Lichen-forming Fungi Research Articles

Chloroidium phycobionts (Watanabeales, Trebouxiophyceae) partner with lecanoralean mycobionts in foliicolous lichen communities of Tenerife (Canary Islands) and Navarra (Iberian Peninsula), Spain

Abstract While the diversity of foliicolous lichen-forming fungi has been explored in substantial depth, relatively little attention has been paid to their algal symbionts. We studied the unicellular green phycobionts of the lecanoralean lichens Bacidina (Ramalinaceae), Byssoloma, Fellhanera and Tapellaria (Pilocarpaceae) and graphidalean Gyalectidium (Gomphillaceae) from two extratropical foliicolous communities in continental Spain and the Canary Islands. We examined the pyrenoids of algal symbionts within thalli using TEM, and obtained several algal nrSSU and rbcL sequences from whole thalli, and also from cultures isolated from some of these lichens. Pyrenoid structure and molecular sequence data provided support for recognizing Chloroidium (Watanabeales, Trebouxiophyceae) as phycobiont in thalli of Byssoloma subdiscordans and Fellhanera bouteillei (Pilocarpaceae) in both communities. Bacidina apiahica (Ramalinaceae) and Tapellaria epiphylla (Pilocarpaceae) likewise appeared to partner with Chloroidium based on the presence of the same pyrenoid type, although we were able to obtain a phycobiont sequence only from a culture isolate of the latter. These results contrast with those obtained previously from a foliicolous lichen community in southern Florida, which revealed only strains of Heveochlorella (Jaagichlorella) as phycobiont of foliicolous Pilocarpaceae and Gomphillaceae. On the other hand, the pyrenoid we observed in the phycobionts associated with Gyalectidium setiferum and G. minus corresponded to that of Heveochlorella (Jaagichlorella). However, the poor quality of the phycobiont sequence data obtained from G. minus, probably due to the presence of epibiontic algae, could not provide additional perspective on the pyrenoid structure observations. Nonetheless, clear differences in pyrenoid ultrastructure can allow Chloroidium and Heveochlorella phycobionts to be distinguished from each other in TEM. Our results indicate a greater diversity of unicellular green-algal symbionts in foliicolous communities from Spain than previously observed in other geographical areas, and suggest that further studies focused on symbiont pairing in these communities might reveal distinctive and varied patterns of phycobiont preference.

Bacterial communities associated with an island radiation of lichen-forming fungi.

Evolutionary radiations are one of the most striking processes biologists have studied in islands. A radiation is often sparked by the appearance of ecological opportunity, which can originate in processes like trophic niche segregation or the evolution of key innovations. Another recently proposed mechanism is facilitation mediated by the bacterial communities associated with the radiating species. Here we explore the role of the bacterial communities in a radiation of lichen-forming fungi endemic to Macaronesia. Bacterial diversity was quantified by high throughput sequencing of the V1-V2 hyper-variable region of 172 specimens. We characterized the taxonomic and phylogenetic diversity of the bacterial communities associated with the different species, tested for compositional differences between these communities, carried out a functional prediction, explored the relative importance of different factors in bacterial community structure, searched for phylosymbiosis and tried to identify the origin of this pattern. The species of the radiation differed in the composition of their bacterial communities, which were mostly comprised of Alphaproteobacteria and Acidobacteriia, but not in the functionality of those communities. A phylosimbiotic pattern was detected, but it was probably caused by environmental filtering. These findings are congruent with the combined effect of secondary chemistry and mycobiont identity being the main driver of bacterial community structure. Altogether, our results suggest that the associated bacterial communities are not the radiation's main driver. There is one possible exception, however, a species that has an abnormally diverse core microbiome and whose bacterial communities could be subject to a specific environmental filter at the functional level.

Multilocus Molecular Phylogeny of the Umbilicaria aprina Group (Umbilicariaceae, Lichenized Ascomycota) Supports Species Level and Neo-Endemic Status of Umbilicaria krascheninnikovii.

