Mycosporine-Like Amino Acids (MAAs) in Time-Series of Lichen Specimens from Natural History Collections.

Mycosporine-like amino acids (MAAs) were quantified in fresh and preserved material of the chlorolichen Dermatocarpon luridum var. luridum (Verrucariaceae/Ascomycota). The analyzed samples represented a time-series of over 150 years. An HPLC coupled with a diode array detector (HPLC-DAD) in hydrophilic interaction liquid chromatography (HILIC) mode method was developed and validated for the quantitative determination of MAAs. We found evidence for substance specific differences in the quality of preservation of two MAAs (mycosporine glutamicol, mycosporine glutaminol) in Natural History Collections. We found no change in average mycosporine glutamicol concentrations over time. Mycosporine glutaminol concentrations instead decreased rapidly with no trace of this substance detectable in collections older than nine years. Our data predict that a screening for MAAs in organism samples from Natural History Collections can deliver results that are comparable to those obtained from fresh collections only for some MAAs (e.g., mycosporine glutamicol). For other MAAs, misleading, biased, or even false negative results will occur as a result of the storage sensitivity of substances such as mycosporine glutaminol. Our study demonstrates the value of pilot studies with time-series based on model taxa with a rich representation in the Natural History Collections.

Freshwater Lichens, Including New Species in the Genera Verrucaria, Placopyrenium and Circinaria, Associated with Lobothallia hydrocharis (Poelt & Nimis) Sohrabi & Nimis from Watercourses of Sardinia.

This work summarizes the results of the exploration of freshwater lichen biota on the island of Sardinia associated with the regional flagship species Lobothallia hydrocharis, a large-sized crustose lichen from the splash zone along mountain streams, so far known from Sardinia only. Molecular data were used to confirm its distinctiveness from other taxa and its systematic placement and to identify critical taxa among its associated lichen biota. We found 25 species of lichenized fungi, including three species new to science in the genera Verrucaria, Placopyrenium, and Circinaria, and seven species new to Sardinia (Hydropunctaria rheithrophila, Ionaspis chrysophana, I. odora, Verrucaria aquatilis, V. collematodes, V. pseudovirescens), or new to Southern Europe (V. devensis). Specific traits for the freshwater lichen biota of Sardinia were identified and compared to those reported from freshwater sites in the Alps and Carpathian mountains, e.g., a relative scarcity of subgelatinous lichens. Parasitic or epilichenic interactions were found frequently but only in the splash zone and not in the permanently submerged zone, i.e., two parasitic Placopyrenium species, and clearly lichenicolous thalli of Kuettlingeria atroflava and Lobothallia hydrocharis. Due to its specific trait profile and the great potential for the discovery of new species, we recommend the inclusion of Sardinian and further Mediterranean sites in continental-scale monitoring programs for freshwater lichens.

Mycobiont-specific primers facilitate the amplification of mitochondrial small subunit ribosomal DNA: a focus on the lichenized fungal genus Melanelia (Ascomycota, Parmeliaceae) in Iceland.

The fungal mitochondrial small subunit (mtSSU) ribosomal DNA is one of the most commonly used loci for phylogenetic analysis of lichen-forming fungi, but their primer specificity to mycobionts has not been evaluated. The current study aimed to design mycobiont-specific mtSSU primers and highlights their utility with an example from the saxicolous lichen-forming fungal genus Melanelia Essl. in Iceland. The study found a 12.5% success rate (3 out of 24 specimens with good-quality mycobiont mtSSU sequences) using universal primers (i.e. mrSSU1 and mrSSU3R), not including off-target amplification of environmental fungi, e.g. Cladophialophoracarrionii and Lichenotheliaconvexa. New mycobiont-specific primers (mt-SSU-581-5' and mt-SSU-1345-3') were designed by targeting mycobiont-specific nucleotide sites in comparison with environmental fungal sequences, and assessed for mycobiont primer specificity using in silico PCR. The new mycobiont-specific mtSSU primers had a success rate of 91.7% (22 out of 24 specimens with good-quality mycobiont mtSSU sequences) on the studied Melanelia specimens. Additional testing confirmed the specificity and yielded amplicons from 79 specimens of other Parmeliaceae mycobiont lineages. This study highlights the effectiveness of designing mycobiont-specific primers for studies on lichen identification, barcoding and phylogenetics.

