This study investigates how anatomical characters can be used to differentiate two morphologically close species, Melanohalea olivacea (L.) O. Blanco et al. and M. septentrionalis (Lynge) O. Blanco et al. Although the two species are morphologically differentiated, mainly based on the position and shape of pseudocyphellae on the lobes, poorly-developed specimens can be difficult to identify based on morphology alone. For example, in Switzerland, both species are at the south-west limit of their distribution range and most specimens are small and not fully developed. In such cases, anatomical characters are particularly useful in separating the two species. Five anatomical characters were studied: spore length and width; hymenium, subhymenium and hypothecium height. A nested ANOVA showed that variation of spore length was 155 times greater between species than between individuals of the same species, itself 5 times higher than within individuals. In a Principal Component Analysis, despite a substantial variation of the anatomical characters within both species, a significant species differentiation appeared distinctly on the first axis (P < 0·001), which accounted for 62·35% of the total variance, and the type specimens nested within each species. Finally, a Discriminant Linear Analysis assigned 100% of the individuals to their species class, the best predictors to discriminate between the two species being subhymenium height and spore length. These characters proved to be powerful in identifying specimens with a poorly-developed morphology and confirmed the presence of M. olivacea in Switzerland.

A corticolous Graphis discovered in the southern part of Venezuela is described as new to science. The new species, Graphis breussii sp. nov., is characterized by rather large, prominent lirellae with striate labia and apically thin complete thalline cover, a completely carbonized exciple, four submuriform ascospores per ascus, and norstictic acid; it belongs in the Graphis acharii group.

Lichen cover and diversity were analysed on the dwarf shrub Helianthemum oelandicum (L.) Dum.Cours. at one site in the calcareous grassland of the area known as the ‘Great Alvar’ on the Island of Öland, Sweden. The age of 22 phorophytes was determined by ring counting and varied from 8 to 41 years and was accurately predicted by the root diameter. A total of 18 lichen species was found, with a range between 0 and 13 species per phorophyte. The number of lichen species on living H. oelandicum were correlated with the phorophyte age. The number of lichen species and their coverage were greater on dead compared with living phorophytes. The species number was also higher on thin branches compared with thick branches and roots and some of the species showed preferences for dead phorophytes, and for thin branches. This study of lichen colonization and growth on dwarf shrubs in relation to phorophyte age is a new application of herbchronology.

Phylogenetic relationships among the hypothetical species pairs Lobaria kurokawae / L. retigera and L. pseudopulmonaria / L. isidiosa were investigated based on TLC techniques and the phylogenetic analysis of the internal transcribed spacers (ITS) nrDNA. Results of TLC demonstrate that L. retigera and L. kurokawae can be chemically distinguished from L. isidiosa and L. pseudopulmonaria by the absence of retigeranic acid. Parsimony analysis of 18 specimens shows two monophyletic clades – a L.retigera lineage and a L. isidiosa lineage—both including their apotheciate counterparts. Unlike the original hypothesis of species pairs, our study reveals transitions from isidiate morphs that have the potential to rarely form apothecia, towards apotheciate morphs that produce no vegetative propagules.

The lichen Usnea longissima was used to examine how distance from forest edge and edge contrast influence growth of pendulous lichens. Thalli of two sizes (12 and 27 cm) were transplanted to the lower canopy of old Picea abies forest at 5, 25 and 100 m distance from cutovers. Sites represented three levels of edge contrast: high (clear-cut), intermediate (3 m tall saplings) and low (6–7 m tall young forest). Lichen growth was assessed as annual length and weight gain. Growth rates of intact thalli were size-dependent, with both growth variables being higher in long than in short thalli. Distance and edge contrast had significant effects on weight gain in long thalli but not in short ones. Weight gain in long thalli was twice as high near the edge (23%) compared to the forest interior (12%). The highest weight gain (31%) occurred at intermediate contrast edges with lower growth at both low (18%) and high contrast edges (20%). Chlorophyll a concentration was highest near the edge and positively correlated with weight gain, suggesting that growth was stimulated by both increased photosynthetic capacity and higher light availability near the edge. The lower part of the canopy in forest edges apparently have favourable growth conditions for U. longissima with growth being influenced by vegetation on adjoining cutovers. Therefore growth responses cannot explain the previously observed decline of pendulous lichens following edge creation. Our results suggest that vegetation buffers can improve conditions for pendulous lichens near forest edges.

