Phylogenetic revision of the lichenized family Gomphillaceae (Ascomycota: Graphidales) suggests post-K-Pg boundary diversification and phylogenetic signal in asexual reproductive structures.

We present the first broad molecular-phylogenetic revision of the lichenized family Gomphillaceae, based on 408 newly generated sequences of the mitochondrial SSU rDNA and nuclear LSU rDNA, representing 342 OTUs. The phylogenetic analysis of 20 out of the 28 currently accepted genera resulted in 48 clades. Twelve genera were resolved as monophyletic: Actinoplaca, Arthotheliopsis, Bullatina, Caleniopsis, Corticifraga, Gomphillus, Gyalectidium, Gyalidea, Jamesiella, Rolueckia, Rubrotricha, and Taitaia. Two genera resulted paraphyletic, namely Aulaxina (including Caleniopsis) and Asterothyrium (including Linhartia). Six genera were in part highly polyphyletic: Aderkomyces, Calenia, Echinoplaca, Gyalideopsis, Psorotheciopsis, and Tricharia. While ascoma morphology and anatomy has traditionally been considered as main character complex to distinguish genera, our study supported the notion that the characteristic asexual anamorph of Gomphillaceae, the so-called hyphophores, are diagnostic for most of the newly recognized clades. As a result, we recognize 26 new genus-level clades, three of which have names available (Microxyphiomyces, Psathyromyces, Spinomyces) and 23 that will require formal description as new genera. We also tested monophyly for 53 species-level names for which two or more specimens were sequenced: 27 were supported as monophyletic and representing a single species, 13 as monophyletic but with an internal topology suggesting cryptic speciation, four as paraphyletic, and nine as polyphyletic. These data suggest that species richness in the family is higher than indicated by the number of accepted names (currently 425); they also confirm that recently refined species concepts reflect species richness better than the broad concepts applied in Santesson's monograph. A divergence time analysis revealed that foliicolous Gomphillaceae diversified after the K-Pg-boundary and largely during the Miocene, a notion supported by limited data available for other common foliicolous lineages such as Chroodiscus (Graphidaceae), Pilocarpaceae, and Porinaceae. This contradicts recent studies suggesting that only macrofoliose Lecanoromycetes exhibit increased diversification rates in the Cenozoic.

A new genus and species of foliicolous lichen in a new family of Strigulales (Ascomycota: Dothideomycetes) reveals remarkable class-level homoplasy.

Phylogenetic analysis of some foliicolous lichens collected in Hainan Province, China, revealed a new lineage morphologically similar to Porina but phylogenetically related to Strigulaceae (Dothideomycetes), differing from the latter in ascus type. The monospecific genus Tenuitholiascus gen. nov. is introduced for the single species, T. porinoides sp. nov., which is placed in the new, monogeneric family Tenuitholiascaceae, sister to Strigulaceae in Strigulales. The new taxon closely resembles the genus Porina in external morphology and ascospore type, as well as the thin-walled asci and unbranched paraphyses. Yet, it is entirely unrelated to the latter, which belongs in class Lecanoromycetes in the order Gyalectales.

Unambiguous identification of fungi: where do we stand and how accurate and precise is fungal DNA barcoding?

True fungi (Fungi) and fungus-like organisms (e.g. Mycetozoa, Oomycota) constitute the second largest group of organisms based on global richness estimates, with around 3 million predicted species. Compared to plants and animals, fungi have simple body plans with often morphologically and ecologically obscure structures. This poses challenges for accurate and precise identifications. Here we provide a conceptual framework for the identification of fungi, encouraging the approach of integrative (polyphasic) taxonomy for species delimitation, i.e. the combination of genealogy (phylogeny), phenotype (including autecology), and reproductive biology (when feasible). This allows objective evaluation of diagnostic characters, either phenotypic or molecular or both. Verification of identifications is crucial but often neglected. Because of clade-specific evolutionary histories, there is currently no single tool for the identification of fungi, although DNA barcoding using the internal transcribed spacer (ITS) remains a first diagnosis, particularly in metabarcoding studies. Secondary DNA barcodes are increasingly implemented for groups where ITS does not provide sufficient precision. Issues of pairwise sequence similarity-based identifications and OTU clustering are discussed, and multiple sequence alignment-based phylogenetic approaches with subsequent verification are recommended as more accurate alternatives. In metabarcoding approaches, the trade-off between speed and accuracy and precision of molecular identifications must be carefully considered. Intragenomic variation of the ITS and other barcoding markers should be properly documented, as phylotype diversity is not necessarily a proxy of species richness. Important strategies to improve molecular identification of fungi are: (1) broadly document intraspecific and intragenomic variation of barcoding markers; (2) substantially expand sequence repositories, focusing on undersampled clades and missing taxa; (3) improve curation of sequence labels in primary repositories and substantially increase the number of sequences based on verified material; (4) link sequence data to digital information of voucher specimens including imagery. In parallel, technological improvements to genome sequencing offer promising alternatives to DNA barcoding in the future. Despite the prevalence of DNA-based fungal taxonomy, phenotype-based approaches remain an important strategy to catalog the global diversity of fungi and establish initial species hypotheses.

