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Variations in stable carbon isotope discrimination (Δ) were investigated across the thalli of several lichen species possessing different photobiont associations. Lichens containing (i) green algae (phycobiont), (ii) green algae in association with cyanobacteria confined in cephalodia, or (iii) cyanobacteria (cyanobiont) as the photobiont partner were studied. Carbon isotope discrimination was analysed in different thallus sections, which varied in distance from the margin and in age. The marginal thallus region is considered to be youngest, while the central region is thought to be oldest. This analysis showed a clear variation in Δ across the thallus related to distance from the growing margin. In most of the species examined, the highest Δ values were found in marginal regions (younger), while the central and basal regions (older) showed significantly lower Δ. To investigate the effects of the historical increase in atmospheric CO2 concentration and the concurrent decrease in the 13C content of atmospheric CO2 on the Δ of lichens, experiments were carried out on herbarium samples of tobaria pulmonaria collected from the mid 19th Century to 1953. The results obtained showed a pattern of variation of Δ consistent with that of freshly collected samples; A decreased substantially with increasing distance from the thallus margin, irrespective of the collection date. Moreover, no consistent variation of discrimination was found among different collection dates. These results demonstrate that the observed variation in Δ is caused by age-related changes in the physiological behaviour of different thallus sections, and that the past 150 years of increasing CO concentration have not had significant effects on Δ in L. pulmonaria. Photosynthesis measurements, chlorophyll analysis and observations using optical microscopy, performed on freshly collected lichens, showed significant changes in morphological and physiological characteristics across the thallus. Particularly, remarkable variations in thickness were found across the thallus. These anatomical changes may be responsible for the variation in Δ, through variations in CO2 transfer resistance and, consequently, in CO2 availability across the thallus. The lack of age-dependent variation in Δ in cyanobiont lichens is possibly attributable to the operation of a CO2 -concentrating mechanism and, therefore, to a more constant CO2 environment across the thallus in this lichen group. [ABSTRACT FROM AUTHOR]

Acacia longifolia is one of the most aggressive invaders worldwide whose invasion is potentiated after a fire, a common perturbation in Mediterranean climates. As a legume, this species establishes symbioses with nitrogen-fixing bacteria inside root nodules; however, the overall microbial diversity is still unclear. In this study, we addressed root nodules' structure and biodiversity through histology and Next-Generation Sequencing, targeting 16S and 25S-28S rDNA genes for bacteria and fungi, respectively. We wanted to evaluate the effect of fire in root nodules from 1-year-old saplings, by comparing unburnt and burnt sites. We found that although having the same general structure, after a fire event, nodules had a higher number of infected cells and greater starch accumulation. Starch accumulated in uninfected cells can be a possible carbon source for the microbiota. Regarding diversity, Bradyrhizobium was dominant in both sites (ca. 77%), suggesting it is the preferential partner, followed by Tardiphaga (ca. 9%), a non-rhizobial Alphaproteobacteria, and Synechococcus , a cyanobacteria (ca. 5%). However, at the burnt site, additional N-fixing bacteria were included in the top 10 genera, highlighting the importance of this process. Major differences were found in the mycobiome, which was diverse in both sites and included genera mostly described as plant endophytes. Coniochaeta was dominant in nodules from the burnt site (69%), suggesting its role as a facilitator of symbiotic associations. We highlight the presence of a large bacterial and fungal community in nodules, suggesting nodulation is not restricted to nitrogen fixation. Thus, this microbiome can be involved in facilitating A. longifolia invasive success.

Biodiversity is declining due to the impact of human activities. However, public awareness of the biodiversity crisis is low, particularly for plants, creating a barrier to engage with conservation programs. In this perspective, we show how citizen science and mobile apps can be used as educational tools to raise awareness about plant biodiversity among students and the general public. We examine the outcomes of three Bachelor of Science activities as well as two informal education initiatives. We discuss the potential of these approaches as educational and outreach tools. Our results show that citizen science and mobile apps are excellent tools for engaging society in biodiversity conservation and environmental issues., (© 2023. The Author(s).)

As climate change increasingly affects forest ecosystems, detailed understanding of major effects is important to anticipate their consequences under future climate scenarios. The Mediterranean region is a prominent climate change hotspot, and evergreen cork oak ( Quercus suber L.) woodlands are particularly climatically sensitive due to cork (bark) harvesting. Cork oak's drought avoidance strategy is well-known and includes structural and physiological adaptations that maximise soil water uptake and transport and limit water use, potentially leading to reduced stem and cork growth. Trees' responses to cope with water-limited conditions have been extensively described based on cork-rings width and, more recently, on cork-rings density, in dendroecological studies. However, so far, tree functional attributes and physiological strategies, namely photosynthetic metabolism adjustments affecting cork formation, have never been addressed and/or integrated on these previous cork-rings-based studies. In this study, we address the relation between carbon and oxygen stable isotopes of cork rings and precipitation and temperature, in two distinct locations of southwestern Portugal-the (wetter) Tagus basin peneplain and the (drier) Grândola mountains. We aimed at assessing whether the two climatic factors affect cork-ring isotopic composition under contrasting conditions of water availability, and, therefore, if carbon and oxygen signatures in cork can reflect tree functional (physiological and structural) responses to stressful conditions, which might be aggravated by climate change. Our results indicate differences between the study areas. At the drier site, the stronger statistically significant negative cork δ C correlations were found with mean temperature, whereas strong positive cork 13 C correlations were found with mean temperature, whereas strong positive cork δ 18 C, indicating, respectively, shallow groundwater as the water source for physiological processes related with biosynthesis of non-photosynthetic secondary tissues, such as suberin, and a weak stomatal regulation under high water availability, consistent with non-existent water availability constrains. In contrast, at the drier site, trees use water from deeper ground layers, depleted in 18 O and depleted in 13 C, indicating, respectively, shallow groundwater as the water source for physiological processes related with biosynthesis of non-photosynthetic secondary tissues, such as suberin, and a weak stomatal regulation under high water availability, consistent with non-existent water availability constrains. In contrast, at the drier site, trees use water from deeper ground layers, depleted in 18 O, and strongly regulate stomatal conductance under water stress, thus reducing photosynthetic carbon uptake and probably relying on stored carbon reserves for cork ring formation. These results suggest that although stable isotopes signatures in cork rings are not proxies for net growth, they may be (fairly) robust indicators of trees' physiological and structural adjustments to climate and environmental changes in Mediterranean environments., Competing Interests: The authors declare there are no competing interests., (©2022 Costa et al.)

