Starting from scratch: Evolution of the lichen thallus in the basidiolichen Dictyonema (Agaricales: Hygrophoraceae)

Phylogenetic studies indicate that the basidiolichen genus Dictyonema s.lat., often thought to represent only a single genus with few species, includes several well-supported genus-level clades, all of which form associations with a unique lineage of obligately lichenized cyanobacteria (Rhizonema). In an attempt to elucidate the evolution and genus- and species-level diversification in Dictyonema s.lat., we generated 68 new sequences of the nuclear large subunit rDNA (nuLSU), the internal transcribed spacer (ITS), and the RNA polymerase II subunit (RPB2), for 29 species-level lineages representing all major clades of Dictyonema s.lat. and most of the species currently known. The multilocus phylogeny obtained via maximum likelihood and Bayesian approaches indicates the presence of five genus-level groups: a basal clade, Cyphellostereum, that is sister to the rest of the species, a paraphyletic grade representing Dictyonema s.str., and three clades representing the genera Acantholichen, Cora, and Corella. To determine the evolutionary transformations of the lichenized thallus in the group, ancestral character state reconstruction was done using six characters (lichenisation, thallus type, cortex type, hyphal sheath and haustorial type, photobiont morphology, and basidiocarp type). Our analysis indicates a progressive development of the lichenized thallus from loosely organized filamentous crusts with separate, cyphelloid basidiocarps in Cyphellostereum, to filamentous crusts with derived hyphal sheath and cyphelloid–stereoid basidiocarps partially incorporated into the lichen thallus in Dictyonema, to squamulose–foliose thalli with corticioid basidiocarps entirely supported by the lichen thallus in Cora. These results indicate a remarkable evolutionary integration of lichenized and reproductive tissues in Dictyonema s.lat., supporting the hypothesis that, at least in this case, lichenized thalli may have evolved from reproductive structures in their nonlichenized ancestors.     

The genus Calycidium Stirt.

Abstract:The lichen genus Calycidium Stirt. (Calycidiaceae,Lecanorales , ‘Caliciales’ s. lat.) contains two species: C. cuneatum Stirt., distributed in Australia (Tasmania) and New Zealand, and C. polycarpum (Colenso) Wedin, comb. nov., distributed in Argentina, Australia (Tasmania), Chile and New Zealand. The morphology, chemistry, ecology and distribution of the two species is discussed, as is the taxonomic position of the genus. Calycidium is reported as new to Argentina.     

Systematics and evolution of the genus Torrubiella (Hypocreales, Ascomycota)

Torrubiella is a genus of arthropod-pathogenic fungi that primarily attacks spiders and scale insects. Based on the morphology of the perithecia, asci, and ascospores, it is classified in Clavicipitaceae s. lat. (Hypocreales), and is considered a close relative of Cordyceps s. 1., which was recently reclassified into three families (Clavicipitaceae s. str., Cordycipitaceae, Ophiocordycipitaceae) and four genera (Cordyceps s. str, Elaphocordyceps, Metacordyceps, and Ophiocordyceps). Torrubiella is distinguished morphologically from Cordyceps s. lat. mainly by the production of superficial perithecia and the absence of a well-developed stipitate stroma. To test and refine evolutionary hypotheses regarding the placement of Torrubiella and its relationship to Cordyceps s. lat., a multi-gene phylogeny was constructed by conducting ML and Bayesian analyses. The monophyly of Torrubiella was rejected by these analyses with species of the genus present in Clavicipitaceae, Cordycipitaceae, and Ophiocordycipitaceae, and often intermixed among species of Cordyceps s. lat. The morphological characters traditionally used to define the genus are, therefore, not phylogenetically informative, with the stipitate stromata being gained and/or lost several times among clavicipitaceous fungi. Two new genera (Conoideocrella, Orbiocrella) are proposed to accommodate two separate lineages of torrubielloid fungi in the Clavicipitaceae s. str. In addition, one species is reclassified in Cordyceps s. str. and three are reclassified in Ophiocordyceps. The phylogenetic importance of anamorphic genera, host affiliation, and stipitate stromata is discussed.     

