The pale yellowish tint of usnic acid in a lichen thallus itself is a commonly used character in identification keys, particularly in the genus Cladonia. Furthermore, the presence of usnic acid is phylogenetically significant in numerous groups of lichens. While the distinctive colour of usnic acid is readily visible when present in high concentrations, it is commonly problematic to discern when in low to moderate concentrations. We explored the use of an anisaldehyde reagent for visualizing usnic acid. Using both usnic acid-containing Cladonia samples and pure usnic acid, this reaction quickly yields a bright magenta colour on HPTLC and TLC plates after heating and directly with crude acetone extracts on glass slides heated with a lighter. The same magenta product was observed whether or not the usnic acid was accompanied by barbatic, fumarprotocetraric, psoromic, squamatic or thamnolic acids, each of which alone did not produce any colour with anisaldehyde reagent. However, the merochlorophaeic acids in C. albonigra also produced a red reaction. Analysis by high resolution LC-MS of the reaction mixture between anisaldehyde and usnic acid revealed several ions at m/z 477.1586 ([M+H]+, C27H25O8) and 463.1385 ([M+H]+, C26H23O8), respectively, consistent with aldol condensation of usnic acid and p-anisaldehyde.

The identity of Lecanora ecorticata has been uncertain due to conflicting reports of the presence of zeorin and the loss of the holotype. Phylogenetic analysis suggests that the species probably belongs in Ramalinaceae, where it is the first leprose species reported. Zeorin was found to be absent in recently collected material of L. ecorticata, including specimens from the type locality, and the report of zeorin in the lost holotype is considered to be due to contamination. A new genus, Lithocalla, is erected to accommodate L. ecorticata and a second closely related species, that occurs in the Falkland Islands, is newly combined as Lithocalla malouina. A lectotype is selected for L. ecorticata. Lithocalla is characterized by a leprose thallus containing usnic acid with fatty acids and terpenoids, but no zeorin; ascomata and conidiomata are unknown.

Usnic acid is a unique lichen metabolite of industrial importance, widely studied to explore its pharmacological potential and valued especially as an antibacterial agent in cosmetics. Although a vast number of papers describe usnic acid extraction from various lichen species, none has so far provided an unequivocal indication of the best extraction procedure for this compound. Thus, the current study was focused on the direct comparison of three commonly used usnic acid extraction methods (heat reflux, shaking, ultrasound-assisted extractions), which were optimized using fractional factorial design. Heat reflux extraction, shaking extraction and ultrasound-assisted extraction were first optimized in a series of experiments using fractional factorial design, with respect to three parameters: the extraction time, the solvent used and the number of extraction repetitions. HPLC was employed for usnic acid quantitative analysis. The best scores for each extraction method were statistically compared and the optimal conditions were indicated. The optimal set of parameters for usnic acid was established to be a single, 60 min heat reflux extraction with acetone. This extraction scheme provided 4.25 ±0.08 mg g−1 d.w. of usnic acid, while for ultrasound-assisted and shaking extractions the amount was two- or even four times lower (2.33 ±0.17 and 0.97 ±0.08 mg g−1 d.w., respectively). The optimal procedure for usnic acid extraction described here may be suitable for effective acquisition of this compound for scientific research purposes, but also for applications in the pharmaceutical or cosmetic industries.

Arctoparmelia collatolica is described as a species new to science based on morphological, chemical and molecular data. The species is similar to the usnic acid-deficient chemotype of A. centrifuga but differs in the grey-brown to brown upper surface in the central part of the thallus and ivory white to pale brown rhizines. The species contains collatolic acid and its derivatives. Seven secondary lichen substances are reported as new for the genus Arctoparmelia. A key to Arctoparmelia taxa is provided.

Leprocaulon calcicola is described as new from walls in SE England; it is leprose, pale to mid blue-grey, and contains zeorin and usnic acid. It differs from L. knudsenii from North America in its habitat on mortared walls rather than non-calcareous rock and in its ITS sequence. ‘Lecanora’ ecorticata differs in the yellower colour, and the presence of unidentified fatty acids and traces of unknown terpenoids (but not zeorin) by thin-layer chromatography. Leprose lichens with usnic acid are still poorly known and sequencing must be used to support morphological and chemical studies.

The corticolous lichen Lecanora subviridis de la Rosa & Messuti is described from temperate forests in south-western Argentina as new to science. It differs from other species of Lecanora occurring on bark and wood by containing usnic or isousnic acid. The relationships with the other taxa are discussed.

Alectorioid and foliose lichens were weighed from full-size branches of Picea abies cut at two canopy heights in 100 trees in an old subalpine forested area in eastern Norway. The mean lichen biomass per branch decreased from 46 to 37 g moving upwards from 2–3 to 5–6 m canopy height. The lichen biomass correlated strongly with branch size variables, branch diameter alone explaining 48 of the variation in lichen biomass per branch (n=200). The alectorioid/foliose biomass ratio increased from 0·149 at 2–3 m to 0·316 at 5–6 m. Site factors reflecting openness of the canopy were computed for the branch at 2–3 m in all trees. The alectorioid/foliose biomass ratio increased significantly with the indirect site factors, suggesting that light, or factors associated with light, determine the balance between these two growth forms in tree canopies. Within the alectorioid biomass component, genera with usnic acid (Alectoria, Usnea) decreased with height, whereas the genus Bryoria with melanic pigments increased. According to the literature, dark melanic pigments have higher visible light screening efficiency than the yellowish usnic acid. Such patterns suggest a functional role for cortical pigments in the niche differentiation of alectorioid lichens.