Comparative Phytotoxicity of Metallic Elements on Duckweed Lemna gibba L. Using Growth- and Chlorophyll Fluorescence Induction-Based Endpoints.

In this study, we exposed a commonly used duckweed species-Lemna gibba L.-to twelve environmentally relevant metals and metalloids under laboratory conditions. The phytotoxic effects were evaluated in a multi-well-plate-based experimental setup by means of the chlorophyll fluorescence imaging method. This technique allowed the simultaneous measuring of the growth and photosynthetic parameters in the same samples. The inhibition of relative growth rates (based on frond number and area) and photochemical efficiency (Fv/Fo and Y(II)) were both calculated from the obtained chlorophyll fluorescence images. In the applied test system, growth-inhibition-based phytotoxicity endpoints proved to be more sensitive than chlorophyll-fluorescence-based ones. Frond area growth inhibition was the most responsive parameter with a median EC50 of 1.75 mg L-1, while Fv/Fo, the more responsive chlorophyll-fluorescence-based endpoint, resulted in a 5.34 mg L-1 median EC50 for the tested metals. Ag (EC50 0.005-1.27 mg L-1), Hg (EC50 0.24-4.87 mg L-1) and Cu (EC50 0.37-1.86 mg L-1) were the most toxic elements among the tested ones, while As(V) (EC50 47.15-132.18 mg L-1), Cr(III) (EC50 6.22-19.92 mg L-1), Se(VI) (EC50 1.73-10.39 mg L-1) and Zn (EC50 3.88-350.56 mg L-1) were the least toxic ones. The results highlighted that multi-well-plate-based duckweed phytotoxicity assays may reduce space, time and sample volume requirements compared to the standard duckweed growth inhibition tests. These benefits, however, come with lowered test sensitivity. Our multi-well-plate-based test setup resulted in considerably higher median EC50 (3.21 mg L-1) for frond-number-based growth inhibition than the 0.683 mg L-1 median EC50 derived from corresponding data from the literature with standardized Lemna-tests. Under strong acute phytotoxicity, frond parts with impaired photochemical functionality may become undetectable by chlorophyll fluorometers. Consequently, the plant parts that are still detectable display a virtually higher average photosynthetic performance, leading to an underestimation of phytotoxicity. Nevertheless, multi-well-plate-based duckweed phytotoxicity assays, combined with chlorophyll fluorescence imaging, offer definite advantages in the rapid screening of large sample series or multiple species/clones. As chlorophyll fluorescence images provide information both on the photochemical performance of the test plants and their morphology, a joint analysis of the two endpoint groups is recommended in multi-well-plate-based duckweed phytotoxicity assays to maximize the information gained from the tests.

Cryptogamic communities on flatroofs in the city of Debrecen (East Hungary).

Cryptogams of ten urban flatroofs, contrasting in their age and size, were studied between 2016 and 2018. Siliceous (bituminous felt, gravel, brick) and calcareous (concrete) substrata occurred at each site. Microclimate (T, RH) at two sites of contrasting shading was monitored from September 2016 to January 2017. Biomass of two differently aged, exposed flatroofs was sampled in October 2018. Taxa of Cladonia and Xanthoparmelia have been identified by spot tests and HPTLC. A total of 61 taxa (25 bryophytes, 36 lichens), mostly widespread synanthropic species, have been detected with an explicit difference of species composition between shaded and exposed sites. Floristically interesting species included acidophilous bryophytes (Hedwigia ciliata, Racomitrium canescens) and lichens (Xanthoparmelia conspersa, Stereocaulon tomentosum) of montane character. The most widespread lichen is Cladonia rei which accounted for a significant part of the biomass at selected sites. Species-area curves for bryophytes at exposed sites have become saturated at 100-150 m2. In contrast, saturation of lichen diversity has not been reached even at the largest sites. Flatroofs with traditional roofing techniques can harbour relatively diverse microhabitats and species-rich synanthropic vegetation. It is urgent to study these sites before renovation with modern roofing techniques eliminates them. Diversification of urban surroundings is possible in the future via application of various substrats in renovated and newly constructed roofs.

