Wanted: virtual or live! How lichens are becoming part of mass internet culture.

While restrictions imposed by the COVID-19 pandemic have strongly limited and affected the work of scientists and communicators, the pandemic has also encouraged the development of new ways of networking and public engagement. People have had to resort to virtual events, with a subsequent proliferation of webinars, online meetings, and digital resources. In this situation we have had to find new ways of measuring the impact of these activities. Using the activities of the Italian Lichen Society (Società Lichenologica Italiana, SLI), Google Trends and colleagues' contributions, we evaluated the performances and impacts of virtual tools on lichenological literacy. We compared the relative success of virtual and in-person events and the effort required; we evaluated followers' appreciation of various categories of posts on SLI Facebook page; and designed a questionnaire to collate information on individual experiences of in-person and virtual events linked to lichens. As expected, online events generally required less effort to put together and deploy than in-person events and engaged more people, especially when recorded and made available online for a long time. Using online searches for the word "lichens" we found an association with national events, and there was a notable increase in membership of SLI over the last 10 years, demonstrating an increasing interest by people in lichens. Without excluding the positive effects of in-person experiences, we believe that online events offer a powerful tool to help increase interest in, and knowledge about lichens. This interest may help to mitigate the impact of anthropogenic activities on this sensitive component of the ecosystem and help human-lichen relationships. SUPPLEMENTARY INFORMATION: The online version contains supplementary material available at 10.1007/s13199-021-00780-6.

Alkaline air: changing perspectives on nitrogen and air pollution in an ammonia-rich world.

Ammonia and ammonium have received less attention than other forms of air pollution, with limited progress in controlling emissions at UK, European and global scales. By contrast, these compounds have been of significant past interest to science and society, the recollection of which can inform future strategies. Sal ammoniac (nushadir, nao sha) is found to have been extremely valuable in long-distance trade (ca AD 600-1150) from Egypt and China, where 6-8 kg N could purchase a human life, while air pollution associated with nushadir collection was attributed to this nitrogen form. Ammonia was one of the keys to alchemy-seen as an early experimental mesocosm to understand the world-and later became of interest as 'alkaline air' within the eighteenth century development of pneumatic chemistry. The same economic, chemical and environmental properties are found to make ammonia and ammonium of huge relevance today. Successful control of acidifying SO2 and NOx emissions leaves atmospheric NH3 in excess in many areas, contributing to particulate matter (PM2.5) formation, while leading to a new significance of alkaline air, with adverse impacts on natural ecosystems. Investigations of epiphytic lichens and bog ecosystems show how the alkalinity effect of NH3 may explain its having three to five times the adverse effect of ammonium and nitrate, respectively. It is concluded that future air pollution policy should no longer neglect ammonia. Progress is likely to be mobilized by emphasizing the lost economic value of global N emissions ($200 billion yr-1), as part of developing the circular economy for sustainable nitrogen management. This article is part of a discussion meeting issue 'Air quality, past present and future'.

The cost of surviving nitrogen excess: energy and protein demand in the lichen Cladonia portentosa as revealed by proteomic analysis.

MAIN CONCLUSION: Different nitrogen forms affect different metabolic pathways in lichens. In particular, the most relevant changes in protein expression were observed in the fungal partner, with NO 3- mostly affecting the energetic metabolism and NH 4+ affecting transport and regulation of proteins and the energetic metabolism much more than NO 3- did. Excess deposition of reactive nitrogen is a well-known agent of stress for lichens, but which symbiont is most affected and how, remains a mystery. Using proteomics can expand our understanding of stress effects on lichens. We investigated the effects of different doses and forms of reactive nitrogen, with and without supplementary phosphorus and potassium, on the proteome of the lichen Cladonia portentosa growing in a 'real-world' simulation of nitrogen deposition. Protein expression changed with the nitrogen treatments but mostly in the fungal partner, with NO3- mainly affecting the energetic metabolism and NH4+ also affecting the protein synthesis machinery. The photobiont mainly responded overexpressing proteins involved in energy production. This suggests that in response to nitrogen stress, the photobiont mainly supports the defensive mechanisms initiated by the mycobiont with an increased energy production. Such surplus energy is then used by the cell to maintain functionality in the presence of NO3-, while a futile cycle of protein production can be hypothesized to be induced by NH4+ excess. External supply of potassium and phosphorus influenced differently the responses of particular enzymes, likely reflecting the many processes in which potassium exerts a regulatory function.

Evaluating green infrastructure in urban environments using a multi-taxa and functional diversity approach.

