Integrative morphological and molecular characterization of Inocybe Species from Yamanlar Mountain (İzmir, Türkiye), with four new national records

Purpose: This study aimed to investigate the diversity of Inocybe species occurring in the Pinus brutia Ten. Forest surrounding Karagöl Nature Park in Yamanlar Mountain (İzmir, western Türkiye) through an integrative taxonomic approach combining detailed morphological examinations and molecular analyses based on ITS rDNA sequences. Although more than 100 Inocybe species have been reported from Türkiye, molecularly supported taxonomic studies of this ectomycorrhizal genus remain limited, particularly in western Anatolia. The study further aimed to document newly recorded taxa and contribute to the knowledge of Turkish fungal biodiversity.Method: Macrofungal specimens were collected from Pinus brutia forest in the Karagöl Nature Park area and examined using detailed macro- and micromorphological characteristics, including basidiospores, basidia, cystidia, and pileipellis structures. Dried basidiomata were used for extracting the genomic DNA and amplifying the ITS rDNA region with primers ITS1F and ITS4. Sequences collected were compared to reference sequences in GenBank and subjected to Bayesian phylogenetic analysis with MrBayes. A combination of morphological and molecular data was analyzed for reliable species delimitation and identification.Findings: Seven Inocybe species were identified: Inocybe amygdaliolens Stangl & J. Veselský, I. geophylla (Bull.) P. Kumm., I. neorufula Cervini, Dovana & Bizio, I. nitidiuscula (Britzelm.) Lapl., I. pinophila Cervini, Bizio & Dovana, I. subnudipes Kühner, and I. tenuicystidiata Esteve-Rav. & Villarreal. The phylogenetic analyses using the Markov Chain Monte Carlo (MCMC) approach recovered all of the specimens examined in their respective species-level clades with moderate to strong support (posterior probability). The new records of Inocybe amigdaliolens, I. neurufula, I. pinophila and I. subnudipes for Türkiye were made by morphological and molecular evidence. The new records significantly extend the country's diversity and distribution of Inocybe.Conclusion: In the present study, the diversity of Inocybe was comprehensively assessed in an integrative manner for the first time in the Yamanlar Mountain – Karagöl region. The results show that both morphological and molecular methods are useful in establishing species limits in fungi, isolated from complex groups. Besides, the new national records identified, accentuate the value of Mediterranean Pinus brutia forest as reservoirs of previously unknown ectomycorrhizal fungal diversity. The study is taxonomically important, as it provides new data on the mycobiota of Turkey, and also adds new data for western Anatolia biogeographically for the future studies of fungal biodiversity.