Biodegradation of C.I. Acid Blue 92 by Nasturtium officinale: Study of Some Physiological Responses and Metabolic Fate of Dye

The present study was conducted to evaluate the potential of aquatic vascular plant, Nasturtium officinale, for degradation of C.I. Acid Blue 92 (AB92). The effect of operational parameters such as initial dye concentration, plant biomass, pH, and temperature on the efficiency of biological decolorization process was determined. The reusability of the plant in long term repetitive operations confirmed the biological degradation process. The by-products formed during biodegradation process were identified by GC-MS technique. The effects of the dye on several plant physiological responses such as photosynthetic pigments content and antioxidant enzymes activity were investigated. The content of chlorophyll and carotenoids was significantly reduced at 20 mg/L of the dye. The activities of superoxide dismutase and peroxidase were remarkably increased in the plant root verifying their importance in plant tolerance to the dye contamination.     

Influence of Temperature on the Effects of Artificially Enhanced UV-B Radiation on Aquatic Bryophytes Under Laboratory Conditions

We examined, under laboratory conditions, the influence of temperature (2 °C vs. 10 °C) on the physiological responses of two aquatic bryophytes from a mountain stream to artificially enhanced UV-B radiation for 82 d. These organisms may be exposed naturally to relatively low temperatures and high levels of UV-B radiation, and this combination is believed to increase the adverse effects of UV-B radiation. In the moss Fontinalis antipyretica, UV-B-treated samples showed severe physiological damages, including significant decreases in chlorophyll (Chl) and carotenoid (Car) contents, Chl a/b and Chl/phaeopigment ratios, Chl a fluorescence parameters F_v/F_m and _PS2, electron transport rate (ETR_max), and growth. In the liverwort Jungermannia cordifolia, UV-B radiation hardly caused any physiological change except for growth reduction. Thus, this liverwort seemed to be more tolerant to UV-B radiation than the moss under the specific experimental conditions used, maybe partly due to the accumulation of UV-B absorbing compounds. The influence of temperature on the effects of UV-B radiation depended on the species: the higher the UV-B tolerance, the lower the influence of temperature. Also, different physiological variables showed varied responses to this influence. Particularly, the lower temperature used in our study enhanced the adverse effects of UV-B radiation on important physiological variables such as F_v/F_m, growth, and Chl/phaeopigment ratios in the UV-B-sensitive F. antipyretica, but not in the more UV-B-tolerant J. cordifolia. Thus, the adverse effects of cold and UV-B radiation were apparently additive in the moss, but this additiveness was lacking in the liverwort. The Principal Components Analyses (PCA) conducted for both species with the physiological data obtained after 36 and 82 d of culture confirmed the above results. Under natural conditions, the relatively high water temperatures in summer might facilitate the acclimation of aquatic bryophytes from mountain streams to high levels of UV-B radiation. This may be relevant to predict the consequences of concomitant global warming and increasing UV-B radiation.     

Lichens as suitable indicators of the biological effects of atmospheric pollutants around a municipal solid waste incinerator (S Italy)

A comprehensive biomonitoring programme should integrate several methods distributed along the biomonitoring chain, allowing to detect exposure, threads and impacts. In the case of a municipal solid waste incinerator (MSWI), biomonitoring of air pollution can contribute to source attribution, detection of ongoing processes and assessment of environmental effects. Three different methods were used to assess the biological effects of air pollution around a MSWI using lichens as biomonitors: (1) lichen diversity; (2) bioaccumulation of trace elements; and (3) physiological status (photosynthetic efficiency, cell membrane damage, viability). The first method takes into account the native lichen flora, while the other two were applied to thalli of the lichen Evernia prunastri transplanted for 6 months in the study area. Lichen diversity and physiological parameters reflected the effects of air pollution around the incinerator and the surrounding industrial area. High frequencies of non-nitrophilous species corresponded to sites with higher environmental quality, while high frequencies of nitrophilous species corresponded to sites with higher level of eutrophication. Transplanted samples showed increased cell membrane damage and reduced vitality respect to control samples. Bioaccumulation of trace elements pointed at the atmospheric origin of Hg depositions in the area. These results suggest that an integrated use of lichen-based methods along the biomonitoring chain can provide useful biological outputs for decision-makers to establish correct sustainable waste management policies.     

