Abstract
In this study, morphological and molecular analyses of three newly collected saddle-like fungi confirm their classification within the genus Helvella L. from the eastern Türkiye. A three-locus phylogeny was constructed using the internal transcribed spacer (ITS) region, the translation elongation factor 1-alpha (TEF1-α), and the large subunit (LSU) of the ribosomal RNA gene. Phylogenetic analyses were performed using Maximum Likelihood (ML) and Bayesian Inference (BI) approaches. The phylogeny inferred from the concatenated dataset revealed three distinct clades. The Turkish Helvella species were distributed across the Elastica, Lacunosa and Acetabulum clades, identified as H. rivularis, H. sublactea, H. arcto-alpina, respectively. Both single-locus and multi-locus data provided informative results. In addition to phylogenetic analysis, detailed macro- and micro- features, along with illustrations, were provided for each studied species.
References
- Abbott, S.P. & Currah, R.S. (1997) The Hevellaceae: systematic revision and occurrence in northern and northwestern North America. Mycotaxon 62: 1–125. https://doi.org/10.7939/R3TD9NG3F
- Aldemir Terman, Ş., Akçay, M.E. & Dizkırıcı, A. (2024) First Record of Helvella capucina in Türkiye: Morphological and Molecular Characterization. Mantar Dergisi 15 (Special issue): 66–72. https://doi.org/10.30708/mantar.1567001
- Carbone, M. (2011) Contributo alla conoscenza di Helvella rivularis, una specie critica e poco conosciuta della sez. Macropodes. Mycologia Montenegrina 13: 7–39.
- Dissing, H. & Sivertsen, S. (1980) Operculate Discomycetes from Rana (Norway). 3. Helvella rivularis sp. nov. Botanisk Tidsskrift 75 (2–3): 101–104.
- Doyle, J.J. & Doyle, J.L. (1990) Isolation of plant DNA from fresh tissue. Focus 12: 13–15. https://doi.org/10.2307/2419362
- Ekanayaka, A.H., Hyde, K.D., Jones, E.B.G. & Zhao, Q. (2018) Taxonomy and phylogeny of operculate discomycetes: Pezizomycetes. Fungal Diversity 90 (1): 161–243. https://doi.org/10.1007/s13225-018-0402-z
- Guindon, S., Dufayard, J.F., Lefort, V., Anisimova, M., Hordijk, W. & Gascuel, O. (2010) New Algorithms and Methods to Estimate Maximum-Likelihood Phylogenies: Assessing the Performance of PhyML 3.0. Systematic Biology 59 (3): 307–21. https://doi.org/10.1093/sysbio/syq010
- Hansen, K. & Pfister, D.H. (2006) Systematics of the Pezizomycetes the operculate discomycetes. Mycologia 98: 1029–1040. https://doi.org/10.3852/mycologia.98.6.1029
- Hansen, K., Schumacher, T., Skrede, I., Huhtinen, S. & Wang, X.H. (2019) Pindara revisited–evolution and generic limits in Helvellaceae. Persoonia 42: 186–204. https://doi.org/10.3767/persoonia.2019.42.07
- Harmaja, H. (1977) A note on Helvella solitaria (syn. H. queletii) and H. confusen. sp. Karstenia 17: 40–44. https://doi.org/10.3767/persoonia.2019.42.07
- Kaygusuz, O., Türkekul, İ. & Battistin, E. (2020) Some noteworthy records of Helvella from Turkey based on morphology and DNA sequence data. Current Research in Environmental & Applied Mycology (Journal of Fungal Biology) 10 (1): 290–299. http://dx.doi.org/10.5943/cream/10/1/27
- Khalid, M., Tan, H., Ali, M., Rehman, A., Liu, X., Su, L., Saeed Ur, R., Zhao, C., Li, X. & Hui, N. (2022) Karst rocky desertification diverged the soil residing and the active ectomycorrhizal fungal communities thereby fostering distinctive extramatrical mycelia. Sci Total Environ 807: 151016. https://doi.org/10.1016/j.scitotenv.2021.151016
