Host Lichen Research Articles

Continental scale comparison of mycobiomes in Parmelia and Peltigera lichens from Turkey and South Korea

BackgroundLichens, traditionally considered as a simple partnership primarily between mycobiont and photobiont, are, in reality, complex holobionts comprised of a multitude of microorganisms. Lichen mycobiome represents fungal community residing within lichen thalli. While it is acknowledged that factors like the host lichen species and environmental conditions influence the structure of the lichen mycobiome, the existing research remains insufficient. To investigate which factor, host genus or location, has a greater impact on the lichen mycobiome, we conducted a comparative analysis of mycobiomes within Parmelia and Peltigera collected from both Turkey and South Korea, using high-throughput sequencing based on internal transcribed spacer region amplification.ResultsOverall, the lichen mycobiome was dominated by Capnodiales (Dothideomycetes), regardless of host or location. At the order level, the taxonomic composition was not significantly different according to lichen genus host or geographical distance. Hierarchical clustering of the top 100 abundant ASVs did not clearly indicate whether the lichen mycobiome was more influenced by host genus or location. Analyses of community similarity and partitioning variables revealed that the structure of the lichen mycobiome is more significantly influenced by location than by host genus. When analyzing the core mycobiome by host genus, the Peltigera mycobiome contained more ASV members than the Parmelia mycobiome. These two core mycobiomes also share common fungal strains, including basidiomycete yeast. Additionally, we used chi-squared tests to identify host genus-specialists and location-specialists.ConclusionsBy comparing lichen mycobiomes of the same genera across different countries, our study advances our comprehension of these microbial communities. Our study elucidates that, although host species play a contributory role, geographic distance exerts a more pronounced impact on the structure of lichen mycobiome. We have made foundational contributions to understanding the lichen mycobiome occupying ecologically crucial niches. We anticipate that broader global-scale investigations into the fungal community structures will provide more detailed insights into fungal residents within lichens.

Exploring the influence of climatic variables on mycobiome composition and community diversity in lichens: insights from structural equation modeling analysis

BackgroundLichens are symbiotic organisms composed of a fungus and a photosynthetic partner, which are key ecological bioindicators due to their sensitivity to environmental changes. The endolichenic fungi (ELF) living inside lichen thalli, are an important but understudied component of playing crucial ecological roles such as nutrient cycling and protection against environmental stressors. Therefore ELF community investigation is vital for fostering sustainable ecosystems and leveraging their ecological benefits. Deciphering the intricate relationships between ELF and their lichen hosts, alongside the influence of environmental factors on these communities, presents a significant challenge in pinpointing the underlying drivers of community structure and diversity.ResultsOur research demonstrated that locational factors were the main drivers of the ELF community structure, rather than host haplotype. Several climatic factors affected the diversity of the ELF community and contributed to the prevalence of different types of fungal residents within the ELF community. A decrease in isothermality was associated with a greater prevalence of pathotrophic and saprotrophic fungi within the ELF community, resulting in an overall increase in community diversity. By conducting a structural equation modeling analysis, we identified a robust link between climatic variables, fungal trophic mode abundance, and the species diversity of the ELF community.ConclusionThis study’s discoveries emphasize the significance of examining climate-related factors when investigating ELF’s structure and function. The connection between fungi and climate is intricate and complex, and can be influenced by various other factors. Investigating the potential for ELF to adapt to changing climatic conditions, as well as the potential effects of changes in ELF communities on lichen function, would be valuable research areas. We anticipate that our research results will establish a basis for numerous future ELF research projects and have a significant impact on the field.

Cylindromonium dirinariae sp. nov. (Ascomycota, Hypocreales), a new nectrioid lichenicolous species on Dirinaria applanata in Japan.

