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.

Preliminary assessment of terrestrial microalgae isolated from lichens as testing species for environmental monitoring: lichen phycobionts present high sensitivity to environmental micropollutants.

Bioassays constitute a tool for pollution analysis providing a holistic approach and high-quality indication of the toxicity. Microbioassays allow evaluating the toxicity of many samples, implying lower costs and enabling routine monitoring and pollution control. But tests conducted so far are limited to the use of a small number of taxa. Lichens are excellent bioindicators of pollution with great ecological significance. Studies show that the phycobiont is more sensitive to pollutants than the mycobiont. Phycobiont have features such as adaptation to anhydrobiosis and relatively rapid growth in vitro, making them suitable for microbioassays. Our aim is to determine the sensitivity of phycobionts to the pharmaceutical micropollutants carbamazepine and diclofenac as a preliminary step for the development of a toxicity microbioassay based on phycobionts. Optical dispersion and chlorophyll autofluorescence were used as endpoints of toxicity on two algal species showing that suspensions present cyclic and taxon specific patterns of aggregation. Trebouxia TR9 suspensions present a very high grade of aggregation while Asterochloris erici cells do not. Both micropollutants alter optical properties of the suspensions of both species. No significant alteration of chlorophyll autofluorescence by carbamazepine is observed. A. erici chlorophyll autofluorescence is extremely sensitive to diclofenac but the effect is not dependent on the drug concentration or on the time of exposure. Differently, TR9 only shows punctual chlorophyll alterations. Fluctuations in optical dispersion may indicate changes in the population structure of the species, including reproductive strategy. A. erici seems more sensitive to micropollutants, is better characterized and is available from commercial collections.

Estimation of Endocarpon pusillum Hedwig carbon budget in the Tengger Desert based on its photosynthetic rate.

This study investigated the photosynthetic rate of the lichen Endocarpon pusillum at the Chinese Academy of Sciences Shapotou Desert Research Station and estimated its annual contribution to the carbon budget in the ecosystem. The software SigmaPlot 10.0 with "Macro-Area below curves" was used to calculate the carbon fixation capacity of the lichen. The total carbon budget (ΣC) of the lichen was obtained by subtracting the respiratory carbon loss (ΣDR) from the photosynthetic carbon gain (ΣNP). Because water from precipitation plays an important role in photosynthesis in this ecosystem, the annual carbon budget of E. pusillum at the station was estimated based on the three-year average precipitation data from 2009 to 2011. Our results indicate that the lichen fixes 14.6 g C m(-2) annually. The results suggest that artificial inoculation of the crust lichen in the Tengger Desert could not only help reduce the sand and dust storms but also offer a significant carbon sink, fixing a total of 438000 t of carbon over the 30000 km(2) of the Tengger Desert. The carbon sink could potentially help mitigate the atmospheric greenhouse effect. Our study suggests that the carpet-like lichen E. pusillum is an excellent candidate for "Bio-carpet Engineering" of arid and semi-arid regions.

Epiphytic lichen diversity in central European oak forests: assessment of the effects of natural environmental factors and human influences.

We investigated lichen diversity in temperate oak forests using standardized protocols. Forty-eight sites were sampled in the Czech Republic, Slovakia and Hungary. The effects of natural environmental predictors and human influences on lichen diversity (lichen diversity value, species richness) were analysed by means of correlation tests. We found that lichen diversity responded differently to environmental predictors between two regions with different human impact. In the industrial region, air pollution was the strongest factor. In the agricultural to highly forested regions, lichen diversity was strongly influenced by forest age and forest fragmentation. We found that several natural factors can in some cases obscure the effect of human influences. Thus, factors of natural gradient must be considered (both statistically and interpretively) when studying human impact on lichen diversity.

Rapid biodiversity assessment in lichen diversity surveys: implications for quality assurance.

Rapid Biodiversity Assessments (RBAs) of lichen communities, obtained by means of simplified sampling lists based on morphospecies, showed good correlations with Lichen Diversity Values (LDVs), based on the complete identification of lichen species only when performed by operators with high levels of taxonomic knowledge. Furthermore, the use of highly simplified sampling lists did not lead to significant advantages in terms of time needed for field operations. This approach proved to be especially unreliable in high diversity ecological contexts where variation of morpho-structural composition within lichen communities is frequent (i.e. co-occurring crustose- and foliose-dominated communities); it may also lead to weak results if applied for conservation purposes. Hence, the use of simplified RBA sampling lists in lichen monitoring has to be carefully evaluated and, in any case, should be based on sound taxonomic knowledge on the part of those in charge of data collection. The proper assessment of descriptors of lichen abundance and/or frequency, however, strictly depends on the skill, taxonomic knowledge, and willingness to learn of the lichenologist-in-training.

Fungus-specific microsatellite primers of lichens: application for the assessment of genetic variation on different spatial scales in Lobaria pulmonaria.

We isolated 12 microsatellite loci for the epiphytic lichen-forming ascomycete Lobaria pulmonaria and studied their patterns of variation within and among populations from Canada and Switzerland. Even though several microsatellites exhibited high levels of variability at different spatial scales, we did not find any evidence for intrathalline variation. Most of the genetic variation was attributed to differences among individuals within populations. High genetic variation was also detected among L. pulmonaria samples taken from individual trees, suggesting that either multiple colonization events had occurred or that local recombination is frequent. The geographically structured distribution of alleles from several microsatellites indicated that L. pulmonaria from Canada and Switzerland represent two distinct evolutionary lineages. The potential to identify multiple alleles, and their transferability to closely related species, make microsatellites an ideal tool to study dispersal, population differentiation, and microevolution in lichens.