Bernardo Duarte

Stock and losses of trace metals from salt marsh plants.

Pools of Zn, Cu, Cd and Co in the leaf, stem and root tissues of Sarcocornia fruticosa, Sarcocornia perennis, Halimione portulacoides and Spartina maritima were analysed for a Tagus estuary (Portugal) salt marsh. Pools of Cu and Cd in the salt marsh were higher in spring/summer, indicating a net uptake of these metals during the growing season. Standing stocks of Zn, Cu, Cd and Co in the leaf and stem biomass of S. fruticosa, S. perennis and H. portulacoides showed a strong seasonal variation, with higher values recorded in autumn. The metal-containing leaves and stems that shed in the autumn become metal-containing detritus. The amount of this material washed out from the total marsh area (200 ha) was estimated as 68 kg of Zn, 8.2 kg of Cu, 13 kg of Co and 0.35 kg of Cd. The high tidal amplitude, a branched system of channels and semi-diurnal tidal cycle greatly favour the export of the organic detritus to adjoining marsh areas.

Accumulation and biological cycling of heavy metal in four salt marsh species, from Tagus estuary (Portugal)

Pools of Zn, Cu, Cd and Co in leaf, stem and root tissues of Sarcocornia fruticosa, Sarcocornia perennis, Halimione portulacoides and Spartina maritima were analyzed on a bimonthly basis, in a Tagus estuary salt marsh. All the major concentrations were found in the root tissues, being the concentrations in the aboveground organs neglectable for sediment budget proposes, as seen by the low root-aboveground translocation. Metal annual accumulation, root turnovers and cycling coefficients were also assessed. S. maritima showed the higher root turnovers and cycling coefficients for most of the analyzed metals, making this a phytostabilizer specie. By contrast the low root turnover, cycling coefficient and low root necromass generation makes S. perennis the most suitable specie for phytoremediation processes. Although the high amounts of metal return to the sediments, due to root senescence, salt marshes can still be considered sinks of heavy metals, cycling heavy metals mostly between sediment and root.

Seasonal variation of extracellular enzymatic activity (EEA) and its influence on metal speciation in a polluted salt marsh

The influence of salt marsh sediment extracellular enzymatic activity (EEA) on metal fractions and organic matter cycling was evaluated on a seasonal basis, in order to study the relation between organic matter cycles and the associated metal species. Metals in the rhizosediment of Halimione portulacoides were fractioned according to the Tessiers scheme and showed a similar pattern regarding the organic-bound fraction, being always high in Autumn, matching the season when organic matter presented higher values. Both organic-bound and residual fractions were always dominant, being the seasonal variations due to interchanges between these two fractions. Phenol oxidase and beta-N-acetylglucosaminidase had higher activities during the Spring and Summer, contrarily to peroxidase which had higher activity during Winter. Protease showed high activities in both Spring and Winter. These different periods of high organic matter hydrolysis caused two periods of organic metal bound decrease. Sulphatase peaks (Spring and Winter) matched the depletion of exchangeable metal forms, probably due to sulphides formation and consequent mobilization. This showed an interaction between several microbial activities affecting metal speciation.

Halophyte anti-oxidant feedback seasonality in two salt marshes with different degrees of metal contamination: search for an efficient biomarker

Salt marshes can be affected by metal contamination when near a polluted area, and this excessive concentration of metals is a source of stress in plants. Production of proteins, flavonoids, phenolic compounds and anti-oxidant feedback can be used as biomarkers, as well to assess the suitability of halophytes to function as a biomonitors. Through monitoring the anti-oxidative feedback in Halimione portulacoides (L.) Aellen, Sarcocornia fruticosa (L.) A.J.Scott and Spartina maritima (Curtis) Fernald in a contaminated and non-contaminated marsh, S. maritima seems to have potential as a bioindicator species, showing different biochemical characteristics according to the degree of contamination to which it is exposed. The evident biochemical separation between individuals from contaminated and non-contaminated salt marshes is mostly due to differences in the activity of SOD as well APx and GPx. Without neglecting the need for further works, the present study suggests that S. maritima enzymatic defences as good candidates for efficient biomarkers for estuarine sediment quality assessment studies.

