Russula albonigra is considered a well-known species, morphologically delimited by the context of the basidiomata blackening without intermediate reddening, and the menthol-cooling taste of the lamellae. It is supposed to have a broad ecological range and a large distribution area. A thorough molecular analysis based on four nuclear markers (ITS, LSU, RPB2 and TEF1-α) shows this traditional concept of R. albonigra s. lat. represents a species complex consisting of at least five European, three North American, and one Chinese species. Morphological study shows traditional characters used to delimit R. albonigra are not always reliable. Therefore, a new delimitation of the R. albonigra complex is proposed and a key to the described European species of R. subgen. Compactae is presented. A lectotype and an epitype are designated for R. albonigra and three new European species are described: R. ambusta, R. nigrifacta, and R. ustulata. Different thresholds of UNITE species hypotheses were tested against the taxonomic data. The distance threshold of 0.5% gives a perfect match to the phylogenetically defined species within the R. albonigra complex. Publicly available sequence data can contribute to species delimitation and increase our knowledge on ecology and distribution, but the pitfalls are short and low quality sequences.

Institute of Botany Plant Science and Biodiversity Center Slovak Academy of Sciences Dúbravská cesta 9 845 23 Bratislava Slovakia

Institute of Forest Ecology Slovak Academy of Sciences Akademická 2 949 01 Nitra Slovakia

Institute of Geology of the Czech Academy of Sciences Rozvojová 269 165 00 Prague 6 Czech Republic

Meise Botanic Garden Research Department Nieuwelaan 38 1860 Meise Belgium

Nuclear Physics Institute of the Czech Academy of Sciences Hlavní 130 250 68 Husinec Řež Czech Republic

Research Group Mycology Department of Biology Ghent University K L Ledeganckstraat 35 9000 Ghent Belgium

Erratum v

IMA Fungus. 2021 Oct 4;12(1):28. doi: 10.1186/s43008-021-00079-7. PubMed

Zobrazit více v PubMed

Adamčík S, Buyck B. Type studies in Russula subsection Nigricantes from the eastern United States. Cryptogamie Mycologie. 2014;35(3):293–309.

Adamčík S, Caboň M, Eberhardt U, Saba M, Hampe F, Slovak M, Kleine J, Marxmüller H, Jančovičová S, Pfister DH, Khalid AN, Kolarik M, Marhold K, Verbeken A. A molecular analysis reveals hidden species diversity within the current concept of Russula maculata (Russulaceae, Basidiomycota) Phytotaxa. 2016;270(2):71–88.

Adamčík S, Looney B, Caboň M, Jančovičová S, Adamčíková K, Avis PG, Barajas M, Bhatt RP, Corrales A, Das K, Hampe F, Ghosh A, Gates G, Kälviäinen V, Khalid AN, Kiran M, De Lange R, Lee H, Lim YW, Kong A, Manz C, Ovrebo C, Saba M, Taipale T, Verbeken A, Wisitrassameewong K, Buyck B. The quest for a globally comprehensible Russula language. Fungal Diversity. 2019;99(1):369–449.

Adamčík S, Slovak M, Eberhardt U, Ronikier A, Jairus T, Hampe F, Verbeken A. Molecular inference, multivariate morphometrics and ecological assessment are applied in concert to delimit species in the Russula clavipes complex. Mycologia. 2016;108(4):716–730. PubMed

Badotti F, de Oliveira FS, Garcia CF, Vaz ABM, Fonseca PLC, Nahum LA, Oliveira G, Goes-Neto A. Effectiveness of ITS and sub-regions as DNA barcode markers for the identification of Basidiomycota (Fungi) BMC Microbiology. 2017;17:12. PubMed PMC

Biodiversity Heritage Library. n.d. https://www.biodiversitylibrary.org/.