Going extinct before being discovered? New lichen fungi from a small fragment of the vanishing Atlantic Rainforest in Brazil / Indo extinto antes de ser descoberto? Novos fungos liquenizados da evanescente Mata Atlântica no Brasil

Abstract In the frame of an ongoing lichen inventory of Atlantic Rainforest remnants in Northeast Brazil, five new species of Graphidaceae were discovered in a small forest fragment, Mata do Cipó, in Sergipe state, the smallest state of Brazil and among those with the highest deforestation rate in the country. An additional new species had already been collected in Panama before and was now also found in the Mata do Cipó and is described here as well. In total, 40 species of Graphidaceae are reported for this remnant, including a large number of taxa indicative of well-preserved rainforest. The new species are: Fissurina atlantica T.A. Pereira, M. Cáceres & Lücking, sp. nov., Graphis subaltamirensis Passos, M. Cáceres & Lücking, sp. nov., Ocellularia cipoensis L.A. Santos, M. Cáceres & Lücking, sp. nov., O. sosma T.A. Pereira, M. Cáceres & Lücking, sp. nov., O. submordenii Lücking, sp. nov. (also known from Panama), and Pseudochapsa aptrootiana M. Cáceres, T.A. Pereira & Lücking, sp. nov. The findings are discussed in the context of the strong fragmentation of the Atlantic Rainforest, with individual remnants apparently serving as refugia for residual populations of rare species of lichen fungi that were more widely distributed in the past, but currently seem to occur only in isolated fragments.

Resumo No âmbito de um inventário atual de liquens dos remanescentes da Mata Atlântica, no Nordeste do Brasil, cinco espécies novas de Graphidaceae foram descobertas em um pequeno fragmento florestal, Mata do Cipó, no estado de Sergipe, o menor estado do Brasil, o qual está entre aqueles com maior taxa de desmatamento no país. Uma nova espécie adicional já havia sido coletada no Panamá, anteriormente, e agora também foi encontrada na Mata do Cipó, e está sendo descrita aqui também. No total, 40 espécies de Graphidaceae são relatadas para este remanescente, incluindo um grande número de táxons indicadores de floresta bem preservada. As novas espécies são: Fissurina atlantica T.A. Pereira, M. Cáceres & Lücking, sp. nov., Graphis subaltamirensis Passos, M. Cáceres & Lücking, sp.nov., Ocellularia cipoensis L.A. Santos, M. Cáceres & Lücking, sp. nov., O. sosma T.A. Pereira, M. Cáceres & Lücking, sp. nov., O. submordenii Lücking, sp. nov. (também conhecido do Panamá), e Pseudochapsa aptrootiana M. Cáceres, T.A. Pereira & Lücking, sp. nov. Os achados são discutidos no contexto da forte fragmentação da Mata Atlântica, com remanescentes aparentemente servindo de refúgio para populações residuais de espécies raras de fungos liquenizados que, provavelmente, tiveram uma distribuição mais amplia no passado, mas que atualmente parecem ocorrer apenas em fragmentos isolados.

Ecogeografía del género Sticta (Ascomycota liquenizados: Lobariaceae) en Colombia / Ecogeography of the genus Sticta (lichenized Ascomycota: Lobariaceae) in Colombia

Colombia is a megadiverse country, but with a substantial gap in the taxonomic and ecological knowledge of fungi and lichens. Thus, the objective of the present study was an ecogeographical analysis of the Sticta species in Colombia. The data included macrodistribution (with respect to mountain ranges and life zones) and microhabitat preferences (light, substrate) and were obtained from field work and herbarium collection labels (completed by comparison with modern geographic and vegetation maps), for 103 species of Sticta currently recognized in Colombia (plus seven additional biotypes with different photobionts or reproductive mode). Using non-metric multidimensional scaling (NMS), correlations between ecogeographical variables and species were established, and ecotypes were delimited. Colombian species of Sticta showed distinct distribution patterns relative to altitude, mountain ranges, life zones, as well as light exposure and substrate, forming several distinct groups. Cyanobacterial species tend to have wider distribution ranges than green algal species. We concluded that the orogeny of the Northern Andes substantially affected speciation of the genus Sticta and its ecogeographical differentiation.