The Northeast Asian endemic species of lichen-forming fungus Umbilicaria krascheninnikovii is herein discussed in the global context of biogeography and phylogeny of the U. aprina group. The name U. krascheninnikovii has been erroneously used by lichenologists for Umbilicaria spp. from high latitudes or altitudes worldwide, as there are omphalodisc apothecia and rough "crystals" of a necral layer on the upper surface. To test the monophyly and phylogenetic relationships within the U. aprina group, four independent DNA regions (nrITS/5.8S, RPB2, mtLSU, and mtSSU) were used for six rare species, including a dozen specimens of U. krascheninnikovii from its locus classicus in Kamchatka. The study is based on the phylograms obtained using maximum likelihood and a Bayesian phylogenetic inference framework. As a result of phylogenetic and biogeographic analyses, it was shown that U. krascheninnikovii is a neo-endemic of the areas of modern volcanism in Kamchatka, Japan, as well as in the Kurile Islands, where this species was recorded for the first time. The morphology of U. krascheninnikovii is herein described and illustrated. Increasing the role of the sexual process and reducing asexual thalloconidiogenesis are shown to be apomorphic traits in the U. aprina group. The combination of sexual and asexual reproduction provides adaptive advantages in changing environmental conditions.

Two novel species of arctic-alpine lichen-forming fungi (Ascomycota, Megasporaceae) from the Deosai Plains, Pakistan.

Members of the lichen-forming fungal genus Oxneriaria are known to occur in cold polar and high altitudinal environments. Two new species, Oxneriariacrittendenii and O.deosaiensis, are now described from the high altitude Deosai Plains, Pakistan, based on phenotypic, multigene phylogenetic and chemical evidence. Phenotypically, O.crittendenii is characterised by orbicular light-brown thalli 1.5-5 cm across, spot tests (K, C, KC) negative, apothecia pruinose, hymenium initially blue then dark orange in response to Lugol's solution. Oxneriariadeosaiensis is characterised by irregular areolate grey thalli 1.5-2 cm across, K test (light brown), KC test (dark brown), apothecia epruinose, hymenium initially blue then dark blue in response to Lugol's solution. Both species share the same characters of thalli with black margins and polarilocular ascospores. The closest previously reported species, O.pruinosa, differs from O.crittendenii and O.deosaiensis in having non-lobate margins, thin thalline exciple (45-80 μm thick), short asci (55-80 × 25-42 μm) and K positive (yellow) and KC negative tests and divergent DNA sequence in the ITS, LSU and mtSSU regions. The newly-described Oxneriaria species add to growing evidence of the Deosai Plains as a region of important arctic-alpine biodiversity.

Patterns of Endemism in Lichens: Another Paradigm-Shifting Example in the Lichen Genus Xanthoparmelia from Macaronesia.

It has long been assumed that lichen-forming fungi have very large distribution ranges, and that endemic species are rare in this group of organisms. This is likely a consequence of the "everything small is everywhere" paradigm that has been traditionally applied to cryptogams. However, the description of numerous endemic species over the last decades, many of them in oceanic islands, is challenging this view. In this study, we provide another example, Xanthoparmelia ramosae, a species that is described here as new to science on the basis of morphological, chemical, and macroclimatic data, and three molecular markers (ITS rDNA, nuLSU rDNA, and mtSSU). The new species is endemic to the island of Gran Canaria but clusters into a clade composed exclusively of specimens collected in Eastern Africa, a disjunction that is here reported for the first time in lichen-forming fungi. Through the use of dating analysis, we have found that Xanthoparmelia ramosae diverged from its closely related African taxa in the Pliocene. This result, together with the reproductive strategy of the species, points to the Relict theory as a likely mechanism behind the disjunction, although the large gap in lichenological knowledge in Africa makes this possibility hard to explore any further.

New microsatellite loci to quantify genetic diversity of the photosymbiodeme-forming lichen Sticta canariensis

AbstractSticta canariensis is a lichen which is rare in all parts of its range in Atlantic Europe and Macaronesia, where it occurs in laurisilva forests, a habitat highly threatened by global change. Thus, this species is of high priority for inclusion in conservation programmes where genetic diversity should be considered. We have established new microsatellite loci and generated a dataset that demonstrates the genetic diversity of the lichen-forming fungus S. canariensis from eight locations across its disjunct range, in Macaronesia, Norway and England. We genotyped 25 microsatellite loci for 65 specimens and detected five genetic clusters which resemble major geographical divisions, specifically among the Macaronesian archipelagos. The total number of observed alleles ranged from 2 to 22. These are the first microsatellite markers developed for S. canariensis and they will be useful for population genetic studies and for conservation assessments.