Species- and Metal-Specific Responses of the Ionome of Three Duckweed Species under Chromate and Nickel Treatments.

In this study, growth and ionomic responses of three duckweed species were analyzed, namely Lemna minor, Landoltia punctata, and Spirodela polyrhiza, were exposed for short-term periods to hexavalent chromium or nickel under laboratory conditions. It was found that different duckweed species had distinct ionomic patterns that can change considerably due to metal treatments. The results also show that, because of the stress-induced increase in leaf mass-to-area ratio, the studied species showed different order of metal uptake efficiency if plant area was used as unit of reference instead of the traditional dry weight-based approach. Furthermore, this study revealed that µXRF is applicable in mapping elemental distributions in duckweed fronds. By using this method, we found that within-frond and within-colony compartmentation of metallic ions were strongly metal- and in part species-specific. Analysis of duckweed ionomics is a valuable approach in exploring factors that affect bioaccumulation of trace pollutants by these plants. Apart from remediating industrial effluents, this aspect will gain relevance in food and feed safety when duckweed biomass is produced for nutritional purposes.

Chlorophyll Fluorescence Imaging-Based Duckweed Phenotyping to Assess Acute Phytotoxic Effects.

Duckweeds (Lemnaceae species) are extensively used models in ecotoxicology, and chlorophyll fluorescence imaging offers a sensitive and high throughput platform for phytotoxicity assays with these tiny plants. However, the vast number of potentially applicable chlorophyll fluorescence-based test endpoints makes comparison and generalization of results hard among different studies. The present study aimed to jointly measure and compare the sensitivity of various chlorophyll fluorescence parameters in Spirodela polyrhiza (giant duckweed) plants exposed to nickel, chromate (hexavalent chromium) and sodium chloride for 72 h, respectively. The photochemistry of Photosystem II in both dark- and light-adapted states of plants was assessed via in vivo chlorophyll fluorescence imaging method. Our results indicated that the studied parameters responded with very divergent sensitivity, highlighting the importance of parallelly assessing several chlorophyll fluorescence parameters. Generally, the light-adapted parameters were more sensitive than the dark-adapted ones. Thus, the former ones might be the preferred endpoints in phytotoxicity assays. Fv/Fm, i.e., the most extensively reported parameter literature-wise, proved to be the least sensitive endpoint; therefore, future studies might also consider reporting Fv/Fo, as its more responsive analogue. The tested toxicants induced different trends in the basic chlorophyll fluorescence parameters and, at least partly, in relative proportions of different quenching processes, suggesting that a basic distinction of water pollutants with different modes of action might be achievable by this method. We found definite hormetic patterns in responses to several endpoints. Hormesis occurred in the concentration ranges where the applied toxicants resulted in strong growth inhibition in longer-term exposures of the same duckweed clone in previous studies. These findings indicate that changes in the photochemical efficiency of plants do not necessarily go hand in hand with growth responses, and care should be taken when one exclusively interprets chlorophyll fluorescence-based endpoints as general proxies for phytotoxic effects.

Retrospective analyses of archive phytotoxicity test data can help in assessing internal dynamics and stability of growth in laboratory duckweed cultures.

High growth potential of duckweed species (Lemnaceae family) has been utilized in wide range of research and practical applications. Based on literature data, however, it can be assumed that duckweed populations maintain constant growth rates only when short periods are considered but can vary over longer time scales. This intrinsic instability in growth can affect the interpretation of growth data. Duckweed phytotoxicity tests are usually performed according to highly standardized protocols. Therefore the archive data provide an opportunity for retrospective comparisons. In the present study we collected growth (frond number- and frond area-based relative growth rates) and morphology (average frond and colony sizes) data from control treatments of phytotoxicity tests. All the analyzed tests were carried out with the same Spirodela polyrhiza (L.) Schleid. (giant duckweed) clone (RDSC ID No. 5501) under the same experimental conditions over more than four years. We aimed to assess the overall variability of the above parameters and to test if intrinsic growth patterns affect growth data in short-term. In general, the results reflected high stability of the measured parameters in long term but also indicated that some temporal variability is inevitable which can bias the comparability of growth tests. The frond area-based relative growth rate resulted in smaller coefficient of variation than the usually preferred frond number-based one. The results also revealed a negative correlation between mean growth rates and their coefficients of variation. Therefore, it would be advisable to introduce higher minimal growth rates and/or maximized tolerable coefficients of variation for control cultures into the standard duckweed growth inhibition tests. Analyses of growth data aggregated on seasonal basis indicated faster growth and larger mean frond size in laboratory duckweed cultures from mid-autumn till mid-spring than during summer and early autumn. But, in shorter term (∼50 days) we did not observe distinct trends in growth suggesting that the successive frond generations have no effect on growth traits within this time-scale. Our results point to the importance of assessing intrinsic growth dynamics in duckweed cultures and also to the re-usability of the already collected phytotoxicity data in addressing new research questions.

