δ15N of lichens reflects the isotopic signature of ammonia source.

Although it is generally accepted that δ15N in lichen reflects predominating N isotope sources in the environment, confirmation of the direct correlation between lichen δ15N and atmospheric δ15N is still missing, especially under field conditions with most confounding factors controlled. To fill this gap and investigate the response of lichens with different tolerance to atmospheric N deposition, thalli of the sensitive Evernia prunastri and the tolerant Xanthoria parietina were exposed for ten weeks to different forms and doses of N in a field manipulation experiment where confounding factors were minimized. During this period, several parameters, namely total N, δ15N and chlorophyll a fluorescence, were measured. Under the experimental conditions, δ15N in lichens quantitatively responded to the δ15N of released gaseous ammonia (NH3). Although a high correlation between the isotopic signatures in lichen tissue and supplied N was found both in tolerant and sensitive species, chlorophyll a fluorescence indicated that the sensitive species very soon lost its photosynthetic functionality with increasing N availability. The most damaging response to the different N chemical forms was observed with dry deposition of NH3, although wet deposition of ammonium ions had a significant observable physiological impact. Conversely, there was no significant effect of nitrate ions on chlorophyll a fluorescence, implying differential sensitivity to dry deposition versus wet deposition and to ammonium versus nitrate in wet deposition. Evernia prunastri was most sensitive to NH3, then NH4+, with lowest sensitivity to NO3-. Moreover, these results confirm that lichen δ15N can be used to indicate the δ15N of atmospheric ammonia, providing a suitable tool for the interpretation of the spatial distribution of NH3 sources in relation to their δ15N signal.

Using nitrogen concentration and isotopic composition in lichens to spatially assess the relative contribution of atmospheric nitrogen sources in complex landscapes.

Reactive nitrogen (Nr) is an important driver of global change, causing alterations in ecosystem biodiversity and functionality. Environmental assessments require monitoring the emission and deposition of both the amount and types of Nr. This is especially important in heterogeneous landscapes, as different land-cover types emit particular forms of Nr to the atmosphere, which can impact ecosystems distinctively. Such assessments require high spatial resolution maps that also integrate temporal variations, and can only be feasibly achieved by using ecological indicators. Our aim was to rank land-cover types according to the amount and form of emitted atmospheric Nr in a complex landscape with multiple sources of N. To do so, we measured and mapped nitrogen concentration and isotopic composition in lichen thalli, which we then related to land-cover data. Results suggested that, at the landscape scale, intensive agriculture and urban areas were the most important sources of Nr to the atmosphere. Additionally, the ocean greatly influences Nr in land, by providing air with low Nr concentration and a unique isotopic composition. These results have important consequences for managing air pollution at the regional level, as they provide critical information for modeling Nr emission and deposition across regional as well as continental scales.

Geostatistical uncertainty of assessing air quality using high-spatial-resolution lichen data: A health study in the urban area of Sines, Portugal.

In most studies correlating health outcomes with air pollution, personal exposure assignments are based on measurements collected at air-quality monitoring stations not coinciding with health data locations. In such cases, interpolators are needed to predict air quality in unsampled locations and to assign personal exposures. Moreover, a measure of the spatial uncertainty of exposures should be incorporated, especially in urban areas where concentrations vary at short distances due to changes in land use and pollution intensity. These studies are limited by the lack of literature comparing exposure uncertainty derived from distinct spatial interpolators. Here, we addressed these issues with two interpolation methods: regression Kriging (RK) and ordinary Kriging (OK). These methods were used to generate air-quality simulations with a geostatistical algorithm. For each method, the geostatistical uncertainty was drawn from generalized linear model (GLM) analysis. We analyzed the association between air quality and birth weight. Personal health data (n=227) and exposure data were collected in Sines (Portugal) during 2007-2010. Because air-quality monitoring stations in the city do not offer high-spatial-resolution measurements (n=1), we used lichen data as an ecological indicator of air quality (n=83). We found no significant difference in the fit of GLMs with any of the geostatistical methods. With RK, however, the models tended to fit better more often and worse less often. Moreover, the geostatistical uncertainty results showed a marginally higher mean and precision with RK. Combined with lichen data and land-use data of high spatial resolution, RK is a more effective geostatistical method for relating health outcomes with air quality in urban areas. This is particularly important in small cities, which generally do not have expensive air-quality monitoring stations with high spatial resolution. Further, alternative ways of linking human activities with their environment are needed to improve human well-being.

Tools for determining critical levels of atmospheric ammonia under the influence of multiple disturbances.

Critical levels (CLEs) of atmospheric ammonia based on biodiversity changes have been mostly calculated using small-scale single-source approaches, to avoid interference by other factors, which also influence biodiversity. Thus, it is questionable whether these CLEs are valid at larger spatial scales, in a multi- disturbances context. To test so, we sampled lichen diversity and ammonia at 80 sites across a region with a complex land-cover including industrial and urban areas. At a regional scale, confounding factors such as industrial pollutants prevailed, masking the CLEs. We propose and use a new tool to calculate CLEs by stratifying ammonia concentrations into classes, and focusing on the highest diversity values. Based on the significant correlations between ammonia and biodiversity, we found the CLE of ammonia for Mediterranean evergreen woodlands to be 0.69 µg m(-3), below the previously accepted value of 1.9 µg m(-3), and below the currently accepted pan-European CLE of 1.0 µg m(-3).