Discovering neglected lichen diversity with DNA-based inventories: metabarcoding lichen-forming fungi in Bryce Canyon National Park, Utah, USA

National parks and other federally designated natural areas play critical roles in preserving unique habitats, communities, and biodiversity. However, in the United States, it is estimated that 80–90% of species diversity in national parks is presently unknown. Therefore, contemporary biodiversity inventories are critical for conservation, management and establishing baselines for future comparisons. Ongoing efforts to characterize lichen diversity highlight diverse and robust communities in a number of national parks in the USA. In arid regions of the western USA facing ecological transformations, lichens can play a pivotal role for monitoring these changes. Lichen diversity in Bryce Canyon National Park (BRCA) in southern Utah, USA remains nearly completely uncharacterized, despite nearly 100 years as a federally protected area. Our study aims to provide a critical perspective into the lichen diversity of BRCA. Using a metabarcoding community sampling approach, we documented 215 candidate lichen-forming fungal species distributed across ecologically distinct sites in BRCA. At each sampled site, species richness ranged from 104 to 133 species, with no more than 20% shared species among the three sites. The limited overlap between collection sites suggests that BRCA harbors greater diversity than initially thought. We document a number of sensitive lichens, particularly Usnea spp. and Ramalina sinensis, that should be monitored as air pollution, land use, and impacts of climate change affect biological communities in the park. The inventory also includes unknown species and other species that have not been documented in the western USA. While our DNA-based inventory highlights strikingly rich lichen diversity, future voucher-based collections will be essential for robust taxonomic determinations.

Reference-Based Restriction-Site-Associated DNA Sequencing Data Are Useful for Species Delineation in a Recently Diverged Asexually Reproducing Species Complex (Parmeliaceae, Ascomycota).

Cryptic species are common in lichen-forming fungi and have been reported from different genera in the most speciose family, Parmeliaceae. Herein, we address species delimitation in a group of mainly asexually reproducing Parmelina species. The morphologically distinct P. pastillifera was previously found nested within a morphologically circumscribed P. tiliacea based on several loci. However, these studies demonstrated a relatively high genetic diversity within P. tiliacea sensu lato. Here, we revisit the species delimitation in the group by analyzing single-nucleotide polymorphisms (SNPs) through genome-wide assessment using Restriction-Site-Associated sequencing and population genomic methods. Our data support previous studies and provide further insight into the phylogenetic relationships of the four clades found within the complex. Based on the evidence suggesting a lack of gene flow among the clades, we recognize the four clades as distinct species, P. pastillifera and P. tiliacea sensu stricto, and two new species, P. clandestina sp. nov. and P. mediterranea sp. nov.

Myrmecia, Not Asterochloris, Is the Main Photobiont of Cladonia subturgida (Cladoniaceae, Lecanoromycetes).

This study explores the diversity of photobionts associated with the Mediterranean lichen-forming fungus Cladonia subturgida. For this purpose, we sequenced the whole ITS rDNA region by Sanger using a metabarcoding method for ITS2. A total of 41 specimens from Greece, Italy, France, Portugal, and Spain were studied. Additionally, two specimens from Spain were used to generate four cultures. Our molecular studies showed that the genus Myrmecia is the main photobiont of C. subturgida throughout its geographic distribution. This result contrasts with previous studies, which indicated that the main photobiont for most Cladonia species is Asterochloris. The identity of Myrmecia was also confirmed by ultrastructural studies of photobionts within the lichen thalli and cultures. Photobiont cells showed a parietal chloroplast lacking a pyrenoid, which characterizes the species in this genus. Phylogenetic analyses indicate hidden diversity within this genus. The results of amplicon sequencing showed the presence of multiple ASVs in 58.3% of the specimens studied.

Wide divergence of fungal communities inhabiting rocks and soils in a hyper-arid Antarctic desert.

