MEASUREMENT OF IN SITU SPECIFIC GROWTH RATES OF MICROCYSTIS (CYANOBACTERIA) FROM THE FREQUENCY OF DIVIDING CELLS1

Diel changes in the frequency of dividing cells (FDC) of three Microcystis species were investigated in a small eutrophic pond from July to October 2005. The representative species was M. aeruginosa (Kütz.) Kütz., constituting 57%–86% of the Microcystis population throughout the study period, and the remainder were M. viridis (A. Braun) Lemmerm. and M. wesenbergii (Komárek) Komárek. The FDC of M. aeruginosa and M. wesenbergii increased in the daytime and fell in the nighttime in July and August, but this regular variation was not observed in September or October. The in situ specific growth rates of Microcystis species were estimated based on the assumption that the specific growth rate can be given as an absolute value of the derivative of FDC with respect to time. The calculated values were similar among species—0.15–0.38 · d^?1 for M. aeruginosa, 0.14–0.63 · d^?1 for M. viridis, and 0.18–0.61 · d^?1 for M. wesenbergii. The specific growth rates in July and August slightly exceeded those in September and October. The analysis of the in situ specific growth rate of Microcystis indicated that recruitment of the benthic population or morphological change, rather than massive growth, was at least partly responsible for the dominance of M. aeruginosa in the study pond.     

INVOLVEMENT OF MICROCYSTINS AND COLONY SIZE IN THE BENTHIC RECRUITMENT OF THE CYANOBACTERIUM MICROCYSTIS (CYANOPHYCEAE)1

The benthic recruitment of Microcystis was simulated in vitro in order to characterize the colonies of Microcystis recruited and to study the impact of intracellular and extracellular microcystins (MCs), and the influence of colony size on the recruitment process. We observed recruitment dynamics consisting of a lag phase followed by a peak and then a return to low recruitment rates, mainly controlled by passive resuspension throughout the experiment, and by physiological processes during the recruitment peak. Ninety‐seven percent of the Microcystis colonies recruited were <160 μm in maximum length, and their cells contained much greater amounts of MCs (0.26 ± 0.14 pg eq microcystin leucine‐arginine variant [MC‐LR] · cell^?1) than those in benthic colonies (0.021 ± 0.004 pg eq MC‐LR · cell^?1). The MC content of recruited Microcystis varied significantly over time and was not related to changes in the proportion of potentially toxic genotypes, determined using real‐time PCR. On the other hand, the changes in MC content in the potentially toxic Microcystis recruited were closely and negatively correlated with recruitment dynamics; the lowest MC contents corresponded to high recruitment rates, and the highest MC contents corresponded to low recruitment rates. Thus, depending on temperature and light conditions, these variations are thought to result from the selection of various subpopulations from among the smallest and the most toxic of the initial benthic population. Adding purified MC‐LR to experimental treatments led to a decreased recruitment of Microcystis and more specifically of mcyB genotypes.     

CHANGES IN THE MORPHOLOGY AND POLYSACCHARIDE CONTENT OF MICROCYSTIS AERUGINOSA (CYANOBACTERIA) DURING FLAGELLATE GRAZING1

To investigate the changes in the morphology and polysaccharide content of Microcystis aeruginosa (Kütz.) Kütz. during flagellate grazing, cultures of M. aeruginosa were exposed to grazing Ochromonas sp. for a period of 9 d under controlled laboratory conditions. M. aeruginosa responded actively to flagellate grazing and formed colonies, most of which were made up of several or dozens of cells, suggesting that flagellate grazing may be one of the biotic factors responsible for colony formation in M. aeruginosa. When colonies were formed, the cell surface ultrastructure changed, and the polysaccharide layer on the surface of the cell wall became thicker. This change indicated that synthesis and secretion of extracellular polysaccharide (EPS) of M. aeruginosa cells increased under flagellate grazing pressure. The contents of soluble extracellular polysaccharide (sEPS), bound extracellular polysaccharide (bEPS), and total polysaccharide (TPS) in colonial cells of M. aeruginosa increased significantly compared with those in single cells. This finding suggested that the increased amount of EPS on the cell surface may play a role in keeping M. aeruginosa cells together to form colonies.     