Evolutionary relationships among narrow-headed rats (genus Stenocephalemys, muridae, rodentia) inferred from complete cytochrome b gene sequences

Using the data on complete sequences of cytochrome b gene, phylogenetic relationships were studied among the Stenocephalemys s. lat. (Stenocephalemys spp. + Praomys albipes) murine rodents, inhabiting adjacent altitudinal belts of the isolated Ethiopian mountain massifs, and among the related Praomys s. lat. species. Extremely low resolution of the relationships among the main Praomys s. lat. lineages hampered identification of the nearest sister group for the Stenocephalemys s. lat. “Ethiopian” clade, monophyly of which was strongly supported. Sister relationships between P. albipes and S. griseicauda (implying “accelerated” morphological and chromosomal evolution upon the formation of the former species), as well as between S. albocaudata and the recently described novel chromosomal form of Stenocephalemus sp. A (2n = 50; NFa = 56) were demonstrated. Definite discordance between the rates of their molecular, chromosomal, and morphological evolution was revealed. Based on phylogenetic reconstructions and the estimates of the divergence time, obtained by use of molecular clock method, an attempt to draw an evolutionary scenario for the group examined was made. The obtained data were compared to those for Sigmodontinae species complexes, distributed across a marked altitudinal gradient on the Andean slopes. It was shown that molecular genetic data on the rodents from mountain tropics did not support the gradient model of diversification, based on the possibility of diversification of the forms up to their achievement of the species rank (without interruption of the gene flow between them) due to differently directed selection across a strong environmental gradient.     

Lichen substance concentrations in the lichen Hypogymnia physodes are correlated with heavy metal concentrations in the substratum

Lichen substances (i.e. lichen-specific carbon-based secondary compounds) are known to be involved in the uptake and immobilization of metal ions, though the biochemical mechanisms of this interaction are largely unexplained. Previous research on potential effects of lichen substances on heavy metal uptake and tolerance mostly focused on lichens in heavily polluted areas with exceptionally high metal concentrations. In the present study, we aimed at gathering information as to whether lichen substances might be involved in the fine-tuning of metal uptake even at not or low-polluted sites. Therefore, we studied lichen substance concentrations in the epiphytic lichen Hypogymnia physodes and metal concentrations in its substratum in a montane spruce forest of Germany. H. physodes produces two depsides and five depsidones, which had been shown to be involved in metal homeostasis, namely in Cu and Mn uptake, in previous laboratory experiments. The amount of lichen substances increased with increasing heavy metal concentration in the substratum, though the latter varied only in the range of a few μmol g⁻¹ between the sample trees. Variability of lichen substance concentrations in H. physodes within the individual trees was low. Among the different lichen substances of H. physodes, the amount of the depsidone physodalic acid relative to the total of lichen substances was most closely correlated to the concentrations of Cu and Mn in the substratum, whereas the amount of the depsidone 3-hydroxyphysodic acid decreased both with increasing concentrations of these two metals and physodalic acid. Thus, our data suggest that lichen substances contribute to metal homeostasis not only in heavy metal-rich habitats, but also at not or low-polluted sites where the lichen substances apparently help to maintain constant intracellular metal concentrations despite of spatially varying availabilities of metal ions.     

Evaluation of Antimicrobial Properties of Lichen Substances against Plant Pathogens

Plant pathogens pose major threats on agriculture and horticulture, causing significant economic loss worldwide. Due to the continuous and excessive use of synthetic pesticides, emergence of pesticide resistant pathogens has become more frequent. Thus, there is a growing needs for environmentally-friendly and selective antimicrobial agents with a novel mode of action, which may be used in combination with conventional pesticides to delay development of pesticide resistance. In this study, we evaluated the potentials of lichen substances as novel biopesticides against eight bacterial and twelve fungal plant pathogens that have historically caused significant phytopathological problems in South Korea. Eight lichen substances of diverse chemical origins were extracted from axenic culture or dried specimen, and further purified for comparative analysis of their antimicrobial properties. Usnic acid and vulpinic acid exhibited strong antibacterial activities against Clavibacter michiganensis subsp. michiganensis. In addition, usnic acid and vulpinic acid were highly effective in the growth inhibition of fungal pathogens, such as Diaporthe eres, D. actinidiae, and Sclerotinia sclerotiorum. Intriguingly, the growth of Rhizoctonia solani was specifically inhibited by lecanoric acid, indicating that lichen substances exhibit some degrees of selectivity to plant pathogens. These results suggested that lichen substance can be used as a selective biopesticide for controlling plant disease of agricultural and horticultural significance, minimizing possible emergence of pesticide resistant pathogens in fields.     