Highly simplified microbial communities colonise rocks and soils of continental Antarctica ice-free deserts. These two habitats impose different selection pressures on organisms, yet the possible filtering effects on the diversity and composition of microbial communities have not hitherto been fully characterised. We hence compared fungal communities in rocks and soils in three localities of inner Victoria Land. We found low fungal diversity in both substrates, with a mean species richness of 28 across all samples, and significantly lower diversity in rocks than in soils. Rock and soil communities were strongly differentiated, with a multinomial species classification method identifying just three out of 328 taxa as generalists with no affinity for either substrate. Rocks were characterised by a higher abundance of lichen-forming fungi (typically Buellia, Carbonea, Pleopsidium, Lecanora, and Lecidea), possibly owing to the more protected environment and the porosity of rocks permitting photosynthetic activity. In contrast, soils were dominated by obligate yeasts (typically Naganishia and Meyerozyma), the abundances of which were correlated with edaphic factors, and the black yeast Cryomyces. Our study suggests that strong differences in selection pressures may account for the wide divergences of fungal communities in rocks and soils of inner Victoria Land.

New records of the genus Cladonia from Algeria

Based on our studies of lichen-forming fungi in Kala National Park, Northeastern Algeria, four species, C. cyathomorpha, C. dimorpha, C. monomorpha and C. subturgida, were new reported for Algeria. Descriptions and taxonomic notes are provided; and a key is presented for all the Cladonia species known from Algeria.

The first records of the lichen-forming, lichenicolous and allied fungi from Ukraine

Fifteen species of lichen-forming, lichenicolous and allied fungi are reported for the first time from Ukraine. These species are Buellia ocellata, Catillaria fungoides, Cecidonia xenophana, Cladonia subturgida, Celothelium lutescens, Endococcus protoblasteniae, Kuettlingeria areolata, Lahmia kunzei, Lecanora microloba, Lecidea nylanderi, Myriolecis zosterae, Pyrenodesmia helygeoides, Sagedia zonata, Verrucula arnoldaria, Zahlbrucknerella calcarea. The genera Cecidonia, Celothelium, Lahmia, Sagedia, Zahlbrucknerella, familia Celotheliaceae, Lahmiaceae, and orders Lahmiales and Phaemoniliales are also new to the country.

Regulation of symbiotic interactions and primitive lichen differentiation by UMP1 MAP kinase in Umbilicaria muhlenbergii

AbstractLichens are of great ecological importance but mechanisms regulating lichen symbiosis are not clear. Umbilicaria muhlenbergii is a lichen-forming fungus amenable to molecular manipulations and dimorphic. Here, we established conditions conducive to symbiotic interactions and lichen differentiation and showed the importance of UMP1 MAP kinase in lichen development. In the initial biofilm-like symbiotic complexes, algal cells were interwoven with pseudohyphae covered with extracellular matrix. After longer incubation, fungal-algal complexes further differentiated into primitive lichen thalli with a melanized cortex-like and pseudoparenchyma-like tissues containing photoactive algal cells. Mutants deleted of UMP1 were blocked in pseudohyphal growth and development of biofilm-like complexes and primitive lichens. Invasion of dividing mother cells that contributes to algal layer organization in lichens was not observed in the ump1 mutant. Overall, these results showed regulatory roles of UMP1 in symbiotic interactions and lichen development and suitability of U. muhlenbergii as a model for studying lichen symbiosis.

Beetles and lichens: tracing the origins and evolution of lichenophagy within the darkling beetle tribe Helopini (Coleoptera: Tenebrionidae)

Abstract Although the complex evolutionary history of lichen-forming fungi has gained considerable attention, particularly regarding the long-debated role of these organisms in shaping early terrestrial ecosystems, the evolution of lichenivory and its potential impact on the diversification of lichenophages have been largely neglected. With > 800 described species worldwide and a broad geographical distribution, the tribe Helopini (Coleoptera: Tenebrionidae) represents a diverse, yet poorly studied, group of predominantly lichenophagous beetles. Using a dataset of 52 ingroup taxa and five gene fragments, a first phylogenetic hypothesis of the tribe was generated, which was subsequently used for reconstructing the ancestral state of the trophic and habitat associations of the beetles and for estimating a time frame of diversification. Our phylogenetic reconstruction sheds light on the higher-level systematics of the tribe, supporting the current subtribal division of the group while also providing a framework for understanding the intergeneric relationships within subtribes. The results also indicate an Early Cretaceous origin of the tribe, highlighting the close association between Helopini and lichen-forming fungi since the emergence of the group. Nevertheless, at least seven independent switches from lichenophagy to alternative feeding habits have occurred since the middle Eocene, which can be linked temporally to transitions from forests to open habitats.

