Lichens as spatially transferable bioindicators for monitoring nitrogen pollution.

Excess nitrogen is a pollutant and global problem that harms ecosystems and can severely affect human health. Pollutant nitrogen is becoming more widespread and intensifying in the tropics. There is thus a requirement to develop nitrogen biomonitoring for spatial mapping and trend analysis of tropical biodiversity and ecosystems. In temperate and boreal zones, multiple bioindicators for nitrogen pollution have been developed, with lichen epiphytes among the most sensitive and widely applied. However, the state of our current knowledge on bioindicators is geographically biased, with extensive research effort focused on bioindicators in the temperate and boreal zones. The development of lichen bioindicators in the tropics is further weakened by incomplete taxonomic and ecological knowledge. In this study we performed a literature review and meta-analysis, attempting to identify characteristics of lichens that offer transferability of bioindication into tropical regions. This transferability must overcome the different species pools between source information - drawing on extensive research effort in the temperate and boreal zone - and tropical ecosystems. Focussing on ammonia concentration as the nitrogen pollutant, we identify a set of morphological traits and taxonomic relationships that cause lichen epiphytes to be more sensitive, or more resistant to this excess nitrogen. We perform an independent test of our bioindicator scheme and offer recommendations for its application and future research in the tropics.

Phytase activity in lichens.

Phytase activity was investigated in 13 lichen species using a novel assay method. The work tested the hypothesis that phytase is a component of the suite of surface-bound lichen enzymes that hydrolyse simple organic forms of phosphorus (P) and nitrogen (N) deposited onto the thallus surface. Hydrolysis of inositol hexaphosphate (InsP6 , the substrate for phytase) and appearance of lower-order inositol phosphates (InsP5 -InsP1 ), the hydrolysis products, were measured by ion chromatography. Phytase activity in Evernia prunastri was compared among locations with contrasting rates of N deposition. Phytase activity was readily measurable in epiphytic lichens (e.g. 11.3 µmol InsP6 hydrolysed g(-1)  h(-1) in Bryoria fuscescens) but low in two terricolous species tested (Cladonia portentosa and Peltigera membranacea). Phytase and phosphomonoesterase activities were positively correlated amongst species. In E. prunastri both enzyme activities were promoted by N enrichment and phytase activity was readily released into thallus washings. InsP6 was not detected in tree canopy throughfall but was present in pollen leachate. Capacity to hydrolyse InsP6 appears widespread amongst lichens potentially promoting P capture from atmospheric deposits and plant leachates, and P cycling in forest canopies. The enzyme assay used here might find wider application in studies on plant root-fungal-soil systems.

Metabolite profile shifts in the heathland lichen Cladonia portentosa in response to N deposition reveal novel biomarkers.

The heathland lichen Cladonia portentosa was collected from sites in mainland Britain differing either in rates of wet N deposition or in annual mean N concentration in rainfall based on a modelled data set. Methanolic extracts of thalli were analyzed by liquid chromatography time-of-flight mass spectrometry to yield metabolic profiles. Differences between sites in metabolite concentration were quantified using multivariate statistical tools and used to identify potential biomarker molecules. The abundances of three structurally related betaine lipids showed an increase with increasing modelled N deposition to a threshold of 22.3 kg ha(-1) year(-1) after which they remained constant. In contrast, the abundance of a phosphatidylcholine (PC) lipid showed concomitant decrease. Correlations of the identified biomarkers with N deposition and precipitation were stronger than those with N concentrations. The results presented in this study clearly show that N enrichment associated with tissue P limitation changes lipid composition, leading to shifts from PCs to betaine lipids, and that these lipids identified have the potential to be used as biomarkers for nitrogen enrichment.

Alterations in the metabolic fingerprint of Cladonia portentosa in response to atmospheric nitrogen deposition.

Nitrogen availability has profound ecological consequences in nutrient-limited systems. In terrestrial settings these would include the upland heaths, sand dunes and blanket bogs of temperate latitudes. Understanding the physiological consequences of nitrogen enrichment is a first critical step in predicting possible consequences. Results are presented from a metabolic fingerprinting study using Fourier transform-infrared spectroscopy (FTIR) to detect biochemical differences in the lichen Cladonia portentosa collected from 25 sites across mainland Britain varying in their nitrogen input. Partial least-squares regression analysis of the FTIR data demonstrated that changes in broad biochemical classes were consistently correlated with mean annual wet inorganic nitrogen deposition loads. These results demonstrated a direct coupling of a broad range of metabolic processes in C. portentosa to nitrogen deposition.

Connectivity, non-random extinction and ecosystem function in experimental metacommunities.

