Consequences of the presence of 24-epibrassinolide, on cultures of a diatom, Asterionella formosa

Addition of the plant hormone 24-epibrassinolide to culture media stimulated the growth of a freshwater diatom, Asterionella formosa. The hormone stimulated activity of glyceraldehyde-3-phosphate dehydrogenase (GAPDH), a key enzyme from Calvin cycle, by 6-fold. Other key metabolic enzymes, phosphofructokinase and malate dehydrogenase were also stimulated but to a lesser extent. The activity of glucose-6-phosphate dehydrogenase, involved in the oxidative pentose phosphate pathway, also increased in the presence of the hormone but only under non reducing conditions. In cells stimulated by epibrassinolide, activated enzymes were sensitive to oxidized-DTT. GAPDH purified from cells grown in the presence of the hormone was not associated with a small protein of 8.5 kDa shown to be similar to CP12. Consequently the activity of GAPDH was no longer regulated by either oxidizing or reducing conditions. Among enzymes that, like GAPDH, responded positively to reducing agent were fructose-1,6-bisphosphatase (FBPase) and glucose-6-phosphate dehydrogenase (G6PDH). These enzymes were also sensitive to, and were negatively regulated by, oxidized-DTT. The activities in extracts from illuminated cells differed from those from darkened cells: FBPase, G6PDH and GAPDH, that were activated by DTT in darkened cells were no more activated in illuminated cells, but were oxidized by oxidized-DTT. Thus, oxidizing or reducing conditions mimic the conditions in dark and light, respectively. Unlike the other enzymes, phosphofructokinase (PFK) was inhibited by DTT but oxidized-DTT reversed this effect. The enzymes shown to be redox regulated in vitro by reduction/oxidation are very likely candidates for regulation in vivo by thioredoxins.     

Changes in the elemental composition of Asterionella formosa during the diatom spring bloom

Changes in the elemental composition of the diatom Asterionellaformosa within mixed phytoplankton samples were determined overthe spring bloom period using energy dispersive X-ray microanalysis.Using a 10 kV electron microprobe, X-ray information from thetop 1–2 µm of the cell revealed overall mean concentrationsof: Si (4636 mmol kg^–1 dry wt), P (82), S (54), Cl (71)K (94) and Ca (90). Concentrations of all elements showed widevariation within each date sample, with unimodal frequency distributionsapproximating to a Normal distribution. Correlation of the entiredata set demonstrated clear statistical associations betweenelements, including Si, P and K. Uptake of Si during the courseof the diatom bloom led to a major fall in lake water concentration(1.0 to 0.07 mg l^–1), which correlated with a decreasein the mean cell Si concentration (6735 to 3107 mmol kg^–1dry wt) during the final phase of the bloom. Mean cell concentrationsof P and K also showed marked decreases with time, in closeparallel with cell Si. These changes in P and K were attributedto the high level of internal correlation with Si and not toany significant decrease in P and K availability in the environment.     

Multiple asexual reproduction of Asterionella formosa Hass. from Lake Baikal and Angara reservoirs

The living plankton diatom Asterionella formosa from Lake Baikal and the Irkutsk Water Reservoir was studied using light microscopy. New facts were obtained which provide evidence of the occurrence of multiple asexual reproduction of Asterionella formosa.     

Molecular characterization of cyanobacterial diversity in a shallow eutrophic lake

We have studied the diversity of pelagic cyanobacteria in Lake Loosdrecht, The Netherlands, through recovery and analysis of small subunit ribosomal RNA gene sequences from lake samples and cyanobacterial isolates. We used an adapted protocol for specific amplification of cyanobacterial rDNA for denaturing gradient gel electrophoresis (DGGE) analysis. This protocol enabled direct comparison of cyanobacterial community profiles with overall bacterial profiles. The theoretical amplification specificity of the primers was supported by sequence analysis of DNA from excised DGGE bands. Sequences recovered from these bands, in addition to sequences obtained by polymerase chain reaction (PCR) and cloning from lake DNA as well as from cyanobacterial isolates from the lake, revealed a diverse consortium of cyanobacteria, among which are representatives of the genera Aphanizomenon, Planktothrix, Microcystis and Synechococcus. One numerically important and persistent cyanobacterium in the lake, Prochlorothrix hollandica, appeared to co-occur with an unknown but related species. However, the lake is dominated by filamentous species that originally have been termed 'Oscillatoria limnetica-like'. We show that this is a group of several related cyanobacteria, co-occurring in the lake, which belong to the Limnothrix/Pseudanabaena group. The available variation among the coexisting strains of this group can explain the persistent dominance of the group under severe viral pressure.     

