The species–area relationship (SAR) states that species richness increases with the increase of the sampled area, although other factors can influence the pattern. SARs have been tested on many different organisms, but only rarely on lichens. We aimed to test the SAR, across a wide range of area sizes, for three main substratum-related guilds of lichens, namely epiphytic, epilithic and epigaeic. The test was performed using data from lichen inventories carried out in 44 protected areas of various sizes across Italy. We found a positive correlation of species richness with area size for all three guilds, better fitted by the logarithmic function for epilithic lichens and by the power function for epiphytic and epigaeic lichens. Our results support the fundamental role of area size as the main driver for lichen diversity, suggesting that in an area-based conservation framework, larger protected areas are fundamental to support high lichen species richness. However, finer scale investigations are also required to better elucidate whether and how other environmental factors could interact with area size and modify SAR patterns. Exhaustive lichen inventories could be useful information sources to more robustly test such relationships, and therefore better inform conservation practices.

Environmental control strategies are commonly practised to limit biodeterioration issues threatening indoor cultural heritage objects, while they are still poorly exploited for the conservation of outdoor stone heritage surfaces, including rock art. In this study, we evaluated the environmental factors driving the diversity and abundance of lithobiontic communities in the Rock Engravings National Park of Naquane (UNESCO WHS n. 94, Italy). The survey considered 23 rocks that had been cleaned in the last three (3YC) or twelve (12YC) years or more than 40 years ago (NRC). A cyanobacteria-dominated biofilm and lichens (37 taxa) were the most widespread and abundant lithobiontic components, prevailing on 3YC–12YC and NRC rocks, respectively. On the latter, a turnover of xerophytic and meso-hygrophytic lichen communities was observed. On 3YC–12YC rocks lichen colonization, if present, was limited to nitrophytic species, including common epiphytes from surrounding trees and a small number of meso-hygrophytic species, with a prevalence of asexual reproductive strategies. Multivariate analyses including environmental parameters (canonical correspondence analyses) indicated that tree cover and the presence of bare or vegetated ground upstream of the rocks, probably prolonging wetness and providing nutrients by water transport, are the factors mostly related to the microbial and lichen recolonization of 3YC–12YC surfaces. On this basis, an experiment on preventive conservation was conducted, consisting of a new cleaning of a strongly recolonized 3YC surface combined with the building of a small wall to protect part of the rock from prolonged water fluxes. The fluorimetric and colorimetric monitoring of the rock surface, carried out 40 months after this new cleaning intervention, displayed recolonization on the unprotected area only, indicating the potential of preventive conservation strategies in outdoor environments.

Peltigera globulata Miadl. & Magain, a new species in the P. ponojensis/monticola species complex of section Peltigera, is formally described. This clade was previously given the interim designation Peltigera sp. 17. It is found in sun-exposed and xeric habitats at high altitudes in Peru and Ecuador. Peltigera globulata can be easily recognized by its irregularly globulated margins covered mostly by thick, white pruina, somewhat resembling the sorediate thallus margins of P. soredians, another South American species from section Peltigera. The hypervariable region of ITS1 (ITS1-HR), which is in general highly variable among species of section Peltigera, does not have diagnostic value for species identification within the P. ponojensis/monticola complex. Nevertheless, no significant level of gene flow was detected among eight lineages representing a clade of putative species (including P. globulata) within this complex. ITS sequences from the holotype specimens of P. monticola Vitik. (collected in 1979) and P. soredians Vitik. (collected in 1981) and lectotype specimens of P. antarctica C. W. Dodge (collected in 1941) and P. aubertii C. W. Dodge (collected in 1952) were successfully obtained through Sanger and Illumina metagenomic sequencing. BLAST results of these sequences revealed that the type specimen of P. monticola falls within the P. monticola/ponojensis 7 clade, which represents P. monticola s. str., and confirmed that the type specimen of P. aubertii falls within a clade identified previously as P. aubertii based on morphology. The ITS sequence from the type specimen of P. soredians, which superficially resembles P. globulata, confirms its placement in the P. rufescens clade. Finally, we discovered that the name P. antarctica was erroneously applied to a lineage in the P. ponojensis/monticola clade. The ITS sequence from the type specimen of P. antarctica represents a lineage within the P. rufescens clade, which is sister to the P. ponojensis/monticola clade.

Porina 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.