Woody invasive species pose a big threat to ecosystems worldwide. Among them, Acacia longifolia is especially aggressive, fundamentally changing ecosystem structure through massive biomass input. This biomass is rarely harvested for usage; thus, these plants constitute a nuisance for stakeholders who invest time and money for control without monetary return. Simultaneously, there is an increased effort to valorise its biomass, e.g., for compost, growth substrate or as biofuel. However, to incentivise A. longifolia harvest and usage, stakeholders need to be able to estimate what can be obtained from management actions. Thus, the total biomass and its quality (C/N ratio) need to be predicted to perform cost-benefit analyses for usage and determine the level of invasion that has already occurred. Here, we report allometric biomass models for major biomass pools, as well as give an overview of biomass quality. Subsequently, we derive a simplified volume-based model (BM ~ 6.297 + 0.982 × Vol; BM = total dry biomass and Vol = plant volume), which can be applied to remote sensing data or with in situ manual measurements. This toolkit will help local stakeholders, forest managers or municipalities to predict the impact and valorisation potential of this invasive species and could ultimately encourage its management.

The pinewood nematode (PWN), Bursaphelenchus xylophilus , responsible for the pine wilt disease (PWD), is a major threat to pine forests worldwide. Since forest mortality due to PWN might be exacerbated by climate, the concerns regarding PWD in the Mediterranean region are further emphasized by the projected scenarios of more drought events and higher temperatures. In this context, it is essential to better understand the pine species vulnerability to PWN under these conditions. To achieve that, physiological responses and wilting symptoms were monitored in artificially inoculated Pinus pinaster ( P. pinaster ), Pinus pinea ( P. pinea ), and Pinus radiata ( P. radiata ) saplings under controlled temperature (25/30°C) and water availability (watered/water stressed). The results obtained showed that the impact of PWN is species-dependent, being infected P. pinaster and P. radiata more prone to physiological and morphological damage than P. pinea . For the more susceptible species ( P. pinaster and P. radiata ), the presence of the nematode was the main driver of photosynthetic responses, regardless of their temperature or water regime conditions. Nevertheless, water potential was revealed to be highly affected by the synergy of PWN and the studied abiotic conditions, with higher temperatures ( P. pinaster ) or water limitation ( P. radiata ) increasing the impact of nematodes on trees' water status. Furthermore, water limitation had an influence on nematodes density and its allocation on trees' structures, with P. pinaster revealing the highest nematode abundance and inner dispersion. In inoculated P. pinea individuals, nematodes' population decreased significantly, emphasizing this species resistance to PWN. Our findings revealed a synergistic impact of PWN infection and stressful environmental conditions, particularly on the water status of P. pinaster and P. radiata , triggering disease symptoms and mortality of these species. Our results suggest that predicted drought conditions might facilitate proliferation and exacerbate the impact of PWN on these two species, through xylem cavitation, leading to strong changes in pine forests of the Mediterranean regions., Competing Interests: FC was employed by company Infarm. The remaining authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2022 Estorninho, Chozas, Mendes, Colwell, Abrantes, Fonseca, Fernandes, Costa, Máguas, Correia and Antunes.)

Background and Aims: Invasive species may undergo rapid evolution despite very limited standing genetic diversity. This so-called genetic paradox of biological invasions assumes that an invasive species has experienced (and survived) a genetic bottleneck and then underwent local adaptation in the new range. In this study, we test how often Australian acacias (genus Acacia), one of the world's most problematic invasive tree groups, have experienced genetic bottlenecks and inbreeding., Methods: We collated genetic data from 51 different genetic studies on Acacia species to compare genetic diversity between native and invasive populations. These studies analysed 37 different Acacia species, with genetic data from the invasive ranges of 11 species, and data from the native range for 36 species (14 of these 36 species are known to be invasive somewhere in the world, and the other 22 are not known to be invasive)., Key Results: Levels of genetic diversity are similar in native and invasive populations, and there is little evidence of invasive populations being extensively inbred. Levels of genetic diversity in native range populations also did not differ significantly between species that have and that do not have invasive populations., Conclusion: We attribute our findings to the impressive movement, introduction effort and human usage of Australian acacias around the world., (© The Author(s) 2021. Published by Oxford University Press on behalf of the Annals of Botany Company. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com.)