Bouckaert R, Vaughan TG, Barido-Sottani J, Duchene S, Fourment M, Gavryushkina A, Heled J, Jones G, Kuhnert D, De Maio N, Matschiner M, Mendes FK, Muller NF, Ogilvie HA, du Plessis L, Popinga A, Rambaut A, Rasmussen D, Siveroni I, Suchard MA, Wu CH, Xie D, Zhang C, Stadler T, Drummond AJ. BEAST 2.5: an advanced software platform for Bayesian evolutionary analysis. PLoS Computational Biology. 2019;15:28. PubMed PMC

Buyck B, Wang XH, Adamčíková K, Caboň M, Jančovičová S, Hofstetter V, Adamčík S. One step closer to unravelling the origin of Russula: subgenus Glutinosae subg. Nov. Mycosphere. 2020;11(1):285–305.

Buyck, B. 1989. Valeur taxonomique du bleu de crésyl pour le genre Russula. Bull Soc Mycol Fr 105:1-6.

Caboň M, Li GJ, Saba M, Kolarik M, Jančovičová S, Khalid AN, Moreau PA, Wen HA, Pfister DH, Adamčík S. Phylogenetic study documents different speciation mechanisms within the Russula globispora lineage in boreal and arctic environments of the northern hemisphere. IMA Fungus. 2019;10:16. PubMed PMC

Cabon M, Eberhardt U, Looney B, Hampe F, Kolarik M, Jancovicova S, Verbeken A, Adamcik S. 2017. New insights in Russula subsect. Rubrinae: phylogeny and the quest for synapomorphic characters. Mycol Prog 16:877-892

Chalange R. Utilisation du gaïac pour une aide à la détermination des russules sur le terrain. Bulletin de la Société Mycologique de France. 2014;130:39–55.

Chernomor O, von Haeseler A, Minh BQ. Terrace aware data structure for Phylogenomic inference from Supermatrices. Systematic Biology. 2016;65(6):997–1008. PubMed PMC

De Crop E, Nuytinck J, Van de Putte K, Lecomte M, Eberhardt U, Verbeken A. Lactifluus piperatus (Russulales, Basidiomycota) and allied species in Western Europe and a preliminary overview of the group worldwide. Mycological Progress. 2014;13(3):493–511.

De Lange R, De Crop E, Delgat L, Tibuhwa D, Baribwegure D, V. A. Lactifluus kigomaensis and L. subkigomaensis: two look-alikes in Tanzania. Mycoscience. 2018;59(5):371–378.

Delgat L, De Crop E, Njouonkou AL, Verbeken A. Lactifluus persicinus sp. nov. from the gallery forests of West Cameroon. Mycotaxon. 2017;132(3):471–483.

Delgat L, Dierickx G, De Wilde S, Angelini C, De Crop E, De Lange R, Halling R, Manz C, Nuytinck J, Verbeken A. Looks can be deceiving: the deceptive milkcaps (Lactifluus, Russulaceae) exhibit low morphological variance but harbour high genetic diversity. IMA Fungus. 2019;10(1):14. PubMed PMC

Drummond AJ, Rambaut A. BEAST: Bayesian evolutionary analysis by sampling trees. BMC Evolutionary Biology. 2007;7:8. PubMed PMC

Drummond AJ, Suchard MA, Xie D, Rambaut A. Bayesian Phylogenetics with BEAUti and the BEAST 1.7. Molecular Biology and Evolution. 2012;29(8):1969–1973. PubMed PMC

Durkin L, Jansson T, Sanchez M, Khomich M, Ryberg M, Kristiansson E, Nilsson RH. When mycologists describe new species, not all relevant information is provided (clearly enough) MycoKeys. 2020;72:109–128. PubMed PMC

Fries EM. Hymenomycetes europaei. Upsaliae. 1874.

Gardes M, Bruns TD. ITS primers with enhanced specificity for Basidiomycetes - application to the identification of mycorrhizae and rusts. Molecular Ecology. 1993;2(2):113–118. PubMed

Guindon S, Dufayard JF, Lefort V, Anisimova M, Hordijk W, Gascuel O. New algorithms and methods to estimate maximum-likelihood phylogenies: assessing the performance of PhyML 3.0. Systematic Biology. 2010;59(3):307–321. PubMed

Hesler LR. Study of Schaeffer, J Russula’s. Lloydia. 1961;24:182–198.