- Kumar, S., Tamura, K., Jajobsen, I.B. & Nei, M. (2001) MEGA: molecular evolutionary genetic analysis software. Bioinformatics 17: 1244–1245. https://doi.org/10.1093/bioinformatics/17.12.1244
- Laessoe, T. & Hansen, K. (2007) Truffle trouble: what happened to the Tuberales? Mycological Research 111: 1075–1099. https://doi.org/10.1016/j.mycres.2007.08.004
- Landeros, F., Iturriaga, T., Rodríguez, A., Vargas-Amado, G. & Guzmán-Dávalos, L. (2015) Advances in the phylogeny of Helvella (Fungi: Ascomycota), inferred from nuclear ribosomal LSU sequences and morphological data. Revista Mexicana de Biodiversidad 86: 856–871. https://doi.org/10.1016/j.rmb.2015.09.005
- Landeros, F., Ferrusca, M., Dávalos, L., Esquivel-Naranjo, E., Matias, N., Burrola-Aguilar, C., Viurcos-Martínez, G.A. & Garibay-Orijel, R. (2021) Helvella jocatoi sp. nov. (Pezizales, Ascomycota), a new species from H. lacunosa complex with cultural importance in central Mexico Abies religiosa forests. Phytotaxa 498 (1): 1–11. https://doi.org/10.11646/phytotaxa.498.1.1
- Lu, J.R., Yu, F.M., Lei, L., Zhang, Y. & Zhao, Q. (2023) A new species of Helvella (Helvellaceae, Pezizomycetes) within the H. macropus group from China. Phytotaxa 601 (3): 212–222. https://doi.org/10.11646/phytotaxa.601.3.2
- Løken, S.B., Skrede, I. & Schumacher, T. (2020) The Helvella corium species aggregate in Nordic countries phylogeny and species delimitation. Fungal Syst Evolut 5: 169–186. https://doi.org/10.3114/fuse.2020.05.11
- Mao, N., Xu, Y.Y., Zhang, Y.X., Zhou, H., Huang, X.B., Hou, C.L. & Fan, L. (2023) Phylogeny and species diversity of the genus Helvella with emphasis on eighteen new species from China. Fungal Systematics and Evolution 12: 111–152. https://doi.org/10.3114/fuse.2023.12.08
- Nouripour-Sisakht, S., Ahmadi, B., Makimura, K., Hoog, S., Umeda, Y., Alshahni, M.M. & Mirhendi, H. (2017) Characterization of the translation elongation factor 1-α gene in a wide range of pathogenic Aspergillus species. Journal of Medical Microbiology 66: 419–429. https://doi.org/10.1099/jmm.0.000450
- Park, S.Y., Kim, S.H., Kim, E.J., Eo, J.K. & Lee, H.Y. (2025) Taxonomic Revision of Korean Saddle Fungi (Helvella, Helvellaceae). Mycobiology 53 (1): 79–112. https://doi.org/10.1080/12298093.2024.2440172
- Rambaut, A. (2018) FigTree–Tree Figure Drawing Tool Version v. 1.4. 4. Institute of Evolutionary Biology, University of Edinburgh.
- Ranjard, L., Poly, F., Lata, J.-C., Mougel, C., Thioulouse, J. & Nazaret, S. (2001) Characterization of bacterial and fungal soil communities by automated ribosomal intergenic spacer analysis fingerprints: biological and methodo-logical variability. Appl. Environ. Microbiol. 67: 4479–4487. https://doi.org/10.1128/AEM.67.10.4479-4487.2001
- Rehner, S.A. & Buckley, E. (2005) A Beauveria phylogeny inferred from nuclear ITS and EF1-α sequences: evidence for cryptic diversification and links to Cordyceps teleomorphs. Mycologia 97: 84–89. https://doi.org/10.1080/15572536.2006.11832842
- Ronquist, F., Teslenko, M., van der Mark, P., Ayres, D.L., Darling, A., Höhna, S., Larget, B., Liu, L., Suchard, M.A. & Huelsenbeck, J.P. (2012) MrBayes 3.2: efficient Bayesian phylogenetic inference and model choice across a large model space. Systematic Biology 61: 539–542. https://doi.org/10.1093/sysbio/sys029
- Skrede, I., Carlsen, T. & Schumacher, T. (2017) A synopsis of the saddle fungi (Helvella: Ascomycota) in Europe–species delimitation, taxonomy and typification. Persoonia 39: 201–253. https://doi.org/10.3767/persoonia.2017.39.09
