Mycological Progress

, Volume 18, Issue 1–2, pp 147–161 | Cite as

Cryptic species within Ophiocordyceps myrmecophila complex on formicine ants from Thailand

  • Artit Khonsanit
  • Janet Jennifer Luangsa-ard
  • Donnaya Thanakitpipattana
  • Noppol Kobmoo
  • Onuma PiasaiEmail author


The Ophiocordyceps myrmecophila complex is composed of pathogens specific to ants, found on the leaf litter or buried in soil and produce Hymenostilbe asexual morph. Species in this complex are morphologically highly similar and can hardly be distinguished macroscopically. To date, it has only been observed on formicine ants of the genera Polyrhachis and Camponotus. In this study, observations were conducted in three sites at Khao Yai National Park of Thailand where three new species are proposed. Molecular phylogenies based on the large subunit of the ribosomal DNA (LSU), partial sequences of translation elongation factor 1-α (TEF), and the largest and second largest subunits of the RNA polymerase Π (PRB1 and RPB2) revealed distinct clades separating these new species, namely Ophiocordyceps megacuculla, Ophiocordyceps khaoyaiensis, and Ophiocordyceps granospora. The morphological features of these three new species are clearly different from Ophiocordyceps thanathonensis, a recently described species of the complex, but mostly overlap between these three. However, they are proposed as distinct species based on molecular phylogenetic relationships, genetic distance, and minor morphological characters related to sexual structures.


Ophiocordycipitaceae Taxonomy Phylogenetic Entomogenous fungi Multigene New species 



The authors would like to express their appreciation to the Graduate School, Kasetsart University, for the Graduate Scholarship for the Fiscal Year 2015 for the first author. We are grateful to the Microbe Interaction and Ecology Laboratory (BMIE), National Center for Genetic Engineering and Biotechnology (BIOTEC), Thailand, for the use of the facilities; the Royal Forest Department of Thailand for the permission to survey and collect fungi in Khao Yai National Park. We would like to thank Dr. Sasithorn Hasin from Valaya Alongkorn Rajabhat University for her help in the identification of the ant hosts.