Contrasting changes in palatability following senescence of the lichenized fungi Lobaria pulmonaria and L. scrobiculata

Epiphytic lichens can contribute significantly to ecosystem nutrient input because they efficiently accumulate atmospheric mineral nutrients and, in the case of cyanolichens, also fix nitrogen. The rate at which carbon and other nutrients gained by lichens enters the ecosystem is determined by lichen litter decomposability and by invertebrate consumption of lichen litter. In turn, these processes are driven by the secondary compounds present in senesced lichens. Therefore, we explored how lichen palatability and concentrations of secondary compounds change with tissue senescence for Lobaria pulmonaria, a green-algal lichen with cyanobacterial cephalodia, and Lobaria scrobiculata, a cyanobacterial lichen. During senescence both lichens lost 38–48 % of their stictic acid chemosyndrome, while m-scrobiculin and usnic acid in L. scrobiculata remained unchanged. Snails preferred senesced rather than fresh L. pulmonaria, while senesced L. scrobiculata were avoided. This provides evidence that species with labile secondary compounds will have higher turnover rates, through consumption and decomposition, than those producing more stable secondary compounds.     

Antimitotic activity of lobaric acid and a new benzofuran,sakisacaulon A from Stereocaulon sasakii

Lobaric acid (1) has been isolated from lichen, Stereocaulon sasakii together with a new benzofuran, sakisacaulon A (2). Lobaric acid (1) inhibited the polymerization of tubulin. Structure–activity relationship of lobaric acid and its derivatives on inhibitory activity of tubulin polymerization was discussed.     

The occurrence of biomineralization products in four lichen species growing on sandstone in western Norway

High performance thin-layer chromatography/thin-layer chromatography, X-ray diffraction, and scanning electron microscopy analysis of thallus and lichen-rock interface samples, were undertaken to characterize biomineralization products in Fuscidea cyathoides, Ochrolechia tartarea, Ophioparma ventosa, and Pertusaria corallina, growing on sandstone in western Norway. Whewellite (monohydrate form of Ca oxalate) was found in the thallus of all species, but not in any of the weathering rinds beneath the species. A significantly higher amount of whewellite was detected in the thalli of F. cyathoides and O. ventosa than in the other two species. There were only a few differences in whewellite occurrence between the thallus edge and centre samples in the four species. HPTLC/TLC and SEM analysis indicate that lichen compounds occur within the rock beneath some of the lichen specimens. Only divaricatic acid was observed within the weathering rind beneath O. ventosa. No lichen substances were found in the weathering rind beneath F. cyathoides and P. corallina, whereas gyrophoric and lecanoric acids were found in the weathering rind beneath O. tartarea.     

3-Chlordivaricatsäure,ein neues depsid aus Thelomma-arten

The structure of 3-chlorodivaricatic acid, a new depside from the lichen Thelomma mammosum was elucidated by ¹H NMR, ¹³C NMR, mass and UV spectroscopy. The synthesis of 5-chlorodivaricatic acid is described.     

(+)-Isorangiformic acid,a lichen substance from Lecanora stenotropa

(+)-Isorangiformic acid from the lichen Lecanora stenotropa has been shown to be (+)-2S-methoxycarbonyl-3S-heptadecanedicarboxylic acid.     

Krogia coralloides,a new lichen genus and species from Mauritius

Abstract:The new corticolous lichen genus Krogia and species K. coralloides are described from humid forest in Mauritius. The genus shows affinities with Phyllopsora, but differs in characters of the ascus, ascospores, and thallus anatomy. The ascal characters make its inclusion in the Bacidiaceae orPhyllopsoraceae problematic. The lichen contains boninic acid and an apparently related compound.     