Key Roles of Dipterocarpaceae, Bark Type Diversity and Tree Size in Lowland Rainforests of Northeast Borneo-Using Functional Traits of Lichens to Distinguish Plots of Old Growth and Regenerating Logged Forests.

Many lowland rainforests in Southeast Asia are severely altered by selective logging and there is a need for rapid assessment methods to identify characteristic communities of old growth forests and to monitor restoration success in regenerating forests. We have studied the effect of logging on the diversity and composition of lichen communities on trunks of trees in lowland rainforests of northeast Borneo dominated by Dipterocarpaceae. Using data from field observations and vouchers collected from plots in disturbed and undisturbed forests, we compared a taxonomy-based and a taxon-free method. Vouchers were identified to genus or genus group and assigned to functional groups based on sets of functional traits. Both datasets allowed the detection of significant differences in lichen communities between disturbed and undisturbed forest plots. Bark type diversity and the proportion of large trees, particularly those belonging to the family Dipterocarpaceae, were the main drivers of lichen community structure. Our results confirm the usefulness of a functional groups approach for the rapid assessment of tropical lowland rainforests in Southeast Asia. A high proportion of Dipterocarpaceae trees is revealed as an essential element for the restoration of near natural lichen communities in lowland rainforests of Southeast Asia.

Functional Traits in Lichen Ecology: A Review of Challenge and Opportunity.

Community ecology has experienced a major transition, from a focus on patterns in taxonomic composition, to revealing the processes underlying community assembly through the analysis of species functional traits. The power of the functional trait approach is its generality, predictive capacity such as with respect to environmental change, and, through linkage of response and effect traits, the synthesis of community assembly with ecosystem function and services. Lichens are a potentially rich source of information about how traits govern community structure and function, thereby creating opportunity to better integrate lichens into 'mainstream' ecological studies, while lichen ecology and conservation can also benefit from using the trait approach as an investigative tool. This paper brings together a range of author perspectives to review the use of traits in lichenology, particularly with respect to European ecosystems from the Mediterranean to the Arctic-Alpine. It emphasizes the types of traits that lichenologists have used in their studies, both response and effect, the bundling of traits towards the evolution of life-history strategies, and the critical importance of scale (both spatial and temporal) in functional trait ecology.

Lichens and associated fungi from Glacier Bay National Park, Alaska.

Lichens are widely acknowledged to be a key component of high latitude ecosystems. However, the time investment needed for full inventories and the lack of taxonomic identification resources for crustose lichen and lichenicolous fungal diversity have hampered efforts to fully gauge the depth of species richness in these ecosystems. Using a combination of classical field inventory and extensive deployment of chemical and molecular analysis, we assessed the diversity of lichens and associated fungi in Glacier Bay National Park, Alaska (USA), a mixed landscape of coastal boreal rainforest and early successional low elevation habitats deglaciated after the Little Ice Age. We collected nearly 5000 specimens and found a total of 947 taxa, including 831 taxa of lichen-forming and 96 taxa of lichenicolous fungi together with 20 taxa of saprotrophic fungi typically included in lichen studies. A total of 98 species (10.3% of those detected) could not be assigned to known species and of those, two genera and 27 species are described here as new to science: Atrophysma cyanomelanos gen. et sp. nov., Bacidina circumpulla, Biatora marmorea, Carneothele sphagnicola gen. et sp. nov., Cirrenalia lichenicola, Corticifraga nephromatis, Fuscidea muskeg, Fuscopannaria dillmaniae, Halecania athallina, Hydropunctaria alaskana, Lambiella aliphatica, Lecania hydrophobica, Lecanora viridipruinosa, Lecidea griseomarginata, L. streveleri, Miriquidica gyrizans, Niesslia peltigerae, Ochrolechia cooperi, Placynthium glaciale, Porpidia seakensis, Rhizocarpon haidense, Sagiolechia phaeospora, Sclerococcum fissurinae, Spilonema maritimum, Thelocarpon immersum, Toensbergia blastidiata and Xenonectriella nephromatis. An additional 71 'known unknown' species are cursorily described. Four new combinations are made: Lepra subvelata (G. K. Merr.) T. Sprib., Ochrolechia minuta (Degel.) T. Sprib., Steineropsis laceratula (Hue) T. Sprib. & Ekman and Toensbergia geminipara (Th. Fr.) T. Sprib. & Resl. Thirty-eight taxa are new to North America and 93 additional taxa new to Alaska. We use four to eight DNA loci to validate the placement of ten of the new species in the orders Baeomycetales, Ostropales, Lecanorales, Peltigerales, Pertusariales and the broader class Lecanoromycetes with maximum likelihood analyses. We present a total of 280 new fungal DNA sequences. The lichen inventory from Glacier Bay National Park represents the second largest number of lichens and associated fungi documented from an area of comparable size and the largest to date in North America. Coming from almost 60°N, these results again underline the potential for high lichen diversity in high latitude ecosystems.

