Two Basidiomycete Fungi in the Cortex of Wolf Lichens.

Since the late 1800s, mycologists have been detecting fungi above and beyond the assumed single fungus in lichen thalli [1-6]. Over the last century, these fungi have been accorded roles ranging from commensalists to pathogens. Recently, Cyphobasidiales yeasts were shown to be ubiquitous in the cortex layer of many macrolichens [7], but for most species, little is known of their cellular distribution and constancy beyond visible fruiting structures. Here, we demonstrate the occurrence of an additional and distantly related basidiomycete, Tremella, in 95% of studied thalli in a global sample of one of the most intensively studied groups of lichens, the wolf lichens (genus Letharia). Tremella species are reported from a wide range of lichen genera [8], but until now, their biology was deduced from fruiting bodies (basidiomata) formed on lichen thalli. Based on this, they have been thought to be uncommon to rare, to occur exclusively in a hyphal form, and to be parasitic on the dominant fungal partner [9, 10]. We show that, in wolf lichens, Tremella occurs as yeast cells also in thalli that lack basidiomata and infer that this is its dominant stage in nature. We further show that the hyphal stage, when present in Letharia, is in close contact with algal cells, challenging the assumption that lichen-associated Tremella species are uniformly mycoparasites. Our results suggest that extent of occurrence and cellular interactions of known fungi within lichens have historically been underestimated and raise new questions about their function in specific lichen symbioses.

Morphophysiological variation and metal concentration in the thallus of Parmotrema tinctorum (Despr. ex Nyl.) Hale between urban and forest areas in the subtropical region of Brazil.

Anthropic activities such as the emission of pollutants resulting from industrial and agropastoral activities promote several changes in urban and forest areas. Lichens are organisms that are used in air quality evaluations due to their sensitivity to these changes. The aim of this study is to analyze the presence of morphophysiological damages and the metal concentration in samples of the lichen Parmotrema tinctorum, in urban and forest areas, checking for possible parameter variations between these areas, in the different matrices and seasons in the Southern region of Brazil. Six areas were selected (urban and forest), assigned to the rural/urban and urban/industrial matrices belonging to the watershed of the Rio dos Sinos, Brazil. The following parameters were analyzed: Index of photobiont vitality (IPV), photosynthetic pigments (chlorophyll, carotene, and pheophytin) and metals (Cu, Cr, Zn, Pb, and Ni) in the thallus of the lichen. The data were analyzed by an ANOVA one way, Pearson correlation test, and principal component analysis (PCA). Variations in the morphophysiological parameters were recorded in all the areas revealing significant differences. The lowest IPV values and highest concentration of metals were recorded in the urban environment, while low chlorophyll levels were found in the forest areas. The PCA showed a distinction between the areas and the season. The level of urbanization, vehicle traffic, and the weather conditions might have influenced the results. The use of P. tinctorum, the index of photobiont vitality and chlorophyll and pheophytin content, has proved to be an efficient tool to diagnose the air quality in the areas analyzed, allowing its use as a model in air monitoring studies, both in urban and forest areas, as well as in distinct matrices in the subtropical region.

Sharing of photobionts in sympatric populations of Thamnolia and Cetraria lichens: evidence from high-throughput sequencing.

In this study, we explored the diversity of green algal symbionts (photobionts) in sympatric populations of the cosmopolitan lichen-forming fungi Thamnolia and Cetraria. We sequenced with both Sanger and Ion Torrent High-Throughput Sequencing technologies the photobiont ITS-region of 30 lichen thalli from two islands: Iceland and Öland. While Sanger recovered just one photobiont genotype from each thallus, the Ion Torrent data recovered 10-18 OTUs for each pool of 5 lichen thalli, suggesting that individual lichens can contain heterogeneous photobiont populations. Both methods showed evidence for photobiont sharing between Thamnolia and Cetraria on Iceland. In contrast, our data suggest that on Öland the two mycobionts associate with distinct photobiont communities, with few shared OTUs revealed by Ion Torrent sequencing. Furthermore, by comparing our sequences with public data, we identified closely related photobionts from geographically distant localities. Taken together, we suggest that the photobiont composition in Thamnolia and Cetraria results from both photobiont-mycobiont codispersal and local acquisition during mycobiont establishment and/or lichen growth. We hypothesize that this is a successful strategy for lichens to be flexible in the use of the most adapted photobiont for the environment.

Panmixia and dispersal from the Mediterranean Basin to Macaronesian Islands of a macrolichen species.

The Mediterranean region, comprising the Mediterranean Basin and the Macaronesian Islands, represents a center of diversification for many organisms. The genetic structure and connectivity of mainland and island microbial populations has been poorly explored, in particular in the case of symbiotic fungi. Here we investigated genetic diversity and spatial structure of the obligate outcrossing lichen-forming fungus Parmelina carporrhizans in the Mediterranean region. Using eight microsatellite and mating-type markers we showed that fungal populations are highly diverse but lack spatial structure. This is likely due to high connectivity and long distance dispersal of fungal spores. Consistent with low levels of linkage disequilibrium and lack of clonality, we detected both mating-type idiomorphs in all populations. Furthermore we showed that the Macaronesian Islands are the result of colonization from the Mediterranean Basin. The unidirectional gene flow, though, seemed not to be sufficient to counterbalance the effects of drift, resulting in comparatively allelic poor peripheral populations. Our study is the first to shed light on the high connectivity and lack of population structure in natural populations of a strictly sexual lichen fungus. Our data further support the view of the Macaronesian Islands as the end of the colonization road for this symbiotic ascomycete.

Ecophysiological and ultrastructural effects of dust pollution in lichens exposed around a cement plant (SW Slovakia).

The study investigated the ecophysiological and ultrastructural effects of dust pollution from a cement industry in the lichen species Evernia prunastri and Xanthoria parietina, which were exposed for 30, 90 and 180 days around a cement mill, two quarries, and inhabited and agricultural sites in SW Slovakia. The results showed that dust deposition from quarrying activities and cement works at the cement mill (mainly enriched in Ca, Fe and Ti) significantly affected the photosynthetic apparatus of E. prunastri (sensitive to dust and habitat eutrophication), while X. parietina (tolerant to dust and habitat eutrophication) adapted to the new environment. The length of the exposure strongly affected the vitality of the mycobiont (measured as dehydrogenase activity) in transplanted lichens. Dust deposition led to ultrastructural alterations, including lipid droplets increase, swelling of cellular components, thylakoid degeneration and sometimes plasmolysis, which, on the whole, gave the cells an aged appearance. Photosynthetic parameters deserve further attention as potential indicators for monitoring early biological symptoms of the air pollution caused during cement production.