Effect of thallus melanisation on the sensitivity of lichens to heat stress.

Extreme climatic phenomena such as heat waves, heavy rainfall and prolonged droughts are one of the main problems associated with ongoing climate change. The global increase in extreme rainfalls associated with summer heatwaves are projected to increase in amplitude and frequency in the near future. However, the consequences of such extreme events on lichens are largely unknown. The aim was to determine the effect of heat stress on the physiology of lichen Cetraria aculeata in a metabolically active state and to verify whether strongly melanised thalli are more resistant than poorly melanised thalli. In the present study, melanin was extracted from C. aculeata for the first time. Our study showed that the critical temperature for metabolism is around 35 °C. Both symbiotic partners responded to heat stress, manifested by the decreased maximum quantum yield of PSII photochemistry, high level of cell membrane damage, increased membrane lipid peroxidation and decreased dehydrogenase activity. Highly melanised thalli were more sensitive to heat stress, which excludes the role of melanins as compounds protecting against heat stress. Therefore, mycobiont melanisation imposes a trade-off between protection against UV and avoidance of damage caused by high temperature. It can be concluded that heavy rainfall during high temperatures may significantly deteriorate the physiological condition of melanised thalli. However, the level of membrane lipid peroxidation in melanised thalli decreased over time after exposure, suggesting greater efficiency of antioxidant defence mechanisms. Given the ongoing climate changes, many lichen species may require a great deal of plasticity to maintain their physiological state at a level that ensures their survival.

Towards understanding the effect of heavy metals on mycobiont physiological condition in a widespread metal-tolerant lichen Cladonia rei.

Heavy metals present in the environment can cause a variety of injury symptoms in various organisms including lichens. Most studies examined metal-induced stress under controlled laboratory conditions, and little is known about actual response of lichens in their natural habitat. This study aims to recognize the effect of heavy metal accumulation (total and intracellular) on lichen physiological and biochemical parameters specifically related to the functioning of fungal component. Cladonia rei was used as a model species due to its common occurrence both in unpolluted and extremely polluted sites. We observed a decline in the fungal metabolism which was expressed by a decrease in ergosterol content and an increase in cell membrane damage as a result of increased Zn, Cd, Cu and Ni accumulation. Additionally, the results indicated that increased accumulation of xenobiotics (Pb and As) caused reduction of glutathione (GSH) concentrations and increased membrane lipid peroxidation. Therefore, we conclude that GSH does not provide high oxidative stress protection in C. rei which is somewhat against its insensitivity to pollution. The reduced pool of GSH could be explained by its oxidation to glutathione disulphide induced by heavy metal stress or its use for phytochelatin (PC) synthesis. The content of secondary metabolites was not related to heavy metal accumulation and remained at a relatively stable level. This indicates that the decline in the physiological condition did not weaken the mycobiont of C. rei enough to inhibit the synthesis of secondary metabolites and their precursors were supplied at a sufficient level. Thus, the potential function of main secondary metabolites as extracellular metal immobilizers and antioxidants is still possible even in individuals growing at extremely polluted sites. Despite the evident heavy metal stress, C. rei copes well and spreads easily through extremely polluted environments, which underlines its unique pioneering abilities in highly disturbed sites.

Metabolic processes involved with sugar alcohol and secondary metabolite production in the hyperaccumulator lichen Diploschistes muscorum reveal its complex adaptation strategy against heavy-metal stress.

The synthesis of various unique secondary metabolites by lichens is the result of mutualistic symbiotic association between the mycobiont and autotrophic photobiont. The function of these compounds and causal factors for their production are not fully understood. This paper examines the effect of heavy-metal bioaccumulation and physiological parameters related to photosynthesis and carbon metabolism on the production of lichen substances in hyperaccumulator Diploschistes muscorum. The obtained model of secondary metabolite concentrations in the thalli demonstrates that the carbon source provided by the photobiont and associated polyols produced by the mycobiont have positive impact on the production; on the contrary, the increased intracellular load of heavy metals and excessive loss of cell membrane integrity adversely affected secondary metabolite contents. Additionally, the production of secondary metabolites appears to be more dependent on intracellular metal concentrations than on soil pollution level. To compensate for metal stress, both efficient functioning of algal component and sufficient production of secondary metabolites are required. The balanced physiological functioning of mycobiont and photobiont constitutes the complex protective mechanism to alleviate the harmful effects of heavy metal stress on primary and secondary metabolism of lichens.

Heavy-metal tolerance of photobiont in pioneer lichens inhabiting heavily polluted sites.

Heavy metals are known for their negative impact on the physiological processes of lichen photobiont. In spite of this, certain lichens are known to be effective pioneers of polluted sites. Cladonia cariosa, C. rei, and Diploschistes muscorum are prominent examples of lichens that spontaneously colonise post-industrial wastes. We examined the effect of total and intracellular Zn, Pb, Cd, As, Cu, and Ni accumulation in the thalli of these species on the physiological parameters of photobiont. Increased accumulation of Zn, Cd, Cu, and Ni in D. muscorum and of Zn and Ni in C. rei negatively affected contents of photosynthetic pigments, whereas concentrations of Pb had a positive effect in all lichen species. Moreover, pigment contents were positively associated with the concentrations of most examined elements in C. cariosa. The results indicate that even if chlorophyll contents reduced, its degradation does not progress. This suggests that metal stress may exert a negative effect on the synthesis rather than on the integrity of chlorophyll. Most importantly, lichen samples of each of the species from polluted sites proved to possess significantly higher FV/FM ratios than those from a reference site; moreover, the contents of elements of lichen thalli positively influenced this parameter. The efficient functioning of the algal component under heavy-metal stress conditions indicates that the examined lichens are well adapted to extremely contaminated substrates.

Secondary metabolites of the lichen Hypogymnia physodes (L.) Nyl. and their presence in spruce (Picea abies (L.) H. Karst.) bark.

Lichen species typically have a characteristic profile of secondary metabolites. Dense populations of Hypogymnia physodes growing frequently as epiphytes on tree branches have harmful effects on the host, likely due to their secondary compounds, which were undetected in tree tissues until now. The aim of the present study was to re-characterise the suite of secondary metabolites of H. physodes thalli and to estimate their translocation into spruce (Picea abies) bark. Thallus and bark extracts were compared using ultra-performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS). The compounds were identified based on their UV, MS and MS/MS spectra as well as retention factors of their TLC analysis. In addition to the previously described secondary metabolites (protocetraric, physodalic, 3-hydroxyphysodic, physodic, and 2'-O-methylphysodic acids, atranorin and chloroatranorin) of H. physodes, further three were identified in its thalli: conphysodalic, 4-O-methylphysodic and α-alectoronic acids. Fragmentation patterns from the negative ionisation of each compound were proposed, some of which were described for the first time. Among all of the detected lichen substances, a few, e.g., physodalic, 3-hydroxyphysodic, physodic acids and atranorin, were present in the bark of spruce branches that were abundantly colonised by lichen. The newly identified compounds of H. physodes thalli may belong to its constant or accessory secondary metabolites. These compounds may be useful in the chemotaxonomic classification of this species. The presence of some lichen substances in spruce bark confirmed their ability to penetrate host tissues. These data suggest that H. physodes compounds may cause long-term effects on spruces in nature.