A novel lichen species occurring on rocks was collected from three different localities within Deosai National Park, Gilgit-Baltistan, Pakistan. Phylogenetic analyses of the nrDNA ITS and nuLSU regions revealed that it clustered within the genus Anamylopsora. Further chemical and morpho-anatomical analyses confirmed its uniqueness, and it is described here as a new species under the name A. pakistanica. The distinguishing characters are: an irregularly squamulose appressed thallus on rocks without rhizines; an epinecral layer up to 25 μm thick; ascospores that are hyaline, simple, thick-walled with a smooth surface; septate paraphyses with a pigmented apical cell in a gel-like matrix; globose to subglobose pycnidia with hyaline and bacilliform pycnidiospores. In particular, the species is distinguished from other members of the genus by morpho-anatomical features including the coloration of the thalli, the presence of a thick lower cortex (up to 100 μm), and the presence of simple, thick-walled ascospores. Specimens were found at altitudes up to 4587 m, the highest elevation yet reported for Anamylopsora. A key and comparison to all existing species of the genus Anamylopsora is also given.

Lecanora caledonica is described as new to science. Molecular analyses show that it belongs to the L. intumescens group. It is also rather similar in appearance to L. intumescens, but differs mainly chemically in containing only atranorin and an unknown UV+ ice blue substance. There are also anatomical and morphological differences to the other species of the group. The new species has a pronounced oceanic distribution and is so far known only from western Norway and Scotland.

Microsatellite markers can provide valuable information about gene flow and population history. We developed and tested new microsatellites for the nitrophilic lichenized fungus Xanthoria parietina and studied its genetic diversity and structure within the urban area of Munich, Bavaria. We compared its local genetic pattern with that of its photobiont partner Trebouxia decolorans, for which existing microsatellites were applied. For comparison, a reference site with clean air was included in the sampling. We found support for three genetic clusters in the fungus X. parietina, which occurred intermingled in collecting sites. There was a high degree of admixture within fungal populations and individuals, and analysis of molecular variance revealed a lack of population structure in the mycobiont. The Trebouxia photobiont, in contrast, exhibited structured populations which grouped into two to five genetic clusters, and individuals showed less admixture than in the mycobiont. This indicates that the two lichen partners differ in their ability to move around in the landscape. The microsatellite markers we report are polymorphic and are suitable for population genetic studies.

An investigation of the genus Usnea, in the biodiversity hotspot of the Republic of São Tomé and Príncipe in tropical West Africa, is presented here. Fifteen species, or species aggregates, were recorded for the islands: Usnea articulata aggr., Usnea baileyi (Stirt.) Zahlbr., Usnea beckeri P. Clerc & Nadel, Usnea bicolorata Motyka, Usnea erinacea aggr., Usnea exasperata (Müll. Arg.) Motyka, Usnea firmula (Stirt.) Motyka, Usnea krogiana P. Clerc, Usnea longiciliata P. Clerc & Nadel, Usnea nodulosa Swinscow & Krog, Usnea pectinata aggr., Usnea sorediosula Motyka, Usnea submollis J. Steiner, and two undetermined species. Two species of lichen are described as new to science: U. beckeri and U. longiciliata. Both species are characterized by a dense and brittle, dark green thallus, the presence of apothecia surrounded by long cilia-like fibrils, a lack of soredia, and the presence of two unknown substances; however, whereas U. beckeri has a pendant growth form and can reach 25 cm in length, U. longiciliata is differentiated by a shrubby growth form of less than 6 cm. These two species also have different ascospore dimensions. A molecular phylogenetic analysis is presented that lends support to their description as new species. Additionally, Usnea krogiana is noted as new to continental Africa and nine Usnea species or aggregates are noted as new to the Islands of São Tomé and Príncipe in the Gulf of Guinea.