(1) Background: the Miombo woodlands comprise the most important vegetation from southern Africa and are dominated by tree legumes with an ecology highly driven by fires. Here, we report on the characterization of bacterial communities from the rhizosphere of Brachystegia boehmii in different soil types from areas subjected to different regimes. (2) Methods: bacterial communities were identified through Illumina MiSeq sequencing (16S rRNA). Vigna unguiculata was used as a trap to capture nitrogen-fixing bacteria and culture-dependent methods in selective media were used to isolate plant growth promoting bacteria (PGPB). PGP traits were analysed and molecular taxonomy of the purified isolates was performed. (3) Results: Bacterial communities in the Miombo rhizosphere are highly diverse and driven by soil type and fire regime. Independent of the soil or fire regime, the functional diversity was high, and the different consortia maintained the general functions. A diverse pool of diazotrophs was isolated, and included symbiotic (e.g., Mesorhizobium sp., Neorhizobium galegae , Rhizobium sp., and Ensifer adhaerens), and non-symbiotic (e.g., Agrobacterium sp., Burkholderia sp., Cohnella sp., Microvirga sp., Pseudomonas sp., and Stenotrophomonas sp.) bacteria. Several isolates presented cumulative PGP traits. (4) Conclusions: Although the dynamics of bacterial communities from the Miombo rhizosphere is driven by fire, the maintenance of high levels of diversity and functions remain unchanged, constituting a source of promising bacteria in terms of plant-beneficial activities such as mobilization and acquisition of nutrients, mitigation of abiotic stress, and modulation of plant hormone levels.

It is often overlooked that even food production is linked to the ecology of plants and animals. Living organisms respond to environmental short-and long-term variability: acknowledging this may help in the ultimate goal of valorizing a territory/product. We investigated acorns of the two main Quercus species of the Portuguese Montado , a main feed of the renown black Iberian pig. We tested their responses to an aridity gradient by morphological parameters and isotopic signature. Q. rotundifolia and Q. suber acorns did not differ morphologically, even if a higher variability in all parameters was observed in acorns of Q. suber . According to the site-specific Aridity Index, correlations are indicative to higher weight and length only in Q. suber acorns from more arid sites. As for isotopic composition, there were no differences in nitrogen or carbon (δ 15 N and δ 13 C) between the two species. However, combining the samples and testing for association with the Aridity Index, we found that more arid sites lead to a 15 N enrichment. This result, combined with the positive correlation between AI and acorns length, support the use of acorns as a tool, their isoscapes of nitrogen being a stepping stone for the provenance of the black Iberian pig.

Tree ring synthesis is a key process in wood production; however, little is known of the origin and fate of the carbon involved. We used natural 13C abundance to investigate the carbon-use process for the ring development in a temperate deciduous (Quercus petraea (Matt.) Liebl.) and a Mediterranean evergreen (Quercus ilex L.) oak. The sapwood carbon reserves, phloem sucrose contents, stem respired CO2 efflux and their respective carbon isotope compositions (δ13C) were recorded over 1 year, in the native area of each species. The seasonal δ13C variation of the current year ring was determined in the total ring throughout the seasons, as well as in slices from the fully mature ring after the growth season (intra-ring pattern). Although the budburst dates of the two oaks were similar, the growth of Quercus ilex began 50 days later. Both species exhibited growth cessation during the hot and dry summer but only Q. ilex resumed in the autumn. In the deciduous oak, xylem starch storage showed clear variations during the radial growth. The intra-ring δ13C variations of the two species exhibited similar ranges, but contrasting patterns, with an early increase for Q. petraea. Comparison between δ13C of starch and total ring suggested that Q. petraea (but not Q. ilex) builds its rings using reserves during the first month of growth. Shifts in ring and soluble sugars δ13C suggested an interspecific difference in either the phloem unloading or the use of fresh assimilate inside the ring. A decrease in ring δ13C for both oaks between the end of the radial growth and the winter is attributed to a lignification of ring cell walls after stem increment. This study highlighted the differences in carbon-use during ring growth for evergreen and deciduous oaks, as well as the benefits of exploring the process using natural 13C abundance., (© The Author(s) 2019. Published by Oxford University Press. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com.)

Although it is generally accepted that δ 15 N in lichen reflects predominating N isotope sources in the environment, confirmation of the direct correlation between lichen δ 15 N and atmospheric δ 15 N 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, δ 15 N and chlorophyll a fluorescence, were measured. Under the experimental conditions, δ 15 N in lichens quantitatively responded to the δ 15 N of released gaseous ammonia (NH 3 ). 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 NH 3 , 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 NH 3 , then NH 4 + , with lowest sensitivity to NO 3 - . Moreover, these results confirm that lichen δ 15 N can be used to indicate the δ 15 N of atmospheric ammonia, providing a suitable tool for the interpretation of the spatial distribution of NH 3 sources in relation to their δ 15 N signal., (Copyright © 2018. Published by Elsevier B.V.)

Predicted droughts and anthropogenic water use will increase groundwater lowering rates and intensify groundwater limitation, particularly for Mediterranean semi-arid ecosystems. These hydrological changes may be expected to elicit differential functional responses of vegetation either belowground or aboveground. Yet, our ability to predict the impacts of groundwater changes on these ecosystems is still poor. Thus, we sought to better understand the impact of falling water table on the physiology of woody vegetation. We specifically ask (a) how is woody vegetation ecophysiological performance affected by water table depth during the dry season? and (b) does the vegetation response to increasing depth to groundwater differ among water-use functional types? We examined a suite of physiological parameters and water-uptake depths of the dominant, functionally distinct woody vegetation along a water-table depth gradient in a Mediterranean semi-arid coastal ecosystem that is currently experiencing anthropogenic groundwater extraction pressure. We found that groundwater drawdown did negatively affect the ecophysiological performance of the woody vegetation. Across all studied environmental factors, depth to groundwater was the most important driver of ecophysiological adjustments. Plant functional types, independent of groundwater dependence, showed consistent declines in water content and generally reduced C and N acquisition with increasing depths to groundwater. Functional types showed distinct operating physiological ranges, but common physiological sensitivity to greater water table depth. Thus, although differences in water-source use exist, a physiological convergence appeared to happen among different functional types. These results strongly suggest that hydrological drought has an important impact on fundamental physiological processes, constraining the performance of woody vegetation under semi-arid conditions. By disentangling the functional responses and vulnerability of woody vegetation to groundwater limitation, our study establishes the basis for predicting the physiological responses of woody vegetation in semi-arid coastal ecosystems to groundwater drawdown., (© 2018 John Wiley & Sons Ltd.)

