Abstract

Van Yüzüncü Yıl Üniversitesi Fungaryumu’nda (VANF) muhafaza edilen Dissingia confusa türü morfolojik ve moleküler karakterler ile Türkiye'nin doğusundan yeni bir kayıt olarak tanımlanmıştır. Şapka, himenyum, sap, spor, askus ve parafiz yapıları morfolojik veri olarak, büyük alt ünite (LSU) ve translasyon uzama faktörü 1-alfa (TEF1-α) bölgelerinin nükleotid dizileri moleküler veri olarak kullanılmıştır. Filogenetik analiz Maksimum parsimoni ve Bayesian çıkarım yöntemi ile gerçekleştirilmiştir. Analizlere, iki cins arasındaki filogeniyi göstermek için GenBank veritabanından indirilen 15 (TEF1-α için 7) Dissingia ve 27 (TEF1-α için 14) Helvella örnekleri de eklenmiştir. Morfolojik çalışmalar ve genetik kanıtlar, Dissingia cinsinin cins düzeyinde Helvella cinsinden filogenetik olarak ayrıldığını ve Dissingia confusa türünün Türkiye'de ilk kez tanımlandığını doğrulamıştır. Detaylı deskripsiyon, taze ve kuru örneklerin renkli fotoğrafları ile mikroskobik karakterlerin görüntüleri ve filogenetik ağaçlar verilmiştir.

Project Number

2022-FHD-10113

Morphological and molecular identification of Dissingia confusa based on the first record of the species in Türkiye

Abstract

In the present study, specimens kept in Fungarium of Van Yüzüncü Yıl University - Türkiye (VANF) were evaluated based on morphological and molecular characters, and identified as Dissingia confusa as a new record from Eastern Türkiye. Ascocarp, hymenium, stipe, asco-spores, ascus and paraphyses were considered as diagnostic morphological characters and the nucleotide sequences of the large subunit (LSU) ribosomal DNA and the translation elonga-tion factor 1-alpha (TEF1-α) were referred as molecular data. A phylogenetic analysis was performed with Maximum parsimony and Bayesian inference methods. The DNA sequences of 15 (7 for TEF1-α) Dissingia and 27 (14 for TEF1-α) Helvella specimens downloaded from the GenBank database were included in the analysis to estimate phylogeny between the two close genera. Morphological evaluations and genetic evidences confirmed that Dissingia is phylogenetically separated from Helvella at genus level and helped to identify the studied specimens as D. confusa as a new record in Türkiye. Detailed description, colour images of fresh and dried ascomata, along with photographs of microscopic characters and the obtained phylogenetic trees are given.

Keywords

• Dissingia
• Helvella
• molecular taxonomy
• new record
• phylogeny

Supporting Institution

Van Yüzüncü Yıl University (Scientific Research Project Foundation)

Project Number

2022-FHD-10113

Ethical Statement

Since the article does not contain any studies with human or animal subject, its approval to the ethics committee was not required.

References

1. 1. Abbott, S.P & Currah, R.S. 1997. The Helvellaceae: systematic revision and occurrence in northern and northwestern North America. Mycotaxon, 62: 1-125. https://doi.org/10.7939/R3TD9NG3F
2. 2. Altschul, S.F., Gish, W., Miller, W., Myers, E.W. & Lipman, D.J. 1990. Basic local alignment search tool. Journal of Molecular Biology, 215(3): 403-410. https://doi.org/10.1016/S0022-2836(05)80360-2
3. 3. Bonito, G., Smith, M.E., Nowak, M, Healy, R.A., Guevara, G., Cazares, E., Kinoshita, A., Nouhra, E.R., Dominguez, L.S., Tedersoo, L., Murat, C., Wang, Y., Moreno, B.A., Pfister, D.H., Nara, K., Zambonelli, A., Trappe, J.M. & Vilgalys, R. 2013. Historical biogeography and diversification of truffles in the Tuberaceae and their newly identified southern hemisphere sister lineage. PLoS One, 8: e52765. https://doi.org/10.1371/journal.pone.0052765
4. 4. Carbone, M., Agnello, C. & Alvarado, P. 2021. Type study of Helvella pedunculata (Ascomycota, Pezizales), a synonym of Dissingia confusa. Ascomycete.org, 13(3): 125-132. https://doi.org/10.25664/ART-0327
5. 5. Doyle, J.J. & Doyle, J.L. 1987. A rapid DNA isolation procedure for small quantities of fresh leaf tissue. Phytochemical Bulletin, 19:11-15.
6. 6. Häffner, J. 1987. Die gattung Helvella, morphologie und taxonomie. Beihefte zur Zeitschrift für. Mykologie, 7: 1-165.
7. 7. Hansen, K., Perry, B.A, Dranginis, A.W. & Pfister, D.H. 2013. A phylogeny of the highly diverse cup-fungus family Pyronemataceae (Pezizomycetes, Ascomycota) clarifies relationships and evolution of selected life history traits. Molecular Phylogenetics and Evolution, 67: 311-335. https://doi.org/10.1016/j.ympev.2013.01.014
8. 8. 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

