István Bácsi

Cylindrospermopsin induces alterations of root histology and microtubule organization in common reed (Phragmites australis) plantlets cultured in vitro.

We aimed to study the histological and cytological alterations induced by cylindrospermopsin (CYN), a protein synthesis inhibitory cyanotoxin in roots of common reed (Phragmites australis). Reed is an ecologically important emergent aquatic macrophyte, a model for studying cyanotoxin effects. We analyzed the histology and cytology of reed roots originated from tissue cultures and treated with 0.5-40 microg ml(-1) (1.2-96.4 microM) CYN. The cyanotoxin decreased root elongation at significantly lower concentrations than the elongation of shoots. As general stress responses of plants to phytotoxins, CYN increased root number and induced the formation of a callus-like tissue and necrosis in root cortex. Callus-like root cortex consisted of radially swollen cells that correlated with the reorientation of microtubules (MTs) and the decrease of MT density in the elongation zone. Concomitantly, the cyanotoxin did not decrease, rather it increased the amount of beta-tubulin in reed plantlets. CYN caused the formation of double preprophase bands; the disruption of mitotic spindles led to incomplete sister chromatid separation and disrupted phragmoplasts in root tip meristems. This work shows that CYN alters reed growth and anatomy through the alteration of MT organization.

Appearance of Planktothrix rubescens Bloom with [D-Asp3, Mdha7]MC–RR in Gravel Pit Pond of a Shallow Lake-Dominated Area

Blooms of toxic cyanobacteria are well-known phenomena in many regions of the world. Microcystin (MC), the most frequent cyanobacterial toxin, is produced by entirely different cyanobacteria, including unicellular, multicellular filamentous, heterocytic, and non-heterocytic bloom-forming species. Planktothrix is one of the most important MC-producing genera in temperate lakes. The reddish color of cyanobacterial blooms viewed in a gravel pit pond with the appearance of a dense 3 cm thick layer (biovolume: 28.4 mm3 L−1) was an unexpected observation in the shallow lake-dominated alluvial region of the Carpathian Basin. [d-Asp3, Mdha7]MC–RR was identified from the blooms sample by MALDI-TOF and NMR. Concentrations of [d-Asp3, Mdha7]MC–RR were measured by capillary electrophoresis to compare the microcystin content of the field samples and the isolated, laboratory-maintained P. rubescens strain. In analyzing the MC gene cluster of the isolated P. rubescens strain, a deletion in the spacer region between mcyE and mcyG and an insertion were located in the spacer region between mcyT and mcyD. The insertion elements were sequenced and partly identified. Although some invasive tropical cyanobacterial species have been given a great deal of attention in many recent studies, our results draw attention to the spread of the alpine organism P. rubescens as a MC-producing, bloom-forming species.

Isolation of viable cell mass from frozen Microcystis viridis bloom containing microcystin-RR.

Cyanobacterial species commonly occur in the phytoplankton of freshwater lakes and sometimes develop as toxin-producing blooms. Microcystis is one of the most common genera of freshwater cyanobacteria and is often the dominating phytoplankton of eutrophic lakes all over the world. In eutrophic lakes, large amounts of Microcystis may overwinter in the sediment and re-inoculate the water column in spring. In most cases, the overwintering pelagic population—if it exists—is small, and its role in re-inoculation has not been clear yet. In December 2005, we found large amounts of Microcystis on the surface, frozen in the ice cover in a eutrophic pond (Pond Hármashegy, Hungary). We identified the Microcystis species and investigated the viability and the toxicity of the frozen cells. The dominant species in the bloom samples was Microcystis viridis. Viability tests showed that the colonies isolated from the ice cover were composed of living cells. The isolated strain was found toxic, we analyzed the microcystin composition in the frozen planktonic Microcystis mass; in the investigated samples microcystin-RR was the main cyanotoxin.

Functional diversity supports the biomass–diversity humped-back relationship in phytoplankton assemblages

Summary Modelling the relationship between biomass and diversity in phytoplankton assemblages provides new insights into the mechanisms responsible for the coexistence of species, even in terrestrial ecosystems. We tested the biomass–diversity relationship in lake phytoplankton along a wide biomass gradient using functional species groups. We hypothesized that changes in the taxonomic diversity of the phytoplankton along a biomass gradient are associated with altered functional diversity. For the analyses, in total 768 samples were collected from 30 oxbows, reservoirs and lakes in the Hungarian Lowland Region and analysed between 1992 and 2002. We found that the diversity and also the number of functional species groups showed a humped-back curve similar to the species richness. The changes in functional group composition act as a good proxy for phytoplankton species responses. We found that the peak of the number of strategy groups and their Shannon diversity was at a much lower biomass than that of species richness. We revealed the fine-scale effects of increasing the dominance of respective species or species groups with increasing biomass. This increase was well reflected by the changes in the functional characteristics: first, the species evenness; then, the Shannon diversity; and finally, the species richness started to decrease with increasing biomass. Cyanoprokaryota were positively correlated with increasing biomass and negatively with the increase in species richness; thus, the high increase both in their abundance and biomass can be responsible for the abruptly decreasing part of the humped-back curve. We detected a humped-back curve between biomass and diversity, where the peak compared to terrestrial plant communities tended to be towards high biomass scores, that is, >60% instead of the 20–60% of the biomass range typical for terrestrial plant communities. Marked differences in the structural and dynamic features of phytoplankton assemblages and terrestrial plant communities are likely responsible for this difference.