Forested areas within cities host a large number of species, responsible for many ecosystem services in urban areas. The biodiversity in these areas is influenced by human disturbances such as atmospheric pollution and urban heat island effect. To ameliorate the effects of these factors, an increase in urban green areas is often considered sufficient. However, this approach assumes that all types of green cover have the same importance for species. Our aim was to show that not all forested green areas are equal in importance for species, but that based on a multi-taxa and functional diversity approach it is possible to value green infrastructure in urban environments. After evaluating the diversity of lichens, butterflies and other-arthropods, birds and mammals in 31 Mediterranean urban forests in south-west Europe (Almada, Portugal), bird and lichen functional groups responsive to urbanization were found. A community shift (tolerant species replacing sensitive ones) along the urbanization gradient was found, and this must be considered when using these groups as indicators of the effect of urbanization. Bird and lichen functional groups were then analyzed together with the characteristics of the forests and their surroundings. Our results showed that, contrary to previous assumptions, vegetation density and more importantly the amount of urban areas around the forest (matrix), are more important for biodiversity than forest quantity alone. This indicated that not all types of forested green areas have the same importance for biodiversity. An index of forest functional diversity was then calculated for all sampled forests of the area. This could help decision-makers to improve the management of urban green infrastructures with the goal of increasing functionality and ultimately ecosystem services in urban areas.

Hormetic Effect of Wood Distillate on Hydroponically Grown Lettuce.

The addition of biostimulants to nutrient solutions of hydroponically grown crops to speed up plant growth and improve plant yield and quality has been attracting more and more attention. This study investigated the effects of wood distillate (WD) addition to hydroponically grown lettuce (Lactuca sativa L.) plants. Two concentrations of WD, 0.2% and 0.5%, were added to the nutrient solution, and biometric (i.e., leaf fresh weight, root fresh weight, root length and root surface area), photosynthetic (i.e., chlorophyll a, chlorophyll b, and carotenoid content) and biochemical (i.e., electrolyte leakage, total polyphenols, total flavonoids, and total antioxidant power content) parameters were evaluated. The effects of WD were hormetic, as the 0.2% concentration stimulated biometric and biochemical parameters, while the 0.5% concentration inhibited plant growth. Based on these results, it can be suggested that the addition of 0.2% WD to the nutrient solution has a stimulating effect on the growth of lettuce plants, and could be a successful strategy to boost the yield of crops grown hydroponically.

Integration of ecological indicators to assess a multitemporal impact of cement industries.

The present study evaluated an integrated biomonitoring approach based on three different bioindicators: tree rings, lichens, and beetles in a complex environment (urban-industrial-forest). In Central Italy, four sampling sites were selected to assess the anthropogenic impact of cement plants taking into account (1) long-term exposure (1988-2020) through the analysis of trace elements in tree rings of Quercus pubescens; (2) medium-term exposure (2020-2021) through the analysis of trace elements in thalli (outermost portions) of the lichen Xanthoria parietina; (3) short-term exposure in spring 2021 through the bioaccumulation and evaluation of sample vitality in transplants of the lichen Evernia prunastri and a periodic survey of entomological biodiversity carried out during spring and summer 2021. Trace elements of industrial origin were found in tree rings, with different levels of accumulation between 1988 and 2020 and a maximum in 2012. Native thalli of the lichen X. parietina showed an overall low bioaccumulation of trace elements except for Cr, probably reflecting the influence of national lockdown measures. The transplants of E. prunastri showed a weak stress response in the industrial and urban sites, but not in the forest, and identified Tl and V as the main elements contributing to atmospheric contamination, with peaks at the industrial sites. Concerning the beetles, a significantly lower number of species was found at the Semonte industrial site.

Non-Toxic Increases in Nitrogen Availability Can Improve the Ability of the Soil Lichen Cladonia rangiferina to Cope with Environmental Changes.

Climate change and atmospheric nitrogen (N) deposition on drylands are greatly threatening these especially vulnerable areas. Soil biocrust-forming lichens in drylands can provide early indicators of these disturbances and play a pivotal role, as they contribute to key ecosystem services. In this study, we explored the effects of different long-term water availability regimes simulating climate changes and their interaction with N addition on the physiological response of the soil lichen Cladonia rangiferina. Three sets of this lichen were subjected to control, reduced watering, and reduced watering and N addition (40 kg NH4NO3 ha-1 year-1) treatments for 16 months. Finally, all samples were subjected to daily hydration cycles with N-enriched water at two levels (40 and 80 kg NH4NO3 ha-1 year-1) for 23 days. We found that reduced watering significantly decreased the vitality of this lichen, whereas N addition unexpectedly helped lichens subjected to reduced watering to cope with stress produced by high temperatures. We also found that long-term exposure to N addition contributed to the acclimation to higher N availability. Overall, our data suggest that the interactions between reduced watering and increased N supply and temperature have an important potential to reduce the physiological performance of this soil lichen.