AN “ENVIRO‐INFORMATIC” ASSESSMENT OF SAGINAW BAY (LAKE HURON,USA) PHYTOPLANKTON: DATA‐DRIVEN CHARACTERIZATION AND MODELING OF MICROCYSTIS (CYANOPHYTA)1

Phytoplankton and Microcystis aeruginosa (Kütz.) Kütz. biovolumes were characterized and modeled, respectively, with regard to hydrological and meteorological variables during zebra mussel invasion in Saginaw Bay (1990–1996). Total phytoplankton and Microcystis biomass within the inner bay were one and one‐half and six times greater, respectively, than those of the outer bay. Following mussel invasion, mean total biomass in the inner bay decreased 84% but then returned to its approximate initial value. Microcystis was not present in the bay during 1990 and 1991 and thereafter occurred at/in 52% of sample sites/dates with the greatest biomass occurring in 1994–1996 and within months having water temperatures >19°C. With an overall relative biomass of 0.03 ± 0.01 (mean + SE), Microcystis had, at best, a marginal impact upon holistic compositional dynamics. Dynamics of the centric diatom Cyclotella ocellata Pant. and large pennate diatoms dominated compositional dissimilarities both inter‐ and intra‐annually. The environmental variables that corresponded with phytoplankton distributions were similar for the inner and outer bays, and together identified physical forcing and biotic utilization of nutrients as determinants of system‐level biomass patterns. Nonparametric models explained 70%–85% of the variability in Microcystis biovolumes and identified maximal biomass to occur at total phosphorus (TP) concentrations ranging from 40 to 45 μg · L^?1. From isometric projections depicting modeled Microcystis/environmental interactions, a TP concentration of <30 μg · L^?1 was identified as a desirable contemporary “target” for management efforts to ameliorate bloom potentials throughout mussel‐impacted bay waters.     

THE ACCLIMATIVE CHANGES IN PHOTOCHEMISTRY AFTER COLONY FORMATION OF THE CYANOBACTERIA MICROCYSTIS AERUGINOSA1

Microcystis aeruginosa (Kütz.) Kütz. commonly occurs as single cells at early recruitment but forms large colonies in summer. Colony formation will induce many acclimative changes. In this study, we demonstrated the photochemical changes before and after colony formation. In the laboratory, light curves showed that colonies were more responsive to high light than single cells. The values of the maximal slope of electron transport rate (ETR)—light curve (α), relative maximal electron transport rate (rETR_max), and onset of light saturation (I_k) of colonies were significantly higher than those of single cells (P < 0.05), indicating that colonies have higher photosynthetic capability than single cells, especially in high light, where values of rETR_max and I_k of colonies were 2.32 and 2.41 times those of single cells. Moreover, the dark‐light experiments showed that colonial cells can more effectively resist darkness damage. In addition, pigments of colonial cells were higher than those of single cells (P < 0.05). The higher pigment contents probably contribute to higher photosynthetic capability. In the field, the inhibition rate of F_v/F_m in single cells increased significantly faster than that of colonies as light increased (P < 0.05), but nonphotochemical quenching (NPQ) value of colonies was higher (32.4%) than that of single cells at noon, which indicated colonial cells can more effectively resist high‐light inhibition than single cells (P < 0.05). Polysaccharides of colonies were significantly higher compared to those in unicellular cells (P < 0.05) based on their contents and ultrastructural characteristics. This finding implies that colonies could not effectively decrease photoinhibition by negative buoyancy regulation. In fact, NPQ may be an important mechanism for avoiding photodamage. All of these phenomena can help explain the ecological success of colonial M. aeruginosa in eutrophic water.     

Seasonal variations of Microcystis populations in sediments of Lake Biwa, Japan

Seasonal variations of colony numbers of Microcystis aeruginosa(Kütz.) Kütz. and M. wesenbergii(Komárek) Komárek in N. V. Kondrat. in sediments of Lake Biwa were investigated over a period of 1 year. At two stations located in the shallow South Basin of Lake Biwa (ca. 4 m water depth), the colony number of Microcystisfluctuated seasonally. The number had a tendency to gradually decrease from winter to early summer, while it increased through mid-summer and autumn. Since the Microcystispopulation in sediment was rather small, intensive growth and accumulation in the water column should be important for the formation of Microcystisblooms in Lake Biwa. Microcystiscolonies in the sediment samples after June were observed to be floating in a counting chamber under a microscope. The observation suggests that the recruitment of Microcystis colonies into the water column mostly occurs in early summer. The number of Microcystiscolonies in the deep North Basin of Lake Biwa (70 – 90 m water depth) was larger than in the South Basin. Because the seasonal variation of colony numbers was not observed in the North Basin, and Microcystiscells do not have gas vesicles, these colonies will not return into the water column. The colonies isolated from the sediment of the North Basin were able to grow in cultured conditions, in the same way as those from the sediment of the South Basin. Therefore, Microcystiscolonies may survive for a long time under stable conditions of low temperature (ca. 8 °C) and darkness, in the sediment of the deep North Basin, accumulating gradually each year.     