A nectrioid fungus forming a pinkish colony with mainly solitary phialides producing ellipsoid, aseptate conidia in mucoid packets was isolated from Dirinaria applanata. Our taxonomic study based on morphology and phylogenetic analysis using ITS rDNA sequences revealed that the isolates represented a member of the genus Cylindromonium. Based on further morphological examination, nucleotide sequence comparison, and phylogenetic analysis based on LSU rDNA, tef1, and rpb2 in addition to the phylogenetic analysis using the ITS rDNA sequences, the fungus from Dirinaria represents a new species, which is described here as Cylindromonium dirinariae sp. nov. Furthermore, inoculation experiments revealed that this species can also produce perithecia when inoculated on the host lichen in laboratory environments. Citation: Ohmaki A, Okane I, Crous PW, Verkley GJM (2023). Cylindromonium dirinariae sp. nov. (Ascomycota, Hypocreales), a new nectrioid lichenicolous species on Dirinaria applanata in Japan. Fungal Systematics and Evolution 11: 1-10. doi: 10.3114/fuse.2023.11.01.

Lichen and Lichenicolous Fungal Communities Tested as Suitable Systems for the Application of Cross-Taxon Analysis

Lichens are outstanding examples of fungal symbioses that form long-lived structures, the lichen thalli, in which a multiplicity of other microorganisms are hosted. Among these, microfungi seem to establish diverse trophic relationships with their lichen hosts. The most specialised of these fungi are the parasitic lichenicolous fungi, of which the diversity has hardly been explained as a proxy for the diversity of lichen species. Here, we used an exemplar dataset of a well-studied alpine lichen community composed of 63 lichen and 41 lichenicolous fungal species and tested it to verify the strength of the co-occurrences of the two species groups with predictive co-correspondence analyses. The results showed that the distribution of lichen abundances affects the abundance and variation of lichenicolous fungi and supports our hypothesis to use lichens as surrogates for lichenicolous fungi in surrogacy analysis.

An annotated catalogue of the lichenicolous fungi of Jammu and Kashmir and Ladakh, India with new records and identification key

This article enumerates 72 species of lichenicolous fungi belonging to 21 families and 43 genera from Jammu and Kashmir and Ladakh. The study is based on published literature and specimens recently collected from some localities of Kishtwar High Altitude National Park, Jammu and Kashmir. Four species viz., Bachmanniomyces santessonii Etayo , Intralichen lichenum (Diederich) D. Hawksw., and M.S. Cole, Milospium graphideorum (Nyl.) D. Hawksw., and Roselliniella oxyspora Matzer and Hafellner are reported as new to India, while Milospium and Roselliniella are new generic records. Brief descriptions of the newly recorded species are presented. The geographical distribution, host lichens, and key to all the species of lichenicolous fungi presently known from Jammu and Kashmir and Ladakh are also provided.

An acetonic extract and secondary metabolites from the endolichenic fungus Nemania sp. EL006872 exhibit immune checkpoint inhibitory activity in lung cancer cell.

Background: Endolichenic fungi (ELF), which live the inside the lichen thallus, contain many secondary metabolites that show various biological activities. Recent studies show that lichen and ELF secondary metabolites have antioxidant, antibacterial, antifungal, cytotoxic, and anticancer activities. Purpose: Here, the effects of an ELF extract and its bioactive compounds were investigated on the H1975 cell line focusing on immune checkpoint marker inhibition. Methods: An ELF was isolated from the host lichen Bryoria fuscescens (Gyelnik) Brodo and D. Hawksw and identified the species as Nemania sp. EL006872. The fungus was cultured on agar medium and acetonic extracts were obtained. Secondary metabolites radianspenes C and D, and dahliane D, were isolated from the crude extract. The biological effects of both the crude extract and the isolated secondary metabolites were evaluated in cell viability, qRT-PCR assays, flow cytometry analysis and western blotting. Results: The cell viability assay revealed that extracts from Nemania sp. EL006872 and the isolated secondary compounds had low cytotoxicity. The crude extract, radianspenes C and D, and dahliane D, suppressed expression of mRNA encoding PD-L1 and aromatic hydrocarbon receptor (AhR), and surface expression of PD-L1 protein by cells exposed to benzo[a] pyrene. Radianspenes C and D, and dahliane D, reduced expression of AhR, PD-L1, ICOSL, and GITRL proteins by H1975 lung cancer cells, as well as exerting anti-proliferative effects. Conclusion: Radianspenes C and D, and dahliane D, bioactive compounds isolated from Nemania sp. EL006872 ELF, have the potential for use as immunotherapy and immunoncology treatments.

