Pykälä & Leena Myllys: Lichen genus Protoblastenia in Finland Protoblastenia geitleri has globose spores (Zahlbruckner 1936), a feature not present in any of the Finnish species. Protoblastenia szaferi is characterised by an endolithic thallus, mainly flat apothecia immersed in rock and relatively distinct apothecial margins (Hafellner 2006). Such a combination of morphological characteristics was not observed in the Finnish material. The number of new species is surprisingly high, particularly compared to the global number of accepted species. Even though the new species are genetically distinct, their morphological identification may be difficult. On average, all species may differ morphologically, but for several species, more material is needed to confirm this. In addition, there is considerable overlap in morphological characteristics between the species. Most of the species may be semi-cryptic, a feature that may be common in lichens (e.g. Orange (2012); Pykälä et al. (2020)). Some specimens can be identified by morphology, while identification of other specimens remains uncertain. Table 3 presents the most important apothecial characters between the new species. Numerous studies have shown that DNA barcoding is efficient in species delimitation and in finding morphologically cryptic or semi-cryptic species (e.g. Divakar et al. (2016); Lücking et al. (2020a); Zhang et al. (2022); Printzen et al. (2023)). However, in several lichen groups, lack of resolution in recently evolving species complexes may occur (Lücking et al. 2020a, b). Thus, lineages receiving high support values in the ITS phylogeny and showing a barcode gap can be described as new species even if they are morphologically cryptic or semi-cryptic. Our results show that ITS is well suitable for species delimitations of Protoblastenia. Table 3. The most important apothecial characters between the new species of Protoblastenia. Mean spore length and width (μm) Size of hypothecium (min and max) (μm) Width of apothecia (min and max) (mm) P. arupii 12.8 × 6.8 50–60 0.2–0.5 P. borealis 11.2 × 6.2 40–160 0.2–1.0 P. compressa 11.4 × 5.9 40–100 0.2–0.8 P. dolomitica 12.8 × 5.4 40–100 0.15–0.7 P. ekmanii 10.2 × 5.9 120–200 0.4–0.8 P. fennoarctica 10.0 × 5.3 60–280 0.3–0.9 P. minuta 7.9 × 4.5 70–100 0.2–0.4 P. oulankaensis 13.9 × 7.4 60–220 0.3–0.9 P. pseudocompressa 10.6 × 5.5 50–120 0.2–0.7 P. pseudoterricola 9.9 × 5.5 60–100 0.3–0.8 P. remota 11.4 × 5.6 100–200 0.2–0.6 P. rikkinenii 9.6 × 5.1 80–300 0.3–0.8 P. saanaensis 8.4 × 4.8 40–100 0.3–0.5 P. timdalii 9.2 × 4.8 60–100 0.3–0.8 P. violacea 9.2 × 4.6 100–280 0.3–1.0 P. westbergii 12.0 × 6.1 60–150 0.2–1.1 Figure 1. Phylogenetic relationships of the studied Protoblastenia species, based on a Maximum Likelihood (ML) analysis of the ITS dataset. Maximum likelihood bootstrap values > 50% are shown at nodes. Thickened lines indicate bootstrap values > 70%. New species described in this study are indicated with shadowed boxes. GenBank accession numbers for sequences downloaded from GenBank and collection numbers for the specimens sequenced for this study are shown after the taxon names. Holotype specimens of the new species are shown in bold. 208 MycoKeys 124: 193–226 (2025), DOI: 10.3897/mycokeys.124.162802 Juha Pykälä & Leena Myllys: Lichen genus Protoblastenia in Finland Habitat and distribution. This species has been found only in the parishes of Salla (in the biogeographical Province Ks) and Enontekiö (biogeographical Province EnL) in N Finland. It grows on dolomite rock outcrops, once collected from dolomite stones and pebbles. The species may prefer sun-exposed and half-shady sites on fells and in gorges. Companion species include, for example, Acarospora glaucocarpa, Hymenelia rhodopis (Sommerf.) Lutzoni, Opegrapha dolomitica (Arnold) Clauzade & Cl. Roux ex Torrente & Egea, Polyblastia spp., Protoblastenia fennoarctica, Synalissa ramulosa (Bernh.) Körb., Thelidium auruntii (A. Massal.) Kremp. and T. declivum Pykälä & Myllys. Etymology. The species grows on dolomite rocks. Notes. Protoblastenia dolomitica is closely related to P. westbergii, which has slightly larger apothecia, broader spores and sometimes a thicker hypothecium. The specimens have been previously identified as P. siebenhaariana in Finland. However, this species