Depsidone constituents from the quintaria group of Nephroma species

A new lichen depsidone was isolated, in the form of its triacetate derivative from the acetylated extracts of Nephroma antarcticum and has been demonstrated to be hypoconstictic acid-triacetate. Two related depsidones, hypostictic acid hyposalazinic acid, were isolated from N. australe.     

Do lichen secondary compounds play a role in highly specific fungal parasitism?

Chemical interactions between highly host-specific lichenicolous fungi and their lichen hosts have been little studied. In an allometric study, we quantified carbon-based secondary compounds (CBSCs) in a mixed natural Lobarina scrobiculata population (N = 147) of the normal and the stictic acid-deficient chemotypes, both with and without galls of Plectocarpon scrobiculatae. We assessed the correlation between the presence/abundance of parasite galls and the lichen CBSCs contents, and quantified size-dependent contents of CBSCs. The parasite produced galls similarly in both chemotypes, indicating that the stictic acid complex does not deter Plectocarpon. Within both chemotypes, thalli with Plectocarpon had half the contents of all individual CBSCs than those without galls. There was a significant size-dependent increase in CBSC contents in thalli without galls, but not in those with. This study shows that lichen chemistry is involved in highly host-specific fungal parasitism, and widens our knowledge of specialized biotrophic fungal interactions.     

Biocidal effect of lichen secondary metabolites against rock-dwelling microcolonial fungi,cyanobacteria and green algae

The use of commercial biocides in outdoor environments is increasingly discouraged because of their ecotoxicity, new methods being thus invoked to control patinas of biological origin on the stone cultural heritage. The effects of secondary metabolites (usnic acid, norstictic acid, parietin) produced by saxicolous lichens, natural competitors of rock dwelling microorganisms, were investigated in vitro against microcolonial fungi (MCF: Coniosporium apollinis, Coniosporium perforans, Coniosporium uncinatum, Phaeococcomyces-like sp.), coccoid cyanobacteria (Chroococcus minutus) and green algae (Scenedesmus ecornis) which commonly occur on stonework. An acetone/water 10/90 vol/vol mixture was screened as suitable to solubilise the lichen metabolites and to not affect the bioassay results. Benzalkonium chloride 1% was used as positive control.All the three metabolites (approx. 10⁻² mM) inhibited the growth of the assayed MCF species, displaying the same effect of benzalkonium chloride. Chroococcus and Scenedesmus exhibited sensibility to the lichen metabolites when exposed to high incubation temperatures (35 °C), chemicals and temperature synergically yielding percentage decreases of intact cells with red chlorophyll epifluorescence. These findings suggest lichen secondary metabolites as allelopathic agents against rock dwelling microorganisms and as potential natural sources for their control on stone materials in restoration and conservation programmes. In this perspective, the detection of a negligible chromatic alteration (ΔE<0.5) caused by LSM to the white Carrara marble is reported as the first step of the necessary extensive evaluation of the LSM-stone material interactions.     

Copper uptake and copper-induced physiological changes in the epiphytic lichen Evernia prunastri

The lichen Evernia prunastri is very frequently employed in lichen biomonitoring studies and its sensitivity to air pollution has been confirmed at the community level many times. However, studies focused on the physiological responses of this lichen to pollutants are underrepresented. The degree of total as well as intracellular Cu uptake by the lichen after 4 and 24?h prolonged exposure was compared with selected physiological markers including levels of assimilation pigments, chlorophyll a fluorescence, ergosterol, soluble proteins, hydrogen peroxide, superoxide, amino acids, reducing sugars and total soluble phenols. Although the degree of total as well as intracellular Cu uptake by the lichen after 4 and 24?h prolonged exposure were comparable, metabolic responses after 4 and 24?h were very different. Carotenoids, ergosterol and soluble proteins decreased due to Cu exposure and these processes were followed with an increase of superoxide content. Excess of Cu in thalli of E. prunastri after 24?h was toxic and altered almost all tested parameters, including depletion of reducing sugars. We did not observe stimulation of free amino acid synthesis in Cu treated lichen, in fact the content of many particular amino acids decreased. Content of total soluble phenols was unaltered by Cu excess.     