- Skrede, I., Gonzalvo, L.B., Mathiesen, C. & Schumacher, T. (2020) The genera Helvella and Dissingia (Ascomycota: Pezizomycetes) in Europe–Notes on species from Spain. Fungal Syst Evolut 6: 65–93. https://doi.org/10.3114/fuse.2020.06.05
- Skrede, I., Løken, S.B., Mathiesen, C. & Schumacher, T. (2023) Additions to the knowledge of the genus Helvella in Europe. New records and de novo description of five species from the Nordic region. Fungal Systematics and Evolution 11: 71–84. https://doi.org/10.3114/fuse.2023.11.06
- Tedersoo, L., Hansen, K., Perry, B.A. & Kjoller, R. (2006) Molecular and morphological diversity of pezizalean ectomycorrhiza. New Phytologist 170: 581–596. https://doi.org/10.1111/j.1469-8137.2006.01678.x
- Terman, S.Ş., Dizkirici, A., Akçay, M.E. & Sadullahoğlu, C. (2024) Morphological and molecular identification of Dissingia confusa based on the first record of the species in Türkiye. Trakya University Journal of Natural Sciences 25 (1): 65–72. https://doi.org/10.23902/trkjnat.1379302
- Thompson, J.D., Higgins, D.G. & Gibson, T.J. (1994) CLUSTAL W: Improving the Sensitivity of Progressive Multiple Sequence Alignment through Sequence Weighting, Position-Specific Gap Penalties and Weight Matrix Choice. Nucleic Acids Research 22: 4673–4680. https://doi.org/10.1093/nar/22.22.4673
- Uzun, Y. (2023) The checklist of Turkish Pezizales species. Anatolian Journal of Botany 7 (1): 1–20. https://doi.org/10.30616/ajb.1230337
- Vilgalys, R. & Hester, M. (1990) Rapid genetic identification and mapping of enzymatically amplified ribosomal DNA from several Cryptococcus species. Journal of Bacteriology 172: 4239–4246. https://doi.org/10.1128/jb.172.8.4238-4246.1990
- Wang, M., Zhao, Y.C., Zhao, Q. & Zhou, D.Q. (2016) Helvella sublactea sp. nov. (Helvellaceae) from southwestern China. Phytotaxa 253 (2): 131–138. https://doi.org/10.11646/phytotaxa.253.2.2
- Wang, X.C., Liu, T.Z., Chen, S.L., Li, Y. & Zhuang, W.Y. (2019) A four-locus phylogeny of rib-stiped cupulate species of Helvella (Helvellaceae, Pezizales) with discovery of three new species. MycoKeys 60: 45–67. https://doi.org/10.3897/mycokeys.60.38186
- Wang, X.C., Zhuang, W.Y. & Zhao, R.L. (2023) Species Diversity of Helvella lacunosa Clade (Pezizales, Ascomycota) in China and Description of Sixteen New Species. Journal of Fungi (Basel) 9: 697. https://doi.org/10.3390/jof9070697
- White, T.J., Bruns, T., Lee, S. & Taylor, J.W. (1990) Amplification and direct sequencing of fungal ribosomal RNA genes for phylogenetics. In: Innis, M.A., Gelfand, D.H., Sninsky, J.J. & White, T.J. (Ed.) PCR Protocols: A Guide to Methods and Applications. Academic Press Inc, New York, pp. 315–322. https://doi.org/10.1016/B978-0-12-372180-8.50042-1
- Xu, R.J., Li, L. & Zhao, Q. (2022) Helvella cystidiata sp. nov. (Helvellaceae, Ascomycota) from Tibetan Plateau, China. Phytotaxa 560 (1): 82–92. https://doi.org/10.11646/phytotaxa.560.1.6
- Yu, F.M., Lei, L., Luangharn, T., Zhao, Q. & Zhu, Y.A. (2023) Four new additions to Helvella (Helvellaceae, Pezizales) from Northern Thailand. Front Microbiol 14: 1182025. https://doi.org/10.3389/fmicb.2023.1182025
- Zhuang, W.Y., Zheng, H.D. & Zeng, Z.Q. (2018) Species Catalogue of China. Volume 3 Fungi. Cup—Fungi. Science Press, Beijing, China, pp. 1–142.
- Zhao, Q., Sulayman, M., Zhu, X.T., Zhao, Y.C., Yang, Z.L. & Hyde, K.D. (2016) Species clarification of the culinary Bachu mushroom in western China. Mycologia 108 (4): 828–836. https://doi.org/10.3852/16-002