The number of alien plants escaping from cultivation into native ecosystems is increasing steadily. We provide an overview of the historical, contemporary and potential future roles of ornamental horticulture in plant invasions. We show that currently at least 75% and 93% of the global naturalised alien flora is grown in domestic and botanical gardens, respectively. Species grown in gardens also have a larger naturalised range than those that are not. After the Middle Ages, particularly in the 18th and 19th centuries, a global trade network in plants emerged. Since then, cultivated alien species also started to appear in the wild more frequently than non-cultivated aliens globally, particularly during the 19th century. Horticulture still plays a prominent role in current plant introduction, and the monetary value of live-plant imports in different parts of the world is steadily increasing. Historically, botanical gardens - an important component of horticulture - played a major role in displaying, cultivating and distributing new plant discoveries. While the role of botanical gardens in the horticultural supply chain has declined, they are still a significant link, with one-third of institutions involved in retail-plant sales and horticultural research. However, botanical gardens have also become more dependent on commercial nurseries as plant sources, particularly in North America. Plants selected for ornamental purposes are not a random selection of the global flora, and some of the plant characteristics promoted through horticulture, such as fast growth, also promote invasion. Efforts to breed non-invasive plant cultivars are still rare. Socio-economical, technological, and environmental changes will lead to novel patterns of plant introductions and invasion opportunities for the species that are already cultivated. We describe the role that horticulture could play in mediating these changes. We identify current research challenges, and call for more research efforts on the past and current role of horticulture in plant invasions. This is required to develop science-based regulatory frameworks to prevent further plant invasions., (© 2018 Cambridge Philosophical Society.)

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 δ 15 N. 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.

The effects of dissolved inorganic carbon (DIC) availability on photosynthesis were studied in two estuarine intertidal microphytobenthos (MPB) communities and in the model diatom species Phaeodactylum tricornutum. Kinetics of DIC acquisition, measured with a liquid-phase oxygen electrode, showed higher K(1/2)(DIC) (0.31 mM) and Vm (7.78 nmol min(-1) µg (Chl a)(-1)) for MPB suspensions than for P. tricornutum (K(1/2)(DIC) = 0.23 mM; Vm = 4.64 nmol min(-1) µg (Chl a)(-1)), suggesting the predominance of species with lower affinity for DIC and higher photosynthetic capacity in the MPB. The net photosynthetic rate of the MPB suspensions reached saturation at a DIC concentration of 1-1.5 mM. This range was lower than the concentrations found in the interstitial water of the top 5-mm sediment layer, suggesting no limitation of photosynthesis by DIC in the MPB communities. Accordingly, carbon isotope discrimination revealed a moderate activity of CO2-concentrating mechanisms in the MPB. However, addition of NaHCO3 to intact MPB biofilms caused a significant increase in the relative maximum photosynthetic electron transport rate (rETR max) measured by imaging pulse-amplitude modulated chlorophyll a fluorescence. These results suggest local depletion of DIC at the photic layer of the sediment (the first few hundred µm), where MPB cells accumulate during diurnal low tides. This work provides the first direct experimental evidence of DIC limitation of photosynthesis in highly productive intertidal MPB communities.

The tropical coffee crop has been predicted to be threatened by future climate changes and global warming. However, the real biological effects of such changes remain unknown. Therefore, this work aims to link the physiological and biochemical responses of photosynthesis to elevated air [CO2 ] and temperature in cultivated genotypes of Coffea arabica L. (cv. Icatu and IPR108) and Coffea canephora cv. Conilon CL153. Plants were grown for ca. 10 months at 25/20°C (day/night) and 380 or 700 μl CO2 l(-1) and then subjected to temperature increase (0.5°C day(-1) ) to 42/34°C. Leaf impacts related to stomatal traits, gas exchanges, C isotope composition, fluorescence parameters, thylakoid electron transport and enzyme activities were assessed at 25/20, 31/25, 37/30 and 42/34°C. The results showed that (1) both species were remarkably heat tolerant up to 37/30°C, but at 42/34°C a threshold for irreversible nonstomatal deleterious effects was reached. Impairments were greater in C. arabica (especially in Icatu) and under normal [CO2 ]. Photosystems and thylakoid electron transport were shown to be quite heat tolerant, contrasting to the enzymes related to energy metabolism, including RuBisCO, which were the most sensitive components. (2) Significant stomatal trait modifications were promoted almost exclusively by temperature and were species dependent. Elevated [CO2 ], (3) strongly mitigated the impact of temperature on both species, particularly at 42/34°C, modifying the response to supra-optimal temperatures, (4) promoted higher water-use efficiency under moderately higher temperature (31/25°C) and (5) did not provoke photosynthetic downregulation. Instead, enhancements in [CO2 ] strengthened photosynthetic photochemical efficiency, energy use and biochemical functioning at all temperatures. Our novel findings demonstrate a relevant heat resilience of coffee species and that elevated [CO2 ] remarkably mitigated the impact of heat on coffee physiology, therefore playing a key role in this crop sustainability under future climate change scenarios., (© 2015 John Wiley & Sons Ltd.)