has only 1–10 apothecia / cm2 (Kainz and Rambold 2004) and the Finnish specimens are not closely related to a GenBank specimen of P. siebenhaariana. Protoblastenia dolomitica is characterised by rather small, often dirty olive-yellow apothecia and a thin hypothecium. Protoblastenia arupii, based on the inspection of only one known specimen, has orange apothecia, on average, broader spores and an only slightly thinner rimose thallus. More material from the morphologically related species is needed to determine whether P. dolomitica can be identified by morphology only. Other specimens examined. Finland • Koillismaa, Salla, Oulanka National Park, Pikkuköngäs, bank of River Oulankajoki, high cliff, calciferous (dolomite) schistose rock, on overhanging SW-facing wall, 180 m a.s.l., 66°25'N, 29°09'E, 10.8.2009, J. Pykälä 36072 (H); • Salla, Oulanka National Park, Pikkuköngäs, bank of River Oulankajoki, high cliff, calciferous (dolomite) schistose rock outcrop, on SE-facing wall, 182 m a.s.l., 66°25'N, 29°08'E, 4.8.2010, J. Pykälä 39217 (H); • Salla, Oulanka National Park, Pikkuköngäs, bank of River Oulankajoki, high cliff, calciferous (dolomite) schistose rock outcrop, beneath SW-facing wall, on pebble, 180 m a.s.l., 66°25'N, 29°08'E, 13.8.2010, J. Pykälä 39851 (H); • Enontekiön Lappi, Enontekiö, Porojärvet, Toskalharji, Toskaljärvi N, fell, dolomite rock outcrop, gentle SE-slope, on dolomite pebbles, 730 m a.s.l., 69°12'N, 21°26'E, 2.8.2011, J. Pykälä 43340 (H); • Enontekiö, Porojärvet, Toskalharji, Toskaljärvi N, lake shore, dolomite scree, on dolomite stones, 705 m a.s.l., 69°11'N, 21°26'E, 7.8.2011, J. Pykälä 43855 (H); • Enontekiö, Kilpisjärvi, Saana, fell, steep NE-slope, dolomite rock outcrop, on NE-facing wall, 820 m a.s.l., 69°02'N, 20°51'E, 11.8.2011, J. Pykälä 44115 (H). Protoblastenia ekmanii Pykälä & Myllys, sp. nov. MycoBank No: 859630 Fig. 3A Diagnosis. Differs from P. borealis in having a thicker hypothecium. Type. Finland • Varsinais-Suomi, Salo (Kisko), Kirkonkylä, 450 m NW of Kolko, shore of Määrjärvi Lake, calcareous rock outcrop, on W-slope, 43 m a.s.l., 60°12'N, 23°31'E, 20 May 2010, J. Pykälä 37508 (H9224071 – holotype, GenBank accession number: PV766674). Description. Thallus white to greyish-white, rimose to areolate, areoles 0.3–0.6 mm, ca. 0.05–0.2 mm thick, K-, UV+ bluish-white, algal cells 5–8 μm. 209 MycoKeys 124: 193–226 (2025), DOI: 10.3897/mycokeys.124.162802 Juha Pykälä & Leena Myllys: Lichen genus Protoblastenia in Finland Apothecia dirty brown-orange, 0.4–0.8 mm, convex to strongly convex, superficial; ca. 30–50 apothecia / cm2. Epihymenium dark orange-brown, 10– 20 μm thick, K+ violet. Hymenium 50–70 μm thick. Hypothecium yellowish-white, ca. 120–200 μm thick, in some apothecia base patchily red-violet. Paraphyses ca. 2–2.5 μm thick, apex not or slightly thickened. Ascospores 0-septate, (8.1–)8.7–10.2–11.6(–12.5) × (5.0–)5.1–5.9–6.6(–7.2) (n = 12). Habitat and distribution. Only one specimen is known from a calcareous rock outcrop on a lake shore in SW Finland. The sun-exposed site suggests that the species may have a southern distribution. The species is apparently very rare in Finland. Companion species include Acarospora glaucocarpa, Clauzadea monticola, Placynthium nigrum (Huds.) Gray and Thelidium incavatum. Etymology. The name is in honour of Dr Stefan Ekman (Uppsala) for his many important contributions on the phylogeny and taxonomy of lichens in Fennoscandia and elsewhere. Notes. The species is closely related to P. borealis (only 2.5% difference in ITS), but seems to differ in having a thicker hypothecium. The hypothecium of P. borealis is very variable, but in no specimens does it reach the upper limit of the thickness in the P. ekmanii specimen. The Finnish specimens of P. calvella have orange-yellow to orange apothecia and thus resemble P. ekmanii. More material is needed to determine whether the species can be unambiguously morphologically distinguished from P. calvella. Protoblastenia fennoarctica Pykälä & Myllys, sp. nov. MycoBank No: 859631 Fig. 3B Diagnosis. Differs from P. calvella in having thinner hymenium, from P. minuta in having rimose to areolate thallus and from P. rikkinenii in usually having more densely occurring apothecia. Type. Finland • Enontekiön Lappi, Enontekiö, Porojärvet, Toskalharji, 