  1. Araújo JPM, Hughes DP (2017) The fungal spore: Myrmecophilous Ophiocordyceps as a case study. In: Dighton J, White JM (eds) The fungal community: its organization and role in the ecosystem, 4th edn. CRC Press, USA, pp 359–367Google Scholar
  2. Araújo JPM, Evans HC, Geiser DM, Mackay WP, Hughes DP (2015) Unravelling the diversity behind the Ophiocordyceps unilateralis (Ophiocordycipitaceae) complex: three new species of zombie-ant fungi from the Brazilian Amazon. Phytotaxa 220(3):224–238. CrossRefGoogle Scholar
  3. Araújo JPM, Evans HC, Kepler RM, Hughes DP (2018) Zombie-ant fungi across continents: 15 new species and new combinations within Ophiocordyceps. I. Myrmecophilous hirsutelloid species. Stud Mycol 90:119−160. CrossRefPubMedPubMedCentralGoogle Scholar
  4. Castlebury LA, Rossman AY, Sung G-H, Hyten AS, Sapatafora JW (2004) Multigene phylogeny reveals new lineage for Stachybotrys chatarum, the indoor air fungus. Mycol Res 108:1–9. CrossRefGoogle Scholar
  5. Chao SC, Hsieh IL, Lee LW, Cheng KT, Su CH (2004) Culture of Cordyceps myrmecophila Cessati. Fung Sci 19(1,2):53–59Google Scholar
  6. Crous PW, Gams W, Stalpers JA, Robert V, Stegehuis G (2004) MycoBank: an online initiative to launch mycology into 21st century. Stud Mycol 50:12–22Google Scholar
  7. Edgar RC (2004) MUSCLE: multiple sequence alignment with high accuracy and high throughput. Nucleic Acids Res 32(5):1792−1797. CrossRefPubMedCentralGoogle Scholar
  8. Evans HC, Samson RA (1982) Cordyceps species and their anamorphs pathogenic on ants (Formicidae) in tropical forest ecosystem. I. The Cephalotes (Myrmicinae) complex. Transaction of the British Mycological Society 79:431–453. CrossRefGoogle Scholar
  9. Evans HC, Samson RA (1984) Cordyceps species and their anamorphs pathogenic of ants (Formicidae) in tropical forest ecosystem. II. The Camponotus (Formicinae) complex. Trans Br Mycol Soc 82:127–150. CrossRefGoogle Scholar
  10. Evans HC, Elliot SL, Hughes DP (2011) Hidden Diversity Behind the Zombie-Ant Fungus Ophiocordyceps unilateralis: Four New Species Described from Carpenter Ants in Minas Gerais, Brazil. PLoS One 6(3):e17024. CrossRefPubMedPubMedCentralGoogle Scholar
  11. Hall TA (2013) BioEdit: a user-friendly biological sequences alignment editor analysis program for windows 95/98/NT. Nucleic Acids Symp Ser 41:95–98CrossRefGoogle Scholar
  12. Hywel-Jones NL (1996) Cordyceps myrmecophila-like fungi infecting ants in the leaf litter of tropical forest in Thailand. Mycol Res 100(5):613−619CrossRefGoogle Scholar
  13. Kepler RM, Kaitsu Y, Tanaka E, Shimano S, Spatafora JW (2011) Ophiocordyceps pulvinata sp. nov., a pathogen of ants with a reduced stroma. Mycoscience 52:39–47. CrossRefGoogle Scholar
  14. Kepler RM, Sung G-H, Ban S, Nakagiri A, Chen MJ, Huang B, Li Z, Spatafora JW (2012) New sexual morph combinations in the entomopathogenic genus Metacordyceps. Mycologia 104(1):182–197. CrossRefPubMedGoogle Scholar
  15. Kobayasi Y (1941) The genus Cordyceps and its allies. Science Reports of the Tokyo Bunrika Daigaku. 5:53–260Google Scholar
  16. Kobayasi Y, Shimizu D (1981) The genus Cordyceps and its allies from Taiwan (Formosa). Bull Nati Sci Mus Tokyo. 7:113–122Google Scholar
  17. Kobmoo N, Mongkolsamrit S, Tasanathai K, Thanakitpipattana D, Luansa-ard JJ (2012) Molecular phylogenies reveal host-specific divergence of Ophiocordyceps unilateralis sensu lato following its host ants. Mol Ecol 21:3022–3031. CrossRefPubMedGoogle Scholar
  18. Kobmoo N, Mongkolsamrit S, Wutikhun T, Tasanathai K, Khonsanit A, Thanakitpipattana D, Luansa-ard JJ (2015) Ophiocordyceps unilateralis, an ubiquitous pathogen of ants from Thailand. Fungal Biol 119:44–52. CrossRefPubMedGoogle Scholar
  19. Kornerup A, Wanscher JH (1963) Methuen handbook of colour. Methuen, LondonGoogle Scholar
  20. Luangsa-ard JJ, Tasanathai K, Mongkolsamrit S, Hywel-Jones NL (2007) Atlas of Invertebrate-Pathogenic Fungi of Thailand Volume 1. National Center for Genetic Engineering and Biotechnology, National Science and Technology Development Agency, Pathum Thani, pp 64–65Google Scholar
  21. Luangsa-ard JJ, Tasanathai K, Mongkolsamrit S, Hywel-Jones NL (2008) Atlas of Invertebrate-Pathogenic Fungi of Thailand Volume 2. National Center for Genetic Engineering and Biotechnology, National Science and Technology Development Agency, Pathum Thani, pp 70–71Google Scholar
  22. Luangsa-ard JJ, Ridkaew R, Tasanathai K, Thanakitpipattana D, Hywel-Jones NL (2011) Ophiocordyceps halabalaensis: a new species of Ophiocordyceps pathogenic to Camponotus gigas in Hala Bala Wildlife Sanctuary, Southern Thailand. Fungal Biol 115:608–614. CrossRefPubMedGoogle Scholar
  23. Luangsa-ard JJ, Mongkolsamrit S, Thanakitpipattana D, Khonsanit A, Tasanathai K, Noisripoom W, Humber RA (2017) Clavicipitaceous entomopathogens: new species in Metarhizium and a new genus Nigelia. Mycol Progress 16:369–391CrossRefGoogle Scholar