Hibbett DS, Ohman A, Glotzer D, Nuhn M, Kirk P, Nilsson RH. Progress in molecular and morphological taxon discovery in Fungi and options for formal classification of environmental sequences. Fungal Biology Reviews. 2011;25(1):38–47.

Hofstetter V, Buyck B, Eyssartier G, Schnee S, Gindro K. The unbearable lightness of sequenced-based identification. Fungal Diversity. 2019;96(1):243–284.

Huelsenbeck JP, Ronquist F. MRBAYES: Bayesian inference of phylogeny. Bioinformatics. 2001;17(8):754–755. PubMed

Jones G. Algorithmic improvements to species delimitation and phylogeny estimation under the multispecies coalescent. Journal of Mathematical Biology. 2017;74(1-2):447–467. PubMed

Jones G, Aydin Z, Oxelman B. DISSECT: an assignment-free Bayesian discovery method for species delimitation under the multispecies coalescent. Bioinformatics. 2014;31:991–998. PubMed

Kalyaanamoorthy S, Minh BQ, Wong TKF, von Haeseler A, Jermiin LS. ModelFinder: fast model selection for accurate phylogenetic estimates. Nature Methods. 2017;14(6):587–589. PubMed PMC

Katoh K, Toh H. Recent developments in the MAFFT multiple sequence alignment program. Briefings in Bioinformatics. 2008;9(4):286–298. PubMed

Kibby G. Key to blackening species of Russula of section Compactae. Field Mycology. 2001;2(3):95–97.

Kibby G, Fatto R. Keys to the species of Russula in Northeastern North America. 3. Somerville: Kibby-Fatto Enterprises; 1990.

Kõljalg U, Larsson K-H, Abarenkov K, Nilsson RH, Alexander IJ, Eberhardt U, Erland S, Høiland K, Kjøller R, Larsson E, Pennanen T, Sen R, Taylor AFS, Tedersoo L, Vrålstad T. UNITE: a database providing web-based methods for the molecular identification of ectomycorrhizal fungi. New Phytologist. 2005;166(3):1063–1068. PubMed

Kõljalg U, Nilsson HR, Schigel D, Tedersoo L, Larsson KH, May TW, Taylor AFS, Jeppesen TS, Froslev TG, Lindahl BD, Poldmaa K, Saar I, Suija A, Savchenko A, Yatsiuk I, Adojaan K, Ivanov F, Piirmann T, Pohonen R, Zirk A, Abarenkov K. The taxon hypothesis paradigm-on the unambiguous detection and communication of taxa. Microorganisms. 2020;8:25. PubMed PMC

Kõljalg U, Nilsson RH, Abarenkov K, Tedersoo L, Taylor AFS, Bahram M, Bates ST, Bruns TD, Bengtsson-Palme J, Callaghan TM, Douglas B, Drenkhan T, Eberhardt U, Duenas M, Grebenc T, Griffith GW, Hartmann M, Kirk PM, Kohout P, Larsson E, Lindahl BD, Luecking R, Martin MP, Matheny PB, Nguyen NH, Niskanen T, Oja J, Peay KG, Peintner U, Peterson M, Poldmaa K, Saag L, Saar I, Schuessler A, Scott JA, Senes C, Smith ME, Suija A, Taylor DL, Telleria MT, Weiss M, Larsson KH. Towards a unified paradigm for sequence-based identification of fungi. Molecular Ecology. 2013;22(21):5271–5277. PubMed

Kornerup A, Wanscher JH. Methuen handbook of colour. 3. London: Methuen; 1978.

Krombholz JV v. Naturgetreue Abbildungen und Beschreibungen der Schwämme. 1845. p. 9.

