Cristina Branquinho

The cellular location of Cu in lichens and its effects on membrane integrity and chlorophyll fluorescence

Abstract The data in the present study supported the hypothesis that Na 2 -EDTA at pH 4.5 is an efficient chelating agent for extracellular Cu without causing cell membrane damage, and that it can be used in a sequential elution procedure to determine the cellular location of Cu in lichens. The patterns of extracellular uptake vs. time or concentration were anticipated from conventional kinetics studies with other organisms and heavy metals. Supplied Cu replaced the naturally acquired extracellular Mg and Ca and induced alterations in the passage of K across the cell membrane. For Usnea spp. intracellular uptake saturated at low supplied Cu concentrations (0.0157 mM) and was toxic, as indicated by chlorophyll fluorescence measurements. In Ramalina fastigiata intracellular Cu concentrations above ca. 4.0 μmol g −1 were linked to a decline in chlorophyll fluorescence. The fluorescence parameter F v F m was shown to be useful in determining the sensitivity of the lichens to Cu uptake. Usnea spp. were the lichens most sensitive to Cu uptake, since physiological changes occurred for lower supplied Cu concentrations than for R. fastigiata . With this work we were able to determine and quantify the cellular location of Cu in lichens. We were also able to evaluate through membrane integrity and fluorescence measurements the physiological state and the relative sensitivity of different lichen species to Cu uptake.

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.

Nitrogen Deposition Effects on Mediterranean-type Ecosystems: An Ecological Assessment

We review the ecological consequences of N deposition on the five Mediterranean regions of the world. Seasonality of precipitation and fires regulate the N cycle in these water-limited ecosystems, where dry N deposition dominates. Nitrogen accumulation in soils and on plant surfaces results in peaks of availability with the first winter rains. Decoupling between N flushes and plant demand promotes losses via leaching and gas emissions. Differences in P availability may control the response to N inputs and susceptibility to exotic plant invasion. Invasive grasses accumulate as fuel during the dry season, altering fire regimes. California and the Mediterranean Basin are the most threatened by N deposition; however, there is limited evidence for N deposition impacts outside of California. Consequently, more research is needed to determine critical loads for each region and vegetation type based on the most sensitive elements, such as changes in lichen species composition and N cycling.

Lead (Pb) uptake and its effects on membrane integrity and chlorophyll fluorescence in different lichen species

Abstract We report an investigation into the effect on membrane integrity and chlorophyll fluorescence of extra- and intracellular lead (Pb) concentrations in lichens possessing different photosynthetic symbionts and thallus growth forms (Lobaria pulmonaria, Ramalina farinacea, Parmelia caperata and Peltigera canina). Pb uptake, particularly in cyanobiont lichens, caused a decrease in photosystem II (PSII) photochemical reactions, measured as a change in the ratio variable fluorescence:maximal fluorescence ( F v F m ). In the lichens L. pulmonaria and Parmelia caperata, the effect on PSII photochemical reactions could only be observed 48 h after Pb uptake, confirming that prolonged exposure to Pb caused an additional decrease in F v F m . We confirmed that Pb enters the cells of the lichens. We found identical maximal extracellular Pb concentrations among the different lichens; however, the maximal intracellular Pb concentrations were not identical in range. Lichen PSII photochemical reactions were more sensitive to Pb than cell membrane damage since, with increasing Pb concentrations, no clear alterations in lichen membrane permeability, as measured by the loss of intracellular K and Mg, were observed. The effects of Pb in lichens were shown to be dependent only on the nature of the photobiont. We confirmed that Pb displaced extracellular exchangeable Ca and Mg.

Spatial modeling of PAHs in lichens for fingerprinting of multisource atmospheric pollution.

PAHs are toxic compounds emitted by several anthropogenic sources, which have a great impact on human health. We show, for the first time, how spatial models based on PAHs intercepted by lichens can be used for fingerprinting multisource atmospheric pollution in a regional area. Urban-industrial areas showed the highest atmospheric deposition of PAHs followed by urban > industrial > agricultural > forest Multivariate analysis of lichen data showed, for the first time, a clear distinction between various sources of PAHs in the same area: urban are dominated by 4-ring PAHs, forest by 3-ring PAHs, and industrial by 5- and 6-ring PAHs or by 2-ring PAHs (petrogenic or pyrogenic, respectively). Heavy metals were also used for supporting the fingerprinting of PAH sources, reinforcing the industrial origin of 5- and 6-ring PAHs and revealing their particular nature. The spatial structure of the models for different PAHs seems to be dependent on the following factors: size and hydrophilic character of different PAHs, type of emission sources (point or nonpoint), and dispersion associated with particulates of different sizes. Based on the long-term integration of PAHs in lichens, these spatial models will significantly improve our knowledge on the impact of PAH chronic-exposure to humans and ecosystems.

Guidelines for biomonitoring persistent organic pollutants (POPs), using lichens and aquatic mosses--a review.

During the last decades, awareness regarding persistent organic pollutants (POPs), such dioxins and furans (PCDD/Fs) and polycyclic aromatic hydrocarbons (PAHs), has become a cutting-edge topic, due to their toxicity, bioaccumulation and persistency in the environment. Monitoring of PCDD/Fs and PAHs in air and water has proven to be insufficient to capture deposition and effects of these compounds in the biota. To overcome this limitation, environmental biomonitoring using lichens and aquatic mosses, have aroused as promising tools. The main aim of this work is to provide a review of: i) factors that influence the interception and accumulation of POPs by lichens; ii) how lichens and aquatic bryophytes can be used to track different pollution sources and; iii) how can these biomonitors contribute to environmental health studies. This review will allow designing a set of guidelines to be followed when using biomonitors to assess environmental POP pollution.