Cylindrospermopsin inhibits growth and modulates protease activity in the aquatic plants Lemna minor L. and Wolffia arrhiza (L.) Horkel

The toxic effects of cylindrospermopsin (cyanobacterial toxin) on animals have been examined extensively, but little research has focused on their effects on plants. In this study cylindrospermopsin (CYN) caused alterations of growth, soluble protein content and protease enzyme activity were studied on two aquatic plants Lemna minor and Wolffia arrhiza in short-term (5 days) experiments. For the treatments we used CYN containing crude extracts of Aphanizomenon ovalisporum (BGSD-423) and purified CYN as well. The maximal inhibitory effects on fresh weight of L. minor and W. arrhiza caused by crude extract were 60% and 54%, respectively, while the maximum inhibitory effects were 30% and 43% in the case of purified CYN at 20 μg ml(-1) CYN content of culture medium. In CYN-treated plants the concentration of soluble protein showed mild increases, especially in W. arrhiza. Protease isoenzyme activity gels showed significant alterations of enzyme activities under the influence of CYN. Several isoenzymes were far more active and new ones appeared in CYN-treated plants. Treatments with cyanobacterial crude extract caused stronger effects than the purified cyanobacterial toxins used in equivalent CYN concentrations.

Effects of Cylindrospermopsin Producing Cyanobacterium and Its Crude Extracts on a Benthic Green Alga-Competition or Allelopathy?

Cylindrospermopsin (CYN) is a toxic secondary metabolite produced by filamentous cyanobacteria which could work as an allelopathic substance, although its ecological role in cyanobacterial-algal assemblages is mostly unclear. The competition between the CYN-producing cyanobacterium Chrysosporum (Aphanizomenon) ovalisporum, and the benthic green alga Chlorococcum sp. was investigated in mixed cultures, and the effects of CYN-containing cyanobacterial crude extract on Chlorococcum sp. were tested by treatments with crude extracts containing total cell debris, and with cell debris free crude extracts, modelling the collapse of a cyanobacterial water bloom. The growth inhibition of Chlorococcum sp. increased with the increasing ratio of the cyanobacterium in mixed cultures (inhibition ranged from 26% to 87% compared to control). Interestingly, inhibition of the cyanobacterium growth also occurred in mixed cultures, and it was more pronounced than it was expected. The inhibitory effects of cyanobacterial crude extracts on Chlorococcum cultures were concentration-dependent. The presence of C. ovalisporum in mixed cultures did not cause significant differences in nutrient content compared to Chlorococcum control culture, so the growth inhibition of the green alga could be linked to the presence of CYN and/or other bioactive compounds.

Halophilic diatom taxa are sensitive indicators of even short term changes in lowland lotic systems

Abstract The occurrence and spread of halophilic diatom taxa in freshwater lotic ecosystems are influenced both by natural processes and anthropogenic pollution. Diatom assemblages were regularly monitored in lowland lotic systems in Hungary (Central Europe) during the unusually dry year of 2012. Highly pronounced changes in diatom composition were observed from spring to autumn. Halophilic taxa (especially Nitzschia sensu lato species) appeared in the dry autumn. In addition, the total relative abundances of halophilic species also increased up to autumn. Abundance of Nitzschia cf. lorenziana and Nitzschia tryblionella showed a positive correlation with chloride and phosphate concentration, while that of other taxa like Tryblionella apiculata or Tryblionella calida showed a positive correlation with the concentration of nitrate. Our findings clearly demonstrated that these halophilic and mesohalophilic diatom taxa were sensitive indicators of even short-term changes in lowland lotic ecosystems, such as the increasing salt concentration from spring to autumn caused by the lack of rainfall and/or environmental loads.

Application of external micro-spectrophotometric detection to improve sensitivity on microchips

The goal of this work was to increase the sensitivity of a UV–Vis spectrophotometer by decreasing the background noise and lengthening the optical path. A microphotometer has been modified to precisely select very small parts of a microfluidic channel pattern of a chip and to measure light absorbance on a magnified area of the selected part of the channel. The viability of combining a projection microscope and a spectrophotometer for external absorbance measurements on disposable PDMS chips was studied. Besides the external direct detection above a microfluidic channel, the optical pathlength was lengthened by detecting in the region of the perpendicular exit port. Increasing the cross-sectional area of the zone of irradiation improved the signal-to-noise ratio and the limits of detection (LOD).

Colonisation processes in benthic algal communities are well reflected by functional groups

Single-trait analyses are used to select the most appropriate species characteristics for an effective indication of changes in multiple stressors, but they are robust to detect fine-scale functional changes in biofilms. The combination of single traits may appropriately reflect ecological properties of changing benthic assemblages. We studied colonisation processes of benthic algal assemblages focusing on the changes in trait composition using life forms, type of attachments, cell size and mobility as single traits in a small lowland stream. We tested the descriptive power of single-trait groups (STGs) and also combined trait groups (CTGs). We assumed that STGs would be significantly affected by environmental factors, but compositional changes in biofilms would be described more easily by using CTGs rather than STGs. Our hypotheses were confirmed by the results. While some STGs correlated positively to environmental factors indicating disturbances, others correlated to environmental factors indicating the stable conditions. The fast settlement of large-sized groups was also relevant determining the compositional changes in the studied benthic community. Despite the strong correlation between STGs and environmental variables, CTGs analyses revealed important functional relations in the ecosystem, since CTGs display more sophisticated functional features of the organisms, which may provide more realistic responses.