Colombia es un país megadiverso, pero con una ausencia significativa en el conocimiento de la taxonomía y ecología de hongos y líquenes. Por lo tanto, el objetivo del presente estudio era un análisis ecogeográfico de las especies de Sticta de Colombia. Los datos incluyeron la macrodistribución (con respeto a las cordilleras y zonas de vida) y preferencias de microhabitat (luz, sustrato) y fueron obtenidos mediante trabajo de campo y en el herbario (complemetados con mapas modernos de geografía y vegetación), para 103 especies actualmente reconocidas en Colombia (más siete biotipos con diferentes fotobiontes o modo de reproducción). Se elaboró un análisis de ordenación usando la técnica de escalamiento multidimensional no métrico (NMS), para establecer correlaciones entre variables ecogeográficas y las especies y para definir ecotipos. Las especies de Sticta en Colombia presentan patrones de distribución relativos a la altitud, las zonas de vida y la ubicación en las cordilleras, tanto como luz y sustrato, formando varios grupos distintos. Las especies con ciano-bionte tienen un rango de distribución más amplio que las especies con fotobionte verde. Concluimos que la orogenia de los Andes afectó significativamente la especiación del género Sticta y su diferenciación ecogeográfica.

Phylogenetic classification at generic level in the absence of distinct phylogenetic patterns of phenotypical variation: a case study in graphidaceae (ascomycota).

Molecular phylogenies often reveal that taxa circumscribed by phenotypical characters are not monophyletic. While re-examination of phenotypical characters often identifies the presence of characters characterizing clades, there is a growing number of studies that fail to identify diagnostic characters, especially in organismal groups lacking complex morphologies. Taxonomists then can either merge the groups or split taxa into smaller entities. Due to the nature of binomial nomenclature, this decision is of special importance at the generic level. Here we propose a new approach to choose among classification alternatives using a combination of morphology-based phylogenetic binning and a multiresponse permutation procedure to test for morphological differences among clades. We illustrate the use of this method in the tribe Thelotremateae focusing on the genus Chapsa, a group of lichenized fungi in which our phylogenetic estimate is in conflict with traditional classification and the morphological and chemical characters do not show a clear phylogenetic pattern. We generated 75 new DNA sequences of mitochondrial SSU rDNA, nuclear LSU rDNA and the protein-coding RPB2. This data set was used to infer phylogenetic estimates using maximum likelihood and Bayesian approaches. The genus Chapsa was found to be polyphyletic, forming four well-supported clades, three of which clustering into one unsupported clade, and the other, supported clade forming two supported subclades. While these clades cannot be readily separated morphologically, the combined binning/multiresponse permutation procedure showed that accepting the four clades as different genera each reflects the phenotypical pattern significantly better than accepting two genera (or five genera if splitting the first clade). Another species within the Thelotremateae, Thelotrema petractoides, a unique taxon with carbonized excipulum resembling Schizotrema, was shown to fall outside Thelotrema. Consequently, the new genera Astrochapsa, Crutarndina, Pseudochapsa, and Pseudotopeliopsis are described here and 39 new combinations are proposed.

Especificidad de forófito y preferencias microambientales de los líquenes cortícolas en cinco forófitos del bosque premontano de finca Zíngara, Cali, Colombia / Phorophyte specificity and microenvironmental preferences of corticolous lichens in five phorophyte species from premontane forest of Finca Zíngara, Cali, Colombia)