Response of a eutrophic, shallow subtropical lake to reduced nutrient loading

1. Lake Apopka (FL, U.S.A.) was subjected to decades of high nutrient loading from farms developed in the 1940s on converted riparian wetlands. Consequences included perennially high densities of cyanobacteria, low water transparency, elimination of submerged vegetation, modified fish community, and deposition of nutrient‐rich, flocculent sediments. 2. Initial steps were taken to reduce phosphorus (P) loading. Through strengthened regulation and purchase of farms for restoration, external P loading was reduced on average from 0.56 to 0.25 g P m^?2 year^?1 (55%) starting in 1993. The P loading target for the lake is 0.13 g P m^?2 year^?1. 3. For the first 6 years of P loading reduction the annual sedimentation coefficient (σ) averaged 13% less than the prior long‐term value (0.97 versus 1.11 year^?1). The sedimentation coefficient, σ, was lower in the last 3 years of the study, but this period included extreme low‐water conditions and may not be representative. Annual σ was negative (net P flux to the water column) only 1 year. 4. Wind velocity explained 43% of the variation in σ during the period before reductions in total phosphorus (TP) concentration of lake water, but this proportion dropped to 6% after TP reductions. 5. Annual mean TP concentrations differed considerably from values predicted from external loading and hydraulic retention time using the Vollenweider–Organization for Economic Co‐operation and Development relationship. Reductions in lake water TP concentration fit model predictions better when multiyear (3‐year) mean values were used. 6. Evidence available to date indicates that this shallow, eutrophic lake responded to the decrease in external P loading. Neither recycling of sediment P nor wind‐driven resuspension of sediments prevented improvements in water quality. Reductions in TP concentration were evident about two TP‐resident times (2 × 0.9 year) after programmes began to reduce P loading. Improvements in concentrations of chlorophyll a and total suspended solids as well as in Secchi transparency lagged changes in lake‐water TP concentration but reached similar magnitudes during the study.     

Mineralization of dissolved organic phosphorus from a shallow eutrophic lake

Release of soluble reactive phosphorus (SRP) from dissolved organic phosphorus (DOP), concentrated by reverse osmosis of water samples from Lough Neagh Northern Ireland, was measured in the presence of enzymes and cultures of lake water bacteria in a basal liquid medium adjusted to the pH of lake water (7.6). No hydrolysis of unfractionated DOP was observed in the presence of alkaline phosphatase but a combination of alkaline phosphatase and phosphodiesterase mineralized 14% of DOP in a 30 day incubation period at 15 °C. A similar amount of mineralization was attained by phytase. Phytase induced the same degree of mineralization in a range of DOP fractions varying from MW > 100 000 to c. 500. A mixed culture of lake water bacteria mineralized 12% of unfractionated DOP. Single cultures of lake water bacteria displayed low mineralizing activity (mean of 49 cultures = 5% DOP hydrolysed). Results indicate that DOP from Lough Neagh in the above molecular weight range is predominantly recalcitrant to bacterial mineralization under natural lake conditions.     

Responses of the planktonic diatom Asterionella formosa Hassall to abiotic environmental factors in a reservoir complex (south-eastern France)

The River Durance and its main tributary, the Verdon, are both highly regulated rivers flowing in south-eastern France. The course of both rivers is interrupted by a series of reservoirs with quite different geographical, morphometric, climatic, hydrodynamic and chemical characteristics. The planktonic diatom Asterionella formosa Hassall, which has undesirable cyclic effects from the water management point of view, was studied in this complex of reservoirs located in the Mediterranean region. The results indicate that only the monomictic calcareous reservoirs show a bimodal pattern of Asterionella formosa abundance. The population dynamics of this algal species was found to depend on both the morphometric features and the hydraulic mode of management (retention time) used at each reservoir, as well as on the physical (temperature, suspended matter) and chemical (nitrogen, silica, calcium) characteristics of the water.     