Early-Warning Signals of Individual Tree Mortality Based on Annual Radial Growth.

Tree mortality is a key driver of forest dynamics and its occurrence is projected to increase in the future due to climate change. Despite recent advances in our understanding of the physiological mechanisms leading to death, we still lack robust indicators of mortality risk that could be applied at the individual tree scale. Here, we build on a previous contribution exploring the differences in growth level between trees that died and survived a given mortality event to assess whether changes in temporal autocorrelation, variance, and synchrony in time-series of annual radial growth data can be used as early warning signals of mortality risk. Taking advantage of a unique global ring-width database of 3065 dead trees and 4389 living trees growing together at 198 sites (belonging to 36 gymnosperm and angiosperm species), we analyzed temporal changes in autocorrelation, variance, and synchrony before tree death (diachronic analysis), and also compared these metrics between trees that died and trees that survived a given mortality event (synchronic analysis). Changes in autocorrelation were a poor indicator of mortality risk. However, we found a gradual increase in inter-annual growth variability and a decrease in growth synchrony in the last ∼20 years before mortality of gymnosperms, irrespective of the cause of mortality. These changes could be associated with drought-induced alterations in carbon economy and allocation patterns. In angiosperms, we did not find any consistent changes in any metric. Such lack of any signal might be explained by the relatively high capacity of angiosperms to recover after a stress-induced growth decline. Our analysis provides a robust method for estimating early-warning signals of tree mortality based on annual growth data. In addition to the frequently reported decrease in growth rates, an increase in inter-annual growth variability and a decrease in growth synchrony may be powerful predictors of gymnosperm mortality risk, but not necessarily so for angiosperms.

A synthesis of radial growth patterns preceding tree mortality.

Tree mortality is a key factor influencing forest functions and dynamics, but our understanding of the mechanisms leading to mortality and the associated changes in tree growth rates are still limited. We compiled a new pan-continental tree-ring width database from sites where both dead and living trees were sampled (2970 dead and 4224 living trees from 190 sites, including 36 species), and compared early and recent growth rates between trees that died and those that survived a given mortality event. We observed a decrease in radial growth before death in ca. 84% of the mortality events. The extent and duration of these reductions were highly variable (1-100 years in 96% of events) due to the complex interactions among study species and the source(s) of mortality. Strong and long-lasting declines were found for gymnosperms, shade- and drought-tolerant species, and trees that died from competition. Angiosperms and trees that died due to biotic attacks (especially bark-beetles) typically showed relatively small and short-term growth reductions. Our analysis did not highlight any universal trade-off between early growth and tree longevity within a species, although this result may also reflect high variability in sampling design among sites. The intersite and interspecific variability in growth patterns before mortality provides valuable information on the nature of the mortality process, which is consistent with our understanding of the physiological mechanisms leading to mortality. Abrupt changes in growth immediately before death can be associated with generalized hydraulic failure and/or bark-beetle attack, while long-term decrease in growth may be associated with a gradual decline in hydraulic performance coupled with depletion in carbon reserves. Our results imply that growth-based mortality algorithms may be a powerful tool for predicting gymnosperm mortality induced by chronic stress, but not necessarily so for angiosperms and in case of intense drought or bark-beetle outbreaks.