Genetic variability and its relationship to substratum preferences within and among populations of the sorediate foliose lichen Hypogymnia physodes was investigated using sequence variation in the complete nrDNA internal transcribed spacer (ITS) region. A few samples of the putatively closely related, sorediate, H. tubulosa were also included. Samples were collected from each tree species in study sites in Estonia, Finland, and Sweden. In total, DNA sequences from 104 individuals of H. physodes and 16 of H. tubulosa were obtained. A group 1 intron situated at the end of the small subunit (SSU) of the nrDNA was detected in both species. Within-species variability was observed in both species: fifteen haplotypes were found for H. physodes and seven for H. tubulosa for the combined alignment of the intron and the ITS. Possible recombination within the total gene fragment was detected and hence the different regions (intron, ITS1, 5.8S, ITS2) were analysed separately. They show a different degree of variability both between each other and between the species. The number of haplotypes of H. physodes in the four regions are 5, 5, 1, and 5 and for H. tubulosa 5, 2, 1 and 2, respectively. A statistical parsimony estimation resulted in two unconnected networks; one containing all the samples of H. physodes and one containing all H. tubulosa samples. It was not possible to show different potentials of the different haplotypes for establishment on different substrata as the network of H. physodes indicates recombination within the ITS region which may be frequent enough to make this primarily clonally reproducing species to behave like a sexual species.

The relationship between thallus size and apothecium density was studied in two umbilicate lichens, Umbilicaria cylindrica and Lasallia pensylvanica, growing in alpine rock surface environments. Samples of the two lichens were collected from the Daisetsuzan National Park, northern Japan. Umbilicaria cylindrica produced many smaller apothecia, whereas L. pensylvanica had larger but fewer apothecia. To explain the differences in reproductive efforts between these two species, we analyzed the lichen data using a hierarchical Bayesian model in which the density of apothecia for each species was predicted by the thallus area and the sky openness index (SOI). The Bayesian model including unobservable effects, i.e. the random effects of individual lichens and rock faces, predicted that the apothecium density for both species increased as the thallus area increased. The estimated coefficient of thallus area to apothecium density for U. cylindrica was larger than that for L. pensylvanica. We also detected a probable positive relationship between SOI and apothecium density for U. cylindrica, whereas none was detected for L. pensylvanica.

The crustose lichen Lecanora conizaeoides has declined markedly around London with progressively diminishing sulphur dioxide pollution of the air since the 1960s. To identify the immediate causes of its decline, we applied S in the form of bisulphite (0·2 & 2 mM) and sulphate (2 mM), and N as nitrate (2 mM) to relict colonies of the lichen on beech trunks in a plantation in Windsor Forest. Growth of the lichen was monitored by estimating changes in percentage cover. By the end of a 25-month period of two-weekly treatments, all the chemical treatments had resulted in significant decreases in cover of L. conizaeoides compared to distilled water controls, with the 2 mM bisulphite causing the greatest loss. Bark surface pH was also lowered by the chemical treatments, but most by the 2 mM bisulphite applications. Similar results were obtained in two laboratory experiments where the nutrient applications were repeated under a controlled environment and thallus area monitored photographically. No evidence was obtained to support the hypothesis that growth of L. conizaeoides is stimulated by an elevated sulphur supply. We conclude that the disappearance of the lichen is linked to gradual increase in bark pH caused by the combined effects of a marked reduction in SO2 emissions and rising emissions of NH3.

The suitability of image analysis by colour-based pixel classification to quantify lichen colonization on the surface of and within marble, travertine and mortar stonework has been investigated. High resolution images of lichenized stonework surfaces were acquired at different field sites using a scanner, thus avoiding invasive surveys, and the percentage cover of lichen species was subsequently measured in the laboratory using dedicated software. Furthermore, microphotographs of polished cross-sections of lichenized marble, travertine and mortar, stained using the periodic acid-Schiff (PAS) method to visualize hyphae, were produced by the same software to quantify hyphal spread within the substratum, a parameter which can be used more successfully than the commonly used depth of hyphal penetration to quantify how much the lichen has affected the conservation of a stone substratum. Significant statistical differences in hue, saturation and intensity (HSI) of the lichen thalli and PAS-stained hyphae, with respect to the lithic substrata, allowed the software to discriminate and quantify the lichen species cover on, and hyphal spread within, the three investigated lithotypes. Since such a quantitative approach highlights the volume of influence of lichens on stonework, where bioweathering processes are likely to develop, it could be used to support decisions on the preservation of our stone cultural heritage.