Bioprospecting of an Endolichenic Fungus Phanerochaete sordida Isolated from Mangrove-Associated Lichen Bactrospora myriadea

Bioassay-guided fractionation of the ethyl acetate extract of Phanerochaete sordida, an endolichenic fungus (ELF) isolated from the host lichen Bactrospora myriadea, collected from Negombo lagoon, Sri Lanka, led to the isolation of a bioactive compound. Following the identification of the fungus using morphological and DNA barcoding techniques, the pure compound was isolated using column chromatography, preparative TLC, and semipreparative HPLC. The structure elucidation was carried out using IR, HR-ESI-MS and 1H, 13C & 2D NMR spectroscopic methods. The in vitro bioassays conducted revealed that compound 1 has a high antioxidant activity with ABTS•+ (IC50 58.91 ± 1.35 μ M ), moderate anti-inflammatory activity (IC50 254.79 ± 1.41 μ M ), comparable antibacterial activity against the oral-bacterial strain Streptococcus mutans (MIC 898.79 μM and MLC 1797.58 μM), moderate tyrosinase inhibition (IC50 1713.69 ± 8.65 μ M ), and moderate cytotoxicity against oral cancer (IC50 13.65 ± 0.02 μ M ), in comparison with respective positive controls. The in silico experiments conducted for tyrosinase inhibition and cytotoxicity using Schrödinger revealed results in line with the in vitro results, thus confirming the bioactivities. The molecule also satisfies the key features of drug likeliness according to pharmacokinetic studies.

Endolichenic fungi from common Usnea lichens found in a montane forest in Malaysia: a study on diversity and bioactivity profiling

This study evaluated the diversity of endolichenic fungi (ELF) isolated from three common lichen species (Usnea baileyi, U. bismolliuscula and U. pectinata) found in Bukit Larut, Peninsular Malaysia. Diversity indices and phylogenetic analysis were performed to identify the various species found in lichens, and their bioactivity profiles were established based on antimicrobial and antioxidant activities. Sixty-two (62) ELF isolates were identified, mostly belonging to the family Xylariaceae (88.7%, 55 isolates), followed by Hypoxylaceae (6.5%, four isolates), Nectriaceae (3.2%, two isolates) and Sporocadaceae (1.6%, one isolate). The common genera were Nemania (45.2%, 28 isolates) and Xylaria (27.4%, 17 isolates). In this study, ELF isolates have shown both host-generalist and host-specific behaviours towards their lichen hosts. The ELF also have relatively strong antimicrobial (minimum inhibitory concentration:0.0625-10.00mg/ml) and antioxidant (IC50:5.72-17.8mg/ml) properties. This study highlights that a diverse ELF collection rich in bioactive compounds can be found in lichens in the montane forest in Malaysia.

Antibacterial Polyketide from Lasiodiplodia theobromae and Pyrenula bahiana on Mangrove Ecosystems in Puttalam Lagoon, Sri Lanka

Lasiodiplodia theobromae is one of the frequently isolated fast growing endolichenic fungus. This fungus was isolated from the lichen host, Pyrenula bahiana collected from the mangrove ecosystems in Puttlam lagoon and its identification was confirmed based on rDNA-ITS sequence homology. Secondary metabolites of L. theobromae were extracted into ethyl acetate and subjected to antibacterial assay against Escherichia coli (ATCC25922), Staphylococcus aureus (ATCC25923) and Bacillus subtilis (ATCC6051). Crude extract at a concentration of 6.8 μg/mL showed good antibacterial activity against the bacterial strain S. aureus compared with the activity of the standard azithromycin at a concentration of 5.0 μg/mL. Active crude extract was partitioned to obtain methanol, hexane and chloroform fractions. Chloroform fraction showed the highest activity to S. aureus out of three fractions. This fraction was subjected to bioassay-guided separation on silica gel column chromatography to isolate bioactive pure compounds. The bioactive pure compound was identified as (3R)-de-O-methyllasiodiplodin using LC-MS, 1D and 2D NMR spectroscopy.

A New Lichenicolous Fungus from Garhwal Himalayan Region of Uttarakhand, India

Sphinctrina pertusa Falswal A. is described as a new species of lichenicolous fungus from India. It is characterised by eight non-septate, broadly ellipsoidal, uniseriate, ascospores with olive green tint and irregular ridge like ornamentations. This fungus is growing on known lichen host Pertusaria but differs from five closely related species i.e., S. anglica, S. leucopoda, S. tubaeformis, S. turbinata, S. paramerae with different taxonomic characters. A detailed comparative analysis with non-septate closely related species is provided.