Towards a common methodology for developing logistic tree mortality models based on ring-width data.

Tree mortality is a key process shaping forest dynamics. Thus, there is a growing need for indicators of the likelihood of tree death. During the last decades, an increasing number of tree-ring based studies have aimed to derive growth-mortality functions, mostly using logistic models. The results of these studies, however, are difficult to compare and synthesize due to the diversity of approaches used for the sampling strategy (number and characteristics of alive and death observations), the type of explanatory growth variables included (level, trend, etc.), and the length of the time window (number of years preceding the alive/death observation) that maximized the discrimination ability of each growth variable. We assess the implications of key methodological decisions when developing tree-ring based growth-mortality relationships using logistic mixed-effects regression models. As examples, we use published tree-ring datasets from Abies alba (13 different sites), Nothofagus dombeyi (one site), and Quercus petraea (one site). Our approach is based on a constant sampling size and aims at (1) assessing the dependency of growth-mortality relationships on the statistical sampling scheme used, (2) determining the type of explanatory growth variables that should be considered, and (3) identifying the best length of the time window used to calculate them. The performance of tree-ring-based mortality models was reasonably high for all three species (area under the receiving operator characteristics curve, AUC > 0.7). Growth level variables were the most important predictors of mortality probability for two species (A. alba, N. dombeyi), while growth-trend variables need to be considered for Q. petraea. In addition, the length of the time window used to calculate each growth variable was highly uncertain and depended on the sampling scheme, as some growth-mortality relationships varied with tree age. The present study accounts for the main sampling-related biases to determine reliable species-specific growth-mortality relationships. Our results highlight the importance of using a sampling strategy that is consistent with the research question. Moving towards a common methodology for developing reliable growth-mortality relationships is an important step towards improving our understanding of tree mortality across species and its representation in dynamic vegetation models.

Incorporation of iron into chloroplasts triggers the restoration of cadmium induced inhibition of photosynthesis.

Photosynthetic symptoms of acute Cd stress can be remedied by elevated Fe supply. To shed more light on the most important aspects of this recovery, the detailed Fe trafficking and accumulation processes as well as the changes in the status of the photosynthetic apparatus were investigated in recovering poplar plants. The Cd-free, Fe-enriched nutrient solution induced an immediate intensive Fe uptake. The increased Fe/Cd ratio in the roots initiated the translocation of Fe to the leaf with a short delay that ultimately led to the accumulation of Fe in the chloroplasts. The chloroplast Fe uptake was directly proportional to the Fe translocation to leaves. The accumulation of PSI reaction centers and the recovery of PSII function studied by Blue-Native PAGE and chlorophyll a fluorescence induction measurements, respectively, began in parallel to the increase in the Fe content of chloroplasts. The initial reorganization of PSII was accompanied by a peak in the antennae-based non-photochemical quenching. In conclusion, Fe accumulation of the chloroplasts is a process of prime importance in the recovery of photosynthesis from acute Cd stress.

Stress hardening under long-term cadmium treatment is correlated with the activation of antioxidative defence and iron acquisition of chloroplasts in Populus.

Cadmium (Cd), a highly toxic heavy metal affects growth and metabolic pathways in plants, including photosynthesis. Though Cd is a transition metal with no redox capacity, it generates reactive oxygen species (ROS) indirectly and causes oxidative stress. Nevertheless, the mechanisms involved in long-term Cd tolerance of poplar, candidate for Cd phytoremediation, are not well known. Hydroponically cultured poplar (Populus jacquemontiana var. glauca cv. 'Kopeczkii') plants were treated with 10 µM Cd for 4 weeks. Following a period of functional decline, the plants performed acclimation to the Cd induced oxidative stress as indicated by the decreased leaf malondialdehyde (MDA) content and the recovery of most photosynthetic parameters. The increased activity of peroxidases (PODs) could have a great impact on the elimination of hydrogen peroxide, and thus the recovery of photosynthesis, while the function of superoxide dismutase (SOD) isoforms seemed to be less important. Re-distribution of the iron content of leaf mesophyll cells into the chloroplasts contributed to the biosynthesis of the photosynthetic apparatus and some antioxidative enzymes. The delayed increase in photosynthetic activity in relation to the decline in the level of lipid peroxidation indicates that elimination of oxidative stress damage by acclimation mechanisms is required for the restoration of the photosynthetic apparatus during long-term Cd treatment.