Do lichens have "memory" of their native nitrogen environment?

This study aimed to deepen the knowledge about intraspecific mechanisms regulating nitrogen tolerance in lichens to wet nitrogen deposition. Thalli of the nitrophilous lichen Xanthoria parietina were collected from environments with different nitrogen availabilities and immersed in 80 mL of ammonium sulphate (NH4)2SO4 solutions with distinct concentrations (0, 0.025, 0.05 and 0.25 M) for 5 h per day during 3 days in a week. After each soaking event, lichens were air dried. After each treatment, maximal PSII efficiency, localization of ammonium ions, concentrations of K+ and Mg²+ and thalli buffer capacity were determined. Our results show that lichens are marked by their native nitrogen environment, since there were important differences between the physiological responses of X. parietina thalli previously grown in an area with high nitrogen deposition (nitrogen emissions of ca. 13,000 t/year) and those previously grown in an unpolluted area (nitrogen emissions of ca. 500 t/year). Greater N availability seems to enable X. parietina to cope better with the effects of nitrogen pollution.

Resilience of Epiphytic Lichens to Combined Effects of Increasing Nitrogen and Solar Radiation.

Lichens are classified into different functional groups depending on their ecological and physiological response to a given environmental stressor. However, knowledge on lichen response to the synergistic effect of multiple environmental factors is extremely scarce, although vital to get a comprehensive understanding of the effects of global change. We exposed six lichen species belonging to different functional groups to the combined effects of two nitrogen (N) doses and direct sunlight involving both high temperatures and ultraviolet (UV) radiation for 58 days. Irrespective of their functional group, all species showed a homogenous response to N with cumulative, detrimental effects and an inability to recover following sunlight, UV exposure. Moreover, solar radiation made a tolerant species more prone to N pollution's effects. Our results draw attention to the combined effects of global change and other environmental drivers on canopy defoliation and tree death, with consequences for the protection of ecosystems.

Time- and dose-dependency of the effects of nitrogen pollution on lichens.

The present work aims at testing if exposure time and dose play a role in the response of lichen species to nitrogen (N) pollution. To this purpose, samples of the N-sensitive Evernia prunastri and the N-tolerant Xanthoria parietina were treated for 5 weeks either with solutions of NH(4)NO(3) 0.05 and 1 M, or (NH(4))(2)SO(4) 0.025 and 0.5 M. Photosynthetic efficiency was measured as an indicator of sample vitality. The results showed that the lowest concentrations were ineffective at the beginning, but after several supplies both compounds inhibited photosynthetic activity of E. prunastri. The highest concentrations had a deleterious effect, but with a temporal trend. For X. parietina no effect was found for the lowest concentrations, while the same trend shown by E. prunastri was instead observed following treatments with the highest concentrations. It was concluded that the response of lichens to N supply is not only species-specific, but also time- and dose-dependent. The results give a clue on field studies on the relationships between lichens and N pollution.

Chlorophyll degradation and inhibition of polyamine biosynthesis in the lichen Xanthoria parietina under nitrogen stress.

This study investigated if some nitrogen (N) compounds commonly used as fertilizers (KNO3, NH4NO3, (NH4)2SO4) cause chlorophyll degradation in the N-tolerant lichen Xanthoria parietina and if polyamines are responsible for the N-tolerance of this species. The results showed that N excess does not cause chlorophyll degradation and suggested the absence of kinetics in the mode of action of the N compounds tested. External supply of inhibitors of polyamine biosynthesis prior to N treatments did not cause any change in the response of chlorophyll integrity, suggesting that at least chlorophyll integrity is not controlled by polyamines.

When the exception becomes the rule: An integrative approach to symbiosis.

