Mycological Progress

, 14:78 | Cite as

Taxonomic relationships among non-macrocystidiate taxa of Lactarius subg. Russularia from Europe with special reference to species from Greece

  • Marina Triantafyllou
  • Elias Polemis
  • Zacharoula Gonou-Zagou
  • Dimitris M. Dimou
  • Panayiotis Delivorias
  • Georgios I. Zervakis
Original Article


Members of the ectomycorrhizal genus Lactarius form symbiotic relationships with a wide range of plant species. For the purposes of this study, 29 specimens originally assigned to L. atlanticus, L. serifluus and L. subumbonatus were examined by including all pertinent available material from Greece. The detailed study of macroscopic and microscopic characteristics by using compound optical, differential interference contrast (DIC) and scanning electron microscopy in combination with molecular approaches (ITS1-5.8S-ITS2 rDNA sequencing) permitted a clear separation between L. atlanticus and L. subumbonatus, while their basidiospore shape and height of ornamentation were found to be of diagnostic value. Moreover, the morphological delimitation between the specimens identified as L. serifluus and L. subumbonatus is not clear, and these specimens were intermixed into the same phylogenetic cluster with high support. Thus, the latter name should be considered as a synonym of the former. As concerns other related taxa of L. subg. Russularia, the American L. rubidus and L. subserifluus demonstrated relatively high evolutionary affinity with European collections of L. serifluus/L. subumbonatus and L. atlanticus, respectively.


Russulaceae Phylogeny Fungal taxonomy Milkcaps ITS-5.8S gene Basidiomycota 



This research has been co-financed by the European Union (European Social Fund—ESF) and Greek national funds through the Operational Program “Education and Lifelong Learning” of the National Strategic Reference Framework (NSRF) Research Funding Program entitled “Contribution of mycorrhizae to the sustainability of marginal Mediterranean ecosystems—development of mycorrhizal inocula (SALTYMYC)” (THALIS – AUA – MIS 380233). We would like to thank our colleagues C. Fasseas (Agricultural University of Athens) for helping with SEM observations, and Z. Athanassiou, Α. Verbeken (Ghent University), G. Venturella (University of Palermo) and G. Lalli (University of L'Aquila) for kindly providing Lactarius specimens. The helpful comments of two anonymous reviewers are gratefully acknowledged.

Conflict of interest

The authors declare that they have no conflict of interest.