Assessment of Giant Duckweed (Spirodela polyrhiza L. Schleiden) Turions as Model Objects in Ecotoxicological Applications.

In this study germination of Spirodela polyrhiza (L.) Schleiden (giant duckweed) turions was assessed under cadmium exposure to test applicability of a novel turion-based ecotoxicology method. Floating success of germinating turions, protrusion of the first and subsequent fronds as test endpoints were investigated and compared to results of standard duckweed growth inhibition tests with fronds of the same species. Our results indicate that turions can be used to characterize effects of toxic substances. Initial phase of turion germination (floating up and appearance of the first frond) was less sensitive to Cd treatments than the subsequent frond production. The calculated effective concentrations for growth rates in turion and normal frond tests were similar. Single frond area produced by germinating turions proved to be the most sensitive test endpoint. Single frond area and colony disintegration as additionally measured parameters in normal frond cultures also changed due to Cd treatments but the sensitivity of these parameters was lower than that of growth rates.

Characterization of the aodA, dnmA, mnSOD and pimA genes in Aspergillus nidulans.

Mitochondria play key roles in cellular energy generation and lifespan of most eukaryotes. To understand the functions of four nuclear-encoded genes predicted to be related to the maintenance of mitochondrial morphology and function in Aspergillus nidulans, systematic characterization was carried out. The deletion and overexpression mutants of aodA, dnmA, mnSOD and pimA encoding alternative oxidase, dynamin related protein, manganese superoxide dismutase and Lon protease, respectively, were generated and examined for their growth, stress tolerances, respiration, autolysis, cell death, sterigmatocystin production, hyphal morphology and size, and mitochondrial superoxide production as well as development. Overall, genetic manipulation of these genes had less effect on cellular physiology and ageing in A. nidulans than that of their homologs in another fungus Podospora anserina with a well-characterized senescence. The observed interspecial phenotypic differences can be explained by the dissimilar intrinsic stabilities of the mitochondrial genomes in A. nidulans and P. anserina. Furthermore, the marginally altered phenotypes observed in A. nidulans mutants indicate the presence of effective compensatory mechanisms for the complex networks of mitochondrial defense and quality control. Importantly, these findings can be useful for developing novel platforms for heterologous protein production, or on new biocontrol and bioremediation technologies based on Aspergillus species.

Comparative study on the sensitivity of turions and active fronds of giant duckweed (Spirodela polyrhiza (L.) Schleiden) to heavy metal treatments.

Standard ecotoxicological test procedures use only active forms of aquatic plants. The potential effects of toxicants on vegetative propagules, which play an important role in the survival of several aquatic plant species, is not well understood. Because turion-like resting propagules overwinter on the water bottom in temperate regions, they could be exposed to contaminants for longer periods than active plants. Due to its turion producing capability, giant duckweed (Spirodela polyrhiza) is widely used in studying morphogenesis, dormancy, and activation mechanisms in plants. It is also suitable for ecotoxicological purposes. The present work aims to compare the growth inhibition sensitivity of active (normal frond) and overwintering (turion) forms of S. polyrhiza to concentrations of nickel (Ni), cadmium (Cd) and hexavalent chromium (Cr) ranging from 0 to 100mgL(-1). The results indicated that in general, resting turions have higher heavy metal tolerance than active fronds. Cd proved to be the most toxic heavy metal to S. polyrhiza active frond cultures because it induced rapid turion formation. In contrast, the toxicity of Ni and Cr were found to be similar but lower than the effects of Cd. Cr treatments up to 10mgL(-1) did not result in any future negative effects on turion activation. Turions did not survive heavy metal treatments at higher concentrations of Cr. Cd and Ni treatments affected both the floating-up and germination of turions but did not significantly affect the vigor of sprouts. Higher concentrations (of 100mgL(-1)) Cd completely inhibited germination.

