Morphological and Molecular Characterization of Hebeloma Subtortum (Hymenogastraceae), a New Record Macrofungus from Bingöl Province, Turkey
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Kastamonu Uni., Orman Fakültesi Dergisi, 2019, 19(1): 1-10 Research Article Kastamonu Univ., Journal of Forestry Faculty Doi:10.17475/kastorman.543375 Morphological and Molecular Characterization of Hebeloma subtortum (Hymenogastraceae), a New Record Macrofungus from Bingöl Province, Turkey Ayten DİZKIRICI1* , İsmail ACAR2 , Ayşenur KALMER1 , Yusuf UZUN2 1Department of Molecular Biology and Genetics, Van Yüzüncü Yıl University, 65080, Van, Turkey 2Department of Organic Agriculture, Başkale Vocational High School, Van Yüzüncü Yıl University, 65080, Van, Turkey 3Department of Pharmaceutical Sciences, Faculty of Pharmacy, Van Yüzüncü Yıl University, 65080, Van, Turkey *Corresponding author: [email protected] Received Date: 05.07.2018 Accepted Date: 08.10.2018 Abstract Aim of Study: The specimen identified as Hebeloma subtortum based on both morphological and molecular characterizations. Area of Study: Samples were collected from Bingöl province and the study was conducted at the Department of Molecular Biology and Genetics in Van Yüzüncü Yıl University. Material and Methods: Characters of pileus, stipe, lamellae and basidia, cystidia, spores were used as macroscopic and microscopic features, respectively. DNA sequences of two loci including the internal transcribed spacer (ITS) region and the large subunit (LSU) of nuclear ribosomal RNA gene were used to show the evolutionary relationship and taxonomic position of the species within Hebeloma genus. The DNA sequences of the above-mentioned regions of H. subtortum were compared to those of the same and different species of the genus downloaded from NCBI. Main results: In phylogenetic analyses, H. subtortum distinctly clustered with its representatives retrieved from NCBI with high bootstrap value. The ITS tree was more informative compared to LSU. Hebeloma subtortum closely grouped with H. mesophaeum in the ITS tree. Research highlights: Hebeloma subtortum has been described and illustrated as a new record from Turkey. Keywords: Hebeloma, ITS, mycogenetics, phylogeny Bingöl ilinden yeni bir kayıt olarak Hebeloma subtortum (Hymenogastraceae) türünün morfolojik ve moleküler karakterizasyonu Öz Çalışmanın Amacı: Hebeloma subtortum türü hem morfolojik hem de moleküler karakterizasyonlara dayanarak tanımlanmıştır. Çalışma Alanı: Örnekler Bingöl ilinden toplanmış ve çalışma Van Yüzüncü Yıl Üniversitesi Moleküler Biyoloji ve Genetik Bölümü'nde yapılmıştır. Gereç ve Yöntemler: Şapka, sap, lamel yapıları ve bazidya, sistidya, spor yapıları sırasıyla makroskopik ve mikroskopik özellikler olarak kullanılmıştır. Nükleer ribozomal iç aralayıcı bölge (nrITS) ve ribozomal en büyük alt birim (LSU) olmak üzere iki DNA bölgesi çalışılan türün evrimsel ilişkisini ve Hebeloma cinsi içindeki taksonomik konumunu göstermek için kullanılmıştır. Çalışılan türün DNA dizileri, NCBI very tabanından indirilen cinse ait aynı ve farklı türlerle karşılaştırılmıştır. Ana sonuçlar: Filogenetik analizlerde, H. subtortum, yüksek bootstrap değeri ile NCBI'den alınan temsilcileri ile belirgin bir şekilde kümelenmiştir. ITS ağacı, LSU'ya kıyasla daha bilgilendirici olmuştur. H. subtortum, ITS ağacında H. mesophaeum ile yakın gruplanmıştır. Araştırma konuları: Hebeloma subtortum, Türkiye'den yeni bir kayıt olarak tanımlanmış ve gösterilmiştir. Anahtar kelimeler: Hebeloma, ITS, mikogenetik, filogeni Dizkırıcı, A., Acar, İ., Kalmer, A. & Uzun, Y. This work is licensed under a Creative Commons Citation (Atıf): (2019). Morphological and molecular characterization of Hebeloma Attribution-NonCommercial 4.0 International License. subtortum (Hymenogastraceae), a new record macrofungus from 1 Bingöl Province, Turkey. Kastamonu University Journal of Forestry Faculty, 19 (1), 1-10. Kastamonu Uni., Orman Fakültesi Dergisi, 2019, 19(1): 1-10 Dizkırıcı et. al. Kastamonu Univ., Journal of Forestry Faculty Introduction Vesterh., H. alpinum (J. Favre) Bruchet, H. avellaneum Kauffman, H. birrus (Fr.) Gillet, Hebeloma (Fr.) P. Kumm. H. cavipes Huijsman (H. album Peck, H. (Hymenogastraceae Vittad., Agaricales vejlense Vesterh.), H. celatum Grilli, H. Underw.) is a genus of ectomycorrhizal fungi circinans (Quél.) Sacc., H. cistophilum Maire, comprising approximately 530 worldwide H. clavulipes Romagn. (H. candidipes published species names with its main Bruchet), H. crustuliniforme (Bull.) Quél., H. distribution in the temperate zones of the dunense L. Corb. & R. Heim (H. collariatum northern hemisphere (Mycobank.org). In the Bruchet), H. eburneum Malençon, H. field, Hebeloma is easily separated from fragilipes Romagn., H. laterinum (Batsch) related genera based on several special Vesterh., H. leucosarx P.D. Orton, H. characters