Symbiosis, mainly due to the advances in -omics technology and to the microbiome revolution, is being increasingly acknowledged as fundamental to explain any aspect of life existence. Previously considered an exception, a peculiar characteristic of few systems like lichens, corals and mycorrhizas, symbiosis is nowadays recognized as the rule, with the microbiome being part of all living entities and systems. However, our knowledge of the ecological meaning and functioning of many symbiotic systems is still limited. Here, we discuss a new, integrative approach based on current findings that looks at commonalities among symbiotic systems to produce theoretical models and conceptual knowledge that would allow a more efficient exploitation of symbiosis-based biotechnologies. The microbiome recruitment and assemblage processes are indicated as one of the potential targets where a holistic approach could bring advantages. Finally, we reflect on the potential socio-economic and environmental consequences of a symbiotic view of the world, where co-dependence is the matrix of life.

The influence of growth form and substrate on lichen ecophysiological responses along an aridity gradient.

In this paper, we investigated whether growth form and substrate in lichens influence their physiological responses along an aridity gradient. Thalli of the foliose lichen Parmotrema perlatum and the fruticose lichen Ramalina canariensis were transplanted in selected rural/forested sites of Southern Portugal characterized by a different aridity index. Physiological parameters including photosynthetic performances, assimilation pigments, ergosterol content and sample viability were measured prior to exposure (winter) and after 6-month exposure (summer). Photosynthetic performances were also investigated in common native foliose and fruticose epiphytic lichens and in fruticose terricolous species. Both transplanted and native lichens showed a decrease in photosynthetic performances in summer and lower performances in sites classified as drier and higher performances in humid forested sites. No relevant differences occurred between epiphytic foliose and fruticose growth forms. However, terricolous fruticose samples showed a significant difference in humid and drier sites and between winter and summer, probably due to microclimatic conditions similarly to other biological crusts.

The role of forest in mitigating the impact of atmospheric dust pollution in a mixed landscape.

Atmospheric dust pollution, especially particulate matter below 2.5 µm, causes 3.3 million premature deaths per year worldwide. Although pollution sources are increasingly well known, the role of ecosystems in mitigating their impact is still poorly known. Our objective was to investigate the role of forests located in the surrounding of industrial and urban areas in reducing atmospheric dust pollution. This was tested using lichen transplants as biomonitors in a Mediterranean regional area with high levels of dry deposition. After a multivariate analysis, we have modeled the maximum pollution load expected for each site taking into consideration nearby pollutant sources. The difference between maximum expected pollution load and the observed values was explained by the deposition in nearby forests. Both the dust pollution and the ameliorating effect of forested areas were then mapped. The results showed that forest located nearby pollution sources plays an important role in reducing atmospheric dust pollution, highlighting their importance in the provision of the ecosystem service of air purification.

Intra- and inter-specific variations in chitin in lichens along a N-deposition gradient.

The mechanisms of nitrogen (N) tolerance in lichens are not yet fully understood. Here, we investigated how the increase of chitin content is related with N excess at inter- and intra-specific levels, by using species with differing ecological N tolerances (the tolerant Xanthoria parietina and Parmotrema hypoleucinum and the sensitive Evernia prunastri and Usnea sp.) and thalli of X. parietina and P. hypoleucinum from sites with different availabilities of N of agricultural origin (livestock), as confirmed by lichen N content and δ15N. Nitrogen, chitin (N-containing compound), and ergosterol contents were measured in lichen thalli. Nitrogen and chitin contents were higher in tolerant species than those in sensitive ones (inter-specific level) and in thalli collected from the N-polluted site than in thalli from the clean site (intra-specific level). We suggest that chitin contributes to N stress tolerance in lichens, and that excess N can be partially stored as chitin (non-toxic form) in the cell walls of tolerant species.

Contrasting effects of nitrogen addition on soil respiration in two Mediterranean ecosystems.

Increased atmospheric nitrogen (N) deposition is known to alter ecosystem carbon source-sink dynamics through changes in soil CO2 fluxes. However, a limited number of experiments have been conducted to assess the effects of realistic N deposition in the Mediterranean Basin, and none of them have explored the effects of N addition on soil respiration (R s ). To fill this gap, we assessed the effects of N supply on R s dynamics in the following two Mediterranean sites: Capo Caccia (Italy), where 30 kg ha-1 year-1 was supplied for 3 years, and El Regajal (Spain), where plots were treated with 10, 20, or 50 kg N ha-1 year-1 for 8 years. Results show a complex, non-linear response of soil respiration (R s ) to N additions with R s overall increasing at Capo Caccia and decreasing at El Regajal. This suggests that the response of R s to N addition depends on dose and duration of N supply, and the existence of a threshold above which the N introduced in the ecosystem can affect the ecosystem's functioning. Soil cover and seasonality of precipitations also play a key role in determining the effects of N on R s as shown by the different responses observed across seasons and in bare soil vs. the soil under canopy of the dominant species. These results show how increasing rates of N addition may influence soil C dynamics in semiarid ecosystems in the Mediterranean Basin and represent a valuable contribution for the understanding and the protection of Mediterranean ecosystems.