Can ammonia tolerance amongst lichen functional groups be explained by physiological responses?

Ammonia (NH3) empirical critical levels for Europe were re-evaluated in 2009, based mainly on the ecological responses of lichen communities without acknowledging the physiological differences between oligotrophic and nitrophytic species. Here, we compare a nitrogen sensitive lichen (Evernia prunastri) with a nitrogen tolerant one (Xanthoria parietina), focussing on their physiological response (Fv/Fm) to short-term NH3 exposure and their frequency of occurrence along an NH3 field gradient. Both frequency and Fv/Fm of E. prunastri decreased abruptly above 3 µg m(-3) NH3 suggesting direct adverse effects of NH3 on its photosynthetic performance. By contrast, X. parietina increased its frequency with NH3, despite showing decreased capacity of photosystem II above 50 µg m(-3) NH3, suggesting that the ecological success of X. parietina at ammonia-rich sites might be related to indirect effects of increased nitrogen (NH3) availability. These results highlight the need to establish NH3 critical levels based on oligotrophic lichen species.

How does an Al-hyperaccumulator plant respond to a natural field gradient of soil phytoavailable Al?

The physiological ability of plants to cope with Al-toxicity has attracted considerable attention. In this study we used an endemic Al-hyperaccumulator plant, Plantago almogravensis, which is the only known representative of the Plantaginaceae with this trait growing under a field gradient of Al, to understand the root and shoot patterns of Al accumulation and tolerance in its natural environment. We analysed phytoavailable elements in the soil and their accumulation in the plant. For the first time under field conditions, the accumulation pattern of an Al-hyperaccumulator showed a saturation curve with a maximum accumulation capacity being reached (ca. 3.0 mg g(-1)). The Al toxicity was not associated with the expected reduction in the Ca and Mg uptake by the plant. Iron was accumulated in a more linear pattern. The magnitude and the proportion of the elements found in the apoplastic fraction of the root, compared to the soil and plant internal fractions, suggested that the control of uptake occurs at the rhizospheric level. Unlike the majority of the Al-hyperaccumulator plants that are found in tropical humid areas, this plant is described from a sub-arid Mediterranean climate, subject to drought conditions which give it a unique status that deserves to be studied further.

Causes of change in nitrophytic and oligotrophic lichen species in a Mediterranean climate: impact of land cover and atmospheric pollutants.

With the aim of determining the main drivers of changes in nitrophytic and oligotrophic macro-lichen communities in an industrial region with a Mediterranean climate, we considered both land-cover types and atmospheric pollutants. We determined the relation between the abundance of nitrophytic and oligotrophic species with environmental factors considering the distance of influence of land-cover types. The results showed that oligotrophic species decreased in the proximity of artificial areas, barren land and agricultural areas, associated with higher concentrations of NO2 and Zn, and Ti, probably dust of industrial and agricultural origin. Nitrophytic species were positively related to all the mentioned land-cover types, and with higher concentrations of Fe and N. Magnesium, probably from ocean aerosols, was negatively related to oligotrophic species and positively to nitrophytic.

Impact of neighbourhood land-cover in epiphytic lichen diversity: analysis of multiple factors working at different spatial scales.

The objective of this work was to determine the impact of neighbourhood land-cover in epiphytic lichen diversity. We used geostatistics to analyse the spatial structure of lichen-indicators (number of lichen species and Lichen Diversity Value) and correlate them to land-cover considering different distances from the observed data. The results showed that lichen diversity was influenced by different environmental factors that act in the same territory but impact lichens at different distances from the source. The differences in the distance of influence of the several land-cover types seem to be related to the size of pollutants/particles that predominantly are dispersed by each land-cover type. We also showed that a local scale of analysis gives a deeper insight into the understanding of lichen richness and abundance in the region. This work highlighted the importance of a multiple spatial scale of analysis to deeply interpret the relation between lichen diversity and the underling environmental factors.

Improving the use of lichens as biomonitors of atmospheric metal pollution.

The data reported on this study supported the hypothesis that the absence of the lichen Ramalina fastigiata near a copper mine site located on the south of Portugal was related to toxic levels of Cu-dust near the centre of the mine. Lichen biodiversity reflected the impact of the copper-mine dust emissions which were more widespread towards the east, correlated with wind direction and frequency. The chemical analysis of R. fastigiata collected at different distances and in different directions from the mine showed that Cu, K and Mg were derived from the centre of the mine site, confirming thus as the major source of atmospheric dust. Total inhibition of PSII photochemical reactions occurred in R. fastigiata both under field and controlled conditions, when intracellular Cu concentrations exceeded a threshold of approximately 2.0 mumol g-1. No samples of this species were found under field conditions beyond the Cu threshold. Thus, the fluorescence parameter Fv/Fm proved to be a good estimator of the survival capacity of R. fastigiata under field conditions and thus a useful parameter in determining the sensitivity of the lichens (photobiont) to Cu pollution. The intracellular location of Cu allowed an explanation of the physiological changes and the survival of the species in the surroundings of the copper-mine.