Despite reductions in atmospheric sulfur (S) concentrations due to abatement policies in some countries, modeling the dispersion of this pollutant and disentangling anthropogenic sources from natural ones is still of great concern. Lichens have been used as biomonitors of the impacts of S for over 40 years, but their potential as source-tracers of specific sources, including natural ones, remains unexplored. In fact, few attempts have been made to try to distinguish and spatially model different sources of S using lichens. We have measured S concentrations and isotopic values in lichens within an industrial coastal region where different sources of S, natural and anthropogenic, interplay. We detected a prevailing influence of natural sea-originated S that mixed with anthropogenic sources of S. We were then able to disentangle the sources of S, by removing the ocean influence on S isotopic values, enabling us to model the impact of different anthropogenic sources on S deposition and highlighting the potential use of lichens to evaluate the weight of different types of anthropogenic sources.

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., (Copyright © 2013 Elsevier Ltd. All rights reserved.)

One of the main drawbacks of using lichens to monitor atmospheric PAHs has been reported as the inexistence of studies aiming to translate PAH values in lichens into the atmospheric equivalents ones, in order to use this information for regulatory purposes. In this work, PAH concentrations in lichens were compared with PAH concentrations measured in a conventional active sampler in an outdoor environment for a 9-month span. Significant positive correlations between HMW-PAHs, Σ16 EPA-PAHs, and BaP equivalent concentrations in lichens and those in air (TSP) were found. Concentrations of Σ16 EPA-PAHs in lichens and air showed a seasonal variation, with highest values during winter and lowest values during summer. Meteorological variables - temperature, atmospheric pressure, relative humidity, and wind speed - showed to significantly influence PAH concentrations in both lichens and air. Based on the significant linear correlations, equations for translating PAH concentrations measured in lichens into equivalent ones for air were proposed for the first time, allowing a broader use of lichens' information regarding PAHs in monitoring schemes and decision-making., (Copyright © 2013 Elsevier Ltd. All rights reserved.)

Polycyclic aromatic hydrocarbons (PAH) are toxic compounds that have been classified by the International Agency for Research on Cancer as probable or possible human carcinogens. Human exposure to PAH is usually assessed by considering data from a single air monitoring station as being representative of a large region; however, air pollution levels change on small spatial scales and thus also affect environmental exposure. The use of environmental biomonitors is a useful tool to assess the levels of PAH with high spatial resolution. The aims of this study were to (1) assess human exposure to PAH in a petrochemical region in Portugal, integrating data from environmental biomonitors (lichens), air, and soil in a regional area, and (2) determine the health risks associated with exposure to PAH with high spatial resolution. Bearing this in mind, benzo[a]pyrene (BaP) equivalent concentrations in samples of soil, air, and lichens collected in the study region were used to assess human exposure through different pathways, including inhalation of air and soil particles, ingestion of soil, and dermal contact with soil. Human health risk was calculated through the Incremental Lifetime Cancer Risk (ILCR). BaP equivalent concentrations found in the region ranged from 6.9 to 46.05 ng BaPeq/g in lichens, from 16.45 to 162.02 ng BaPeq/g in soils, and from 0.02 to 0.16 ng BaPeq/m³ in air, indicative of high variability in this regional area. Human exposure to PAH varied between 976 and 42,877 ng BaPeq/d. When considering all exposure pathways, ILCR values were between 10⁻⁴ and 10⁻³. Considering only inhalation, ILCR values were between 10⁻⁶ and 10⁻⁵. The main risk seemed to arise from soil (either ingestion or inhalation of resuspended soil particles). The high spatial resolution of our environmental data allowed for detection of critical exposure levels at unexpected sites. Our results identified important areas where health studies on local populations need to be focused, and where environmental levels of PAH need to be monitored over time in order to protect human health.

Stable carbon and nitrogen isotopes (δ¹³C, δ¹⁵N) were used to analyse food web dynamics of two of the main estuaries of the Portuguese coast: Tejo and Mira. The ultimate sources of organic matter supporting production of some of the most abundant and commercially important fish species were determined; and seasonal, inter- and intra- estuarine differences in the trophic relations among producers and consumers were identified. Stable isotope analysis was performed in different producers, primary consumers (main prey items for fish) and fish species (Solea solea, Solea senegalensis, Pomatoschistus microps, Dicentrarchus labrax, Liza ramada, Diplodus vulgaris and Atherina presbyter) of two areas in each estuary, in July and October 2009. Model calculations showed that the main prey for the fish species in the Tejo estuary used mostly salt marsh-derived organic matter as nutritional sources, with no marked differences between the sampled months. Trophic levels of fish species from the same estuary differed at multiple scales: inter-species, seasonally and spatially (both between and within estuaries). Significant differences in isotopic composition of fish species were more pronounced spatially (between the two sampled areas in the estuary) than seasonally (between sampled months). Trophic relationships in both estuaries demonstrated that organic matter is transferred to higher trophic positions mainly through benthic pathways. This shows the flexibility of these species to share resources and to exploit temporary peaks in prey populations. The present results showed that extensive disturbance in intertidal habitats from both estuaries may potentially change the balance of organic matter in the base of these complex food webs., (Copyright © 2011 Elsevier Ltd. All rights reserved.)

Green coffee bean isotopes have been used to trace the effects of different climatic and geological characteristics associated with the Hawaii islands. Isotope ratio mass spectrometry (IRMS) and inductively coupled plasma mass spectrometry ((MC)-ICP-SFMS and ICP-QMS) were applied to determine the isotopic composition of carbon (δ13C), nitrogen (δ15N), sulfur (δ34S), and oxygen (δ18O), the isotope abundance of strontium (87Sr/86Sr), and the concentrations of 30 different elements in 47 green coffees. The coffees were produced in five Hawaii regions: Hawaii, Kauai, Maui, Molokai, and Oahu. Results indicate that coffee plant seed isotopes reflect interactions between the coffee plant and the local environment. Accordingly, the obtained analytical fingerprinting could be used to discriminate between the different Hawaii regions studied.