The seasonal periodicity of planktonic diatoms in a shallow eutrophic lake

The seasonal periodicity of four species of planktonic diatoms in a small eutrophic lake in the Shropshire-Cheshire Plain, England, is examined. Diatoms typically dominate the spring increase; a second period of growth follows in the summer months. The growth phases are considered in relation to environmental factors in the mere. Of these, light levels appear to be critical in determining the onset of growth and the size of the population maxima, whilst stratification and turbulence play a leading role in the vertical distribution of the algae, and hence, of the growth conditions to which they are exposed. The thermocline is believed to provide a reservoir of diatoms in summer, maintaining them in a position where they are able to gain maximum advantage from increased wind-induced turbulence. Relative specific differences in growth requirements and in behaviour under varying physical conditions are important in determining which species dominate. Dominance may be modified by the effect of attacks by fungal parasites. It is also shown that, generally, nutrients are present in relative abundance, and only rarely does their availability become a limiting factor. Tt is concluded that diatom growth in this lake is typically subject to physical rather than chemical control.     

Adaptation of the fungal parasite Zygorhizidium planktonicum during 200 generations of growth on homogeneous and heterogeneous populations of its host, the diatom Asterionella formosa

We followed adaptation of the chytrid parasite Zygorhizidium planktonicum during 200 generations of growth on its host, the freshwater diatom Asterionella formosa, in a serial passage experiment. Evolution of parasite fitness was assessed both on a homogenous and heterogeneous host population, consisting of respectively a single new and ten different new host strains. These 10 host strains were genetically different and also varied in their initial susceptibility to the parasite. Parasite fitness increased significantly and rapidly on the new, genetically homogenous host population, but remained unaltered during 200 generations of growth on the heterogeneous host population. Enhanced parasite fitness was the result of faster and more efficient transmission, resulting in higher values of R0 (number of secondary infections). Consequently, parasites that evolved within the uniclonal host population infected significantly more of these hosts than did their ancestors. We thus provide experimental evidence for the widely held view that host genetic diversity restricts evolution of parasites and moderates their harmful effects. Genetically uniform host populations are not only at increased risk from fungal epidemics because they all share the same susceptibility, but also because new parasite strains are able to adapt quickly to new host environments and to improve their fitness.     

Climate forcing of diatom productivity in a lowland,eutrophic lake: White Lough revisited

1. In cultural landscapes, lake response to climate can be masked by land‐use change and nutrient loss from their catchments. Palaeolimnological methods were used to reconstruct the ecological response of diatoms in a eutrophic lowland lake (White Lough, Co. Tyrone, Northern Ireland) to altered nutrient P loading and precipitation variability over c. 100 years. 2. ²¹⁰Pb‐dated sediment cores were analysed to determine diatom assemblage variability, biogenic silica concentration, geochemical phosphorus concentration and accumulation rate. Manure P and agricultural N surplus data were collated from documentary sources. Long‐term trends in annual temperature and precipitation were derived from the Armagh Observatory. 3. Diatom community turnover from 1890 until c. 1960 was limited, and assemblages were dominated by Aulacoseira subarctica; after this date, changes primarily reflected a eutrophication sequence owing to increased diffuse nutrient inputs associated with intensification of land use (external P loading increased by a factor of three). 4. Diatom and biogenic Si profiles were compared with North Atlantic Oscillation (NAO) records, an index of regional weather patterns. Biogenic Si exhibited a c. 7‐year cycle, which tracked a cycle of similar timescale in the Armagh climate record for dry summers. In turn, this cycle was related to the variation in the NAO. 5. Monitoring data from 1971 to 2007 of nitrate exports from the Blackwater River showed that these too followed a roughly 7‐year cycle at least up to 2000, in which dry summers were followed by sharp increases in nitrate export. It is argued that diatom production in White Lough reflects the cyclic behaviour in nitrate loading and the constraints that nitrogen availability places on the spring diatom bloom in a lake that is dominated by cyanobacteria.     