SCANNING ELECTRON MICROSCOPY OBSERVATIONS OF DEFORMITIES IN SMALL PENNATE DIATOMS EXPOSED TO HIGH CADMIUM CONCENTRATIONS1

Different types of malformations are likely to affect the morphology of diatoms when exposed to particularly unstable environmental conditions, the most easily identifiable being distortion of the whole frustule. In the present study, we investigated, by means of SEM, valve abnormalities induced by high cadmium contamination (100 μg · L^?1) in small pennate diatoms. Changes in the shape of Amphora pediculus (Kütz.) Grunow and anomalous sculpturing of the cell wall of many species, such as Encyonema minutum (Hilse) D. G. Mann, Mayamaea agrestris (Hust.) Lange‐Bert., Gomphonema parvulum (Kütz.) Kütz., or Eolimna minima (Grunow) Lange‐Bert., were observed, which were not, or almost not, noticeable in the LM. With consideration to current knowledge of diatom morphogenesis, metal uptake by the cell would induce, directly or indirectly, damage to many cytoplasmic components (e.g., microtubules, cytoskeleton, Golgi‐derived vesicles) involved in the precisely organized silica deposition. This study confirms that many species, whatever their size, are likely to exhibit morphological abnormalities under cadmium stress, and that this indicator may be valuable for the biomonitoring of metal contamination, even if SEM observations are not necessary for routine studies.     

SCHIZOMERIS—A GROWTH FORM OF STIGEOCLONIUM TENUE (CHLOROPHYTA:CHAETOPHORACEAE)1

Schizomeris leibleinii Kütz. was found as an ecophene of Stigeoclonium tenue (Ag.) Kütz. in Centennial Lake, Palmerston North, New Zealand. The Schizomeris form occurred in autumn when the water was polluted by a large number of ducks. Sampling of the plant population of the lake and culture work showed that S. tenue has a high degree of polymorphism. Other forms of it which developed both in the lake and in culture were indistinguishable from Protoderma Kütz. emend. Borzi, from Uronema Lagerheim, or from Pearsoniella Fritsch and Rich, respectively.     

Genetic Diversity： Geographical Distribution and Toxin Profiles of Microcystis Strains （Cyanobacteria） in China

Twenty strains of Microcystis Kütz were isolated from different freshwater bodies in China to analyze the diversity, geographical distribution and toxin profiles. Based on whole-cell polymerase chain reaction of cpcBA-IGS nucleotlde sequence, the derived neighbor-joining （NJ） and maximum parsimony （MP） trees Indicate that these strains of Microcystis can be divided into four clusters. The strains from south, middle and north region of China formed distinct lineages, suggesting high diversity and a geographical distribution from south to north locations. Moreover, the results being indicating high variable genotypes of the strains of the Microcystis strains from the same lake show that there Is high diversity of Microcystis within a water bloom population. Comparing the results of the present study with those reported for compared with 43 strains of Microcystis from other locations, also reveals Chinese strains have high similarity with those from regions in the North Hemispherical. This suggests that the Microcystis strains In the world might have a geographical distribution. Analysis of 30 strains using the primers MCF/TER and TOX2P/TOX2M showed that there was no correlation between the gene of cpcBA-IGS and the presence of racy. Toxic strains were founded to be predominant in different water bodies throughout China.     

COMMUNITY STRUCTURE DYNAMICS OF EPILITHIC AND EPIPHYTIC DIATOMS IN A MISSISSIPPI STREAM1

Epilithic and epiphytic diatoms were collected monthly from 18 June 1979 through 19 May 1980 from both shaded and unshaded sites of Sessums Creek, a shallow, oligotrophic and strongly alkaline stream in northeastern Mississippi. The dominant macroalgae were filamentous green algae belonging to the non-mucilage producing genera Cladophora and Rhizoclonium. Ninety taxa were identified with Navicula minima Grun. and Nitzschia dissipata (Kütz.) Grun. accounting for 50% of all valves counted. Other important taxa were Navicula menisculus var. upsaliensis (Grun.) Grun., N. cryptocephala var. veneta (Kütz.) Rabh., Achnanthes minutissima Kütz. and Cymbella turgida (Greg.) Cl. With few exceptions, the more dominant taxa were equally abundant in the shaded and unshaded sites and also in the epilithon and epiphyton. Species diversity (H') and the number of taxa in a sample in all four habitats showed an identical pattern, being lowest in winter and highest in fall. No one pair of habitats was more structurally similar than any-other pair when compared by a similarity index (SIMI). Apparently, differences in light intensity and substrate type had little effect on diatom distribution in Sessums Creek and it was concluded that the locality sampled supported a single, nearly homogeneous diatom community.     