Additions and corrections are provided for the South African species of Graphidaceae tribe Graphideae with hyaline ascospores. Allographa oldayana I. Medeiros sp. nov. is described as new to science based on morphological, chemical and molecular data. The new species is characterized by lirellae with striate labia and a complete thalline margin, a completely carbonized excipulum, large, muriform ascospores, and the presence of hirtifructic acid. Allographa consanguinea (Müll. Arg.) Lücking, A. leptospora (Vain.) Lücking & Kalb, Diorygma aff. minisporum Kalb et al., Graphis crebra Vain., Gr. dupaxana Vain., Gr. furcata Fée, Gr. handelii Zahlbr., Gr. longula Kremp., Gr. pinicola Zahlbr., Gr. proserpens Vain, Gr. subhiascens (Müll. Arg.) Lücking and Platythecium sp. are reported as new records for South Africa. Allographa striatula (Ach.) Lücking & Kalb, Graphis analoga Nyl. and Gr. scripta (L.) Ach. are shown to be misapplied names that should be removed from the South African checklist. The new combination Mangoldia bylii (Vain.) I. Medeiros comb. nov. (bas. Graphis bylii Vain) is made; this represents an earlier name for M. atronitens (A. W. Archer) Lücking et al. Taxonomic notes are provided for Graphis bylii var. lividula Vain. and Gr. denudans Vain., species that are known only from their South African holotypes. Phylogenetic analyses that include new DNA sequence data from the nrLSU, mtSSU and RPB2 loci confirm the generic placements of several species for which molecular data were lacking: Allographa consanguinea, Glyphis atrofusca (Müll. Arg.) Lücking, Graphis crebra and Gr. subhiascens.

Recent studies have demonstrated that species boundaries among the lichen-forming fungi are in need of revision with the discovery of cryptic species in numerous clades, especially in parmelioid lichens. Here we focus on addressing the species boundaries in Canoparmelia texana, a sorediate species with a pantropical distribution that extends into temperate regions. We extracted DNA sequences of the nuclear ribosomal internal transcribed spacer region (ITS), large subunit (nuLSU) and mitochondrial small subunit (mtSSU) from samples mostly collected in Kenya, and analyzed them in a phylogenetic framework. We illustrate that our samples of the species as currently circumscribed do not form a monophyletic group but fall into two distinct clades, with the apotheciate C. nairobiensis nested within. Both of the discovered lineages have a wide distributional range and are common in Kenya, and Parmelia albaniensis C. W. Dodge is resurrected to accommodate one of the clades; consequently a new combination, Canoparmelia albaniensis (C. W. Dodge) Divakar & Kirika comb. nov., is proposed.

Tropical mountain forests are hotspots of biodiversity that are widely threatened by human population pressure and climate change. However, the cryptogamic species richness of many tropical mountain regions is insufficiently known, the poorly understood biodiversity of tropical African lichens being a prime example. To study the diversity of the genus Peltigera (Ascomycota, Lecanoromycetes) in East Africa, we studied lichens in a wide range of habitats on the slopes of Mt Kilimanjaro in Tanzania. Ranging from savannah to alpine heath vegetation and from natural forests to agricultural environments, 13 habitat types were sampled for lichens, which were then identified based on the nuITS genetic marker and morphology. We found eight Peltigera species on the slopes of Mt Kilimanjaro, including P. alkalicola sp. nov., P. dolichorhiza, P. polydactyloides, P. praetextata, P. rufescentiformis, P. seneca, P. sorediifera and P. ulcerata. Peltigera is most common and species-rich in the subalpine Erica forest zone, and four of the eight detected species were present only in the subalpine and alpine vegetation zones. Peltigera alkalicola was identified as a previously undescribed species, growing on trachybasaltic lava in the subalpine and alpine zones of Mt Kilimanjaro. The species resembles P. lepidophora but differs by possessing smaller thalli and peltate isidia that are distinctly dark on the lighter, tomentose lamina. Based on data from the NCBI GenBank, P. alkalicola probably also occurs in Alaska (USA) and Ningxia (China). This suggests that even though the species might generally be rare, it may have a global distribution in extreme mountain environments. For the first time, we report P. sorediifera from Tanzania and P. seneca from Africa.

The widespread species Parmotrema crinitum (Ach.) M. Choisy and Parmotrema perlatum (Huds.) M. Choisy are mainly distinguished by their reproductive strategies. While P. crinitum propagates by isidia, P. perlatum produces soredia. In this study, we aim to evaluate the phylogenetic relationship between both species and to critically examine their species boundaries. To this purpose, 46 samples belonging to P. crinitum and P. perlatum were used in our analysis, including 22 for which we studied the morphology and chemistry, before extracting their DNA. We used 35 sequences of the internal transcribed spacer region of nuclear ribosomal DNA (ITS) of Parmotrema perlatum from Europe and Africa (20 of which were newly generated), and 11 of Parmotrema crinitum from Europe, North America and North Africa (two newly generated). Additionally, 28 sequences of several species from Parmotrema were included in the ITS dataset. The ITS data matrix was analyzed using different approaches, such as traditional phylogeny (maximum likelihood and Bayesian analyses), genetic distances, automatic barcode gap discovery (ABGD) and the coalescent-based method poisson tree processes (PTP), in order to test congruence among results. Our results indicate that all samples referred to P. crinitum and P. perlatum nested in a well-supported monophyletic clade, but phylogenetic relationships among them remain unresolved. Delimitations inferred from PTP, ABGD and genetic distance analyses were comparable and suggested that P. crinitum and P. perlatum belong to the same lineage. Interestingly, two samples of P. perlatum separate in a different monophyletic clade, which is supported as a different lineage by all the analyses.