Toxic polycyclic aromatic hydrocarbons (PAHs) can be found in wastewaters and sewages released from industries and/or urban areas. When discharged untreated to stream waters, they can be a problem to human health. This work represents the first attempt to use PAH and metal concentrations in aquatic moss transplants together with land-use information to identify water pollution sources in urban areas. To do this, the moss Fontinalis antipyretica was collected from a natural stream and transplanted to four different streams in a densely populated area of Lisbon, Portugal. After three months of exposure, mosses were collected and analyzed for metals and for the 16 priority PAHs recommended by the U.S. EPA. Urban streams seem to have a scattered contamination of 6-ring PAHs. Correlations among land-use, metal concentrations, and PAH concentrations indicated that areas occupied by activities of tertiary and industrial sectors had higher PAH concentrations in transplanted mosses, mainly for the sum of the 16 EPA-PAHs and for the 2-, 3- and 5-ringed PAHs, than areas occupied by urban and wooded areas. These PAHs were associated with enhanced Zn and Cu and land use activities that linked the sites to high traffic density. Industrial land use influences PAH concentration in water up to 1000 m of distance from the stream, whereas tertiary sector land use influences it up to 500 m.

In this study we measured δ¹³C in various carbon pools along the basipetal transport pathway in co-occurring Pinus pinaster and Acacia longifolia trees under Mediterranean climate conditions in the field. Overall, species differences in photosynthetic discrimination resulted in more enriched δ¹³C values in the water-conserving overstory P. pinaster relative to the water-spending understory invasive A. longifolia. Post-photosynthetic fractionation effects resulted in differences in δ¹³C of water-soluble organic matter pools along the plant axis with progressive depletion in δ¹³C from the canopy to the trunk (∼6.5‰ depletion in A. longifolia and ∼0.8‰ depletion in P. pinaster). Regardless of these fractionation effects, phloem sap δ¹³C in both terminal branches and the main stem correlated well with environmental parameters driving photosynthesis for both species, indicating that phloem sap δ¹³C has potential as an integrative tracer of changes in canopy carbon discrimination (Δ¹³C). Furthermore, we illustrate that a simple model based on sap flow estimated canopy stomatal conductance (G(S)) and phloem sap δ¹³C measurements has significant potential as a tool for estimating canopy-level carbon assimilation rates.

This paper presents an approach that may be applied as an accurate and rapid tool for classifying coffee beans on the basis of the specific kahweol content. Using Fourier-transform Raman spectroscopy with 1064 nm excitation it is possible to monitor the characteristic Raman bands of kahweol in green coffee beans without chemical and physical processing of the beans. The procedure was optimized on the basis of 83 and 125 measurements of whole and ground beans, respectively, using coffee samples of two different species, Coffea arabica L. and Coffea canephora L. (var. Robusta), and different origins (Asia, Africa, and South America). The relative contribution of the kahweol in individual beans can be determined quantitatively by means of a component analysis of the spectra, yielding a spectral kahweol index (σka) that is proportional to the relative content of kahweol in a coffee bean. The reproducibility of the spectroscopic measurement and analysis was found to be 3.5%. Individual beans of the same type and origin reveal a scattering of the σka values. Nevertheless, an unambiguous distinction between Arabica and Robusta samples is possible on the basis of single-bean measurements as the σka values are greater than and less than 10 for Arabica and Robusta coffees, respectively. Measurements of whole and ground beans afforded very similar results, despite the heterogeneous distribution of kahweol within a bean. Unlike conventional analytical techniques, the single-bean sensitivity of the present approach may also allow for a rapid detection of unwanted admixtures of low-value Robusta coffee to high-quality and more expensive Arabica coffee.

Combining C flux measurements with information on their isotopic composition can yield a process-based understanding of ecosystem C dynamics. We studied the variations in both respiratory fluxes and their stable C isotopic compositions (delta(13)C) for all major components (trees, understory, roots and soil microorganisms) in a Mediterranean oak savannah during a period with increasing drought. We found large drought-induced and diurnal dynamics in isotopic compositions of soil, root and foliage respiration (delta(13)C(res)). Soil respiration was the largest contributor to ecosystem respiration (R (eco)), exhibiting a depleted isotopic signature and no marked variations with increasing drought, similar to ecosystem respired delta(13)CO(2), providing evidence for a stable C-source and minor influence of recent photosynthate from plants. Short-term and diurnal variations in delta(13)C(res) of foliage and roots (up to 8 and 4 per thousand, respectively) were in agreement with: (1) recent hypotheses on post-photosynthetic fractionation processes, (2) substrate changes with decreasing assimilation rates in combination with increased respiratory demand, and (3) decreased phosphoenolpyruvate carboxylase activity in drying roots, while altered photosynthetic discrimination was not responsible for the observed changes in delta(13)C(res). We applied a flux-based and an isotopic flux-based mass balance, yielding good agreement at the soil scale, while the isotopic mass balance at the ecosystem scale was not conserved. This was mainly caused by uncertainties in Keeling plot intercepts at the ecosystem scale due to small CO(2) gradients and large differences in delta(13)C(res) of the different component fluxes. Overall, stable isotopes provided valuable new insights into the drought-related variations of ecosystem C dynamics, encouraging future studies but also highlighting the need of improved methodology to disentangle short-term dynamics of isotopic composition of R (eco).