9. 9. 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
10. 10. Harmaja, H. 1977. A note on Helvella solitaria (syn. H. queletii) and H. confusa n. sp. Karstenia, 17: 40-44. https://doi.org/10.29203/ka.1977.123
11. 11. Harmaja, H. 1979. Studies on cupulate species of Helvella. Karstenia, 19: 33-45. https://doi.org/10.29203/ka.1979.184
12. 12. Katoh, K. & Standley, M.D. 2013. MAFFT multiple sequence alignment software version 7: improvements in performance and usability. Molecular Biology Evolution, 30(4):772-80. https://doi.org/10.1093/molbev/mst010
13. 13. Laessoe, T. & Hansen, K. 2007. Truffle trouble: what happened to the Tuberales. Mycological Reseach, 111: 1075-1099. https://doi.org/10.1016/j.mycres.2007.08.004
14. 14. Landeros F, Iturriaga T, Rodriguez A, Amado, G.V. & Davalos, L.G. 2015. Advances in the phylogeny of Helvella (Fungi: Ascomycota), inferred from nuclear ribosomal LSU sequences and morphological data. Revista Mexicana de Boversidad, 86: 856-871. https://doi.org/10.1016/j.rmb.2015.09.005
15. 15. Loken, S.B., Skrede, I. & Schumacher, T. 2019. The Helvella corium species aggregate in Nordic countries phylogeny and species delimitation. Fungal Systematics and Evolution, 5: 169-186. http://dx.doi.org/10.3114/fuse.2020.05.11
16. 16. Maddison, W.P. & Maddison, D.R. 2009. Mesquite: a modular system for evolutionary analysis, 2010 version 2.73. Available from: http://mesquiteproject.org/mesquite/mesquite.html (Date accessed: 01.10.2023).
17. 17. 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. http://dx.doi.org/10.3114/fuse.2023.12.08
18. 18. Mycobank online database. 2023. Dissingia confusa https://www.mycobank.org/Simple%20names%20search (Date accessed: 01.10.2023).
19. 19. Nguyen, N.H., Landeros, F., Garibay-Orijel, R., Hansen, K. & Vellinga, E.C. 2013. The Helvella lacunosa species complex in western North America: cryptic species, misapplied names and parasites. Mycologia, 105: 1275-1286. https://doi.org/10.3852/12-391
20. 20. Rambaut, A., 2010. FigTree v1.3.1. Institute of Evolutionary Biology, University of Edinburgh, Edinburgh.
21. 21. 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/10.3852/mycologia.97.1.84
22. 22. Ronquist, F. & Huelsenbeck, J.P. 2003. MRBAYES 3: Bayesian phylogenetic inference under mixed models. Bioinformatics, 19: 1572-1574. https://doi.org/10.1093/bioinformatics/btg180
23. 23. Schoch, C.L., Seifert, K.A., Huhndorf, S., Robert, V. & Spouge, J.L. 2012. Fungal Barcoding Consortium. Nuclear ribosomal internal transcribed spacer (ITS) region as a universal DNA barcode marker for Fungi. PNAS, 109: 6241-6246. https://doi.org/10.1073/pnas.1117018109
24. 24. Skrede, I., Carlsen, T. & Schumacher T. 2017. A synopsis of the saddle fungi (Helvella: Ascomycota) in Europe species delimitations, taxonomy and typification. Persoonia, 39: 201-253. http://dx.doi.org/10.3767/persoonia.2017.39.09
25. 25. 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 Systematics and Evolution, 6: 65-93. https://doi.org/10.3114/fuse.2020.06.05
26. 26. 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
27. 27. Swofford, D.L. 2003. PAUP: Phylogenetic Analysis Using Parsimony. Version 4.0b10. Sinauer Associates, Sunderland, MA, USA.
28. 28. 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
29. 29. Uzun, Y. 2023. The checklist of Turkish Pezizales species. Anatolian Journal of Botany, 7(1): 1-20. https://doi.org/10.30616/ajb.1230337
30. 30. 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
31. 31. Wang, Z., Binder, M. & Hibbett, D.S. 2005. Life history and systematics of the aquatic discomycete Mitrula (Helotiales, Ascomycota) based on cultural, morphological, and molecular studies. American Journal of Botany, 92(9): 1565-1574. https://doi.org/10.3732/ajb.92.9.1565
32. 32. Wang, M., Zhao, Y.C., Zhao, Q. & Zhou, D.Q. 2016. Helvella sublactea sp. Nov. (Helvellaceae) from southwestern China. Phytotaxa, 253(2): 131-138. http://dx.doi.org/10.11646/phytotaxa.253.2.2
33. 33. 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
34. 34. 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, 9: 697. https://doi.org/10.3390/jof9070697
35. 35. Weber, N.S. 1972. The genus Helvella in Michigan. The Michigan Botanist, 11: 147-201.
36. 36. Weber, N.S. 1975. Notes on Western species of Helvella. I. Beihefte. Nova Hedwigia, 51: 25-38.
37. 37. 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. Mycologia, 108(4): 828-836. https://doi.org/10.3852/16-002
38. 38. Zhao, Q., Brooks, S., Zhao, Y.C., Yang, Z.L., Hyde, K.D. & Tolgor, B. 2016. Morphology and phylogenic position of Wynnella subalpina sp. nov. (Helvellaceae) from western China. Phytotaxa, 270(1): 41-48. https://doi.org/10.11646/phytotaxa.270.1.4