The epiphytic lichen species richness and community composition was compared for 600 living trees distributed within the interior of 60 Scots pine and Norway spruce monoculture stands in southern Sweden. A higher species richness, and more unique species, was found on trees of Scots pine than of Norway spruce, and distinctive communities were associated with the two tree species. Lichen species composition also shifted between the 30-, 55- and 80-year-old stands, although there was no significant difference in species richness between the different age classes. Tree species and age of the stand explained most of the variation in community composition (41%), with additional variance explained by lichen proximity to the ground (6%) and aspect (1%) (northern/southern side of trunk). Scots pine and Norway spruce share many attributes, such as both being conifers with acidic bark properties and having a similar geographical distribution in Fennoscandia. However, our study showed that species richness and community composition can nevertheless diverge in stands dominated by these two tree species. Since the occurrence of red-listed species was low in these stands, we suggest that 80-year rotations are not long enough for the occurrence of the many rare and specialized lichen species that require old forest structures and long forest continuity in this region.

Trees have a crucial importance in the functioning of ecosystems on Earth. They are among the largest and longest-living taxa and provide habitat and shelter to numerous species belonging to diverse groups of organisms. Relict trees are of particular interest through their history of survival and adaptation, and because they potentially shelter rare or threatened organisms today. We investigated for the first time the diversity and distribution of epiphytic lichens and bryophytes found on the Cretan (Greek) endemic and relict phorophyte Zelkova abelicea (Ulmaceae). Our results showed that Z. abelicea hosts a high number of epiphytes. The Levka Ori mountain range in western Crete seems to be a hot spot for epiphytic lichens on Z. abelicea. Bryophytes had the highest diversity on Mt Kedros in central Crete but were absent from several other sites. Moreover, 17% of the studied lichens were recorded for the first time for Crete and 5% have never been recorded for Greece. Geographical position and browsing intensity seem to be important factors influencing the epiphytic community encountered. Tree morphology (dwarfed or arborescent) was also significant in influencing community composition although it was not possible to dissociate this factor from the effect of topography. Dwarfed individuals were found to have as much epiphytic diversity as arborescent trees. Ecological indicator values showed that high epiphytic diversity was found in some sites despite signs of eutrophication and disturbance due to pastoral activities and suggest the co-occurrence of both disturbance tolerant and sensitive species. Our results show how little is known about the biodiversity of Cretan phorophytes and highlights the need for further research on the topic.

Gene expression variation can be partitioned into different components (regulatory, genetic and acclimatory effects) but for lichen-forming fungi, the relative importance of each of these effects is unclear. Here, we studied gene expression in the lichen-forming fungus Lobaria pulmonaria in response to thermal stress and parasitism by the lichenicolous fungus Plectocarpon lichenum. Our experimental procedure was to acclimate lichen thalli to 4 °C over three weeks and then expose them to 15 °C and 25 °C for 2 hours each, sampling infected and visually asymptomatic thalli at each temperature. Quantitative Real-Time PCR was utilized to quantify gene expression of six candidate genes, normalizing expression values with two reference genes. We found that two genes encoding heat shock proteins (hsp88 and hsp98), two polyketide synthase genes (rPKS1, nrPKS3) and elongation factor 1-1-α (efa) were upregulated at higher temperatures. Moreover, we observed higher expression of hsp98 at 25 °C in samples infected by P. lichenum than in uninfected samples. Finally, in partial redundancy analyses, most of the explained variation in gene expression was related to temperature treatment; genetic variation and long-term acclimatization to sites contributed far less. Hence, regulatory effects (i.e. direct adjustments of gene expression in response to the temperature change) dominated over genetic and acclimatory effects in the gene expression variability of L. pulmonaria. This study suggests that L. pulmonaria could become a valuable lichen model for studying heat shock protein responses in vivo.