Ecological impacts of atmospheric pollution and interactions with climate change in terrestrial ecosystems of the Mediterranean Basin: Current research and future directions.

Mediterranean Basin ecosystems, their unique biodiversity, and the key services they provide are currently at risk due to air pollution and climate change, yet only a limited number of isolated and geographically-restricted studies have addressed this topic, often with contrasting results. Particularities of air pollution in this region include high O3 levels due to high air temperatures and solar radiation, the stability of air masses, and dominance of dry over wet nitrogen deposition. Moreover, the unique abiotic and biotic factors (e.g., climate, vegetation type, relevance of Saharan dust inputs) modulating the response of Mediterranean ecosystems at various spatiotemporal scales make it difficult to understand, and thus predict, the consequences of human activities that cause air pollution in the Mediterranean Basin. Therefore, there is an urgent need to implement coordinated research and experimental platforms along with wider environmental monitoring networks in the region. In particular, a robust deposition monitoring network in conjunction with modelling estimates is crucial, possibly including a set of common biomonitors (ideally cryptogams, an important component of the Mediterranean vegetation), to help refine pollutant deposition maps. Additionally, increased attention must be paid to functional diversity measures in future air pollution and climate change studies to establish the necessary link between biodiversity and the provision of ecosystem services in Mediterranean ecosystems. Through a coordinated effort, the Mediterranean scientific community can fill the above-mentioned gaps and reach a greater understanding of the mechanisms underlying the combined effects of air pollution and climate change in the Mediterranean Basin.

Nitrogen tolerance in the lichen Xanthoria parietina: the sensitive side of a resistant species.

To investigate the mechanisms of nitrogen (N) tolerance in lichens, we examined the physiological responses to increased N availability in different functional groups. Thalli of the nitrophytic Xanthoria parietina (L.) Th.Fr. previously grown both in an N-poor environment (~2kgNha-1year-1) and in an N-rich environment (~52kgNha-1year-1) were compared with the oligotrophic species Evernia prunastri (L.) Ach. and Usnea sp. Lichens were submitted to ammonium treatments. Maximum PSII efficiency, redistribution of the ions between the intra- and extracellular compartments and potassium and magnesium concentrations were the parameters used to check for the effects of N supply. The buffering capacity of the lichen extracts was also determined in untreated lichen thalli to check if different lichen behaviours were due to their ability to maintain the pH. The results showed a more similar response between X. parietina from the N-poor environment and the N-sensitive species than between X. parietina from the N-poor and N-rich environments, suggesting that X. parietina achieved N-tolerance after long-term exposure to N-rich environment. These results are important in understanding the effects of chronic ammonium pollution on one of the most sensitive components of the ecosystem, linking physiological response and ecological consequences.

Influence of angular exposure and proximity to vehicular traffic on the diversity of epiphytic lichens and the bioaccumulation of traffic-related elements.

This study investigated the influence of angular exposure and distance from vehicular traffic on the diversity of epiphytic lichens and the bioaccumulation of traffic-related elements in a town of central Italy. An Index of Lichen Diversity (ILD) was calculated on the street-facing and the opposite side of road-lining trees and in a urban park 250 m away, and the content of selected trace elements (Al, Ba, Ce, Cd, Cr, Cu, Fe, Mn, Ni, Pb, Sb, V, and Zn) was determined in samples of the lichen Punctelia borreri (Sm.) Krog growing on tree bark, both on the exposed and opposite sides. ILD increases with distance from traffic emissions. However, at the site with vehicle traffic, non-nitrophilous lichens decreased while nitrophilous ones increased. The concentration of the traffic-related elements Ba, Cr, Cu, Mn, Sb, and Zn accumulated in thalli of P. borreri was higher on roadside trees than in trees from the urban park. ILD was not affected by the angular exposure to the road and the bioaccumulation of traffic-related elements was similar in lichens from the side of the bole exposed to traffic emissions and particulate resuspension and from the opposite side. The angular exposure in respect to the traffic source does not influence trace element accumulation. These results are important when using lichens for biomonitoring purposes, both for planning future studies and for the reliability of the interpretation of past surveys that do not report information about the angular exposure of the collected lichen material.