The budget model of ecosystem of a shallow highly eutrophic lake

A model of energy budget of Lake Bolshoi Okunenok ecosystem was based on the data received during field studies from May through November 1986. The model takes into account 36 components including dissolved organic matter, bacteria, phytoplankton, zooplankton, meiobenthos, macrobenthos, fish, suspended and sediment detritus. The growing season has been divided into 16 intervals according to the number of observations. The balance equation for each live component describes the change in its biomass for a time interval between two successive sampling dates. The change is considered as a balance of energy input with assimilation or feeding, and energy loss due to respiration, excretion, predation, natural mortality, fishery catchment or and emergence of imago insects. For non-live components we estimate an increase and a decrease in their mass due to the activity of living organisms, as well as organic matter exchange between water and sediments. Seasonal value of balance elements for each component are equal to sums of appropriate interval value. Comparison of energy flows through different links of a trophic web has shown that the role of a bacterial-detrial link was extremely important in Lake Bolshoi Okunenok for the growth season of 1986. Detritus constituted 58% of seasonal diet of non-predatory zooplankton, 39% of diet of predatory zooplankton, 50% of diet of planktivorous fish (fry of whitefish) and 92% of diet of benthivorous fish (fry of carp). The contribution of bacteria to the total seasonal decomposition amounted to 46%. Approximately 57% of the forage phytoplankton production, 86% of non-predatory benthos production, and 23-38% of the other trophic groups production were consumed by all grazers. "Coefficient of energy transformation" is proposed. It is calculated as: CET(s, k) = Ps(k)/Pk, where Ps(k) is production of consumers "s", built due to consumption of source "k"; Pk is production of source "k" itself. In Lake Bolshoi Okunenok only 14% of energy built by phytoplankton were accumulated in organic matter of zooplankton due to direct consumption.     

Evidence of interference of Asterionella formosa with the feeding of Bosmina longirostris: a field study in a south Andes lake

Intensive sampling of a small temperate lake of the Araucanian region (Argentina), revealed changes in the demography of Bosmina longirostris when the diatom Asterionella formosa density increased (to 54 000 cells ml^–1). Bosmina population behaviour in the field was consistent with the idea that large numbers of Asterionella caused its starvation. Individuals were actually colourless and fecundity dropped to zero, as was evident when Asterionella reached 99.4% of total phytoplankton biovolume. Since cyanophytes, as a whole, are scarce in these lakes, we propose that the flagellate/diatom ratio has a direct effect on cladoceran demography.     

Vertical distribution of major sulfate-reducing bacteria in a shallow eutrophic meromictic lake

The vertical distribution of sulfate-reducing bacteria was investigated in a shallow, eutrophic, meromictic lake, Lake Harutori, located in a residential area of Kushiro, Japan. A steep chemocline, characterized by gradients of oxygen, sulfide and salinity, was found at a depth of 3.5–4.0 m. The sulfide concentration at the bottom of the lake was high (up to a concentration of 10.7 mM). Clone libraries were constructed using the aprA gene, which encodes adenosine-5′-phosphosulfate reductase subunit A, in order to monitor sulfate-reducing bacteria. In the aprA clone libraries, the most abundant sequences were those from the Desulfosarcina–Desulfococcus (DSS) group. A primer set for a DSS group-specific 16S rRNA gene was used to construct another clone library, analysis of which revealed that the uncultured group of sulfate-reducing bacteria, SEEP SRB-1, accounted for nearly half of the obtained sequences. Quantification of the major bacterial groups by catalyzed reporter deposition-fluorescence in situ hybridization demonstrated that the DSS group accounted for 3.2–4.8% of the total bacterial community below the chemocline. The results suggested that the DSS group was one of the major groups of sulfate-reducing bacteria and that these presumably metabolically versatile bacteria might play an important role in sulfur cycling in Lake Harutori.     

Seasonal modulation of bacterioplankton community at a temperate eutrophic shallow lake

At the present time, there is still a lack of information about environmental parameters modulating variations on bacterial diversity in temperate lakes, particularly from Portugal. Fermentelos Lake (Central Portugal) is a shallow water body that sustains an important wetland area. The strong nutrient inputs from agriculture and industrial runoffs have led to its current eutrophic status. The present work aimed to understand which factors modulate the seasonal bacterioplankton diversity at this lake using 16S rRNA PCR-denaturing gradient gel electrophoresis (DGGE) and multivariate analysis. Environmental data demonstrated eutrophic features throughout all samples with nitrate concentrations reaching 12.0 mg N (NO₃ ⁻) l⁻¹ in March 2006, while the highest conductivity (609 μS cm⁻¹), soluble reactive phosphorus (0.37 mg l⁻¹), total suspended solids (87.2 mg l⁻¹) and chlorophyll a (286.6 μg l⁻¹) levels were recorded in August 2007. Over the past two decades there was a general increase in nitrate, pH and conductivity levels at this lake, suggesting the eutrophication process is still in progress. Multivariate analysis showed that summer versus winter DGGE patterns could be established for bacterial assemblages and were mainly defined by water temperature and chlorophyll a. Actinobacteria were dominant throughout the study period although a general preference for higher temperature, pH, total suspended solids, conductivity, soluble reactive phosphorus (SRP) and chlorophyll a levels was observed. The highest concentrations of nitrogen sources were related to Bacteroidetes and phototrophic eukaryote (cryptophycean) dominance. The expansion of Betaproteobacteria, Alphaproteobacteria and Cyanobacteria phylotypes was generally associated to high temperature, pH, conductivity and SRP values.     