Phorophyte specificity and microenvironmental preferences of corticolous lichens in five phorophyte species from premontane forest of Finca Zíngara, Cali, Colombia). Lichenized fungi or lichens are organisms that have been little studied in the tropics and which distribution is affected by microenvironmental factors and substrate characteristics. The present study aimed to identify phorophyte specificity and microenvironmental preferences of corticolous lichens in five phorophyte species from premontane forest of the farm Finca Zingara in Cali, Colombia. For this, five individuals were selected from five tree species (phorophytes). Lichen species present in a 0.50x0.20m² quadrant located in the trunk of each tree at a height of 1.3m were identified. Substrate parameters such as bark pH, Diameter Breast Height (DBH) and bark structure were measured. Also, microenvironmental factors including temperature, humidity and irradiance were determined. In order to detect phorophyte preferences, a non-metric multidimensional scaling (NMS) and indicator species analysis were made. Spearman correlation analysis was used to assert the relationship between environmental variables and groupings found in the NMS. A total of 69 species of lichens were found, of which 37 were identified to species, 18 to genera and 14 were not determined because they were sterile or had no spores. NMS showed that some individuals of the same tree species were grouped alongside the analysis dimensions, and they were related with the factors of light intensity, temperature and DBH. Only three lichens with preference for certain tree species were found (Arthonia microsperma by Meriania sp., Cladonia ceratophylla and sorediado 8 by Clusia sp.), suggesting absence of phorophyte preferences. Thus, it can be concluded that lichens from the study area do not show phorophyte preference, but their distribution is affected by light, temperature and DBH. Rev. Biol. Trop. 60 (2): 843-856. Epub 2012 June 01.

El objetivo de este trabajo fue evaluar la especificidad de forófito y las preferencias microambientales de los líquenes cortícolas en el Bosque de la Finca Zíngara (Cali, Colombia). Asimismo, se seleccionaron cinco individuos de cinco especies de árboles. Además, se identificaron las especies de líquenes presentes en un cuadrante de 0.50x0.20m2 ubicado en el tronco de cada árbol a 1.3 m de altura. También, se midieron parámetros microambientales como pH de la corteza, diámetro a la altura del pecho (DAP), estructura de la corteza, humedad relativa e irradianza. Para detectar las preferencias de forófito, se realizó un escalamiento no métrico multidimensional (NMS) y un análisis de especies indicadoras. Luego se efectúo un análisis de correlación de Spearman para evaluar la relación entre las variables ambientales y los agrupamientos encontrados en el NMS. Se encontraron 69 especies de líquenes, de los cuales 37 fueron determinados hasta especie, 18 hasta género y 14 no fueron determinados. El resultado del NMS mostró que algunos individuos de la misma especie de árbol se agruparon a lo largo de las dimensiones del análisis, y están relacionados con los factores intensidad de luz, temperatura y DAP. Sólo tres especies mostraron preferencia por ciertas especies de árboles (Arthonia microsperma por Meriania sp., Cladonia ceratophylla y sorediado 8 por Clusia sp.), lo cual sugiere ausencia de preferencias de forófito.

High concentration of basidiolichens in a single family of agaricoid mushrooms (Basidiomycota: Agaricales: Hygrophoraceae).

The Agaricales is the largest and most diverse order of mushroom-forming Basidiomycota, with over 100 natural groups recognized in recent Fungal Tree of Life studies. Most agarics are either saprotrophic or ectomycorrhizal fungi, but the family Hygrophoraceae is in part characterized by a unique and remarkable diversity of lichenized forms. The most familiar of these is the chlorolichen genus Lichenomphalia, whose phylogenetic position in the Agaricales has been established. Recent limited evidence suggested that Hygrophoraceae also contains cyanolichens in the genus Dictyonema, which indicates a remarkable concentration and diversity of lichen-formers in a single family of agarics. To demonstrate the relationships of lichen-formers to other fungi in the family, we assembled ribosomal sequences from 52 species representing recognized groups within the Hygrophoraceae, among them new sequences representing Acantholichen and most species and forms of Dictyonema. The molecular data were evaluated using parsimony, likelihood, Bayesian, and distance analyses, including coding of ambiguous regions by means of INAASE and ARC, all of which indicate that Dictyonema and Acantholichen form a monophyletic clade derived from the primarily bryophilous genus Arrhenia and sister to the enigmatic Athelia pyriformis, a species unrelated to the Atheliales for which we are proposing a new genus name Eonema. The chlorolichen genus Lichenomphalia may be polyphyletic. Fungi in the Dictyonema-Acantholichen clade are typically tropical, entirely lichenized, and associate with cyanobacterial photobionts. Our data indicate a transition from agaricoid-omphalinoid basidiomes observed in Arrhenia to stereoid-corticioid forms in Dictyonema, and also support a previous suggestion of a connection between loss of clamp connections and lichenization. The diverse basidiome and thallus morphologies and nutritional ecologies of these fungi indicate a remarkable evolutionary flexibility that appears to have developed in part as a consequence of symbiosis.

The Ascomycota tree of life: a phylum-wide phylogeny clarifies the origin and evolution of fundamental reproductive and ecological traits.