The spatial insurance hypothesis indicates that connectivity is an important attribute of natural ecosystems that sustains both biodiversity and ecosystem function. We tested the hypothesis by measuring the impact of manipulating connectivity in experimental metacommunties of a natural and diverse microecosystem. Isolation led to the extinction of large-bodied apex predators, subsequently followed by increases in prey species abundance. This trophic cascade was associated with significantly altered carbon and nitrogen fluxes in fragmented treatments. The ecosystem impacts were characteristic of a function debt because they persisted for several generations after the initial loss of connectivity. Local extinctions and disruption of ecosystem processes were mitigated, and even reversed, by the presence of corridors in the connected metacommunities, although these beneficial effects were unexpectedly delayed. We hypothesized that corridors maintained grazer movement between fragments, which enhanced microbial activity, and decomposition in comparison to isolated fragments. Our results indicate that knowledge of habitat connectivity and spatial processes is essential to understand the magnitude and timing of ecosystem perturbation in fragmented landscapes.

Perceived and measured levels of environmental pollution: interdisciplinary research in the subarctic lowlands of northeast European Russia.

Using interdisciplinary field research in the Usa Basin, northeast European Russia, we compared local inhabitants' perception of environmental problems with chemical and remote-sensing signatures of environmental pollution and their local impacts. Extensive coal mining since the 1930s around Inta and Vorkuta has left a legacy of pollution, detected by measuring snowpack, topsoil, and lichen chemistry, together with remote-sensing techniques and analysis of lake water and sediments. Vorkuta and its environs suffered the worst impacts, with significant metal loading and alkalization in lakes and topsoils, elevated metals and cations in terricolous (reindeer) lichens, and changes in vegetation communities. Although the coal industry has declined recently, the area boasts a booming oil and gas industry, based around Usinsk. Local perceptions and concerns of environmental pollution and protection were higher in Usinsk, as a result of increased awareness after a major oil spill in 1994, compared with Vorkuta's inhabitants, who perceived air pollution as the primary environmental threat. Our studies indicate that the principal sources of atmospheric emissions and local deposition within 25 to 40 km of Vorkuta were coal combustion from power and heating plants, coal mines, and a cement factory. Local people evaluated air pollution from direct observations and personal experiences, such as discoloration of snow and respiratory problems, whereas scientific knowledge played a minor role in shaping these perceptions.

Breeding systems in the lichen-forming fungal genus Cladonia.

The breeding systems of three species of the lichen-forming fungal genus Cladonia were investigated. Cladonia floerkeana, Cladonia galindezii, and Cladonia portentosa were selected due to their contrasting ecologies and reproductive strategies, and because they belong to the Lecanorales, the major lichen-forming order. Sibling single-spore progeny were collected from apothecia and used to establish axenic cultures. Two experimental approaches were used to determine breeding systems. First, RAPD-PCR and AFLP fingerprinting revealed that spores from the same apothecium were not genetically uniform, indicating heterothallism in each of these species. Second, segregation of a MAT-2 mating-type gene was assessed using degenerate PCR primers designed to amplify the high-mobility group region. A MAT-2 gene occurred in 40-60% of progeny, consistent with a heterothallic breeding system. The PCR product from C. galindezii was cloned and sequenced, and confirmed to have the characteristic motifs of a MAT-2 HMG gene. This is thought to be the first report of the use of segregation of a mating-type gene among ascospore progeny to determine the breeding system of a fungal species. The ecological significance of the results is discussed.

The natural abundance of (15)N in mat-forming lichens.

Natural abundance of (15)N and [N] was studied in thalli of mat-forming lichens collected from tundra and heathland sites in the northern and southern hemispheres. The study includes samples of British Cladonia portentosa from sites in regions of high and low N-loading and in heathland growing both directly on peat and independently of the soil substratum, in a canopy of prostrate gorse ( Ulex minor). In the mat-forming lichens examined, a non-random pattern in [N] and delta(15)N was characterised by a minimum in delta(15)N, which occurred most frequently at 20-40 mm below the thallus apex. Nitrogen concentration increased above this point, towards the apex, though remained invariably low towards the thallus base. We discuss the significance of the pattern in [N] and delta(15)N for current theories describing the uptake and recycling of nitrogen by mat-forming lichens in oligotrophic habitats. Our data are incompatible with the suggested uptake of soil organic-N depleted in (15)N, though are consistent with possible internal recycling and the development of a structural necromass. The study emphasises the internal fractionation of nitrogen isotopes and provides a caveat against the assumption that values of delta(15)N provide an unequivocal indicator of source-sink relationships in nitrogen cycling.