Kumar S, Stecher G, Tamura K. MEGA7: molecular evolutionary genetics analysis version 7.0 for bigger datasets. Molecular Biology and Evolution. 2016;33(7):1870–1874. PubMed PMC

Lanfear R, Calcott B, Ho SYW, Guindon S. PartitionFinder: combined selection of partitioning schemes and substitution models for phylogenetic analyses. Molecular Biology and Evolution. 2012;29(6):1695–1701. PubMed

Lanfear R, Frandsen PB, Wright AM, Senfeld T, Calcott B. PartitionFinder 2: new methods for selecting partitioned models of evolution for molecular and morphological phylogenetic analyses. Molecular Biology and Evolution. 2017;34(3):772–773. PubMed

Le HT, Nuytinck J, Verbeken A, Lumyong S, Desjardin DE. Lactarius in northern Thailand: 1. Lactarius subgenus Piperites. Fungal Diversity. 2007;24:173–224.

Leache AD, Fujita MK. Bayesian species delimitation in West African forest geckos (Hemidactylus fasciatus) Proceedings of the Royal Society B: Biological Sciences. 2010;277(1697):3071–3077. PubMed PMC

Leonhardt T, Borovička J, Sácký J, Šantrůček J, Kameník J, Kotrba P, (2019) Zn overaccumulating Russula species clade together and use the same mechanism for the detoxification of excess Zn. Chemosphere 225:618-626 PubMed

Liu YJJ, Whelen S, Benjamin DH. Phylogenetic relationships among Ascomycetes: evidence from an RNA polymerase II subunit. Molecular Biology and Evolution. 1999;16(12):1799–1808. PubMed

Looney BP, Adamcik S, Matheny PB. Coalescent-based delimitation and species-tree estimations reveal Appalachian origin and Neogene diversification in Russula subsection Roseinae. Molecular Phylogenetics and Evolution. 2020;147:13. PubMed

Lucking R, Aime MC, Robbertse B, Miller AN, Ariyawansa HA, Aoki T, Cardinali G, Crous PW, Druzhinina IS, Geiser DM, Hawksworth DL, Hyde KD, Irinyi L, Jeewon R, Johnston PR, Kirk PM, Malosso E, May TW, Meyer W, Opik M, Robert V, Stadler M, Thines M, Vu D, Yurkov AM, Zhang N, Schoch CL. Unambiguous identification of fungi: where do we stand and how accurate and precise is fungal DNA barcoding? IMA Fungus. 2020;11:32. PubMed PMC

Matheny PB. Improving phylogenetic inference of mushrooms with RPB1 and RPB2 nucleotide sequences (Inocybe; Agaricales) Molecular Phylogenetics and Evolution. 2005;35(1):1–20. PubMed

Matute DR, Sepulveda VE. Fungal species boundaries in the genomics era. Fungal Genetics and Biology. 2019;131:9. PubMed PMC

Moncalvo JM, Lutzoni FM, Rehner SA, Johnson J, Vilgalys R. Phylogenetic relationships of agaric fungi based on nuclear large subunit ribosomal DNA sequences. Systematic-Biology. 2000;49(2):278–305. PubMed

Morehouse EA, James TY, Ganley ARD, Vilgalys R, Berger L, Murphy PJ, Longcore JE. Multilocus sequence typing suggests the chytrid pathogen of amphibians is a recently emerged clone. Molecular Ecology. 2003;12(2):395–403. PubMed

Naranjo-Ortiz MA, Gabaldon T. Fungal evolution: diversity, taxonomy and phylogeny of the Fungi. Biological Reviews. 2019;94(6):2101–2137. PubMed PMC

Nguyen L-T, Schmidt HA, von Haeseler A, Minh BQ. IQ-TREE: a fast and effective stochastic algorithm for estimating maximum-likelihood phylogenies. Molecular Biology and Evolution. 2014;32:268–274. PubMed PMC

Nilsson RH, Hyde KD, Pawlowska J, Ryberg M, Tedersoo L, Aas AB, Alias SA, Alves A, Anderson CL, Antonelli A, Arnold AE, Bahnmann B, Bahram M, Bengtsson-Palme J, Berlin A, Branco S, Chomnunti P, Dissanayake A, Drenkhan R, Friberg H, Froslev TG, Halwachs B, Hartmann M, Henricot B, Jayawardena R, Jumpponen A, Kauserud H, Koskela S, Kulik T, Liimatainen K, Lindahl BD, Lindner D, Liu JK, Maharachchikumbura S, Manamgoda D, Martinsson S, Neves MA, Niskanen T, Nylinder S, Pereira OL, Pinho DB, Porter TM, Queloz V, Riit T, Sanchez-Garcia M, de Sousa F, Stefanczyk E, Tadych M, Takamatsu S, Tian Q, Udayanga D, Unterseher M, Wang Z, Wikee S, Yan JY, Larsson E, Larsson KH, Koljalg U, Abarenkov K. Improving ITS sequence data for identification of plant pathogenic fungi. Fungal Diversity. 2014;67(1):11–19.