We present a 6-gene, 420-species maximum-likelihood phylogeny of Ascomycota, the largest phylum of Fungi. This analysis is the most taxonomically complete to date with species sampled from all 15 currently circumscribed classes. A number of superclass-level nodes that have previously evaded resolution and were unnamed in classifications of the Fungi are resolved for the first time. Based on the 6-gene phylogeny we conducted a phylogenetic informativeness analysis of all 6 genes and a series of ancestral character state reconstructions that focused on morphology of sporocarps, ascus dehiscence, and evolution of nutritional modes and ecologies. A gene-by-gene assessment of phylogenetic informativeness yielded higher levels of informativeness for protein genes (RPB1, RPB2, and TEF1) as compared with the ribosomal genes, which have been the standard bearer in fungal systematics. Our reconstruction of sporocarp characters is consistent with 2 origins for multicellular sexual reproductive structures in Ascomycota, once in the common ancestor of Pezizomycotina and once in the common ancestor of Neolectomycetes. This first report of dual origins of ascomycete sporocarps highlights the complicated nature of assessing homology of morphological traits across Fungi. Furthermore, ancestral reconstruction supports an open sporocarp with an exposed hymenium (apothecium) as the primitive morphology for Pezizomycotina with multiple derivations of the partially (perithecia) or completely enclosed (cleistothecia) sporocarps. Ascus dehiscence is most informative at the class level within Pezizomycotina with most superclass nodes reconstructed equivocally. Character-state reconstructions support a terrestrial, saprobic ecology as ancestral. In contrast to previous studies, these analyses support multiple origins of lichenization events with the loss of lichenization as less frequent and limited to terminal, closely related species.

A five-gene phylogeny of Pezizomycotina.

Pezizomycotina is the largest subphylum of Ascomycota and includes the vast majority of filamentous, ascoma-producing species. Here we report the results from weighted parsimony, maximum likelihood and Bayesian phylogenetic analyses of five nuclear loci (SSU rDNA, LSU rDNA, RPB1, RPB2 and EF-lalpha) from 191 taxa. Nine of the 10 Pezizomycotina classes currently recognized were represented in the sampling. These data strongly supported the monophyly of Pezizomycotina, Arthoniomycetes, Eurotiomycetes, Orbiliomycetes and Sordariomycetes. Pezizomycetes and Dothideomycetes also were resolved as monophyletic but not strongly supported by the data. Lecanoromycetes was resolved as paraphyletic in parsimony analyses but monophyletic in maximum likelihood and Bayesian analyses. Leotiomycetes was polyphyletic due to exclusion of Geoglossaceae. The two most basal classes of Pezizomycotina were Orbiliomycetes and Pezizomycetes, both of which comprise species that produce apothecial ascomata. The seven remaining classes formed a monophyletic group that corresponds to Leotiomyceta. Within Leotiomyceta, the supraclass clades of Leotiomycetes s.s. plus Sordariomycetes and Arthoniomycetes plus Dothideomycetes were resolved with moderate support.

New insights into classification and evolution of the Lecanoromycetes (Pezizomycotina, Ascomycota) from phylogenetic analyses of three ribosomal RNA- and two protein-coding genes.

The Lecanoromycetes includes most of the lichen-forming fungal species (> 13500) and is therefore one of the most diverse class of all Fungi in terms of phenotypic complexity. We report phylogenetic relationships within the Lecanoromycetes resulting from Bayesian and maximum likelihood analyses with complementary posterior probabilities and bootstrap support values based on three combined multilocus datasets using a supermatrix approach. Nine of 10 orders and 43 of 64 families currently recognized in Eriksson's classification of the Lecanoromycetes (Outline of Ascomycota--2006 Myconet 12:1-82) were represented in this sampling. Our analyses strongly support the Acarosporomycetidae and Ostropomycetidae as monophyletic, whereas the delimitation of the largest subclass, the Lecanoromycetidae, remains uncertain. Independent of future delimitation of the Lecanoromycetidae, the Rhizocarpaceae and Umbilicariaceae should be elevated to the ordinal level. This study shows that recent classifications include several nonmonophyletic taxa at different ranks that need to be recircumscribed. Our phylogenies confirm that ascus morphology cannot be applied consistently to shape the classification of lichen-forming fungi. The increasing amount of missing data associated with the progressive addition of taxa resulted in some cases in the expected loss of support, but we also observed an improvement in statistical support for many internodes. We conclude that a phylogenetic synthesis for a chosen taxonomic group should include a comprehensive assessment of phylogenetic confidence based on multiple estimates using different methods and on a progressive taxon sampling with an increasing number of taxa, even if it involves an increasing amount of missing data.