1.2 km NE of Toskaljärvi, fell, SW-slope, scree, on dolomite boulder, 875 m a.s.l., 69°12'N, 21°28'E, 5 August 2011, J. Pykälä 43656 (H9220129 – holotype, GenBank accession number: PV766677). Description. Thallus white to pale greyish-brown, rimose to areolate, areoles 0.15–1.0 mm, ca. 0.03–0.25 mm thick, K-, C-, UV-, algal cells 5–10 μm. Apothecia yellow, orange-yellow to orange, 0.3–0.9 mm, plane to strongly convex, immersed in thallus to superficial; ca. 20–110 apothecia / cm2. Epihymenium orange-yellow to brown, 12–20 μm thick, K+ violet. Hymenium 50–80 μm thick. Hypothecium colourless to yellow, base sometimes patchily reddish-brown, ca. 60–280 μm thick. Paraphyses ca. 2–3 μm thick, cells 5–8 μm long, apex not thickened to slightly thickened, sparingly branched and anastomosing. Ascospores 0-septate, (6.6–)8.7–10.0–11.3(–12.5) × (4.2– )4.8–5.3–5.9(–7.2) μm (n = 93). Habitat and distribution. This species has been found only in the Parish Enontekiö (biogeographical Province EnL in NW Finland). It grows on fells on dolomite rock outcrops, boulders and pebbles. Companion species include Enchylium polycarpon, Farnoldia jurana, Physcia caesia (Hoffm.) Fürnr., Polyblastia spp., Protoblastenia dolomitica and Verrucaria vacillans Pykälä & Myllys. 210 MycoKeys 124: 193–226 (2025), DOI: 10.3897/mycokeys.124.162802 Juha Pykälä & Leena Myllys: Lichen genus Protoblastenia in Finland Etymology. The species occurs in the most northern, almost arctic (oroarctic) sites in Finland (Fennia). Notes. Protoblastenia fennoarctica morphologically resembles P. calvella, P. minuta, P. remota and P. rikkinenii in having a pale hypothecium and small spores. In the ITS phylogeny, the species is most closely related to P. rikkinenii (Fig. 1). Protoblastenia rikkinenii differs in having more sparsely occurring apothecia, but more material is needed to confirm whether the two species can be identified by morphology. Protoblastenia calvella has a thicker hymenium (80–100 μm thick). Protoblastenia minuta and P. remota have an endolithic to thin thallus and P. minuta has smaller spores compared to P. fennoarctica, while P. remota may have slightly larger spores. Other specimens examined. Finland • Enontekiön Lappi, Enontekiö, Porojärvet, Toskalharji, Toskalpahta fell, SW-slope, scree, on dolomite pebbles, 795 m a.s.l., 69°11'N, 21°29'E, 1.8.2011, J. Pykälä 43105 (H); • Enontekiö, Porojärvet, Toskalharji, Toskaljärvi N, fell, dolomite scree, gentle SE-slope, on dolomite boulder, 715 m a.s.l., 69°11'N, 21°26'E, 2.8.2011, J. Pykälä 43194 (H); • Enontekiö, Kilpisjärvi, Saana, fell, steep NE-slope, dolomite rock outcrop, on NE-facing wall, 820 m a.s.l., 69°02'N, 20°51'E, 11.8.2011, J. Pykälä 44116 (H). Protoblastenia lilacina Poelt & Vězda, Čas. slezsk. Mus. Opavě, Ser. A 19: 26 (1970) Description. Thallus white to grey, endolithic, farinose to small areoles, K-, C-, UVto UV+ bluish-white, algal cells 3–8 μm. Apothecia orange to dark orange, 0.2–0.6 mm, slightly convex to strongly convex, 1/4-immersed to superficial, sometimes leaving shallow pits, Kor K+ fairly weakly red; ca. 40–100 apothecia Figure 3. A. Protoblastenia ekmanii (holotype); B. P. fennoarctica (holotype); C. P. minuta (holotype); D. P. oulankaensis (holotype). Scale bars: 0.5 mm. 211 MycoKeys 124: 193–226 (2025), DOI: 10.3897/mycokeys.124.162802 Juha Pykälä & Leena Myllys: Lichen genus Protoblastenia in Finland / cm2. Epihymenium (yellowish) orange-brown, 12–20 μm thick, Kto K+ orange or carmine red, not into solution. Hymenium 50–120 μm thick, with oil droplets. Hypothecium pale yellow, sometimes patchily violet, ca. 60–120 μm thick. Paraphyses ca. 2–3 μm thick, apex not thickened to slightly thickened, sparingly branched and anastomosing. Ascospores 0-septate, (8.4–)10.7–12.4– 14.2(–16.6) × (5.2–)6.0–6.7–7.3(–7.8) μm (n = 30). Habitat and distribution. The species is rare and occurs only on calcareous rock outcrops and boulders in the hemi-boreal zone in SW Finland. This seems to be the northern limit of this predominantly temperate species. Protoblastenia lilacina grows both on fairly sun-exposed and shady habitats. It may prefer rather dry calcareous rocks. Companion species include, for example, Bagliettoa baldensis s.lat, B. calciseda (DC.) Gueidan & Cl. Roux, Catillaria lenticularis (Ach.) Th. Fr., Hymenelia spp., Lecidella stigmatea (Ach.) Hertel & Leuckert, Staurothele rupifraga (A. Massal.) Arnold, Verrucaria deversa Vain., V. foveolata and V. viridula. Notes. The species differs from most species of Protoblastenia in having Kor weakly K+ red apothecia. The closely-related Protoblastenia oulankaensis is the only other species with this characteristic. However, the two species have different distribution areas as Protoblastenia oulankaensis is only known from the Oulanka area in NE Finland. Morphologically, P. oulankaensis differs in having a thicker hypothecium and often more sparsely occurring apothecia (in most specimens 10–30 apothecia / cm2). Specimens examined. Finland • Varsinais-Suomi, Kemiönsaari (Västanfjärd), Vesterillo, Strömmen 200–400 m SE, Pinus sylvestris-dominated forest, calcareous rock outcrop, on 70 cm high E-facing wall, 17 m a.s.l., 60°00'N, 22°44'E, 9.9.2009, J. Pykälä 37012 (H); • Lohja, Moisio, between Lohjanharjuntie and Nummentie roads, NW-slope of Lohjanharju esker, Pinus sylvestris-heath forest, on calcareous boulder, 82 m a.s.l., 60°15'N, 24°06'E, 18.6.2005, J. Pykälä 27009 H); • Lohja, Lohja, Pitkäniemi industrial area, 5 m from shore of Lohjanjärvi Lake, deciduous forest, calcareous rock outcrop, on NW-facing 2 m high rock wall, 35 m a.s.l., 60°15'N, 24°02'E, 19.8.2005, J. Pykälä 27629 (H); • Lohja, Lohja, Pitkäniemi industrial area, deciduous forest ca. 5 m from shore of Lohjanjärvi Lake, calcareous rock outcrop, on 1.7 m high overhanging NW-facing wall, 35 m a.s.l., 60°15'N, 24°02'E, 12.7.2021, J. Pykälä 57976 (H). Protoblastenia minuta Pykälä & Myllys, sp. nov. MycoBank No: 859654 Fig. 3C Diagnosis. Differs from the other Protoblastenia species with a pale hypothecium in having small apothecia and spores and apothecia which leave pits. Type. Finland • Enontekiön Lappi, Enontekiö, Kilpisjärvi, Saana Nature Reserve, E-part, fell, steep SW-slope, dolomite rock outcrop, on S-facing wall, scarce, 715 m a.s.l, 69°02'N, 20°51'E, 12 August 2011, J. Pykälä 44275 (H9250941 – holotype, GenBank accession number: PV766683). Description. Thallus ochraceous, endolithic, K-, UV-. Apothecia orange, 0.2–0.4 mm, slightly convex to strongly convex, immersed in rock to superficial, leaving shallow to fairly deep pits; ca. 80 apothecia / cm2. Epihymenium 212 MycoKeys 124: 193–226 (2025), DOI: 10.3897/mycokeys.124.162802 Juha Pykälä & Leena Myllys: Lichen genus Protoblastenia in Finland brown-yellow, 10–12 μm thick, K+ violet. Hymenium 45–55 μm thick. Hypothecium colourless to yellow, ca. 70–100 μm thick. Paraphyses ca. 2–3 μm thick, apex not thickened to slightly thickened. Ascospores 0-septate, (7.3–)7.6–7.9– 8.3(–8.8) × (4.1–)4.3–4.5–4.7(–4.8) μm (n = 9). Habitat and distribution. Only one specimen has been found on the calcareous Saana fell in NE Finland, on a dolomite rock outcrop. Companion species include Acarospora macrospora (Hepp) A. Massal. ex Bagl., Enchylium polycarpon, Placynthium asperellum (Ach.) Trevis. s.lat, Romjularia lurida (Ach.) Timdal and Thalloidima alutaceum Anzi. Etymology. The name refers to the small size of the apothecia and spores. Notes. The species is rather isolated amongst the sequenced species of Protoblastenia (minimum difference from other species over 9% in ITS). The species has smaller apothecia and spores than most other Protoblastenia species with a pale hypothecium. Furthermore, other Finnish Protoblastenia species do not leave pits or only rarely leave shallow pits. However, only one specimen is known and the species may be expected to be morphologically more variable. More material is needed to evaluate whether the species can be identified by morphology only. Protoblastenia saanaensis has a rimose thallus and the apothecia do not leave pits. Protoblastenia szaferi differs in having larger apothecia and spores (see Hafellner (2006)). Protoblastenia oulankaensis Pykälä & Myllys, sp. nov. MycoBank No: 859655 Fig. 3D Diagnosis. Differs from P. lilacina in often having slightly larger and more sparse apothecia and an often thicker hypothecium. Type. Finland • Koillismaa, Salla, Oulanka National Park, W of Savikoski, cliff, dolomite rock outcrop, on NE-facing wall, 190 m a.s.l., 66°25'N, 29°10'E, 17 August 2010, J. Pykälä 40201 (H9224130 – holotype, UPS – isotype, GenBank accession