Methodological aspects of moss sample preparation. Effects of freezing and duration of washing on the cellular distribution of elements in Fontinalis squamosa Hedw.

Aquatic bryophytes are widely used in biomonitoring studies, mainly because of their capacity to act as bioaccumulators. As different methods can be used to preserve and process moss samples, the results of elemental analyses may also vary, particularly if the contents of different cellular compartments are analyzed separately. In the present study, we compared the total concentrations of some elements and the concentrations of these in different cellular locations in frozen-thawed and fresh samples of the aquatic moss Fontinalis squamosa that were also washed for different lengths of time before analysis. We used the sequential elution technique (SET) to extract the different fractions, and we determined the concentrations of K, Mg, Ca, Cd, Co, Cu, Al and Zn in the extracts. The results varied depending on the element, cellular location and moss sampling site. In the moss samples processed after freezing, the greatest differences were in the intracellular concentrations of K, Mg and Cd, which were lower than in the fresh samples. Minor differences were found in the concentrations of elements in other cellular locations and in the total concentrations of elements. The increase in the duration of the initial washing step, carried out to remove soluble and particulate intercellular elements, may also cause the release of elements (e.g. K and Mg) bound to extracellular exchange sites. The concentrations in the other cellular fractions and the total concentrations were less affected by the washing duration. Neither freezing nor the use of long washing times are recommended for processing moss samples prior to extraction of elements by the SET. Further studies are needed to confirm and clarify the observations.     

Analysis of temporal variability in the concentrations of some elements in the terrestrial moss Pseudoscleropodium purum

The uptake of contaminants by moss is a complex process that depends on the nature and characteristics of the contaminants and on the physicochemical and biological processes that affect the moss. All of these factors in turn depend on environmental variables that may vary within a short time. It can therefore be assumed that the uptake of elements varies in such short periods that it is very difficult to obtain temporally representative results in biomonitoring studies with terrestrial moss. In order to investigate the temporal variability and the implications that it may have on the interpretation of the results obtained in this type of study, the tissue concentrations of 12 elements were determined in samples of the moss Pseudoscleropodium purum collected from two sampling sites in Galicia (NW Spain), during a long term survey with a high frequency of sampling. The results showed that the concentrations of the elements in the moss varied greatly within very short periods, and that the error associated with the temporal variability in the results was high. In accordance with these findings, and taking into account that the technique enabled contaminated and uncontaminated sites to be distinguished, we recommend that the results of biomonitoring studies should be regarded as qualitative or semiqualitative, rather than attempting to provide absolute data, which may not be temporally representative, and may have a high degree of uncertainty associated with them.     

Plant desiccation and protein synthesis: an in vitro system from dry and hydrated mosses using endogenous and synthetic messenger ribonucleic Acid

The conditions and requirements for an in vitro protein synthesizing system from the moss Tortula ruralis are outlined. Using this system the effects of desiccation, achieved quickly or slowly, were studied. Slowly dried moss retained fewer polyribosomes on desiccation but more active ribosomes than rapidly dried moss. Even in the completely desiccated moss the polyribosomes and/or free ribosomes present have retained their synthetic capacities. On rehydration, the slowly dried moss resumed protein synthesis more quickly than moss previously desiccated rapidly. Moss ribosomes are cycloheximide sensitive and chloramphenicol insensitive and thus the major protein synthesis occurs within the cytoplasm on rehydration. Extracted polyribosomes per se can withstand desiccation to a significant extent, suggesting that protection by the cytoplasm might not be necessary. The aquatic moss Hygrohypnum luridum can retain polyribosomal and ribosomal activity during desiccation, but this decreases greatly on rehydration.     

Copper uptake and copper-induced physiological changes in the epiphytic lichen Evernia prunastri

The lichen Evernia prunastri is very frequently employed in lichen biomonitoring studies and its sensitivity to air pollution has been confirmed at the community level many times. However, studies focused on the physiological responses of this lichen to pollutants are underrepresented. The degree of total as well as intracellular Cu uptake by the lichen after 4 and 24?h prolonged exposure was compared with selected physiological markers including levels of assimilation pigments, chlorophyll a fluorescence, ergosterol, soluble proteins, hydrogen peroxide, superoxide, amino acids, reducing sugars and total soluble phenols. Although the degree of total as well as intracellular Cu uptake by the lichen after 4 and 24?h prolonged exposure were comparable, metabolic responses after 4 and 24?h were very different. Carotenoids, ergosterol and soluble proteins decreased due to Cu exposure and these processes were followed with an increase of superoxide content. Excess of Cu in thalli of E. prunastri after 24?h was toxic and altered almost all tested parameters, including depletion of reducing sugars. We did not observe stimulation of free amino acid synthesis in Cu treated lichen, in fact the content of many particular amino acids decreased. Content of total soluble phenols was unaltered by Cu excess.     