Nilsson RH, Larsson K-H, Taylor AFS, Bengtsson-Palme J, Jeppesen TS, Schigel D, Kennedy P, Picard K, Glöckner FO, Tedersoo L, Saar I, Kõljalg U, Abarenkov K. The UNITE database for molecular identification of fungi: handling dark taxa and parallel taxonomic classifications. Nucleic Acids Research. 2018;47:D259–D264. PubMed PMC

Nuytinck J, Verbeken A. Lactarius sanguifluus versus Lactarius vinosus – molecular and morphological analyses. Mycological Progress. 2003;2(3):227–234.

Pauvert C, Buee M, Laval V, Edel-Hermann V, Fauchery L, Gautier A, Lesur I, Vallance J, Vacher C. Bioinformatics matters: the accuracy of plant and soil fungal community data is highly dependent on the metabarcoding pipeline. Fungal Ecology. 2019;41:23–33.

Rambaut A, Drummond AJ, Xie D, Baele G, Suchard MA. Posterior summarization in Bayesian phylogenetics using tracer 1.7. Systematic Biology. 2018;67(5):901–904. PubMed PMC

Romagnesi H. Taxa nova ex genere Russula. Publications de la Société Linnéenne de Lyon. 1962;31:172–177.

Romagnesi H. Les Russules d’Europe et d’Afrique du Nord. Paris: Bordas; 1967.

Ronquist F, Huelsenbeck JP. MrBayes 3: Bayesian phylogenetic inference under mixed models. Bioinformatics. 2003;19(12):1572–1574. PubMed

Ryberg M. Molecular operational taxonomic units as approximations of species in the light of evolutionary models and empirical data from Fungi. Molecular Ecology. 2015;24(23):5770–5777. PubMed

Ryberg M, Matheny PB. Asynchronous origins of ectomycorrhizal clades of Agaricales. Proceedings of the Royal Society B: Biological Sciences. 2012;279(1735):2003–2011. PubMed PMC

Sarnari M. Monografia illustrate del genere Russula in Europa, Tomo Primo. Trento: Associazioni Micologica Bresadola; 1998.

Shaffer RL. The subsection Compactae of Russula. Brittonia. 1962;14(3):254–284.

Singer R. New and interesting species of Basidiomycetes V. Sydowia. 1958;11:141–374.

Stielow JB, Levesque CA, Seifert KA, Meyer W, Irinyi L, Smits D, Renfurm R, Verkley GJM, Groenewald M, Chaduli D, Lomascolo A, Welti S, Lesage-Meessen L, Favel A, Al-Hatmi AMS, Damm U, Yilmaz N, Houbraken J, Lombard L, Quaedvlieg W, Binder M, Vaas LAI, Vu D, Yurkov A, Begerow D, Roehl O, Guerreiro M, Fonseca A, Samerpitak K, van Diepeningen AD, Dolatabadi S, Moreno LF, Casaregola S, Mallet S, Jacques N, Roscini L, Egidi E, Bizet C, Garcia-Hermoso D, Martin MP, Deng S, Groenewald JZ, Boekhout T, de Beer ZW, Barnes I, Duong TA, Wingfield MJ, de Hoog GS, Crous PW, Lewis CT, Hambleton S, Moussa TAA, Al-Zahrani HS, Almaghrabi OA, Louis-Seize G, Assabgui R, McCormick W, Omer G, Dukik K, Cardinali G, Eberhardt U, de Vries M, Robert V. One fungus, which genes? Development and assessment of universal primers for potential secondary fungal DNA barcodes. Persoonia. 2015;35(1):242–263. PubMed PMC

Stubbe D, Nuytinck J, Verbeken A. Critical assessment of the Lactarius gerardii species complex (Russulales) Fungal Biology. 2010;114(2-3):271–283. PubMed

Tel-Zur N, Abbo S, Myslabodski D, Mizrahi Y. Modified CTAB procedure for DNA isolation from epiphytic cacti of the genera Hylocereus and Selenicereus (Cactaceae) Plant Molecular Biology Reporter. 1999;17(3):249–254.