STRUCTURE,LIFE HISTORY AND SYSTEMATICS OF RHOICOSPHENIA (BACILLARIOPHYTA). IV. CORRELATION OF SIZE REDUCTION WITH CHANGES IN VALVE MORPHOLOGY OF RH. GENUFLEXA1

Rhoicosphenia Grun. is a relatively isolated genus among the biraphid diatoms. Morphological changes in an isopolar member of the genus, Rh. genuflexa (Kütz.) Medlin, were investigated using light and scanning electron microscopy. The fully raphid valve showed changes in its flexure that could be correlated with size reduction during its life history from the initial cells to the smallest cells found in the population. Bands showed changes in number (from three to one) related to size reduction. Rh. genuflexa is morphologically similar to Rh. abbreviata (C. Ag.) Lange-Bert. (=Rh. curvata (Kütz.) Grun.), although the two are distinct taxa. These observations support previous contentions that Rhoicosphenia is a natural taxonomic grouping.     

POLYMORPHISM IN THE DESMID MICRASTERIAS PINNATIFIDA AND ITS TAXONOMICAL IMPLICATIONS1

A case of polymorphism in the desmid Micrasterias pinnatifida (Kütz.) Ralfs is described. It is based on material collected from the overflow channel of a dam in Caeté, Minas Gerais State. After careful examination of nearly 500 specimens, 26 different morphological expressions of the alga were found which led the authors to make the following deductions: (1) for delimitation of species and infraspecific taxa in desmids it is absolutely necessary to analyze sample populations; (2) the morphological characteristics presently used, for delimination of some varieties of M. Pinnatifida (Kütz.) Ralfs seem to have no taxonomical value if only isolated individuals are examined; (3) the varieties granulata, inflata, and tridentata must be considered with some caution, until further studies are carried out, because they can be merely distinct morphological expressions of M. pinnatifida (Kütz.) Ralfs var. pinnatifida.     

PERIPHYTON COLONIZATION OF ROCK SURFACES IN A BOREAL FOREST STREAM STUDIED BY SCANNING ELECTRON MICROSCOPY AND TRACK AUTORADIOGRAPHY1

Periphyton colonization of natural rock surfaces and granite tiles was followed experimentally in the Matamek River, an acidic (pH 5.5) sixth order boreal stream in northeast Quebec, Canada. Accumulation of chlorophyll a and freshweight algal biomass was logarithmic over a 25 day colonization period. The major colonizers were Tabellaria flocculosa (Roth) Kütz., T. fenestrata (Lyngb.) Kütz., Mougeotia sp., and Eunotia pectinalis (Kütz) Rabh., and its varieties. The microcolonization sequence on granite tiles, followed over 27 days with scanning electron microscopy, showed an initial accumulation of algal cells on the upstream and downstream margins and in depressions, followed by a gradual filling-in of the flat surfaces. It is hypothesized that the observed slow rate of colonization was due to the high surface tension of the granite substratum and the absence of preconditioning by an organic film. It is further hypothesized that the increase in cellular carbon fixation rates of T. flocculosa measured over a 23 day period using nuclear track autoradiography parallels the development of an algal-detrital microcosm on the granite tile, and is evidence for the establishment of localized tight nutrient spiralling. It is suggested that the periphyton community may be regarded as a nutrient recycling system at a microenvironmental level which may be especially important in oligotrophic systems.     

ABSOLUTE CONFIGURATION ANALYSIS OF THE FLAGELLAR APPARATUS IN CLADOPHORA AND CHAETOMORPHA MOTILE CELLS,WITH AN ASSESSMENT OF THE PHYLOGENETIC POSITION OF THE CLADOPHORACEAE (ULVOPHYCEAE,CHLOROPHYTA)