Diurnal variation in photochemical dynamics and surface reflectance of the desiccation-tolerant moss,Tortula ruralis

Most desiccation-tolerant plants alter shoot structure during drying, making it possible to use changes in surface reflectance as a proxy measure of plant water status. Diurnal courses of surface reflectance (albedo) and chlorophyll fluorescence parameters of the ectohydric moss, Tortula ruralis (Hedw.) Gaertn, were measured to assess the coordination between anatomical and physiological features under field conditions. Albedo showed a sigmoidal relationship with relative humidity and the deviation of moss mat temperature from dew point. Maximum photosynthetic quantum yield (F _v/F _m) also displayed a sigmoidal relationship pooled across three days differing in light, temperature, and relative humidity. Depending on the light conditions and rapidity of drying during the morning, there were distinct differences in the ability of T. ruralis to establish thermal dissipation of excess light energy (NPQ) across a range of light levels following rehydration through the day. These findings suggest that there is a coordinated suite of architectural and physiological characteristics maintaining the photosynthetic integrity of these plants in highly variable arid and semi-arid environments.     

Photosynthesis of two moss crusts from the Tengger Desert with contrasting sensitivity to supplementary UV-B radiation

Predicting the effects of increased ultraviolet-B (UV-B) radiation due to stratospheric ozone depletion on temperate desert ecosystems requires better knowledge of the ecophysiological response of common moss species. The aim of the current work was to determine whether elevated UV-B radiation affected photosynthetic performance and chloroplast ultrastructure of two moss crusts and whether response differences were observed between the crusts. In laboratory experiments, Bryum argenteum and Didymodon vinealis, which show microdistributions and are dominant in soil crusts at the Tengger Desert, Northern China, were subjected to four levels of UV-B radiation of 2.75 (control), 3.08, 3.25, and 3.41 W m^?2 for 10 days, simulating 0, 6, 9, and 12% of stratospheric ozone at the latitude of Shapotou, respectively. The results showed that chlorophyll a fluorescence parameters (i.e., the maximal quantum yield of PSII photochemistry, the effective quantum yield of PSII photochemistry, and photochemical quenching coefficient), pigment contents, soluble protein contents, and the ultrastructure were negatively influenced by elevated UV-B radiation and the degree of detrimental effects significantly increased with the intensity of UV-B radiation. Moreover, results indicated that B. argenteum was probably more sensitive to supplementary UV-B radiation than D. vinealis. Therefore, we propose the use of B. argenteum crusts as a bioindicator of responses to elevated UV-B radiation.     

Acclimation of photosynthetic characteristics of the moss Pleurozium schreberi to among‐habitat and within‐canopy light gradients

Light availability varies strongly among moss habitats and within the moss canopy, and vertical variation in light within the canopy further interacts with the age gradient. The interacting controls by habitat and canopy light gradient and senescence have not been studied extensively. We measured light profiles, chlorophyll (Chl), carotenoid (Car) and nitrogen (N) concentrations, and photosynthetic electron transport capacity (J_max) along habitat and canopy light gradients in the widespread, temperate moss Pleurozium schreberi to separate sources of variation in moss chemical and physiological traits. We hypothesised that this species, like typical feather mosses with both apical and lateral growth, exhibits greater plasticity in the canopy than between habitats due to deeper within‐canopy light gradients. For the among‐habitat light gradient, Chl, Chl/N and Chl/Car ratio increased with decreasing light availability, indicating enhanced light harvesting in lower light and higher capacity for photoprotection in higher light. N and J_max were independent of habitat light availability. Within the upper canopy, until 50–60% above‐canopy light, changes in moss chemistry and photosynthetic characteristics were analogous to patterns observed for the between‐habitat light gradient. In contrast, deeper canopy layers reflected senescence of moss shoots, with pigment and nitrogen concentrations and photosynthetic capacity decreasing with light availability. Thus, variation in chemical and physiological traits within the moss canopy is a balance between acclimation and senescence. This study demonstrates extensive light‐dependent variation in moss photosynthetic traits, but also that between‐habitat and within‐canopy light gradient affects moss physiology and chemistry differently.     