Two new species and new records of lichen-feeding darkling beetles (Coleoptera: Tenebrionidae: Helopini) from Turkey with notes on bionomics and trophic relations.

Turkonalassus mavi M. Nabozhenko B. Keskin, sp. n. and Odocnemis rufocruralis M. Nabozhenko B. Keskin, sp. n., two distinctive darkling beetle species of the tribe Helopini, are described from the Southeastern Anatolia region of Turkey, based on both their external morphology and genital structures. Turkonalassus mavi sp. n. differs from all congeners by the bluish dorsal surface of the body, the pronotum widest before middle, and wide merged baculi of the median lobe of the aedeagus. Odocnemis rufocruralis sp. n. belongs to the praelonga species-group. Odocnemis rufocruralis sp. n. is similar to O. torosica Nabozhenko Keskin, 2016, O. terminassianae (Nabozhenko, 2011) and O. kakunini Nabozhenko Keskin, 2016 but differs from all three taxa by the reddish-brown body with red legs, male apical maxillary palpomere wider than in female, elevate and narrowly separate male protrochanters, and the structure of the very long and narrow apical piece of the aedeagus. New data on distribution, bionomics, and trophic relations of several species of Helopini from Turkey are given. Host lichens are determined for nine species. The majority of studied adult beetles feed on lichens from the families Physciaceae and Parmeliaceae. Some examined taxa feed on Cladoniaceae. Feeding on crustose lichens is registered for the first time for Coleoptera, specifically Odocnemis rufocruralis sp. n. was observed to feed on Pertusaria sp. (on Prunus) in Mardin Province.

Notes on lichenicolous taxa of the asexual fungal genera Intralichen and Trimmatostroma, with a revised key and descriptions of four new species

Diederich, P. 2021. Notes on lichenicolous taxa of the asexual fungal genera Intralichen and Trimmatostroma, with a revised key and descriptions of four new species. – Herzogia 34: 101–126.The genus Intralichen comprises several mainly intrahymenial lichenicolous species of unknown phylogenetic affinities, each reported from a very large host-spectrum, with many reports obviously representing wrong identifications. We suggest restricting the use of each of these species names to collections growing on the same host genus as the type specimen, i.e., all other reports from different hosts are considered uncertain. Intralichen christiansenii is considered as a synonym of I. lichenicola, a species confined to Candelariella species. Intralichen lichenum s. str. is a poorly known and rarely collected species from Arthoniales hosts, while similar populations on Strangospora are treated as I. aff. lichenum. Intralichen baccisporus has not been revised, as sufficient material from the many Teloschistaceae genera was not available. Morphologically similar species growing on the thalli, not in the hymenium, of various lichen hosts are included in the genus Trimmatostroma, although most lichenicolous species do not seem to be related to the generic type. The new species Trimmatostroma acetabuli (on Pleurosticta acetabulum), T. denigrans (on Lecanora horiza), T. rouxii (on Opegrapha durieui and other saxicolous Arthoniales) and T. vandenboomii (on Myriolecis albescens) are described. Fertile material of T. arctoparmeliae has been studied, with the result that this species is combined as Sphaerellothecium arctoparmeliae. A revised key to all known lichenicolous species of Intralichen and Trimmatostroma is given.

Identification of Novel Bioactive Compounds, Neurosporalol 1 and 2 from an Endolichenic Fungus, Neurospora ugadawe Inhabited in the Lichen Host, Graphis tsunodae Zahlbr. from Mangrove Ecosystem in Puttalam Lagoon, Sri Lanka