Osmotic stress responses of individual white oak (Quercus section, Quercus subgenus) genotypes cultured in vitro.

White oaks (Quercus section, Quercus subgenus) are widely distributed in Europe. Quercus petraea (sessile oak), an economically important species is predicted to be affected by climate change. Q. pubescens (pubescent oak) and Q. virgiliana (Italian pubescent oak) are economically less important, drought tolerant species. Frequent hybridization of white oaks was observed and currently the introgression of Q. pubescens and Q. virgiliana in non-mediterranean regions of Europe has been reported. Our goal was to use tissue cultures established from individual trees of the above taxa and their putative hybrids, all present in the forest stand of Síkfokút LTER Research Area (NE Hungary) as simple experimental model systems for studying drought/osmotic stress tolerance. Tissue cultures are more suitable models for such studies, than seedlings, because they are genetically identical to the parent plants. Polyethylene glycol (PEG6000) treatments were used for this purpose. The identification of taxa was based on leaf morphological traits and microsatellite analysis and showed that Q. petraea is genetically distinct to all other taxa examined. We established six callus lines of Quercus. As expected, in Q. petraea cultures PEG6000 induced severe loss of fresh weight and the ability to recover after removal of the osmoticum, which was not characteristic for Q. pubescens and Q. virgiliana. Putative hybrids exhibited an intermediate response to osmotic stress. Activity gels showed the increase of single-strand preferring (SSP) nuclease and no significant change of guaiacol-peroxidase activities in drought-sensitive genotypes/cultures and no significant increase of SSP nuclease activities accompanied with increases of guaiacol-peroxidase activities in drought-tolerant ones. This indicates that drought/osmotic stress tolerance is associated to increased capacity of scavenging reactive oxygen species and hence less susceptibility to DNA damage. Our results confirm that tissue cultures of oak are suitable model systems for studying drought/osmotic stress responses.

Impact of moderate Fe excess under Cd stress on the photosynthetic performance of poplar (Populus jacquemontiana var. glauca cv. Kopeczkii).

Cadmium interference with Fe nutrition has a strong impact on the development and efficiency of the photosynthetic apparatus. To shed more light on the interaction between Fe and Cd, it was studied how iron given in moderate excess under Cd stress affects the development and functioning of chlorophyll-protein complexes. Poplar plants grown in hydroponics up to four-leaf stage were treated with 10 µM Cd(NO3)2 in the presence of 50 µM Fe([III])-citrate as iron supply (5xFe + Cad) for two weeks. Though leaf area growth was inhibited similarly to that of Cad (10 µM Cd(NO3)2 + 10 µM Fe([III])-citrate) plants, chlorophyll content, ¹4CO2 fixation and quenching parameters calculated from PAM fluorescence induction measurements were control-like in 5xFe+Cad leaves. Increased chloroplast iron content (measured photometrically by the bathophenanthroline disulfonate method) without changes in the iron and cadmium content of leaves (determined by inductively coupled plasma mass spectrometry) pointed out that a key factor in the observed protection of photosynthesis is the iron-excess-induced redistribution of iron in the leaf. However, the chlorophyll a/b ratio and the chlorophyll-protein pattern obtained by Deriphat PAGE remained similar to that of Cad leaves. The decreased amount of PSII core and PSI in mature and developing leaves, respectively, refers to developmental stage-dependent remodelling of thylakoids in the presence of Cd. The results underline not only the beneficial effect of iron excess under Cd stress, but also refer to the importance of a proper Fe/Cd ratio and light environment to avoid its possible harmful effects.

Biodiversity differences between managed and unmanaged forests: meta-analysis of species richness in Europe.