Diatom teratologies as biomarkers of contamination: Are all deformities ecologically meaningful?

Contaminant-related stress on aquatic biota is difficult to assess when lethal impacts are not observed. Diatoms, by displaying deformities (teratologies) in their valves, have the potential to reflect sub-lethal responses to environmental stressors such as metals and organic compounds. For this reason, there is great interest in using diatom morphological aberrations in biomonitoring. However, the detection and mostly the quantification of teratologies is still a challenge; not all studies have succeeded in showing a relationship between the proportion of abnormal valves and contamination level along a gradient of exposure. This limitation in part reflects the loss of ecological information from diatom teratologies during analyses when all deformities are considered. The type of deformity, the severity of aberration, species proneness to deformity formation, and propagation of deformities throughout the population are key components and constraints in quantifying teratologies. Before a metric based on diatom deformities can be used as an indicator of contamination, it is important to better understand the “ecological signal” provided by this biomarker. Using the overall abundance of teratologies has proved to be an excellent tool for identifying contaminated and non-contaminated environments (presence/absence), but refining this biomonitoring approach may bring additional insights allowing for a better assessment of contamination level along a gradient. The dilemma: are all teratologies significant, equal and/or meaningful in assessing changing levels of contamination? This viewpoint article examines numerous interrogatives relative to the use of diatom teratologies in water quality monitoring, provides selected examples of differential responses to contamination, and proposes solutions that may refine our understanding and quantification of the stress. This paper highlights the logistical problems associated with accurately evaluating and interpreting teratologies and stimulates more discussion and research on the subject to enhance the sensitivity of this metric in bioassessments.     

Plant Desiccation and Protein Synthesis: II. On the Relationship between Endogenous Adenosine Triphosphate Levels and Protein-synthesizing Capacity

Rehydration of Tortula ruralis in 2,4-dinitrophenol inhibits protein synthesis, polysome formation, and ATP production. Polysomes are conserved intact and are active in vitro in hydrated Tortula placed in this chemical, although in vivo protein synthesis is inhibited. Hydrated moss placed under nitrogen in the dark shows a reduced capacity for ATP and protein synthesis, but polysomes are conserved. During anaerobiosis in light, ATP and protein synthesis are unaffected. Rehydration of slow-dried Tortula in nitrogen in the dark results in reduced in vivo protein synthesis, but not polysome formation; this reduction is much less in the light. Slow-dried moss, but not fast-dried, has a greatly reduced ATP content in the dry state, but this rapidly returns to normal levels on rehydration. The prolonged burst in respiration observed previously on rehydration of Tortula is not paralleled by ATP accumulation. Changes in energy charge in all treatments tested follow the changes in ATP. The aquatic moss, Hygrohypnum luridum, which is intolerant to drought, loses ATP during fast drying and this is not replenished on subsequent rehydration.     

Stress responses and metal exchange kinetics following transplant of the aquatic moss Fontinalis antipyretica

1. We studied stress responses and metal exchange (uptake and loss) kinetics in the aquatic moss Fontinalis antipyretica Hedw. following transplant (in plastic mesh bags) to clean and metal-contaminated river sites and under laboratory conditions. 2. The stress response (estimated on the basis of chlorophyll:phaeophytin ratio) was more pronounced, and the moss took longer to recover, following transplant to heavily contaminated sites. 3. Metal uptake over the 28 day exposure period was predicted well by a two-compartment kinetic model: uptake velocity was initially high and gradually declined, with metal concentration in the moss showing a tendency to reach an equilibrium with metal concentration in the water. Mean uptake rate, time to reach equilibrium and metal concentration in moss at equilibrium all increased with increasing metal concentration in the water. 4. Bioconcentration constants calculated on the basis of our data rank the metals studied in the order Zn < Ni < Co < Cu < Pb < Cd. 5. Following retransplant of mosses to a clean site, metal loss was not predicted well by a passive exchange mode of the above type; instead, two phases (rapid and slow) were apparent. Loss rates during both phases, and proportion of metal load lost during the rapid phase, were proportional to the concentration of metal in the moss at the start of the recuperation trial. 6. We present nomograms to allow estimation of severity of contamination on the basis of metal concentration in moss and duration of transplant.     