number: PV766686). Description. Thallus white to whitish-grey, endolithic, farinose or rimose, ca. 0–0.1 mm thick, K-, C-, UVto UV+ bluish-white, algal cells 5–9 μm. Apothecia orange to dark orange, 0.3–0.9 mm, convex to strongly convex, 1/4-immersed to superficial, often leaving shallow pits, K+ weakly red; ca. 10–50 apothecia / cm2 (in one specimen, ca. 50–80 apothecia / cm2). Epihymenium dark orange-brown, 12–25 μm thick, Kto K+ orange or carmine red. Hymenium 70–120 μm thick, with oil droplets, in one specimen, partly violet in few apothecia. Hypothecium colourless to yellow, more rarely patchily red-brown, ca. 60–220 μm thick. Paraphyses ca. 2–3 μm thick, apex not thickened to slightly thickened, sparingly branched and anastomosing. Ascospores 0-septate, (8.8– )11.8–13.9–16.0(–18.8) × (5.2–)6.6–7.4–8.2(–9.6) μm (n = 92). Habitat and distribution. The species occurs only in the Oulanka area in the parishes of Kuusamo and Salla in NE Finland. It grows on Nand S-facing walls of dolomite rock outcrops. Companion species include, for example, Diploschistes gypsaceus (Ach.) Zahlbr., Gyalecta jenensis (Batsch) Zahlbr., Lempholemma isidioides (Nyl. ex Arnold) H. Magn., Protoblastenia borealis, Sagiolechia protuberans, Scytinium parvum, Verrucaria foveolata and 213 MycoKeys 124: 193–226 (2025), DOI: 10.3897/mycokeys.124.162802 Juha Pykälä & Leena Myllys: Lichen genus Protoblastenia in Finland Xanthocarpia crenulatella (Nyl.) Frödén, Arup & Søchting s.lat. Based on the ITS phylogeny, one GenBank specimen of P. lilacina from northern Italy (AY425629) belongs to P. oulankaensis (Fig. 1). Etymology. In Finland, the species occurs only in the Oulanka area in the northeast. Notes. Protoblastenia oulankaensis is closely related to P. lilacina, but differs clearly from that species in its ITS profile (10% difference). It is also morphologically rather similar to P. lilacina, having a weak K+ reaction of apothecia and large spores. Protoblastenia oulankaensis tends to have slightly larger apothecia, which usually occur more sparsely (in most specimens ca. 10–50 apothecia /cm2) and, on average, a thicker hypothecium. Protoblastenia lilacina is only known from the hemi-boreal zone of SW Finland, ca. 800 kilometres south to south-west. Other specimens examined. Finland • Koillismaa, Salla, Oulanka National Park, Savikoski 300 m N, Pinus sylvestris-forest, steep N-slope, dolomite rock outcrop, on NW-facing wall, st pc – sp, 180 m a.s.l., 66°25'N, 29°10'E, 10.8.2010, J. Pykälä 39629 (H); • Salla, Oulanka National Park, Savilamminniemi, shore of Savilampi Lake, cliff, dolomite rock outcrop, on E-facing wall, sp, 180 m a.s.l., 66°25'N, 29°10'E, 12.8.2010, J. Pykälä 39810 (H); • Salla, Oulanka National Park, Savilampi 1.4 km NE, bank of Savinajoki river, cliff, dolomite rock outcrop, upper slope, on NE-facing wall, 192 m a.s.l., 66°26'N, 29°11'E, 23.8.2011, J. Pykälä 44928 (H); • Salla, Hautajärvi, Kurtinniittykuru, dolomite rock outcrop, on SW-facing wall, 195 m a.s.l., 66°26'N, 29°09'E, 24.8.2011, J. Pykälä 44949 (H); • Salla, Hautajärvi, Kurtinniittykuru, dolomite rock outcrop, on SE-facing wall, 195 m a.s.l., 66°26'N, 29°09'E, 24.8.2011, J. Pykälä 44953 (H); • Kuusamo, Juuma, Myllyniemi, dolomite rock outcrop, on NE-facing wall, 235 m a.s.l., 66°16'N, 29°22'E, 11.8.2020, J. Pykälä 55995 (H). Protoblastenia pseudocompressa Pykälä & Myllys, sp. nov. MycoBank No: 859656 Fig. 4A Diagnosis. Differs from P. compressa in often having convex apothecia. Type. Finland • Koillismaa, Salla, Onkamo, Latvarova SW, clear-cut heath forest, on calcareous boulder, 258 m a.s.l., 66°42'N, 28°055'E, 9 August 2021, J. Pykälä 58457 (H9250942 – holotype, UPS – isotype, GenBank accession number: PV766696). Description. Thallus white to grey, continuous, rimose (usually) to areolate, rarely endolithic, ca. 0–0.15 mm thick, K-, C-, UVto UV+ bluish-white, algal cells 6–12 μm. Apothecia orange-yellow to orange, 0.2–0.7 mm, plane to strongly convex, 1/4–1-immersed in thallus or rock to superficial, sometimes leaving shallow pits; ca. 20–120 apothecia / cm2. Epihymenium yellow-brown to orange-brown, 12–18 μm thick, K+ violet. Hymenium 50–60 μm thick. Hypothecium colourless to pale yellow, rarely pink (in one specimen), ca. 50–120 μm thick. Paraphyses ca. 1.5–2.5 μm thick, sparsely branched