  1. Basso MT (1999) Lactarius Pers. Fungi Europei VIII. Mycoflora, AlassioGoogle Scholar
  2. Betak J (2013) Distribution and ecology of Lactarius rostratus and Lactarius rubrocinctus (Basidiomycota, Russulales) in the Czech Republic. Czech Mycol 65:25–43Google Scholar
  3. Bon M (1975) Agaricales de la côte atlantique française. Doc Mycol 5:1–40Google Scholar
  4. Bon M (1976) Lactarius atlanticus Bon forma strigipes Bon f. nov. Doc Mycol 7:57–58Google Scholar
  5. Bon M (1980) Clé monographique du genre Lactarius (Pers. ex Fr.) S.F. Gray. Doc Mycol 10:1–85Google Scholar
  6. Buyck B, Hofstetter V, Eberhardt U, Verbeken A, Kauff F (2008) Walking the thin line between Russula and Lactarius: the dilemma of Russula subsect. Ochricompactae. Fungal Divers 28:15–40Google Scholar
  7. Buyck B, Hofstetter V, Verbeken A, Walleyn R (2010) Proposal to conserve Lactarius nom. cons. (Basidiomycota) with a conserved type. Taxon 59:295–296Google Scholar
  8. Calonge FD, Martín MP (2003) Zelleromyces hispanicus, the gasteroid phase of Lactarius aurantiacus. Bol Soc Micol Madrid 27:231–236Google Scholar
  9. Clémençon H (1973) Zwei verbesserte Präparierlösungen für die microskopische Untersuchung von Pilze. Z Pilzkd 38:49–53Google Scholar
  10. De Candolle AU (1815) Flore Française 6:45Google Scholar
  11. Eberhardt U, Verbeken A (2004) Sequestrate Lactarius species from tropical Africa: L. angiocarpus sp. nov. and L. dolichocaulis comb. nov. Mycol Res 108:1042–1052CrossRefPubMedGoogle Scholar
  12. Eberhardt U, Oberwinkler F, Verbeken A, Rinaldi AC, Pacioni G, Comandini O (2000) Lactarius ectomycorrhizae on Abies alba: morphological description, molecular characterization, and taxonomic remarks. Mycologia 92:860–873Google Scholar
  13. Felsenstein J (1985) Confidence limits on phylogenies: an approach using the bootstrap. Evolution 39:783–791CrossRefGoogle Scholar
  14. Heilmann-Clausen J, Verbeken A, Vesterholt J (1998) The genus Lactarius. Fungi of Northern Europe Vol. 2. Svampetryk, DenmarkGoogle Scholar
  15. Hesler LR, Smith AH (1979) North American species of Lactarius. University of Michigan, Ann ArborGoogle Scholar
  16. Huang X, Madan A (1999) CAP3: A DNA sequence assembly program. Genome Res 9:868–877PubMedCentralCrossRefPubMedGoogle Scholar
  17. Kimura M (1980) A simple method for estimating evolutionary rate of base substitutions through comparative studies of nucleotide sequences. J Mol Evol 16:111–120CrossRefPubMedGoogle Scholar
  18. Kuo M, Methven AS, Minnis AM, Halling RE (2013) Studies of North American macrofungi, 1. Validation of Lactarius rubidus comb. nov. and Leccinellum quercophilum sp. nov. Mycotaxon 124:323–332CrossRefGoogle Scholar
  19. Lamus V, Montoya L, Aguilar CJ, Bandala VM, Ramos D (2012) Ectomycorrhizal association of three Lactarius species with Carpinus and Quercus trees in a Mexican montane cloud forest. Mycologia 104:1261–1266CrossRefPubMedGoogle Scholar
  20. Lindgren SJ (1845) Notiser från Wenern. Bot Notiser 12:193–202Google Scholar
  21. Montoya L, Bandala VM (2008) A new species and new records of Lactarius (subgenus Russularia) in a subtropical cloud forest from eastern Mexico. Fungal Divers 29:61–72Google Scholar
  22. Moser Μ (1978) Kleine Kryptogamenflora - Die Röhrlinge und Blätterpilze (Agaricales). IIb/2:1–532Google Scholar
  23. Neuhoff W (1956) Die Milchlinge. (Lactarii). Die Pilze Mitteleuropas Bd IIb. Julius Klinkhardt, Bad HeilbrunnGoogle Scholar
  24. Nuytinck J, Verbeken A, Delarue S, Walleyn R (2003) Systematics of European sequestrate lactarioid Russulaceae with spiny spore ornamentation. Belg J Bot 136:145–153Google Scholar
  25. Nygren CM, Edqvist J, Elfstrand M, Heller G, Taylor AF (2007) Detection of extracellular protease activity in different species and genera of ectomycorrhizal fungi. Mycorrhiza 17:241–248CrossRefPubMedGoogle Scholar
  26. Osmundson TW, Robert VA, Schoch CL, Baker LJ, Smith A, Robich G, Mizzan L, Garbelotto MM (2013) Filling gaps in biodiversity knowledge for macrofungi: contributions and assessment of an herbarium collection DNA barcode sequencing project. PLoS ONE 8(4):e62419PubMedCentralCrossRefPubMedGoogle Scholar