The impact of dehydration rate on the production and cellular location of reactive oxygen species in an aquatic moss

BACKGROUND AND AIMS The aquatic moss Fontinalis antipyretica requires a slow rate of dehydration to survive a desiccation event. The present work examined whether differences in the dehydration rate resulted in corresponding differences in the production of reactive oxygen species (ROS) and therefore in the amount of cell damage. METHODS Intracellular ROS production by the aquatic moss was assessed with confocal laser microscopy and the ROS-specific chemical probe 2,7-dichlorodihydrofluorescein diacetate. The production of hydrogen peroxide was also quantified and its cellular location was assessed. KEY RESULTS The rehydration of slowly dried cells was associated with lower ROS production, thereby reducing the amount of cellular damage and increasing cell survival. A high oxygen consumption burst accompanied the initial stages of rehydration, perhaps due to the burst of ROS production. CONCLUSIONS A slow dehydration rate may induce cell protection mechanisms that serve to limit ROS production and reduce the oxidative burst, decreasing the number of damaged and dead cells due upon rehydration.

Lichen traits responding to aridity

Summary 1. Climate change is expected to cause several impacts at the global scale, and drylands will be amongst the most affected areas. Thus, investigating how these changes will affect the composition, structure and functioning of dryland ecosystems has become a priority. From an ecological indicator point of view, several works have shown that functional diversity is better than species richness to understand ecosystem functioning or response to environmental factors. However, most of these works focus on plants, while those of other organisms remain largely unknown. Lichens are amongst the ecosystem components more sensitive to climatic changes due to several physiological and ecological characteristics. Their poikilohydric nature (therefore highly dependent on the atmosphere for water supply) and their ubiquity on terrestrial ecosystems underlie their potential as indicators of climate. Nonetheless, works specifically aiming to identify lichen functional traits that respond to aridity remain poorly explored, particularly in drylands. 2. We proposed to identify lichen functional traits and respective functional groups responding to aridity in a Mediterranean drylands ecosystem. 3. Lichen diversity was sampled in open holm oak woodlands along an aridity gradient in SW Europe (Iberian Peninsula). Lichen functional traits that could be easily identified and related to water uptake were selected to be tested: type of photobiont, growth form and reproduction strategy. 4. Lichen species composition was related to the aridity gradient. The three traits chosen were related with the community’s response to aridity, but with contrasting responses in different functional groups. More specifically crustose and fruticose lichens, isidiate species and the ones with Trentepohlia as photobiont were related to the less arid part of the gradient. Foliose species and cyanolichens, on the contrary, were associated with the most arid areas. 5. Synthesis. We were able to identify lichen traits responding to aridity. Type of photobiont was particularly responsive, with Trentepohlia and cyanobacteria functional groups, responding clearly in contrasting ways to aridity in this drylands ecosystem. This work emphasizes functional diversity role on understanding and assessing the response to environmental factors, namely to climate. It also highlights the potential use of lichen functional groups as ecological indicators of climate change.

Differential proteomics of dehydration and rehydration in bryophytes: evidence towards a common desiccation tolerance mechanism

All bryophytes evolved desiccation tolerance (DT) mechanisms during the invasion of terrestrial habitats by early land plants. Are these DT mechanisms still present in bryophytes that colonize aquatic habitats? The aquatic bryophyte Fontinalis antipyretica Hedw. was subjected to two drying regimes and alterations in protein profiles and sucrose accumulation during dehydration and rehydration were investigated. Results show that during fast dehydration, there is very little variation in protein profiles, and upon rehydration proteins are leaked. On the other hand, slow dehydration induces changes in both dehydration and rehydration protein profiles, being similar to the protein profiles displayed by the terrestrial bryophytes Physcomitrella patens (Hedw.) Bruch and Schimp. and, to what is comparable with Syntrichia ruralis (Hedw.) F. Weber and D. Mohr. During dehydration there was a reduction in proteins associated with photosynthesis and the cytoskeleton, and an associated accumulation of proteins involved in sugar metabolism and plant defence mechanisms. Upon rehydration, protein accumulation patterns return to control values for both photosynthesis and cytoskeleton whereas proteins associated with sugar metabolism and defence proteins remain high. The current results suggest that bryophytes from different ecological adaptations may share common DT mechanisms.

A new framework for selecting environmental surrogates

Surrogate concepts are used in all sub-disciplines of environmental science. However, controversy remains regarding the extent to which surrogates are useful for resolving environmental problems. Here, we argue that conflicts about the utility of surrogates (and the related concepts of indicators and proxies) often reflect context-specific differences in trade-offs between measurement accuracy and practical constraints. By examining different approaches for selecting and applying surrogates, we identify five trade-offs that correspond to key points of contention in the application of surrogates. We then present an 8-step Adaptive Surrogacy Framework that incorporates cross-disciplinary perspectives from a wide spectrum of the environmental sciences, aiming to unify surrogate concepts across disciplines and applications. Our synthesis of the science of surrogates is intended as a first step towards fully leveraging knowledge accumulated across disciplines, thus consolidating lessons learned so that they may be accessible to all those operating in different fields, yet facing similar hurdles.