Is lichen symbiont mutualism a myth?

AbstractLichens are classic examples of symbiosis, but some biologists have questioned whether the algal partner benefits from the relationship. Among the diverse lichen symbioses, the carbon transfer systems show remarkable convergences. When a compatible fungus is encountered, the alga proactively releases large amounts of carbohydrate, suggesting active participation rather than victimhood. Some lichen-related fungus–alga symbioses appear obligatory for the algal partner. Within true lichens, algal symbionts can persist at microsites where they might not otherwise be competitive, because of improved stress tolerance, reduced photoinhibition, protection from herbivores, and the more efficient moisture management and positioning for light interception that fungal structures provide. Algal clones continually disperse from the lichen thallus by diverse means, allowing the genotype to pioneer aposymbiotic colonies from a stable refuge. Because lichen-forming fungi conserve rather than consume their algal symbionts, the mutual self-interests of both partners substantially align in the stressful microhabitats where lichens are successful.

Independent, structurally distinct transitions to microfruticose growth in the crustose genus Porina (Ostropales, Lecanoromycetes): new isidioid species from south-western Florida

AbstractPorina is a widely distributed, species-rich genus of crustose, lichen-forming fungi, some with thalline outgrowths that have been recognized as isidia. We studied three taxa with thalli consisting chiefly of ascending isidioid structures occurring on trunks and branches of Taxodium in southwestern Florida, and provide details of their structure with light and electron microscopy. Two of these taxa we describe as new species: P. microcoralloides and P. nanoarbuscula. Genetic sequences (mtSSU) suggest that they are closely related to each other, yet they differ markedly in the size, morphology and anatomical organization of their isidioid branches as well as in the length of their ascospores. In the three Floridian taxa studied, the crustose portion of the thallus is partly endophloeodic and partly superficial, the latter often patchy, evanescent or inconspicuous, and completely lacks the differentiated anatomical organization characteristic of the isidioid structures arising from it. In Porina microcoralloides, the ascendant thallus consists of branched, coralloid inflated structures with phycobiont (Trentepohlia) unicells arranged at the periphery of a loose central medulla. Sparse fungal cells are interspersed and overlie the algal layer in places, but no differentiated cortex is present, leaving phycobiont cells more or less exposed at the surface. In the closely related Porina nanoarbuscula, the isidioid structures are much finer, more densely branched, and composed of a single, central file of roughly spherical Trentepohlia cells surrounded by a jacket of subglobose fungal cells. The ascospores of P. microcoralloides are more than twice the length of those of P. nanoarbuscula. Although thalli of these two Porina species occur in the same habitats and are sometimes found growing alongside each other, phylogenetic analysis of rbcL sequences suggest that they partner with distinct clades of Trentepohlia phycobionts. A third taxon examined, Porina cf. scabrida, is morphologically rather similar to P. microcoralloides, but the ascendant branches are bright yellow-orange, more cylindrical, and corticated by a thin layer of agglutinated fungal hyphae; perithecia were not seen. Analysis of mtSSU sequences places it distant from P. microcoralloides and P. nanoarbuscula phylogenetically. None of the Floridian taxa studied was particularly close to the European isidiate species Porina hibernica and P. pseudohibernica, which appeared as sister to each other in the analysis. While a particular type of isidiose structure may be reliably characteristic of specific taxa, similarities or differences in these structures do not seem to be useful indicators of phylogenetic proximity or distances among taxa. The morphological trends evident in Porina suggest that multiple transitions from crustose to isidioid or microfruticose growth have arisen repeatedly and in quite different ways within this single genus. At least some of the diverse structures treated within the broad concept of isidia may be representative of the developmental pathways by which fruticose growth forms may arise.