Chemical consequences of oxygenation in a shallow eutrophic lake studied with mesocosms

To assess the effects of oxygenation in a shallow, highly eutrophic lake, an experiment with large enclosures was carried out. From the results two main effects can be expected from the oxygenation of the whole lake: (a) fish mortality and the resulting putrefactive processes would be avoided; (b) an increase in nitrogen and phosphorus concentration and, consequently, in the trophic level of the lake would probably occur.     

Abrupt shift from clear to turbid state in a shallow eutrophic, biomanipulated lake

Monitoring data were used to assess causes behind a recent shift from a clear-water to a turbid-water state in Lake Major, a 10 ha shallow lake in Hungary. In 1999–2000, fish manipulation was conducted in this hypertrophic lake. Reduced fish stock resulted in clearing water and the development of a dense (>80% coverage) submerged vegetation in 2005. During the recent abrupt shift, which occurred in 2007, submerged vegetation subsequently declined after a two-year period of clear water and abundant vegetation. An intense decay of macrophytes within the lake produced a rapid transition between the clear- and turbid-water states. During the clear-water state in 2005–2006, the most important variables predominantly correlating with macrophyte cover were Secchi transparency, temperature and TN, while TN, temperature, Secchi depth and chlorophyll-a were the most significant variables during the turbid-water state in 2007. Nitrogen may play a significant role in the cover of submerged macrophytes when TP is moderate. We argue that several factors in concert are necessary to initiate a shift. Water temperature likely has contributed to triggering shift through inter-year-dependent changes in cover of macrophytes, with fish recruitment having key roles in the dynamics of shallow lakes. Handling editor: Luigi Naselli-Flores     

Estimation of reference conditions for phytoplankton in a naturally eutrophic shallow lake

Classification of waters using biological quality elements and determination of the degree of deviation from reference levels is a key issue in the Water Framework Directive of EU. Lakes in reference conditions with sufficient biological data are available for several boreal lake types with the exception of naturally eutrophic lakes. An empirical approach is one alternative for estimating the reference conditions of such lakes. We used the water transparency of the naturally eutrophic Lake Tuusulanjärvi recorded in August in the early 1910s to estimate reference values for phytoplankton biomass and chlorophyll a concentrations. Three phytoplankton samples during August 2000–2001 corresponded to the estimated reference values for total biomass (<5.6 mg l^?1) and chlorophyll a (<28 μg l^?1), as did the simultaneous Secchi depths. The phytoplankton assemblage in these samples with 24 eutrophy indicators (17% of the total taxa number) corresponded in general the species list from the early 1900s, which as such could be regarded as reference assemblage. Furthermore, in August 2000, 3 years after intensive fish removal a prominent decrease in cyanobacterial biomass and toxin concentration was observed. The costs of the measures and studies in Lake Tuusulanjärvi during 1989–2003 have been approximately 2.5 million euros.     

Bloom formation of Gloeotrichia echinulata and Aphanizomenon flos-aquae in a shallow,eutrophic, Danish lake

Over a period of four years, the seasonal periodicity of dominant phytoplankton species in a shallow, eutrophic Danish lake changed markedly. Cyanophytes prevailed during the summer period of all four years. In the first three years, species of Microcystis, Anabaena and Aphanothece dominated, whereas in the fourth year of investigation, these algae were replaced by Gloeotrichia echinulata (J. E. Smith) Richter and Aphanizomenon flos-aquae (L.) Ralfs. The most striking environmental differences in the fourth year as compared with the previous three years, were an increase in tranparency, from about 0.5 meter in 1989–1991 to more than 2 metres preceding the summer maximum in 1992, and a simultaneous occurrence of low oxygen concentrations. A collapse of the fish population was followed by an increased proportion of large Cladocerans in the zooplankton. Improved light conditions at the bottom and grazing pressure from large Cladocerans favoured growth of the large colony forming blue-green algae, Gloeotrichia echinulata and Aphanizomenon flos-aquae. These species germinate from resting spores in the sediment and are able to sustain some growth there before migration to the lake water. The transfer of algal biomass from the bottom sediment to the water phase was accompanied by a marked increase in concentrations of particulate phosphorus and nitrogen in the entire lake.