  27. Palmer JM, Lindner DL, Volk TJ (2008) Ectomycorrhizal characterization of an American chestnut (Castanea dentata)-dominated community in Western Wisconsin. Mycorrhiza 19:27–36CrossRefPubMedGoogle Scholar
  28. Peay KG, Bruns TD, Kennedy PG, Bergemann SE, Garbelotto M (2007) A strong species-area relationship for eukaryotic soil microbes: island size matters for ectomycorrhizal fungi. Ecol Lett 10:470–480CrossRefPubMedGoogle Scholar
  29. Pickles BJ, Genney DR, Anderson IC, Alexander IJ (2012) Spatial analysis of ectomycorrhizal fungi reveals that root tip communities are structured by competitive interactions. Mol Ecol 21:5110–5123CrossRefPubMedGoogle Scholar
  30. Ravash R, Shiran B, Alavi A-A, Bayat F, Rajaee S, Zervakis GI (2010) Genetic variability and molecular phylogeny of Pleurotus eryngii species-complex isolates from Iran, and notes on the systematics of Asiatic populations. Mycol Prog 9:181–194CrossRefGoogle Scholar
  31. Royal Botanic Garden Edinburgh (1969) Flora of British fungi: colour identification chart. H.M. Stationery Office, EdinburghGoogle Scholar
  32. Suz LM, Barsoum N, Benham S, Dietrich H-P, Fetzer KD, Fischer R, García P, Gehrman J, Kristöfel F, Manninger M, Neagu S, Nicolas M, Oldenburger J, Raspe S, Sánchez G, Schröck HW, Schubert A, Verheyen K, Verstraeten A, Bidartondo MI (2014) Environmental drivers of ectomycorrhizal communities in Europe's temperate oak forests. Mol Ecol 23:5628–5644CrossRefPubMedGoogle Scholar
  33. Tamura K, Stecher G, Peterson D, Filipski A, Kumar S (2013) MEGA6: molecular evolutionary genetics analysis version 6.0. Mol Biol Evol 30:2725–2729PubMedCentralCrossRefPubMedGoogle Scholar
  34. Triantafyllou M, Polemis E, Dimou DM, Gonou-Zagou Z, Delivorias P, Zervakis GI (2011) A reappraisal of existing knowledge on the diversity of the genus Lactarius Pers. in Greece. Proceedings of the XVI Congress of European Mycologists, pp. 122–123 (abstract). European Mycological Association. Porto Karras, GreeceGoogle Scholar
  35. Vellinga EC (1988) Glossary. In: Bas C, Kuyper TW, Noordeloos ME, Vellinga EC (eds) Flora Agaricina Neerlandica, vol 1. AA Balkema, Rotterdam, pp 54–64Google Scholar
  36. Verbeken A, Nuytinck J (2013) Not every milkcap is a Lactarius. Scripta Bot Belgica 51:162–168Google Scholar
  37. Verbeken A, Stubbe D, Van de Putte K, Eberhardt U, Nuytinck J (2014) Tales of the unexpected: angiocarpous representatives of the Russulaceae in tropical South East Asia. Persoonia 32:13–24Google Scholar
  38. Wang XH (2007) Type studies of Lactarius species published from China. Mycologia 99:253–268CrossRefPubMedGoogle Scholar
  39. Wang XH, Liu PG (2002) Lactarius chichuensis and L. hirtipes, two easily confused species. Mycotaxon 84:391–400Google Scholar
  40. White TJ, Buns TD, Lee SB, Taylor JW (1990) Amplication and direct sequencing of fungal ribosomal genes for phylogenetics. In: Innis MA, Gelfand DH, Sninsky JJ, White TJ (eds) PCR protocols. Academic Press, San Diego, pp 315–322Google Scholar
  41. Wisitrassameewong K, Nuytinck J, Hyde KD, Verbeken A (2014) Lactarius subgenus Russularia (Russulaceae) in Southeast Asia: 1. species with very distant gills. Phytotaxa 158:23–42CrossRefGoogle Scholar
  42. Zervakis GI, Ntougias S, Gargano ML, Besi MI, Polemis E, Typas MA, Venturella G (2014) A reappraisal of the Pleurotus eryngii complex – New species and taxonomic combinations based on the application of a polyphasic approach, and an identification key to Pleurotus taxa associated with Apiaceae plants. Fungal Biol 118:814–834CrossRefPubMedGoogle Scholar

Copyright information

© German Mycological Society and Springer-Verlag Berlin Heidelberg 2015

Authors and Affiliations

  • Marina Triantafyllou
    • 1
    • 2
  • Elias Polemis
    • 1
  • Zacharoula Gonou-Zagou
    • 2
  • Dimitris M. Dimou
    • 1
    • 3
  • Panayiotis Delivorias
    • 2
  • Georgios I. Zervakis
    • 1
  1. 1.Laboratory of General and Agricultural MicrobiologyAgricultural University of AthensAthensGreece
  2. 2.Faculty of Biology, Department of Ecology and SystematicsNational and Kapodistrian University of AthensAthensGreece
  3. 3.Agia ParaskeviGreece

Personalised recommendations