Secondary fungal metabolites (mycotoxins) in lichens of different taxonomic groups

Secondary fungal metabolites (mycotoxins) in 22 lichen species of the families Parmeliaceae, Nephromataceae, Umbilicariaceae, Ramalinaceae, Cladoniaceae, Peltigeraceae, and Teloschistaceae were determined by enzyme-linked immunosorbent assay. The following mycotoxins were found in these lichens in a broad concentration range with a frequency of 70–100%: sterigmatocystin (7–2090 ng/g), alternariol (20–6460 ng/g), and emodin (45–94500 ng/g). Mycophenolic acid frequently occurred in 19 lichen species; citrinin, in 17 species; diacetoxyscirpenol, in 11 species; cyclopiazonic acid, in 10 species; and zearalenone, in 9 species. PR toxin was regularly detected in three lichen species; deoxynivalenol, fumonisins, and ochratoxin A, in two species; and T-2 toxin and ergot alkaloids, in one species. Aflatoxin B₁ was detected in only six species with a frequency of 2–42%, whereas roridin A was present in 10% of Hypogymnia physodes samples.     

Effects of Culture Conditions on the Growth of Usneaceae Lichen Tissue Cultures

Conditions influencing the in vitro growth of tissues from Usneaceae(sens, lat.) species were investigated. Thallus segments ofall species except the alpine lichen grew at 20?C on malt-yeastextract agar medium in the dark. Tissues of the subtropicallichen grew at 29?C, but those of other species did not. CulturedU. longissima tissues grew rapidly to about 13 times the initialweight in 12 weeks of culture on Lilly-Barnett medium containing2% mannitol. D-Asparagine promoted the growth of cultured lichentissues, as did the corresponding L-amino acid. Cultured tissuesderived from different lichen species used different sugarsand amino acids. Addition of vitamins, phytohormones, and nucleicacid derivatives did not accelerate growth (Received February 9, 1987; Accepted August 26, 1987)     

LEC-2, a highly variable lectin in the lichen Peltigera membranacea

Lectins are a diverse group of carbohydrate binding proteins often involved in cellular interactions. A lectin gene, lec-2, was identified in the mycobiont of the lichen Peltigera membranacea. Sequencing of lec-2 open reading frames from 21 individual samples showed an unexpectedly high level of polymorphism in the deduced protein (LEC-2), which was sorted into nine haplotypes based on amino acid sequence. Calculations showed that the rates of nonsynonymous versus synonymous nucleotide substitutions deviated significantly from the null hypothesis of neutrality, indicating strong positive selection. Molecular modeling revealed that most amino acid replacements were around the putative carbohydrate-binding pocket, indicating changes in ligand binding. Lectins have been thought to be involved in the recognition of photobiont partners in lichen symbioses, and the hypothesis that positive selection of LEC-2 is driven by variation in the Nostoc photobiont partner was tested by comparing mycobiont LEC-2 haplotypes and photobiont genotypes, as represented by the rbcLX region. It was not possible to pair up the two types of marker sequences without conflicts, suggesting that positive selection of LEC-2 was not due to variation in photobiont partners.     

Epiphytic lichen diversity and its dependence on bark chemistry in the northern Mongolian dark taiga

Epiphytic lichen diversity was studied in a dark taiga forest of Pinus sibirica, Abies sibirica and Picea obovata in the western Khentey Mountains, northern Mongolia. Though most lichen species occurred on all three tree species, lichen diversity was higher on Abies and Picea than on Pinus. On branches, lichen vegetation differed less between tree species than on the trunk. The occurrence of many Parmeliaceae species with a hydrophilic surface and of many species producing the dibenzofuran usnic acid gives evidence of the low deposition of acidic pollutants in the study area. The Mn content of bark, which is known to limit at high values the abundance of epiphytic lichens in coniferous forests of Europe and North America, is apparently not controlling the spatial distribution of epiphytic lichens in the dark taiga of Mongolia. This is attributed to the dry and cold winters in Mongolia, as high Mn is especially leached from the surface of trees under moist conditions at temperatures around the freezing point, when the contact between water droplets and the tree surface is particularly intensive. Such moist and cold weather conditions are frequent in most parts of the northern coniferous forests of Europe and North America, but are rare events in the most continental parts of Asia, i.e. in Mongolia and eastern Siberia.