The aim of this study was to validate lichens as biomonitors of PAH atmospheric deposition; for that, an inter-comparison between the PAH profile and concentrations intercepted in lichens with those of air, soil and pine needles was performed. The study was conducted in a petro-industrial area and the results showed that PAH profiles in lichens were similar to those of the air and pine needles, but completely different from those of soils. Lichens accumulated higher PAH concentrations when compared to the other environmental compartments and its concentrations were significantly and linearly correlated with concentrations of PAHs in soil; we showed that a translation of the lichen PAHs concentrations into regulatory standards is possible, fulfilling one of the most important requirements of using lichens as biomonitors. With lichens we were then able to characterize the air PAHs profile of urban, petro-industrial and background areas., (Copyright (c) 2009 Elsevier Ltd. All rights reserved.)

Sediment descriptors (grain size, total volatile solids, redox potential) and stable carbon and nitrogen isotopes were used to trace the origin of organic matter in a coastal area under multiple organic enrichment sources (urban outfall and a major estuary). The sediments fines content and total volatile solids were similar to outfall pre-operation period (1994), but the incorporation of terrestrial organic matter within the sediments located closer to the outfall was diagnosed by depleted (13)C values (-24.2+/-0.38 per thousand) and (15)N values (2.4+/-0.93 per thousand). Data also indicated depleted nitrogen signature at larger distances from the outfall than the carbon signature, due to confounding sediment grain size properties. Analysis in the bivalve Abra alba gave the same results for both isotopes and thus allowed a coherent interpretation of the spatial extent of the organic enrichment, highlighting the importance of bringing a biological element into the environmental assessment., (Copyright 2009 Elsevier Ltd. All rights reserved.)

The stable isotopic composition of water is routinely used as a tracer to study water exchange processes in vascular plants and ecosystems. To date, no study has focussed on isotope processes in non-vascular, poikilohydric organisms such as lichens and bryophytes. To understand basic isotope exchange processes of non-vascular plants, thallus water isotopic composition was studied in various green-algal lichens exposed to desiccation. The study indicates that lichens equilibrate with the isotopic composition of surrounding water vapour. A model was developed as a proof of concept that accounts for the specific water relations of these poikilohydric organisms. The approach incorporates first their variable thallus water potential and second a compartmentation of the thallus water into two isotopically distinct but connected water pools. Moreover, the results represent first steps towards the development of poikilohydric organisms as a recorder of ambient vapour isotopic composition.

The aim of this study was to compare the performance of two lichen species--Xanthoria parietina and Ramalina canariensis--as biomonitors of the toxic organic compounds dioxins and furans (PCDD/Fs). For that purpose, the concentrations and profiles of PCDD/Fs found in samples of these two lichen species were compared. Results showed that R. canariensis presented higher concentrations than X. parietina and that the PCDD/F homologue profiles were substantially different between these species. Xanthoria parietina appeared to be a more efficient interceptor of more chlorinated PCDD/Fs and also of particles, whereas R. canariensis mainly reflected less chlorinated PCDD/Fs. Results also showed that the PCDD/F profile of X. parietina differed from the one found in other foliose and fruticose lichen species. Despite the differences observed between the profiles of R. canariensis and X. parietina, the calibration of PCDD/F concentrations between the two species was achieved, allowing the biomonitoring of PCDD/Fs at a regional scale using both species simultaneously.

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.

The study presents first data on the delta(18)O performance of poikilohydrous lichen ground cover, and its potential impact on the isotopic composition of water fluxes arising from subjacent soil layers. As a model organism, the globally distributed lichen Cladina arbuscula was studied under laboratory conditions as well as in the field. During a desiccation experiment, delta(18)O of the lichen's thallus water and of its respired CO(2) became enriched by approximately 7 per thousand and followed a similar enrichment pattern to that expected from homoiohydrous, vascular plants. However, the observed degree of enrichment was lower in comparison to vascular plants due to (i) the lichen's inherent lower evaporative resistances; and (ii) a stronger effect of the more depleted surrounding water vapour. In lichens growing in their natural habitat, this specific pattern may show substantial variations depending on prevailing microclimatic conditions. Within a field study, thallus water delta(18)O of lichens principally proved to become more depleted when close to equilibration with the surroundings. It thereby strongly depended on the absorption of surrounding water vapour. Moreover, the results indicate that lichen mats substantially reduce evaporation rates arising from subjacent soil layers, and may alter the isotopic signal of vapour diffusing away from these layers into more depleted values.

The objective of this work was to evaluate the spatial and temporal impact of dust-pollution in the vicinity of a cement industry, located in an area with dry climate. The spatial impact integrated over time was evaluated from the concentrations of Ca, Fe and Mg in in-situ Xanthoria parietina. The temporal pattern was assessed through one-month transplants of the lichen Ramalina canariensis. Four potential sources of atmospheric dust were evaluated: the limestone-quarry; the unpaved roads, the deposit area and the cement mill. Calcium concentration in lichens was considered the best cement-dust indicator. Different types of dust (clinker and grinded-limestone-dust) resulted in different time-patterns of Ca accumulation, which was also related with the different influence that wet and dry periods have in the lichen accumulation process. The dust pollution was found to be deposited locally and dependent on: the nature of dust particles and the volume and frequency of precipitation.