The Rostania occultata species complex (‘Collema occultatum s. lat.’) is revised in Fennoscandia and found to consist of four species, all epiphytes on deciduous trees: Rostania effusa A. Košuth., M. Westb. & Wedin sp. nov., R. occultata (Bagl.) Otálora et al., R. pallida A. Košuth., M. Westb. & Wedin sp. nov. and R. populina (Th. Fr.) A. Košuth., M. Westb. & Wedin comb. nov. Rostania effusa and R. pallida are newly described from humid habitats in old-growth boreal coniferous forests, usually with a mixture of deciduous trees, and from similar areas in the subalpine birch-dominated forests of Fennoscandia. Rostania effusa is characterized by apothecia with red-brown apothecium discs and an excipulum thallinum with a simple pseudocortex and cubic to oblong, muriform spores. Rostania pallida has apothecia with whitish to pale yellowish discs and an excipulum thallinum with a distinct cellular pseudocortex, and ellipsoid, muriform mature spores that are often constricted at the centre. A lectotype is designated for Collema quadratum J. Lahm ex Körb. The new combination Rostania populina is introduced for the species recognized until now as the variety Rostania occultata var. populina (Th. Fr.) Perlmutter & Rivas Plata. A key to the six species in Rostania s. str. is included.

To clarify deep relationships among species lineages within Bacidia s. str., and to investigate the robustness of the deeper branches, we combined data from three traditionally used RNA-coding genes (nrITS, nrLSU and mtSSU) with two protein-coding genes (RPB1 and RPB2). The multigene phylogeny contained 48 newly generated sequences from the Russian Far East and all Bacidia s. str. sequences from GenBank (131 sequences). We subjected the alignments for the single and concatenated data sets to Bayesian inference (BI) and two maximum likelihood (ML) analyses (RAxML and IQ-TREE). The topologies of phylogenetic trees recovered from BI and ML analyses were highly concordant. The multilocus phylogeny of Bacidia s. str. was congruent with previous results based on nrITS sequences from the Russian Far East but with considerably higher support values for most of the deeper branches. A correlation between the recovered clades and apothecial pigments in the upper part of the hymenium and lateral exciple was observed. Based on morphological and molecular evidence, Bacidia obtecta is described as new to science. It was recovered as the sister lineage of B. elongata. The two species are alike in having up to four enlarged lumina cells along the exciple edge, but B. obtecta differs in the abundant crystals found in the upper hymenium and lateral exciple, and by having spores with fewer septa.

To facilitate population-genetic studies, we developed simple sequence repeat (SSR) markers and a molecular species identification assay for Peltigera membranacea (Ascomycota, Peltigerales), a common ground-dwelling lichen of forest and tundra ecosystems. Additional markers were developed for its Nostoc photobiont. Twenty-one fungal markers for P. membranacea were found to be polymorphic, with the number of alleles ranging from 3–21. Nei's unbiased gene diversity ranged from 0.588 to 0.640 in four significantly structured (FST = 0.059) mycobiont populations. For the Nostoc photobiont, 14 polymorphic SSR were developed, yielding 4–14 alleles each, with gene diversity ranging from 0.062 to 0.771 in four populations showing substantial population structure (FST = 0.278). The new markers developed are suitable for population genetic studies of Peltigera membranacea and of its cyanobiont, and at the same time allowed us to distinguish 98.5% of P. membranacea specimens from morphologically similar species of Peltigera.

A review of algal (including cyanobacterial) symbionts associated with lichen-forming fungi is presented. General aspects of their biology relevant to lichen symbioses are summarized. The genera of algae currently believed to include lichen symbionts are outlined; approximately 50 can be recognized at present. References reporting algal taxa in lichen symbiosis are tabulated, with emphasis on those published since the 1988 review by Tschermak-Woess, and particularly those providing molecular evidence for their identifications. This review is dedicated in honour of Austrian phycologist Elisabeth Tschermak-Woess (1917–2001), for her numerous and significant contributions to our knowledge of lichen algae (some published under the names Elisabeth Tschermak and Liesl Tschermak).

The evolution of multicellularity paved the way for significant increases in biological complexity. Although multicellularity has evolved many times independently, we know relatively little about its origins. Directed evolution is a promising approach to studying early steps in this major transition, but current experimental systems have examined only a subset of the possible evolutionary routes to multicellularity. Here we consider egalitarian routes to multicellularity, in which unrelated unicellular organisms evolve to become a multicellular organism. Inspired by microbial syntrophies and lichens, we outline three such routes from a system of different species to an interdependent relationship that replicates. We compare these routes to contemporary experimental systems and consider how physical structure, the threat of invasion, division of labour and co-transmission affect their evolution.