Endolichenic fungi (ELF) have been recognized as a promising source of bioactive secondary metabolites. Sri Lanka, a biodiversity hotspot, harbours a remarkable diversity in mangrove ecosystems. In the present study, endolichenic fungi obtained from mangrove lichens of Puttalam lagoon were studied for their secondary metabolites. An endolichenic fungus, Neurospora ugadawe, isolated from the lichen host Graphis tsunodae Zahlbr. was identified up to the species level. Secondary metabolites of N. ugadawe were extracted into ethyl acetate and crude extract showed promising antioxidant and anti-inflammatory activities. Bioassay guided separation on silica gel column chromatography and preparative TLC were performed to isolate the bioactive pure compounds. Two novel bioactive pure compounds were identified as Neurosporalol 1 and Neurosporalol 2. Neurosporalol 1 showed the highest antioxidant activity compared with the positive control BHT. Further, this compound showed very low anti-inflammatory activity compared to that of the positive control aspirin. Neurosporalol 2 showed comparable antioxidant and moderate anti-inflammatory activities.

Networks of epiphytic lichens and host trees along elevation gradients: Climate change implications in mountain ranges

Abstract Several studies have evaluated lichen responses in terms of shifts in species climate suitability, species richness and community composition. In contrast, patterns of co‐occurrence among species that could be related to complex species interactions have received less consideration. Biotic interactions play a major role in shaping species niches, fitness and adaptation to new environments. Therefore, considering the specific relationships among co‐occurring species is essential to further deepen our knowledge of biodiversity response to climate change. In this perspective, the analysis of lichen ecological networks across elevational gradients may provide a powerful tool to understand how communities are structured and how biotic interactions are modulated by changing climatic conditions. We evaluated the contribution of environmental and species biological attributes to the structure of epiphytic lichen–host tree networks. Specifically, we studied lichen communities considering two different network levels: the whole lichen community, and groups of lichen species that presented similar biological traits. In this framework, we (a) characterized the structure of the epiphytic lichen–host tree networks; (b) assessed how network structure varied with climate, forest attributes and community trait diversity and (c) evaluated the role that biological traits played in the connections established between co‐occurring lichens. On the one hand, results indicate that epiphytic lichen communities are dominated by local segregation, suggesting habitat specialization among lichens within their host tree, and that climatic conditions and, to a lesser extent, lichen diversity are the main drivers of community assemblage. On the other hand, the role of lichen species in the networks depends on their particular biological traits, supporting the hypothesis that biological traits contribute to shape network structure by influencing the ability of the species to interact between each other. These findings warn about the potential impact of climate change on epiphytic lichen communities. Synthesis. This study builds towards a better understanding of lichen community assembly and on biodiversity response to climate change in forest alpine ecosystems. In particular, our results highlight the value of lichen–tree networks to inform about assemblage processes acting at different organizational levels and indicate that lichens might become one of the most threatened groups under global change scenarios.

A Novel Cytotoxic Compound From the Endolichenic Fungus, Xylaria psidii Inhabiting the Lichen, Amandinea medusulina:

The lichen host, Amandinea medusulina, collected from mangrove habitats in Sri Lanka, and its associated endolichenic fungi were isolated and identified by rDNA-ITS sequence analysis and morphologi...

A Novel Cytotoxic Compound From the Endolichenic Fungus, Xylaria psidii Inhabiting the Lichen, Amandinea medusulina

The lichen host, Amandinea medusulina, collected from mangrove habitats in Sri Lanka, and its associated endolichenic fungi were isolated and identified by rDNA-ITS sequence analysis and morphological features. One of the fungal strains frequently isolated from the lichen thalli was identified as Xylaria psidii. This study aimed at the isolation and identification of the cytotoxic compounds present in this fungus. Secondary metabolites of X. psidii were first extracted into ethyl acetate and subsequently subjected to bioassay-guided fractionation to isolate the bioactive compounds. Sulforhodamine B assay against a lung cancer (NCI-H292) cell line was used to determine the differential cytotoxic activity. Bioassay-guided fractionation led to the isolation of an active compound, SS/02/29/08, showing moderate cytotoxicity (IC50 = 27.2 µg/mL). Its structure was elucidated by IR, 1D- and 2D-NMR, and 13C-NMR spectrophotometry and MS, in combination with HRMS, 13C NMR, HSQC, HMBC, and DQF-COSY. The structure of SS/02/29/08 was determined as ( Z)-3-{(3-acetyl-2-hydroxyphenyl)diazenyl}-2,4-dihydroxybenzaldehyde and identified as a new compound. This novel compound has promising differential cytotoxic activity against human lung cancer cell line (NCI-H292).

