Lichens are symbiotic organisms that effectively survive in harsh environments, including arid regions. Maintaining viability with an almost complete loss of water and the rapid restoration of metabolism during rehydration distinguishes lichens from most eukaryotic organisms. The lichen Xanthoria parietina is known to have high stress tolerance, possessing diverse defense mechanisms, including the presence of the bright-orange pigment parietin. While several studies have demonstrated the photoprotective and antioxidant properties of this anthraquinone, the role of parietin in the tolerance of lichens to desiccation is not clear yet. Thalli, which are exposed to solar radiation and become bright orange, may require enhanced desiccation tolerance. Here, we showed differences in the anatomy of naturally pale and bright-orange thalli of X. parietina and visualized parietin crystals on the surface of the upper cortex. Parietin was extracted from bright-orange thalli by acetone rinsing and quantified using HPLC. Although acetone rinsing did not affect PSII activity, thalli without parietin had higher levels of lipid peroxidation and a lower membrane stability index in response to desiccation. Furthermore, highly pigmented thalli possess thicker cell walls and, according to thermogravimetric analysis, higher water-holding capacities than pale thalli. Thus, parietin may play a role in desiccation tolerance by stabilizing mycobiont membranes, providing an antioxidative defense, and changing the morphology of the upper cortex of X. parietina.

Peroxidases are widespread in animal and plant tissues, wherein they perform a variety of functions. Peroxidases have a broad specificity for substrates of various chemical structures. Along with hydrogen peroxide, phenolic compounds, and toxic compounds of aromatic nature, nitrogen-containing compounds are substrates for peroxidases. This work is devoted to the study of the role of wheat extracellular peroxidases in the metabolism of nitrogen-containing compounds. It has been shown that partially purified isozymes differing in peroxidase activity are involved in the metabolism of nitrogen-containing compounds. The formation of primary and secondary phenoxyl radicals during the combined oxidation of chlorogenic acid, nitrite, and H₂O₂ was demonstrated. With cooxidation with purified isoenzymes p˗coumaric acid and nitrite, the formation of 4˗hydroxy˗3˗nitrocinnamic acid was revealed. It is assumed that the same isoforms can participate both in the oxidation of nitrite with the formation of nitrophenol and in the reduction of nitrate. The participation of plant peroxidases in nitrogen metabolism can be represented as a set of reactions for the reduction and/or oxidation of nitrogen of different oxidation states with the formation of active intermediates.

Extremophile organisms can survive under extreme conditions through tolerance mechanisms. Such organisms include lichens, which are symbiotic associations of fungi and algae and/or cyanobacteria. Among other factors, the high stress tolerance of lichens can be attributed to their ability to synthesize a wide range of metabolites, including chemically diverse lipid compounds. Despite their obvious relevance, the biochemical mechanisms of stress tolerance in lichens that are mediated by changes in the lipid profile remain understudied. Peltigerous lichens constitute a separate division of lichens characterized by high growth rates and metabolic activity indices. Taking into account the temperate climate in which these lichens grow, it can be assumed that they may be highly sensitive to temperature fluctuations. These factors led to the choice of Peltigera canina, epigeic lichen, as the subject matter of this study. The present work examined stress-induced changes in the redox status of P. canina, as well as changes in its lipid composition at an elevated temperature (40 °С). The exposure of hydrated lichen thalli to an unfavorable temperature led to an increase in the level of hydrogen peroxide, phenoloxidase activity, and lipid peroxidation, which constitutes an important part of stress response in lichen. The stress-induced changes in the redox status of lichen thalli stimulated an increase in lipophilic antioxidant agents: a dramatic increase in the level of α-tocopherol and an increase in carotenoids, specifically β-carotene. Thus, the accumulation of lipophilic antioxidants constitutes an important part of the lipid-mediated stress response of P. canina to temperature elevation.

Currently, due to the increasing impact of anthropogenic factors and changes in solar activity, the temperature on Earth is rising, posing a threat to biodiversity. Lichens are among the most sensitive organisms to climate change. Elevated ambient temperatures can have a significant impact on lichens, resulting in more frequent and intense drying events that can impede metabolic activity. It has been suggested that the possession of a diverse sterol composition may contribute to the tolerance of lichens to adverse temperatures and other biotic and abiotic stresses. The major sterol found in lichens is ergosterol (ERG); however, the regulation of the ERG biosynthetic pathway, specifically the step of epoxidation of squalene to 2,3-oxidosqualene catalyzed by squalene epoxidase during stress, has not been extensively studied. In this study, we used lichen Lobaria pulmonaria as a model species that is well known to be sensitive to air pollution and habitat loss. Using in silico analysis, we identified cDNAs encoding squalene epoxidase from L. pulmonaria, designating them as LpSQE1 for the mycobiont and SrSQE1 for the photobiont Symbiochloris reticulata. Our results showed that compared with a control kept at room temperature (+20 °C), mild temperatures (+4 °C and +30 °C) did not affect the physiology of L. pulmonaria, assessed by changes in membrane integrity, respiration rates, and PSII activity. An extreme negative temperature (−20 °C) noticeably inhibited respiration but did not affect membrane stability. In contrast, treating lichen with a high positive temperature (+40 °C) significantly reduced all physiological parameters. Quantitative PCR analysis revealed that exposing thalli to −20 °C, +4 °C, +30 °C, and +40 °C stimulated the expression levels of LpSQE1 and SrSQE1 and led to a significant upregulation of Hsps. These data provide new information regarding the roles of sterols and Hsps in the response of lichens to climate change.

Lichens, symbiotic organisms with a high tolerance to harsh environments, possess a greater diversity of sterols than other organisms. Sterols are involved in maintaining membrane integrity, hormone biosynthesis, and signal transduction. (1) Background: A characteristic feature of lichen sterols is a high degree of unsaturation, which influences membrane properties. Desaturases play an important role in the synthesis of unsaturated sterols, in particular, sterol C-5 desaturase (ERG3), which controls the conversion of episterol to ergosterol. Earlier, we demonstrated that the treatment of the lichen Peltigera canina with low and elevated temperatures results in changes in the levels of episterol and ergosterol. (2) Methods: Here, for the first time, we identified ERG3 in P. canina and, using an in silico analysis, we showed that PcERG3 belongs to the superfamily of fatty acid hydrolyases. A phylogenetic analysis was conducted to determine the evolutionary relationships of PcERG3. (3) Results: A phylogenetic analysis showed that PcERG3 clusters with ERG3 from other Peltigeralian and non-Peltigeralian lichens and also with ERG3 from free-living fungi. This suggests that PcERG3 has an ancient evolutionary origin and is related to fungi with lichenized ancestors, e.g., Penicillium. The differential expression of PcERG3 in response to temperature stress, a dehydration/rehydration cycle, and heavy metal exposure suggests that it plays a crucial role in maintaining the balance between more and less saturated sterols and, more generally, in membrane functioning. The multifaceted response of P. canina to abiotic stresses was documented by simultaneously measuring changes in the expression of PcERG3, as well as the genes encoding the heat shock proteins, PcHSP20 and PcHSP98, and PcSOD1, which encodes the antioxidant enzyme superoxide dismutase. (4) Conclusions: These findings suggest that PcERG3 is similar to the sterol C-5 desaturases from related and free-living fungi and plays important roles in the molecular mechanisms underlying the tolerance of lichens to environmental stress.