Hexavalent chromium reduction, uptake and oxidative biomarkers in Halimione portulacoides

The in situ reduction of Cr (VI) to its less toxic form Cr (III) may be a useful detoxification mechanism for phytoremediation. Using a hydroponics mesocosmos approach, we evaluated the ability of Halimione portulacoides to reduce and uptake Cr (VI) and its anti-oxidative feedback and biomarkers. It was found that this specie can, not only reduce large amounts of Cr (VI) in the external medium, but also withdrawn and accumulate this element in its roots and aboveground organs. Both these mechanisms were found to be dose dependent. Jointly with this phytoremediative potential the oxidative feedback was also assessed. Chromium uptake had its major implications on the chlorophyll content and flavonoid content, with potential consequences in the photosynthetic and photo-protective mechanisms. Although the high Cr root accumulation in H. portulacoides, there were no inactivation of the enzymatic defenses, allowing a continuous defense against reactive oxygen species. In fact, GPX and specially SOD revealed to be an excellent dose-related biomarker of Cr induced stress. All these aspects make this specie suitable for Cr (VI) phytoremediation processes, either by phytoextraction or by reduction of Cr (VI) to Cr (III) and also for monitoring programs using SOD and GPX as biomarkers of Cr environmental contamination.

Biophysical and biochemical constraints imposed by salt stress: learning from halophytes

Soil salinization is one of the most important factors impacting plant productivity. About 3.6 billion of the world’s 5.2 billion ha of agricultural dry land, have already suffered erosion, degradation, and salinization. Halophytes are typically considered as plants able to complete their life cycle in environments where the salt concentration is above 200 mM NaCl. Salinity adjustment is a complex phenomenon but essential mechanism to overcome salt stress, with both biophysical and biochemical implications. At this level, halophytes evolved in several directions, adopting different strategies. Otherwise, the lack of adaptation to a salt environment would negatively affect their electron transduction pathways and the entire energetic metabolism, the foundation of every plant photosynthesis and biomass production. The maintenance of ionic homeostasis is in the basis of all cellular counteractive measures, in particular in terms of redox potential and energy transduction. In the present work the biophysical mechanisms underlying energy capture and transduction in halophytes are discussed alongside with their relation with biochemical counteractive mechanisms, integrating data from photosynthetic light harvesting complexes, electron transport chains to the quinone pools, carbon fixation, and energy dissipation metabolism.

Particulate metal distribution in Tagus estuary (Portugal) during a flood episode.

Particulate metal concentrations were assessed before, during and after a flood episode in the Tagus estuary. Particulate metal concentrations showed a decrease during the flood period and very similar values in the months before and after the flood event. Before this period, sampling station characteristics were verified to be homogenous during the peak of the flooding event, as all of the sampling stations assumed very specific characteristics. One of the main consequences from the flood, concurrent with a decrease in particulate metal concentrations, was the high input of SPM into the estuarine area. This finding indicates higher levels of heavy metals in fine-sized particles at low SPM concentration than those present in coarser particles at high SPM levels. These periodic flood events can be considered as estuarine contamination masks and should be interpreted as periods of dilution in heavy metal contamination rather than as an estuarine cleansing process.