Thiers HD. The subgenus Compactae of Russula in California. Mycologia Helvetica. 1994;2:107–120.

Van de Putte K. Hidden diversity exposed: a case study of Lactifluus volemus sensu lato. Ghent: Ghent University; 2012.

Van de Putte K, Nuytinck J, De Crop E, Verbeken A. Lactifluus volemus in Europe: three species in one – revealed by a multilocus genealogical approach, Bayesian species delimitation and morphology. Fungal Biology. 2016;120(1):1–25. PubMed

Van de Putte K, Nuytinck J, Stubbe D, Le HT, Verbeken A. Lactarius volemus sensu lato (Russulales) from northern Thailand: morphological and phylogenetic species concepts explored. Fungal Diversity. 2010;45(1):99–130.

Varga T, Krizsan K, Foldi C, Dima B, Sanchez-Garcia M, Sanchez-Ramirez S, Szollosi GJ, Szarkandi JG, Papp V, Albert L, Andreopoulos W, Angelini C, Antonin V, Barry KW, Bougher NL, Buchanan P, Buyck B, Bense V, Catcheside P, Chovatia M, Cooper J, Damon W, Desjardin D, Finy P, Geml J, Haridas S, Hughes K, Justo A, Karasinski D, Kautmanova I, Kiss B, Kocsube S, Kotiranta H, LaButti KM, Lechner BE, Liimatainen K, Lipzen A, Lukacs Z, Mihaltcheva S, Morgado LN, Niskanen T, Noordeloos ME, Ohm RA, Ortiz-Santana B, Ovrebo C, Racz N, Riley R, Savchenko A, Shiryaev A, Soop K, Spirin V, Szebenyi C, Tomsovsky M, Tulloss RE, Uehling J, Grigoriev IV, Vagvolgyi C, Papp T, Martin FM, Miettinen O, Hibbett DS, Nagy LG. Megaphylogeny resolves global patterns of mushroom evolution. Nature Ecology & Evolution. 2019;3(4):668–678. PubMed PMC

Vu D, Groenewald M, de Vries M, Gehrmann T, Stielow B, Eberhardt U, Al-Hatmi A, Groenewald JZ, Cardinali G, Houbraken J, Boekhout T, Crous PW, Robert V, Verkley GJM. Large-scale generation and analysis of filamentous fungal DNA barcodes boosts coverage for kingdom Fungi and reveals thresholds for fungal species and higher taxon delimitation. Studies in Mycology. 2019;92:135–154. PubMed PMC

Vu D, Groenewald M, Verkley G. Convolutional neural networks improve fungal classification. Scientific Reports. 2020;10:12. PubMed PMC

White TJ, Bruns T, Lee S, Taylor JW. Amplification and direct sequencing of fungal ribosomal RNA genes for phylogenetics. In: Innis MA, Gelfand DH, Sninsky JJ, White TJ, editors. PCR protocols: a guide to methods and applications. New York: Academic Press; 1990. pp. 315–322.

Yang ZH. The BPP program for species tree estimation and species delimitation. Current Zoology. 2015;61(5):854–865.

Yang ZH, Rannala B. Bayesian species delimitation using multilocus sequence data. Proceedings of the National Academy of Sciences of the United States of America. 2010;107(20):9264–9269. PubMed PMC

Yang ZH, Rannala B. Unguided species delimitation using DNA sequence data from multiple loci. Molecular Biology and Evolution. 2014;31(12):3125–3135. PubMed PMC

Correction to: Enlightening the black and white: species delimitation and UNITE species hypothesis testing in the Russula albonigra species complex