such as the chocolate colored mesophaeum, H. nauseosum Sacc. (H. fusipes lamellae, the occasional presence of a Bres., H. gigaspermum Gröger & Zschiesch.), fibrillose veil, often scanty, that when broken H. populinum Romagn., H. porphyrosporum usually leaves remnants on the margin of the Maire (often reported as H. sarcophyllum pileus and/or on the stipe and the presence of (Peck) Sacc. which was incorrectly distinct sterile, often narrow (sometimes synonymised and is a north American species fusoid-ventricose) or greatly elongated cells that as far as we are aware has never been among those along the edges of the gills (the confirmed in Europe), H. pusillum J.E. Lange, cheilocystidia) (Smith, Evenson & Mitchel,. H. radicosum (Bull.) Ricken, H. 1983). Beker, Eberhardt & Vesterholt (2016) sacchariolens Quél. (H. pallidoluctuosum divided the genus into thirteen sections Gröger & Zschiesch.), H. sinapizans (Paulet) (Denudata, Hebeloma, Sinapizantia, Gillet, H. sordescens Vesterh., H. subtortum Sacchariolentia, Velutipes, Theobrominum, (H. mesophaeum var. lacteum Vesterh., H. Naviculospora, Scabrispora, Myxocybe, sordidum Maire), H. syrjense P. Karst., H. Pseudoamarescens, Duracinus, theobrominum Quadr., H. velutipes Bruchet Porphyrospora, Syrjense) using different (H. stenocystis J. Favre ex Quadr.). Apart characters such as habitat, smell, number of from the collections cited by Beker et al. full-length lamellae, presence of cortina, (2016), Hebeloma subtortum has been shape of cheilocystidia and dextrinoid previously reported under the two names H. reaction of the spore. mesophaeum var. lacteum (Yılmaz Ersel, In Turkey, 28 species of Hebeloma have 2005) and H. sordidum (Doğan and Kurt, been reported so far (Sesli and Denchev, 2016). The latter description is unlikely to 2014; Güngör, Solak, & Kalmış, 2015; Sesli, represent this taxon; while the description fits Contu, Vila, Moreau & Battistin, 2015; Solak, reasonably well, the figure (Fig. 6 in Doğan Işıloğlu, Erbil & Allı, 2015; Beker et al. 2016; and Kurt, 2016) showing spores and Doğan and Kurt, 2016; Sesli, Örtücü & Aytaç, cheilocystidia, appears to be more likely to 2018). In Beker et al. (2016), seven Hebeloma represent a species from Hebeloma sect. records were cited for Turkey; these citations Scabrispora, perhaps H. pumilum. The were based on morphological and molecular former description and figure (Fig. 1 in characters, but no morphological details were Yılmaz Ersel, 2005) are representative of this provided for these particular collections. taxon. The present study contributes to the They were: H. celatum Grilli (1), H. documentation of a new record of Hebeloma cistophilum Maire (1), H. dunense L. Corb. & subtortum, supported by a full description and R. Heim (2), H. laterinum (Batsch) Vesterh. phylogenetic results. The species is found in (1), H. mesophaeum (Pers.) Quél. (1), H. section Hebeloma which is one of the largest subtortum P. Karst. (1). We have reviewed the sections of the genus. This section is content of all these papers but not examined differentiated from others by the presence of a all cited collections. If we remove synonyms partial veil and cheilocystidia which are (in brackets) and use the names accepted always lageniform or ventricose. Deciding the within Beker et al. (2016), then the following section where the sample is classified, species have been reported for Turkey (names depends on a few characters such as smell, in bold are names as reported): H. aestivale number of full length lamellae, habitat, the 2 Kastamonu Uni., Orman Fakültesi Dergisi, 2019, 19(1): 1-10 Dizkırıcı et. al. Kastamonu Univ., Journal of Forestry Faculty presence of remnants of veil, the shape and confirmed record for Turkey based on dextrinoidity of the spores and the shape of the microscopic, macroscopic and molecular cheilocystidia (Beker et al. 2016). Since a characters. raphanoid smell is common to several sections of the genus, this character is insufficient to Material and methods determine the section to which a collection may belong. Therefore, a few characters Sampling, macroscopic and should be used together to properly decide the microscopic studies section where the sample is found. For During field work in 2017, samples were instance, if a raphanoid smell is accompanied collected from Bingöl province of Turkey and by the presence of a cortina and ventricose identified based on microscopic, macroscopic cheilocystidia, it is true to say that the and molecular characters. Collected samples specimen belongs to the section Hebeloma were associated with conifer in the field. (Beker et al. 2016). Basidiocarps were photographed, dug out Though identification of the genus in the from the soil with a sharp knife, and carried to field is not difficult, determination at the the laboratory. Collected materials were species level can be confusing