The Microbiomes of Seven Lichen Genera Reveal Host Specificity, a Reduced Core Community and Potential as Source of Antimicrobials.

The High Andean Paramo ecosystem is a unique neotropical mountain biome considered a diversity and evolutionary hotspot. Lichens, which are complex symbiotic structures that contain diverse commensal microbial communities, are prevalent in Paramos. There they play vital roles in soil formation and mineral fixation. In this study we analyzed the microbiomes of seven lichen genera in Colombian Paramos using 16S rRNA gene amplicon sequencing and provide the first description of the bacterial communities associated with Cora and Hypotrachyna lichens. Paramo lichen microbiomes varied in diversity indexes and number of OTUs, but were composed predominantly by the phyla Acidobacteria, Actinobacteria, Bacteroidetes, Cyanobacteria, Proteobacteria, and Verrucomicrobia. In the case of Cora and Cladonia, the microbiomes were distinguished based on the identity of the lichen host. While the majority of the lichen-associated microorganisms were not present in all lichens sampled, sixteen taxa shared among this diverse group of lichens suggest a core lichen microbiome that broadens our concept of these symbiotic structures. Additionally, we identified strains producing compounds active against clinically relevant microbial strains. These results indicate that lichen microbiomes from the Paramo ecosystem are diverse and host-specific but share a taxonomic core and can be a source of new bacterial taxa and antimicrobials.

Contrasting co-occurrence patterns of photobiont and cystobasidiomycete yeast associated with common epiphytic lichen species.

The popular dual definition of lichen symbiosis is under question with recent findings of additional microbial partners living within the lichen body. Here we compare the distribution and co-occurrence patterns of lichen photobiont and recently described secondary fungus (Cyphobasidiales yeast) to evaluate their dependency on lichen host fungus (mycobiont). We sequenced the nuclear internal transcribed spacer (ITS) strands for mycobiont, photobiont, and yeast from six widespread northern hemisphere epiphytic lichen species collected from 25 sites in Switzerland and Estonia. Interaction network analyses and multivariate analyses were conducted on operational taxonomic units based on ITS sequence data. Our study demonstrates the frequent presence of cystobasidiomycete yeasts in studied lichens and shows that they are much less mycobiont-specific than the photobionts. Individuals of different lichen species growing on the same tree trunk consistently hosted the same or closely related mycobiont-specific Trebouxia lineage over geographic distances while the cystobasidiomycete yeasts were unevenly distributed over the study area - contrasting communities were found between Estonia and Switzerland. These results contradict previous findings of high mycobiont species specificity of Cyphobasidiales yeast at large geographic scales. Our results suggest that the yeast might not be as intimately associated with the symbiosis as is the photobiont.

Biodiversity assessment of ascomycetes inhabiting Lobariella lichens in Andean cloud forests led to one new family, three new genera and 13 new species of lichenicolous fungi