Phylogenetic insight into the Lecidea atrobrunnea complex – evidence of narrow geographic endemics and the pressing need for integrative taxonomic revisions

AbstractSpecies of lichen-forming fungi (LFF) display an array of geographical distribution patterns. Among the broadly distributed lichen-forming fungal species, the degree of reproductive isolation and genetic substructure among populations varies widely, in some cases masking unrecognized diversity or meaningful biogeographical patterns. Lecidea atrobrunnea (Raymond ex Lam. & DC.) Schaer. s. lat. (Lecideaceae) is a widespread species complex that has been studied for over two centuries since its initial description. The diversity of the L. atrobrunnea group is highest in western North America, where a dizzying array of morphologies and chemistry can occur at local scales. Here we investigate whether the assumed cosmopolitan distribution of L. atrobrunnea s. lat. is an artifact of taxonomic limitations and masks biogeographical patterns in this species complex. To address these questions, we compiled sequence data from the standard fungal barcoding marker (ITS) for over 100 specimens within this complex, in addition to genome-scale data from a subset of these representing over 1600 single-copy nuclear genes spanning over 3 Mb of the genome. Our study corroborates the perspective that the morphologically and chemically variable Lecidea atrobrunnea group reflects a complex of distinct species-level lineages, with 42–83 candidate species inferred from the ITS region and high levels of diversity inferred from a subset of specimens using genome-scale data. However, both phenotype- and molecular-based species boundaries remained unsettled, with the most common nominal taxa recovered as highly polyphyletic and with conflict among different molecular species delimitation approaches. Our study also highlights the potential for geographically restricted species, with fascinating biogeographical patterns, challenging, in part, the assumed cosmopolitan distribution of L. atrobrunnea s. lat. This study provides valuable direction for future research that will be crucial in understanding diversification and establishing a robust taxonomy for this well-known species complex.

Using morphological, chemical, and molecular data to study the diversity of Xanthoparmelia species from South Africa (Ascomycota, Parmeliaceae)

AbstractThere is still a high diversity of lichen-forming fungi that remains undescribed, especially cryptic lineages at the species level. Integrating morphological, chemical, and DNA sequence data has proved useful in corroborating species descriptions and delimitations. Here we reviewed morphological features, secondary metabolites and the DNA sequences of ITS, mtSSU and nuLSU markers to study the diversity of Xanthoparmelia in southern Africa. A total of 37 species were recorded. Three of these appear undescribed, and we therefore describe them here as new: Xanthoparmelia nimisii, with a sorediate thallus and broad lobes, is well supported as a clade separate from X. annexa; X. pseudochalybaeizans with a white medulla is phylogenetically distinct from the otherwise similar X. chalybaeizans; and X. sipmaniana, well supported as a separate clade from the similar X. hypoprocetrarica. In addition, the separation of Xanthoparmelia capensis and X. tinctina requires further studies.

A Custom Regional DNA Barcode Reference Library for Lichen-Forming Fungi of the Intermountain West, USA, Increases Successful Specimen Identification.

DNA barcoding approaches provide powerful tools for characterizing fungal diversity. However, DNA barcoding is limited by poor representation of species-level diversity in fungal sequence databases. Can the development of custom, regionally focused DNA reference libraries improve species-level identification rates for lichen-forming fungi? To explore this question, we created a regional ITS database for lichen-forming fungi (LFF) in the Intermountain West of the United States. The custom database comprised over 4800 sequences and represented over 600 formally described and provisional species. Lichen communities were sampled at 11 sites throughout the Intermountain West, and LFF diversity was characterized using high-throughput ITS2 amplicon sequencing. We compared the species-level identification success rates from our bulk community samples using our regional ITS database and the widely used UNITE database. The custom regional database resulted in significantly higher species-level assignments (72.3%) of candidate species than the UNITE database (28.3-34.2%). Within each site, identification of candidate species ranged from 72.3-82.1% using the custom database; and 31.5-55.4% using the UNITE database. These results highlight that developing regional databases may accelerate a wide range of LFF research by improving our ability to characterize species-level diversity using DNA barcoding.

Lineages of the lichen-forming fungus Stereocaulon alpinum and their photobionts in southern South America and maritime Antarctica

Lineages of the lichen-forming fungus Stereocaulon alpinum and their photobionts in southern South America and maritime Antarctica