Parasites of flounder (Platichthys flesus L.) from the German Bight, North Sea, and their potential use in biological effects monitoring

In the frame of an integrated biological effect monitoring programme, the parasite community of flounder (Platichthys flesus) was investigated at different locations in the German Bight from 1995 to 2000. In order to assess the impact of environmental contamination caused by anthropogenic activities on the parasite community, selected parasitological parameters that displayed significant differences between the sampling sites were subjected to correlation analyses with site-specific contamination and individual pollution loads of their fish hosts. In addition, correlation analyses were conducted with the responses of selected genetic, biochemical, histopathological, physiological and immunological parameters of fish, used as potential biomarkers. In total, 802 flounder were analysed for these parameters. Information on the chemical background at the sampling sites was derived from sediment samples and from 120 samples of blue mussel (Mytilus edulis) tissue, collected at each of the sampling sites. Based on chemical data available from the sediment and blue mussel samples, a pollution gradient could be established between the sampling sites for individual contaminants. The relative abundance of Acanthochondria cornuta, Cucullanus heterochrous and Zoogonoides viviparus, and the community measures species richness and number of heteroxenous species decreased with increasing concentrations of individual heavy metals or hydrocarbons in sediment and blue mussel samples. Most of the parasitological parameters significantly reflected the established site-specific contamination gradient, when data were pooled over all sampling campaigns. Significant correlations were also found with the contamination level of individual flounder. The parasitological parameters included the parasite species Lepeophtheirus pectoralis and Lernaeocera branchialis, which were not correlated to site-specific contamination. Several biomarkers were significantly correlated to the abundance of parasitic copepods A. cornuta, Lernaeocera branchialis and Lepeophtheirus pectoralis and to parasite community parameters. The results showed that the abundance of several metazoan parasite species, species richness and parasite diversity were reduced in contaminated habitats, and that differences between sites were not only related to natural factors, such as salinity, but also to pollution-induced stress. Thus, it can be concluded that the parasite community of fish responds to the level of pollution at a specific site as well as to residues of xenobiotics in individual fish. These findings give indications that the parasite community of fish is a valuable parameter for the assessment of ecological consequences of chemical contamination in aquatic habitats.     

Biomagnetic monitoring of urban air pollution using moss bags (Sphagnum girgensohnii)

Active moss biomonitoring is widely applied in polluted areas for monitoring of airborne particle-bound trace element pollution. This study explored the suitability of the moss Sphagnum girgensohnii for biomagnetic monitoring in cities. To this end S. girgensohnii moss bags were exposed at three different microenvironments characterized by heavy traffic – street canyons, a city tunnel and parking garages during the summer and autumn of 2011 in the city of Belgrade. The ferro(i)magnetic PM fraction in the moss samples was quantified by Saturated Isothermal Remanent Magnetization (SIRM) and the measured values were compared with the trace element concentration in the moss samples. SIRM values were significantly different across the considered urban microenvironments. Moreover, a high correlation between moss SIRM values and concentrations of Al, Ba, Co, Cr, Cu, Fe, Ni and Pb was found. These results demonstrate that moss bags can be effectively applied for biomagnetic monitoring of the spatio-temporal distribution of road traffic and vehicles derived pollutants in urban areas.     

Impact of changes in natural ultraviolet radiation on pigment composition, physiological and morphological characteristics of the Antarctic moss, Grimmia antarctici

The impact of ambient ultraviolet (UV)‐B radiation on the endemic bryophyte, Grimmia antarctici, was studied over 14 months in East Antarctica. Over recent decades, Antarctic plants have been exposed to the largest relative increase in UV‐B exposure as a result of ozone depletion. We investigated the effect of reduced UV and visible radiation on the pigment concentrations, surface reflectance and physiological and morphological parameters of this moss. Plexiglass screens were used to provide both reduced UV levels (77%) and a 50% decrease in total radiation. The screen combinations were used to separate UV photoprotective from visible photoprotective strategies, because these bryophytes are growing in relatively high light environments compared with many mosses. G. antarctici was affected negatively by ambient levels of UV radiation. Chlorophyll content was significantly lower in plants grown under near‐ambient UV, while the relative proportions of photoprotective carotenoids, especially β‐carotene and zeaxanthin, increased. However, no evidence for the accumulation of UV‐B‐absorbing pigments in response to UV radiation was observed. Although photosynthetic rates were not affected, there was evidence of UV effects on morphology. Plants that were shaded showed fewer treatment responses and these were similar to the natural variation observed between moss growing on exposed microtopographical ridges and in more sheltered valleys within the turf. Given that other Antarctic bryophytes possess UV‐B‐absorbing pigments which should offer better protection under ambient UV‐B radiation, these findings suggest that G. antarctici may be disadvantaged in some settings under a climate with continuing high levels of springtime UV‐B radiation.     