and anastomosing. Ascospores 0-septate, (6.8–)9.1–10.6–12.1(–13.0) × (4.2–)4.7–5.5–6.2(–8.0) μm (n = 51). Habitat and distribution. Protoblastenia pseudocompressa has been verified by ITS only from the Oulanka area (parishes Kuusamo and Salla) in NE Finland. However, a morphologically similar specimen was found in the neighbouring 214 MycoKeys 124: 193–226 (2025), DOI: 10.3897/mycokeys.124.162802 Juha Pykälä & Leena Myllys: Lichen genus Protoblastenia in Finland biogeographical Province of Kainuu. The species cannot be identified with certainty by morphology only and it is likely that more populations occur in the Oulanka area. It mainly grows on dolomite rock outcrops (one specimen was collected from a dolomite boulder) and may prefer half-shady habitats. Typical companion species are Acarospora glaucocarpa, Clauzadea monticola, Farnoldia jurana, Hymenelia prevostii (Duby) Kremp., H. rhodopis, Lathagrium fuscovirens, Placynthium nigrum, Thelidium declivum, Verrucaria deversa, V. muralis, V. kuusamoensis Pykälä, Kantelinen & Myllys and V. subtilis. Etymology. The name refers to the close relationship in ITS and morphological similarity to P. compressa. Notes. Protoblastenia pseudocompressa is closely related to P. compressa and the two species are also morphologically similar. Protoblastenia pseudocompressa has, on average, slightly smaller spores than P. compressa, but there is a wide overlap. Furthermore, P. pseudocompressa specimens with plane to slightly convex apothecia may be confused with P. compressa. However, the apothecia of P. pseudocompressa are often convex, while the apothecia of P. compressa are always plane to slightly convex. Based on the distribution of sequenced specimens, the two species are geographically well separated. Protoblastenia pseudocompressa specimens with convex apothecia may also be confused with P. borealis, which, however, often has a thicker hymenium. Other specimens examined. Finland • Koillismaa, Kuusamo, Oulanka National Park, Pikkukönkäänkuru, dolomite rock outcrop, small W-facing wall, 175 m a.s.l., 66°21'N, 29°19'E, 8.8.2009, J. Pykälä 35901 (H); • Salla, Oulanka National Park, 400 m N of Savilampi, bank of River Savinajoki, dolomite rock outcrop, on NW-facing wall, 178 m a.s.l., 66°25'N, 29°10'E, 13.8.2009, J. Pykälä 36325 (H); • Kuusamo, Oulanka National Park, Kiutaköngäs 400 m N, SW-slope, Pinus sylvestris-dominated forest, small dolomite rock outcrop, Figure 4. A. Protoblastenia pseudocompressa (holotype); B. P. pseudoterricola (holotype); C. P. remota (holotype); D. P. rikkinenii (holotype). Scale bars: 0.5 mm. 215 MycoKeys 124: 193–226 (2025), DOI: 10.3897/mycokeys.124.162802 Juha Pykälä & Leena Myllys: Lichen genus Protoblastenia in Finland on SW-facing wall, scarce, 175 m a.s.l., 66°22'N, 29°19'E, 6.8.2010, J. Pykälä 39367 (H); • Kuusamo, Oulanka National Park, Kiutaköngäs, bank of Oulankajoki River, by the rapids, calciferous (dolomite) schistose rock outcrop, on rather flat rock, 150 m a.s.l., 66°22'N, 29°20'E, 10.8.2010, J. Pykälä 39693 (H); • Kuusamo, Oulanka, Putaanoja, 500 m W-NW of Hautala, dolomite rock outcrop, on E-facing wall, scarce, 232 m a.s.l., 66°22'N, 29°25'E, 15.8.2010, J. Pykälä 39987 (H). Protoblastenia pseudoterricola Pykälä & Myllys, sp. nov. MycoBank No: 859657 Fig. 4B Diagnosis. Differs from P. terricola in having a thinner thallus and hypothecium. Type. Finland • Enontekiön Lappi, Enontekiö, Kilpisjärvi, Saana, fell, steep NE-slope, dolomite rock outcrop, beneath NE-facing wall, on dolomite pebbles, 820 m a.s.l., 69°02'N, 20°51'E, 11 August 2011, J. Pykälä 44157 (H9235068 – holotype, GenBank accession number: PV766697). Description. Thallus white, rimose, ca. 0.05–0.1 mm thick, K-, UV+ bluish-white. Apothecia orange (young apothecia) to dirty orange-brown, 0.3–0.8 mm, convex to strongly convex, superficial; ca. 30 apothecia / cm2. Epihymenium orange-brown, 10–15 μm thick, K+ violet. Hymenium 70 μm thick. Hypothecium dark reddish-brown to violet, ca. 60–100 μm thick. Ascospores 0-septate, (7.9–)8.6–9.9–11.2(–12.4) × (4.5–)4.8–5.5–6.2(–7.0) μm (n = 20). Habitat and distribution. Only one specimen of this species is known. It was collected from dolomite pebbles beneath an NE-facing wall of a dolomite rock outcrop on the Saana fell (NW Finland). Companion species include