Recent insights into fractionation during dark respiration and rapid dynamics in isotope signatures of leaf- and ecosystem-respired CO(2) indicate the need for new methods for high time-resolved measurements of the isotopic signature of respired CO(2) (delta(13)C(res)). We present a rapid and simple method to analyse delta(13)C(res) using an in-tube incubation technique and an autosampler for small septum-capped vials. The effect of storage on the delta(18)O and delta(13)C ratios of ambient CO(2) concentrations was tested with different humidity and temperatures. delta(13)C ratios remained stable over 72 h, whereas delta(18)O ratios decreased after 24 h. Storage at 4 degrees C improved the storage time for delta(18)O. Leaves or leaf discs were incubated in the vials, flushed with CO(2)-free air and respired CO(2) was automatically sampled within 5 min on a microGas autosampler interfaced to a GV-Isoprime isotope ratio mass spectrometer. Results were validated by simultaneous on-line gas-exchange measurements of delta(13)C(res) of attached leaves. This method was used to evaluate the short-term (5-60 min) and diurnal dynamics of delta(13)C(res) in an evergreen oak (Quercus ilex) and a herb (Tolpis barbata). An immediate depletion of 2-4 per thousand from the initial delta(13)C(res) value occurred during the first 30 min of darkening. Q. ilex exhibited further a substantial diurnal enrichment in delta(13)C(res) of 8 per thousand, followed by a progressive depletion during the night. In contrast, T. barbata did not exhibit a distinct diurnal pattern. This is in accordance with recent theory on fractionation in metabolic pathways and may be related to the different utilisation of the respiratory substrate in the fast-growing herb and the evergreen oak. These data indicate substantial and rapid dynamics (within minutes to hours) in delta(13)C(res), which differed between species and probably the growth status of the plant. The in-tube incubation method enables both high time-resolved analysis and extensive sampling across different organs, species and functional types., (Copyright (c) 2007 John Wiley & Sons, Ltd.)

Temporal dynamics in carbon isotope ratios of ecosystem respiration (delta13C(R)) were evaluated on hourly, daily and annual timescales in a Mediterranean woodland. Emphasis was given to the periods of transition from wet to dry season and vice versa, when the system turns from a net carbon sink to a source. The constancy of nocturnal delta13C(R) was tested. The relationship between delta13C(R) (determined through Keeling plots) and environmental factors was evaluated through time-lag analysis. Delta13C(R) exhibited high annual variation (> 7). During the transition periods, delta13C(R) correlated significantly with factors influencing photosynthetic discrimination, soil respiration, and whole-canopy conductance. Time-lags differed between below- and above-ground variables, and between seasons. A shift in regression parameters with environmental factors indicated seasonal differences in ecosystem responsiveness (e.g. temperature acclimation). Delta13C(R) exhibited substantial nocturnal enrichment (> 4) from dusk to dawn. These data indicate pronounced short-term dynamics in delta13C(R) at hourly to daily timescales and a modulated response to environmental drivers. Substantial short-term changes in nocturnal delta13C(R) may have important implications for the sampling protocols of nocturnal Keeling plots.

The aim of this study was to compare the physiological responses to increased nitrogen (N) supply between the nitrophytic lichen Xanthoria parietina (L.) Th. Fr. and the acidophytic lichen Evernia prunastri (L.) Ach. The two lichens were exposed to a weekly dosage of 0.05, 0.1, 0.2, 0.6 or 2.4 g N m(-2) for 2 months, administered as NH(4)NO(3) dissolved in artificial rainwater (1 l m(-2)). After the treatments, in vivo chlorophyll a fluorescence was determined to assess vitality; concentrations of total N, ammonium, nitrate and dominant amino acids, including glutamate, glutamine and arginine, were quantified in order to follow changes in N status; and the polyols ribitol, arabitol and mannitol were quantified to follow changes in the lichens' carbon (C) status. The uptake of N was quantified by labelling the fertiliser with (15)N in the ammonium position; chlorophyll a was used as an indirect marker for algal activity, and ergosterol as an indirect marker of fungal activity. Nitrogen uptake was higher in E. prunastri than in X. parietina, although the latter species may have used the mannitol reserves to obtain C skeletons and energy for N assimilation. Chlorophyll a and ergosterol concentrations remained unaltered in X. parietina irrespective of N dosage while ergosterol decreased with increasing N uptake in E. prunastri. The latter species had accumulated a large pool of ammonium at the highest N dosage, whilst in X. parietina a significant nitrate pool was instead observed. Taken together, these short-term responses to high N supply observed in the two lichens, and the differences between them, can partly explain the higher tolerance of X. parietina towards increased atmospheric N levels.

Exposure to high light induced a quantitatively similar decrease in the rate of photosynthesis at limiting photon flux density (PFD) and of photosystem II (PSII) photochemical efficiency, FV/FM, in both green and blue-green algal lichens which were fully hydrated. Such depressions in the efficiency of photochemical energy conversion were generally reversible in green algal lichens but rather sustained in blue-green algal lichens. This greater susceptibility of blue-green algal lichens to sustained photoinhibition was not related to differences in the capacity to utilize light in photosynthesis, since the light-and CO2-saturated rates of photosynthetic O2 evolution were similar in the two groups. These reductions of PSII photochemical efficiency were, however, largely prevented in lichen thalli which were fully desiccated prior to exposure to high PFD. Thalli of green algal lichens which were allowed to desiccate during the exposure to high light exhibited similar recovery kinetics to those which were kept fully hydrated, whereas bluegreen algal lichens which became desiccated during a similar exposure exhibited greatly accelerated recovery compared to those which were kept fully hydrated. Thus, green algal lichens were able to recover from exposure to excessive PFDs when thalli were in either the hydrated or desiccated state during such an exposure, whereas in blue-green algal lichens the decrease in photochemical efficiency was reversible in thalli illuminated in the desiccated state but rather sustained subsequent to illumination of thalli in the hydrated state.