AbstractNeotropical mountain forests are characterized by having hyperdiverse and unusual fungi inhabiting lichens. The great majority of these lichenicolous fungi (i.e., detectable by light microscopy) remain undescribed and their phylogenetic relationships are mostly unknown. This study focuses on lichenicolous fungi inhabiting the genusLobariella(Peltigerales), one of the most important lichen hosts in the Andean cloud forests. Based on molecular and morphological data, three new genera are introduced:Lawreyellagen. nov. (Cordieritidaceae, forUnguiculariopsis lobariella),Neobaryopsisgen. nov. (Cordycipitaceae), andPseudodidymocyrtisgen. nov. (Didymosphaeriaceae). Nine additional new species are described (Abrothallus subhaleisp. nov.,Atronectria lobariellaesp. nov.,Corticifraga microsporasp. nov.,Epithamnolia rugosopycnidiatasp. nov.,Lichenotubeufia crypticasp. nov.,Neobaryopsis andensissp. nov.,Pseudodidymocyrtis lobariellaesp. nov.,Rhagadostomella hypolobariellasp. nov., andXylaria lichenicolasp. nov.). Phylogenetic placements of 13 lichenicolous species are reported here forAbrothallus,Arthonia,Globonectria,Lawreyella,Monodictys,Neobaryopsis,Pseudodidymocyrtis,Sclerococcum,TrichonectriaandXylaria. The nameSclerococcum ricasoliaecomb. nov. is reestablished for the neotropical populations formerly namedS. lobariellum(Sclerococcales). A key to sexual and asexual states of 40 species of lobariellicolous ascomycetous fungi is provided. Teleomorph-anamorph connections were established for several species using molecular methods and/or visual observations in nature. Additionally, we found that the anamorphic speciesCornutispora ophiurosporainhabitingLobariellawas often accompanied by ascomata ofSpirographa. Results of phylogenetic analyses, including newly generated sequences of severalCornutisporaandSpirographaspecies inhabiting various host lichens, support the conclusion thatCornutisporais a synonym ofSpirographa. Our Maximum Likelihood inference based on multiple loci show that all studiedSpirographa(includingCornutispora) belong to a new lineage withinOstropales. Based on these highly supported phylogenetic placements and the distinct character states of their conidiomata, in comparison with otherLecanoromycetes, a new family is proposed –Spirographaceaefam. nov. This new lineage includes broadly distributed mycoparasites, inhabiting various lichen and fungal hosts, and representing an early diversification event preceding the lichen-forming clade ofFissurinaceae,GomphillaceaeandGraphidaceae. Two lichenicolous species,Asteroglobulus giselaeandPleoscutula arsenii, were found to be nested within theSpirographaclade, and their teleomorph-anamorph connections were confirmed based on genotypic and phenotypic data. This phylogenetic result is corroborated by their highly similar ascomata anatomy. Together these results strongly indicate that both species are congeneric withSpirographa. As a result, four new species (S. aggregatasp. nov.,S. galligenasp. nov.,S. maroneaesp. nov., andS. parmotrematissp. nov.) and 15 new combinations are proposed (Spirographa ascaridiellacomb. nov.,S. arseniicomb. nov.,S. ciliatacomb. nov.,S. giselaecomb. nov.,S. hertelianacomb. nov.,S. hypotrachynaecomb. nov.,S. intermediacomb. nov.,S. lichenicolacomb. nov.,S. limaciformiscomb. nov.,S. ophiurosporacomb. nov.,S. pittiicomb. nov.,S. pyramidaliscomb. nov.,S. triangulariscomb. nov.,S. tricupulatacomb. nov., andS. vermiformiscomb. nov.). Species of the genusSpirographa, as outlined here, are strongly host-specific, mainly at the generic level of their host. Some host genera can harbour more than oneSpirographaspecies.

Muellerella, a lichenicolous fungal genus recovered as polyphyletic within Chaetothyriomycetidae (Eurotiomycetes, Ascomycota)

AbstractMolecular data and culture-dependent methods have helped to uncover the phylogenetic relationships of numerous species of lichenicolous fungi, a specialized group of taxa that inhabit lichens and have developed diverse degrees of specificity and parasitic behaviors. The majority of lichenicolous fungal taxa are known in either their anamorphic or teleomorphic states, although their anamorph-teleomorph relationships have been resolved in only a few cases. The pycnidium-forming Lichenodiplis lecanorae and the perithecioid taxa Muellerella atricola and M. lichenicola were recently recovered as monophyletic in Chaetothyriales (Eurotiomycetes). Both genera are lichenicolous on multiple lichen hosts, upon which they show a subtle morphological diversity reflected in the description of 14 species in Muellerella (of which 12 are lichenicolous) and 12 in Lichenodiplis. Here we focus on the teleomorphic genus Muellerella and investigate its monophyly by expanding the taxon sampling to other species occurring on diverse lichen hosts. We generated molecular data for two nuclear and one mitochondrial loci (28S, 18S and 16S) from environmental samples. The present multilocus phylogeny confirms the monophyletic lineage of the teleomorphic M. atricola and M. lichenicola with their L. lecanorae-like anamorphs, but places the rest of the Muellerella species studied in two different monophyletic lineages with strong support. The first, Muellerella spp. 1, is nested within some new lineages of black fungi isolated from different epilithic lichen thalli, while the second, Muellerella spp. 2, is closely related to the Verrucariales. Based on these results, we reappraise the phylogenetic placement of Muellerella and suggest its polyphyly within Chaetothyriomycetidae.