Farnoldia jurana, Polyblastia inconspicua Savić & Tibell and Thelidium huuskonenii Pykälä & Myllys. Etymology. The name refers to the similarities in morphology and ITS to P. terricola. Notes. The species is closely related to P. terricola and P. violacea. Protoblastenia terricola has rimose to areolate thallus (when growing on rock) mainly exceeding 0.1 mm in thickness and an often thicker hypothecium. Protoblastenia violacea has orange apothecia and smaller spores. More material is needed to confirm whether these three species can be distinguished by morphology only. Protoblastenia westbergii resembles the new species, but has more densely occurring apothecia (ca. 40–160 apothecia / cm2) and larger spores. Protoblastenia remota Pykälä & Myllys, sp. nov. MycoBank No: 859658 Fig. 4C Diagnosis. Differs from P. borealis and P. pseudocompressa by often smaller more convex apothecia and a smaller thallus. Type. Finland • Varsinais-Suomi, Perniö, Lupaja, Alhonmäki, abandoned lime quarry, on W-slope, 9 August 1991, J. Pykälä 8547 (H9250943 – holotype, GenBank accession number: PV766698). 216 MycoKeys 124: 193–226 (2025), DOI: 10.3897/mycokeys.124.162802 Juha Pykälä & Leena Myllys: Lichen genus Protoblastenia in Finland Description. Thallus white to grey, endolithic to sparse small areoles, 0.2– 0.5 mm wide, ca. 0–0.15 mm thick, K-, C-, UVto UV+ bluish-white. Apothecia orange, 0.2–0.6 mm, convex to strongly convex, 1/2-immersed to mainly superficial; ca. 15–80 apothecia / cm2. Epihymenium yellow-brown to dark orange, 12–20 μm thick, K+ violet. Hymenium 50–80 μm thick. Hypothecium colourless to pale yellow, ca. 100–200 μm thick. Paraphyses ca. 2–2.5 μm thick, apex not thickened to thickened to ca. 3–5 μm thick. Ascospores 0-septate, (8.4–)9.8–11.4–12.9(–14.4) × (4.8–)5.0–5.6–6.1(–7.1) μm (n = 36). Habitat and distribution. Two specimens of this species are known: one from a lime quarry in SW Fnland and one from a calciferous schistose cliff in NE Finland. Companion species include Lathagrium fuscovirens, Mycobilimbia tetramera (De Not.) Vitik., Ahti, Kuusinen, Lommi & T. Ulvinen ex Hafellner & Türk and Verrucaria deversa. Furthermore, two closely-related specimens from North America and previously identified as Protoblastenia aff. rupestris (MZ922194, MZ922195) (Munger et al. 2022) may belong to P. remota (Fig. 1). Etymology. The name refers to the large difference in ITS of the species compared to other Protoblastenia species (differs by ca. 15–18% from other species). Notes. Based on ITS sequences, this species is not closely related to any sequenced Protoblastenia species. The species is morphologically a typical Protoblastenia and very difficult to distinguish from several species, particularly from P. borealis and P. pseudocompressa. However, these two species have, on average, less convex apothecia, a thinner hypothecium and often a better developed thallus, but there is a wide overlap. In addition, Protoblastenia borealis often has larger apothecia (up to 1 mm). Other specimens examined. Finland • Koillismaa, Kuusamo, Oulanka National Park, Päähkänänkallio, high cliff, calciferous (dolomite) schistose rock outcrop, on SE-facing wall, st pc, 195 m a.s.l., 66°16'N, 29°31'E, 7.8.2010, J. Pykälä 39455 (H). Protoblastenia rikkinenii Pykälä & Myllys, sp. nov. MycoBank No: 859659 Fig. 4D Diagnosis. Differs from P. fennoarctica in always having convex to strongly convex apothecia and from P. saanaensis and P. timdalii in having sparsely occurring apothecia and a thick hypothecium. Type. Finland • Enontekiön Lappi, Enontekiö, Porojärvet, Toskalharji, Toskaljärvi N, fell, dolomite rock outcrop, on dolomite pebbles, 735 m a.s.l., 69°12'N, 21°26'E, 2 August 2011, J. Pykälä 43252 (H9250944 – holotype, GenBank accession number: PV766700). Description. Thallus white, grey to pale brown, rimose to areolate, ca. 0.05–0.2 mm thick, K-, C-, UV-, algal cells 5–9 μm. Apothecia yellow-orange, 0.3–0.8 mm, convex to strongly convex, 1/2-immersed to superficial; ca. 20–40 apothecia /cm2. Epihymenium dirty yellow to orange-yellow, 12– 18 μm thick, K+ violet. Hymenium 50–70 μm thick. Hypothecium pale yellow, ca. 80–300 μm thick. Paraphyses ca. 2–2.5 μm thick, apex slightly thickened to 3 μm thick. Ascospores 0-septate, (7.4–)8.5–9.6–10.7(–12.6) × (4.4–)4.7– 5.1–5.5(–6.3) μm (n = 52). 