Three new species of Krogia (Ramalinaceae, lichenised Ascomycota) from the Paleotropics.

Krogiaborneensis Kistenich & Timdal, K.isidiata Kistenich & Timdal and K.macrophylla Kistenich & Timdal are described as new species, the first from Borneo and the two latter from New Caledonia. The new species are supported by morphology, secondary chemistry and DNA sequence data. Krogiaborneensis and K.isidiata contain sekikaic and homosekikaic acid, both compounds reported here for the first time from the genus. Krogiamacrophylla contains an unknown compound apparently related to boninic acid as the major compound. DNA sequences (mtSSU and nrITS) are provided for the first time for Krogia and a phylogeny of the genus based on 15 accessions of five of the six accepted species is presented. Krogiaantillarum is reported as new to Brazil, Guatemala and Mexico.

New erythritol derivatives from the fertile form of Roccella montagnei.

Chemical investigation of the methanol extract of the fertile form of Roccella montagnei collected in Vietnam afforded twelve secondary metabolites, including five new montagnetol derivatives, orsellinylmontagnetols A-D and a furanyl derivative together with seven known compounds. Their chemical structures were elucidated by analysis of 1D and 2D NMR and high resolution mass spectroscopic data. The relative stereochemistry of two chiral centers (C-2 and C-3) of orsellinylmontagnetols A and B was elucidated by comparison of their coupling constants and the specific rotation with those reported in the literature while the absolute stereochemistry was determined by the application of a modified Mosher method for the hydroxy group at C-3. The absolute configuration (2R,3S) of the butanetetraol moiety of these compounds is in accordance with that of erythrin, a recognized chemotaxonomic marker of the genus Roccella. Five of these compounds were evaluated for their cytotoxic activities against four cancer cell lines. Only orsellinylmontagnetol A exerted a moderate activity against MCF-7 cell line with an IC50 value of 68.39 ± 3.46 µM.

Early colonization of stone by freshwater lichens of restored habitats: a case study in northern Italy.

This study evaluated the effectiveness and life-strategies of freshwater lichens in colonizing newly constructed stone structures in low-elevation streams in a small nature reserve in northern Italy. Species richness, size of thalli, morphological and ontogenetic traits of the species were related to the age of restored habitats. Lichen colonization was surprisingly rapid, indicating the high potential of these organisms in colonizing restored habitats. However, the species pool found in the restored habitats was different than that found in natural sites in the same study area. The age of newly constructed habitats influenced both species richness and thallus size of the two most frequent Verrucaria species. Verrucaria aquatilis was a rapid colonizer invading the substrate by several small-sized and thin thalli which soon supported a large number of small perithecia whose development began in the earlier phase of thallus formation. V. elaeomelaena, on the contrary, developed according to a different strategy, establishing a thick thallus on which relatively large perithecia were formed much later than in V. aquatilis. As these taxa are important photoautotrophic components of freshwater ecosystems more ecological knowledge is needed to evaluate the effectiveness of different measures of river restoration on lichen communities. The main practical implication of our study is related to the value of small stone structures, such as riffles and ramps, for enhancing the establishment of pioneer freshwater lichens to rapidly colonize newly available substrata.