  24. Luangsa-ard JJ, Tasanathai K, Thanakitpipattana D, Khonsanit A, Stadler M (2018) Novel and interesting Ophiocordyceps spp. (Ophiocordycipitaceae , Hypocreales) with superficial perithecia from Thailand. Stud Mycol 89:125−142. CrossRefPubMedPubMedCentralGoogle Scholar
  25. Mains EB (1940) Species of Cordyceps. Mycologia 32:310–320CrossRefGoogle Scholar
  26. Mongkolsamrit S, Kobmoo N, Tasanathai K, Khonsanit A, Noisripoom W, Srikitrikulchai P, Somnuk R, Luangsa-ard JJ (2009a) Life cycle, host range and temporal variation of Ophiocordyceps unilateralis/Hirsutella formicarum on Formicine ants. J Invertebr Pathol 111:217–224. CrossRefGoogle Scholar
  27. Mongkolsamrit S, Luangsa-ard JJ, Spatafora JW, Sung G-H, Hywel-Jones NL (2009b) A combined ITS rDNA and b-tubulin phylogeny of Thai species of Hypocrella with non-fragmenting ascospores. Mycol Res 113:684–699. CrossRefPubMedGoogle Scholar
  28. Moureau J (1961) Nouveaux Cordyceps du Congo. Lejeunia Mémoires 15:1–38Google Scholar
  29. Quandt CA, Kepler RM, Gams W, Araújo JPM, Ban S, Evans HC, Hughes D, Humber R, Hywel-Jones NL, Li Z, Luangsa-ard JJ, Rehner S, Sanjuan T, Sato H, Shrestha B, Sung GH, Yao Y, Zare R, Spatafora JW (2014) Phylogenetic-based nomenclatural proposals for Ophiocordycipitaceae (Hypocreales) with new combinations in Tolypocladium. IMA Fungus 5:121–134. CrossRefPubMedPubMedCentralGoogle Scholar
  30. Rabenhorst L (1846) Klotzschii Herbarium vivum mycologicum sistens Fungorum per totam Germanium crescentium collectionem perfectam. Centuria unidecima Dresdae. Bot Zeitung (Berlin) 4:876–880Google Scholar
  31. Saccardo PA (1883) Sylloge pyrenomycetum. Vol II. Sylloge Fungorum 2:1-813Google Scholar
  32. Sanjuán T, Franco-Molano AE, Kepler RM, Spatafora JW, Tabima J, Vasco-Palacios AM, Restrepo S (2015) Five new species of entomopathogenic fungi from the Amazon and evolution of neotropical Ophiocordyceps. Fungal Biol 119:901−916. CrossRefPubMedGoogle Scholar
  33. Schoch CL, Seifert KA, Huhndorf S, Robert V, Spouge JL, Levesque CA, Wen Chen W, Fungal Barcoding Consortium (2012) Nuclear ribosomal internal transcribed spacer (ITS) region as a universal DNA barcode marker for Fungi. PNAS 109(16):6241–6246. CrossRefPubMedGoogle Scholar
  34. Shrestha B, Tanaka E, Hyun MW, Han JG, Kim CS, Jo JW, Han SK, Oh J, Sung JM, Sung GH (2017) Mycosphere essay 19. Cordyceps species parasitizing hymenopteran and hemipteran insects. Mycosphere 8(9):1424−1442. CrossRefGoogle Scholar
  35. Spatafora JW, Sung GH, Sung JM, Hywel-Jones NL, White JF Jr (2007) Phylogenetic evidence for an animal pathogen origin of ergot and the grass endophytes. Mol Ecol 16(8):1701–1711. CrossRefPubMedGoogle Scholar
  36. Spatafora JW, Quandt CA, Kepler RM, Sung GH, Shrestha B, Hywel-Jones NL, Luangsa-Ard JJ (2015) New 1F1N species combinations in Ophiocordycipitaceae (Hypocreales). IMA Fungus 6(2):357–362. CrossRefPubMedPubMedCentralGoogle Scholar
  37. Stamatakis A (2014) RAxML version 8: a tool for phylogenetic analysis and post-analysis of large phylogenies. Bioinformatics 30(9):1312–1313. CrossRefPubMedPubMedCentralGoogle Scholar
  38. Sung G-H, Hywel-Jones NL, Sung JM, Luangsa-ard JJ, Shrestha B, Spatafora JW (2007) Phylogenetic classification of Cordyceps and the clavicipitaceous fungi. Stud Mycol 57:5–59. CrossRefPubMedPubMedCentralGoogle Scholar
  39. Swofford DL, 2002. PAUP*: Phylogenetic analysis using parsimony (*and other methods) Version 4.0b10. Sunderland, MassachusettsGoogle Scholar
  40. Vincent R, Vu D, Amor ABH, Nvd W, Brouwer C, Jabas B, Szoke S, Dridi A, Triki M, Daoud S, Chouchen O, Vaas L, Ad C, Stalpers JA, Stalpers D, Verkley GJM, Groenewald M, FBd S, Stegehuis S, Li W, Wu L, Zhang R, Ma J, Zhou M, Gorjón SP, Eurwilaichitr L, Ingsriswang S, Hansen K, Schoch C, Robbertse B, Irinyi L, Meyer W, Cardinali G, Hawksworth DL, Taylor JW, Crous PW (2013) MycoBank gearing up for new horizons. IMA Fungus 4(2):371−379. CrossRefGoogle Scholar
  41. White TJ, Bruns T, Lee S, Taylor J (1990) Amplification and direct sequencing of fungal ribosomal RNA genes for phylogenics. In: Innis MA, Gelfand DH, Sninsky JJ, White TJ (eds) PCR Protocols: a guide to methods and applications. Academic Press, San Diego, pp 315–322Google Scholar
  42. Wongsa P, Tasanatai K, Watts P, Hywel-Jones NL (2005) Isolation and in vitro cultivation of the insect pathogenic fungus Cordyceps unilateralis. Mycol Res 109:936–940. CrossRefPubMedGoogle Scholar
  43. Xiao YP, Wen TC, Hongsanan S, Sun JZ, Hyde KV (2017) Introducing Ophiocordyceps thanathonensis, a new species of entomogenous fungi on ants, a reference specimen for O. pseudiloydii. Phytotaxa 328(2):115–126. CrossRefGoogle Scholar

Copyright information

© German Mycological Society and Springer-Verlag GmbH Germany, part of Springer Nature 2018

Authors and Affiliations

  1. 1.Department of Plant Pathology, Faculty of AgricultureKasetsart UniversityBangkokThailand
  2. 2.Microbe Interaction and Ecology Laboratory (BMIE), BIOTEC, National Science and Technology Development Agency (NSTDA)Khlong LuangThailand

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