217 MycoKeys 124: 193–226 (2025), DOI: 10.3897/mycokeys.124.162802 Juha Pykälä & Leena Myllys: Lichen genus Protoblastenia in Finland Habitat and distribution. The species occurs on dolomite rock outcrops, dolomite stone and dolomite pebbles on the fells. It may prefer sun-exposed sites. Companion species include Halecania alpivaga (Th. Fr.) M. Mayrhofer, Hymenelia heteromorpha (Kremp.) Lutzoni, H. prevostii, H. rhodopis, Lecidea polycocca Sommerf., Polyblastia sp., Rhizocarpon petraeum, Thelidium huuskonenii and Verrucaria lapponica Pykälä. One GenBank sequence (KY266981) originates from northern Norway. Etymology. The name is in honour of Prof Jouko Rikkinen (Helsinki) for his innovative work on various fields of lichenology. Notes. The species is related to P. fennoarctica, which often has more densely occurring, often plane and/or yellow apothecia. However, an ITS sequence may be needed for unambiguous identification of specimens. Protoblastenia saanaensis and P. timdalii have a thinner hypothecium and more densely occurring apothecia. Protoblastenia calvella has more densely occurring apothecia and slightly larger spores. Other specimens examined. Finland • Enontekiön Lappi, Enontekiö, Kilpisjärvi, Saana Nature Reserve, W-part, fell, dolomite rock outcrop, on W-facing wall, 700 m a.s.l., 69°03'N, 20°48'E, 9.8.2011, J. Pykälä 43910 (H); • Enontekiö, Kilpisjärvi, Saana Nature Reserve, fell, steep SW-slope, dolomite rock outcrop, beneath SW-facing wall, on dolomite stone, 840 m a.s.l., 69°02'N, 20°50'E, 14.8.2011, J. Pykälä 44387 (H). Protoblastenia rupestris (Scop.) J. Steiner, Verh. Kaiserl.-Königl. zool.-bot. Ges. Wien 61: 47 (1911) Lichen rupestris Scop., Fl. carniol., Edn 2 (Wien) 2: 363 (1772). Description. Thallus whitish-grey to rarely pale brown, rimose to predominantly areolate, areoles 0.1–0.8 mm, ca. 0.1–0.2 mm thick, K-, C-, UVto UV+ bluish-white, algal cells 4–12 μm. Apothecia yellow-orange to orange, 0.2–0.9 mm, slightly convex to convex, 3/4-immersed in thallus to superficial; ca. 40–100 apothecia / cm2. Epihymenium orange-brown, 12– 20 μm thick, K+ violet. Hymenium 50–80 μm thick. Hypothecium pale yellow, ca. 60–80 μm thick. Paraphyses ca. 1.5–2.5 μm thick, apex not thickened to slightly thickened to 2–3 μm thick, sparingly branched and anastomosing. Ascospores 0-septate, (8.2–)9.1–10.3–11.6(–13.0) × (5.2–)5.6–6.3– 7.1(–7.6) μm (n = 43). Habitat and distribution. Sequenced specimens were collected from rather sun-exposed calcareous rock outcrops in the hemi-boreal zone in Finland. The species may prefer calcareous rock outcrops close to shores. Notes. Protoblastenia rupestris sequences available in GenBank are heterogeneous and distributed in three different groups in our ITS phylogeny (Fig. 1). Specimens similar to those in Kainz and Rambold (2004) are likely to belong to P. rupestris s. stricto as the species was described from Central Europe. The sequence MZ159571 (UK) is related to P. incrustans and the sequence KY266981 (Norway) belongs in P. rikkinenii. Morphologically, the Finnish specimens of P. rupestris may be confused with the closely-related P. borealis. However, the P. rupestris specimens have a predominantly areolate thallus, while the thalli 224 MycoKeys 124: 193–226 (2025), DOI: 10.3897/mycokeys.124.162802 Juha Pykälä & Leena Myllys: Lichen genus Protoblastenia in Finland Use of AI No use of AI was reported. Funding The fieldwork and manuscript preparation were mainly conducted as parts of the research projects “Threatened lichens of calcareous rocks” (Grant number YTB059), “Key habitats for red-listed lichens” (Grant number 700T-MTX003) and “Inventory of calcareous and serpentine rocks” financed by the Finnish Ministry of the Environment. The first two projects were part of the Research Programme of Deficiently Known and Threatened Forest Species (PUTTE). DNA barcoding was supported by the Kone Foundation, the Finnish Cultural Foundation and the Academy of Finland through their grants to the Finnish Barcode of Life (FinBOL) and the Finnish Biodiversity Information Facility (FinBIF). In the project “Key habitats for red-listed lichens”, 32 specimens of Protoblastenia were barcoded. Author contributions All authors have contributed equally. 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