Photochemical and biophysical feedbacks of C3 and C4 Mediterranean halophytes to atmospheric CO2 enrichment confirmed by their stable isotope signatures

According the latest predictions, an increase of about two times in atmospheric CO2 concentrations, is expected to occur by the end of this century. In order to understand the effects of this atmospheric composition changes on two abundant Mediterranean halophytes (Halimione portulacoides and Spartina maritima), mesocosmos trials were performed simulating two atmospheric CO2 environments (380 ppm and 760 ppm of CO2 respectively). The two chosen halophyte species present different metabolic characteristics: H. portulacoides, is a C3 specie while S. maritima is a C4 species. Distinct feedbacks were obtained for each of the studied species. Stable Isotope discrimination showed that both species showed an enhancement of the Rubisco carboxylation capacity and photosynthetic efficiency mostly due to an increase in intracellular [CO2]. In H. portulacoides CO2 fertilization induced an enhancement of ETR and a decrease in non-photochemical quenching and in dissipated energy fluxes. On the other hand the C4 grass S. maritima, already at full capacity, showed no photosynthetic enhancement. In fact this highly productive grass presented lower photosynthetic efficiencies accompanied by increases in dissipated energy fluxes mostly due to reductions in energy flux associated with the transport of reducing power throughout the quinone pool. The accumulation of reducing power led to oxidative stress, and thus the photosynthetic ability of this grass was greatly reduced. Both these feedbacks to realistic future CO2 concentrations are important consideration for in future primary productivity models, indicating a possible reduced abundance of the pioneer S. maritima and an increased biomass spreading of the sediment stabilizer H. portulacoides, inevitably affecting the morphology and function of the salt marshes imposed by these atmospheric changes, both in terms of ecosystem functioning and loss of biodiversity.

Unveiling Zn hyperaccumulation in Juncus acutus: Implications on the electronic energy fluxes and on oxidative stress with emphasis on non-functional Zn-chlorophylls

Juncus acutus arises as possible hyperaccumulator specie, tolerating exogenous Zn concentrations as high as 60 mM. Zinc concentrations here detected in seedlings germinated in the presence high Zn concentrations, were above the described upper toxic levels for higher plants. Even at the highest Zn concentration, growth inhibition only accounted to approximately 30% of control seedlings biomass, presenting an EC₅₀ value in the range of 10-20 mM of metal. PSII quantum yields showed a marked decline, reflection of changes in the thylakoid structure on the PSII electron donor sites. In fact, the electron transport rate was severely affected by Zn in seedlings exposed to higher Zn concentrations leading to a decrease in their maximum electronic transport rate and consequently presenting lower light saturation and lower photosynthetic efficiencies. Although light absorption capacity was not affected by Zn exposure and uptake, energy trapping flux in the photosynthetic apparatus and transport throughout the electronic chain was severely impaired. This lack of efficiency is related with non-functional Zn-chlorophylls formation. There was a strong linear correlation between exogenous Zn concentration applied and the concentration actually verified in the seedlings tissue with the concentration of both ZnChl a and b. There was also a gradual loss of connectivity between the antennae of the PSII units being this more evident at the higher Zn concentrations and thus impairing the energetic transport. The reduction in light harvesting efficiency, leads inevitably to the accumulation of redox energy inside the cells. To counteract ROS generation, all anti-oxidant enzymatic activities (except catalase) showed a proportional response to exogenous and in vivo Zn concentrations. Not only this plant appears to be highly tolerant to high Zn concentrations, but also it can overcome efficiently the damage produced during this uptake by efficiently dissipating the excessive cellular redox potential accumulated, essentially due to Zn incorporation into the chlorophyll molecule.

The role of organic acids in assisted phytoremediation processes of salt marsh sediments

Some plants have high ability to absorb heavy metals in high concentrations. In this study, Spartina maritima was tested in conjunction with low molecular weight organic acids (LMWOA), in order to evaluate the possible use of this plant in phytoremediation processes in salt marshes. Three different LMWOA (citric acid, malic acid and acetic acid) were applied to contaminated intact cores of S. maritima colonized sediment and several heavy metals (Cd, Zn, Pb, Cu, Cr and Ni) were analyzed in sediment and plant parts. Acetic acid application proved to be the most efficient, enhancing greatly the uptake of all metals analyzed. Citric acid also showed good results, while malic acid proved to be very inefficient in most of the cases. The highest enhancement was observed for Cr with a 10-fold increase of the uptake upon application of acetic acid, while improving the Pb uptake proved to be the most difficult, probably due to its low solubility.