Past and present pressures on forest resources have led to a drastic decrease in the surface area of unmanaged forests in Europe. Changes in forest structure, composition, and dynamics inevitably lead to changes in the biodiversity of forest-dwelling species. The possible biodiversity gains and losses due to forest management (i.e., anthropogenic pressures related to direct forest resource use), however, have never been assessed at a pan-European scale. We used meta-analysis to review 49 published papers containing 120 individual comparisons of species richness between unmanaged and managed forests throughout Europe. We explored the response of different taxonomic groups and the variability of their response with respect to time since abandonment and intensity of forest management. Species richness was slightly higher in unmanaged than in managed forests. Species dependent on forest cover continuity, deadwood, and large trees (bryophytes, lichens, fungi, saproxylic beetles) and carabids were negatively affected by forest management. In contrast, vascular plant species were favored. The response for birds was heterogeneous and probably depended more on factors such as landscape patterns. The global difference in species richness between unmanaged and managed forests increased with time since abandonment and indicated a gradual recovery of biodiversity. Clearcut forests in which the composition of tree species changed had the strongest effect on species richness, but the effects of different types of management on taxa could not be assessed in a robust way because of low numbers of replications in the management-intensity classes. Our results show that some taxa are more affected by forestry than others, but there is a need for research into poorly studied species groups in Europe and in particular locations. Our meta-analysis supports the need for a coordinated European research network to study and monitor the biodiversity of different taxa in managed and unmanaged forests.

Cylindrospermopsin and microcystin-LR alter the growth, development and peroxidase enzyme activity of white mustard (Sinapis alba L.) seedlings, a comparative analysis.

This work focuses on the comparative analysis of the effects of two cyanobacterial toxins of different chemical structure cylindrospermopsin (CYN) and microcystin-LR (MC-LR) on the white mustard (Sinapis alba L.) seedlings. Both cyanotoxins reduced significantly the fresh mass and the length of cotyledons, hypocotyls and main roots of seedlings in a concentration dependent manner. For various mustard organs the 50% inhibitory concentration values (IC50) of growth were between 3-5 µg ml(-1) for MC-LR and between 5-10 µg ml-1 for CYN, respectively. Cyanotoxins altered the development of cotyledons, the accumulation of photosynthetically active pigments and anthocyanins. Low MC-LR concentrations (0.01 and 0.1 µg ml(-1)) stimulated anthocyanin formation in the cotyledons but higher than 1 µg ml(-1) MC-LR concentrations strongly inhibited it. The CYN treated chlorotic cotyledons were violet coloured in consequence of high level of anthocyanins, while MC-LR induced chlorosis was accompanied by the appearance of necrotic patches. Necrosis and increases of peroxidase enzyme activity (POD) are general stress responses but these alterations were characteristic only for MC-LR treated mustard plants. These findings provide experimental evidences of developmental alterations induced by protein synthesis and protein phosphatase inhibitory cyanotoxins (CYN and MC-LR) in a model dicotyledonous plant.

Seasonal and diurnal variability in sap flow intensity of mature sessile oak (Quercus petraea (Matt.) Liebl.) trees in relation to microclimatic conditions.

In this study sap flow dynamics of mature sessile oak trees (Quercus petraea) in a marginal sessile oak-turkey oak forest was investigated in 2009. That year spring was dry without significant rain in April and May and the driest month was August. Due to the extreme weather conditions the volumetric soil water content (SWC) of upper 30 cm was low on experimental days in May (0.13-0.14 cm3 cm-3) but it reached the lowest value in August (0.08 cm3 cm-3). Sap flow was measured in a dominant and a co-dominant tree by heat dissipation method from 26 March till 30 October. In the present paper several three-day long periods of the continuous seasonal recordings were chosen to represent the effects of typical weather conditions and different stages of canopy development on sap flow dynamics. The daily maximum sap flow density values of dominant and co-dominant trees were similar (0.30-0.32 cm3 cm-2 min-1) in moist period (July). Rains and transient increase of SWC after proceeding drought resulted in change of diurnal course of sap flow in experimental days of July. In this period dominant trees also showed considerable sap flow (0.19 cm3 cm-2 min-1) during night hours and short sap flow peaks in early morning (6:00 to 8:00 a.m.) indicating the refilling of desiccated tissues. After the progressive drought in August the daily maximum sap flow density decreased to 0.07 cm3 cm-2 min-1 in dominant tree and to 0.12 cm3 cm-2 min-1 in the co-dominant. Both trees exhibited gradual stomatal closure from morning hours.