Physiological Effects of Heavy Metals on the Moss Rhytidiadelphus squarrosus

Experiments on the effect of Cd, Cu, Ni, Pb and Zn, suppliedas single salt solutions between 10 µM and 0·1M, on the moss Rhytidiadelphus squarrosus showed little alterationto respiratory rates but reduced photosynthetic rates, and somemembrane damage as assessed by K leakage. Two distinct morphologicalforms of the moss showed different responses to supplied heavymetal. Storage of material, after 30 mins exposure to heavymetals, resulted in a further decrease in the photosyntheticrate. Expressing the photosynthetic decline relative to thetotal heavy metal recovered from the moss showed a similar pattern.Transfer of metal from extracellular exchange sites into theprotoplast was also demonstrated with storage after exposure.An approximately linear relationship was demonstrated betweenphotosynthetic decline and intracellular heavy-metal concentration,irrespective of the duration of exposure or morphological natureof the material used. Photosynthetic decline on storage is concludedto be a response to additional metal stress rather than a progressivedeterioration of the physiological process. Cadmium, copper, lead, nickel, zinc, photosynthesis, respiration, membrane damage, intracellular metals, Rhytidiadelphus squarrosus (Hedw.) Warnst     

Effects of Irradiance on Crassulacean Acid Metabolism in the Epiphyte Tillandsia usneoides L. (Bromeliaceae)

Spanish moss (Tillandsia usneoides L.) was collected in South Carolina, maintained in a greenhouse, then exposed to five levels of photosynthetic photon flux density (PPFD) for 3 weeks. Following this treatment, plants were sampled for chlorophyll concentrations, nocturnal acid accumulations, and photosynthetic responses to subsequent exposure at a range of PPFD. No acclimation to PPFD was observed; all plants exhibited similar patterns of nocturnal CO₂ uptake and acid accumulation regardless of initial PPFD treatment. These patterns revealed that at a PPFD level of approximately 200 micromoles per square meter per second (daytime integrated PPFD of 10 moles per square meter per day), CAM saturated or, in low-PPFD plants, was optimal. The results of this study indicate that adaptation to high PPFD is not necessarily a requirement of CAM.     

Biochemical responses of the aquatic moss Fontinalis antipyretica to Cd,Cu, Pb and Zn determined by chlorophyll fluorescence and protein levels

Biochemical reactions to Cu, Cd, Zn and Pb in the aquatic moss Fontinalis antipyretica were studied in order to characterize the physiological background of the metal response. Chlorophyll fluorescence and intracellular metal localization and stress protein levels were measured. Exposure to 25 or 100 μM Cu over a 7-day period resulted in a decline of chlorophyll fluorescence to about 70% and 52%, respectively. Up to 100 μM Cd caused a decrease in chlorophyll fluorescence to 75%. With all metals used at 25–100 μM concentrations, the intracellular uptake increased. For all metals investigated at 25–100 μM, the intracellular uptake increased. Maximum values were reached at 100 μM Cu, Pb, Zn or Cd exposure. As shown by analytical electron microscopy (EDX, EELS) moss material treated with 50 μM Cu exhibited reduced sulphur levels in the cytoplasm and an increase in phosphate in vacuolar dense particles. EEL-spectra indicated that Cu is chelated in the cytoplasm by SH-groups and coprecipitated with orthophosphate in vacuoles. To monitor the stress response at the protein level, heavy metal induced heat shock protein 70 (hsp70) was measured. An antibody was raised against conserved plant metallothionein p2 motifs derived from Brassica juncea. In all metal-treated samples the antibody bound to proteins of about 8 kDa. However, sequencing failed to reveal significant homologies to known proteins. These experiments provide for the first time results on protein level after heavy metal stress in the aquatic bioindicator moss.