Absolute configurational analyses of flagellar apparatus components were performed on the motile cells produced by three species of Cladophora, Cl. dalmatica Kütz., Cl. flexuosa (Dillw.) Harv., and Cl. glomerata (L.) Kütz., and by Chaetomorpha aerea (Dillw.) Kütz. There was little variation among the species. All of the flagellar apparatuses demonstrated the ulvophyceous features of 180° rotational symmetry, counterclockwise absolute orientation, and basal body overlap, as well as the alignment of the basal bodies perpendicular to the long axis of the cell. Diagnostic features included the nearly complete absence of C tubules from the basal bodies and the presence of a coarsely striated component dorsal to the two-membered rootlets in all cells, as well as, in quadriflagellate cells, a tetralobate distal fiber, the coaxial arrangement of the lowermost pair of basal bodies, and the presence of a characteristic array of basal-body-associated striated bands. The distal fiber architecture, the presence of a “wing” in the X-membered rootlets, and the “flattening” of the flagellar apparatus components suggests a close relationship of the Cladophoraceae to the Dasycladales, and indicates that these two groups may have shared a common ancestor, possibly ancient in terms of the geological time scale but relatively recent in the context of ulvophyceous evolution. A sizeable phylogenetic gap exists between the Cladophoraceae and uninucleate-celled, presumably primitive members of the Ulvophyceae.     

Morphological,biochemical and phylogenetic assessments of water‐bloom‐forming tropical morphospecies of Microcystis (Chroococcales,Cyanobacteria)

Previous polyphasic analyses of five morphospecies of the water‐bloom‐forming cyanobacterial genus Microcystis, Microcystis aeruginosa (Kützing) Lemmermann (=Microcystis aeruginosa (Kützing) Kützing), Microcystis ichthyoblabe Kützing, Microcystis novacekii (Komárek) Compère, Microcystis viridis (A. Braun) Lemmermann, and Microcystis wesenbergii (Komárek) Komárek in Kondratieva, have shown them to be conspecific and they have been proposed to be included under the binomial Microcystis aeruginosa (Kützing) Lemmermann. However, several morphospecies from tropical regions, such as Microcystis bengalensis Banerji, Microcystis panniformis Komárek, Komárková‐Legnerová, Sant'anna, Azevedo & Senna, Microcystis protocystis Crow, Microcystis pseudofilamentosa Crow, Microcystis ramosa Bharadwaya, and Microcystis robusta (Clark) Nygaard, have never been analyzed biochemically or phylogenetically; consequently, their taxonomic status is uncertain. To resolve this issue, we collected 57 strains of Microcystis from Vietnam for taxonomic analysis using a polyphasic approach. Strains were assigned to the six tropical morphospecies listed above or to four morphospecies with cosmopolitan distributions (M. aeruginosa, M. ichthyoblabe, M. novacekii, and M. wesenbergii). Several strains produced colony variants in different culture media; some of these variants had forms that overlapped with those of other morphospecies. Cell diameters varied widely between strains (2.6–9.3 µm) and were unrelated to morphospecies discrimination criteria. Strains of the 10 morphospecies examined had similar fatty acid compositions and closely similar 16S rRNA gene sequences (>99.2% similar). Phylogenetic analyses using 16S rRNA gene and 16S–23S internal transcribed spacer sequences did not identify any clear separations corresponding to morphospecies concepts or microcystin‐producing abilities. Thus, the six tropical morphospecies (M. bengalensis, M. panniformis, M. protocystis, M. pseudofilamentosa, M. ramosa, and M. robusta) are not natural taxonomic units within the genus Microcystis and should be included under M. aeruginosa.     

PRODUCTION OF NATURAL BUTYLATED HYDROXYTOLUENE AS AN ANTIOXIDANT BY FRESHWATER PHYTOPLANKTON1

Butylated hydroxytoluene (BHT) is one of the synthetic antioxidant agents commonly used for food additives. In the present study, we determined that four freshwater phytoplankton, including a green alga (Botryococcus braunii Kütz.) and three cyanobacteria [Cylindrospermopsis raciborskii (Wol?osz.) Seenaya et Sabba Raju, Microcystis aeruginosa (Kütz.) and Oscillatoria sp.] were capable of producing this compound. Hexane extracts from all the studied species exhibited various degrees of antioxidative properties when they were tested with the β‐carotene‐linoleate (β‐CL) assay and the 2,2‐diphenyl‐1‐picrylhydrazyl (DPPH) free‐radical‐scavenging assay. The highest antioxidant activity was observed in the crude extracts of M. aeruginosa and B. braunii, which displayed a similar activity to synthetic BHT. Gas chromatography/mass spectroscopy (GC‐MS) analysis of the purified fractions revealed that the active compound was identical to synthetic BHT. Culturing under various irradiances gave rise to different magnitudes of BHT production in cyanobacterial cells, showing that more BHT was produced in the cells irradiated with a higher light intensity, and its production was irradiance dependent. Moreover, the quantity of cellular BHT displayed a positive correlation with the antioxidative activity of the tested species. The present study confirms the production of BHT in all four of the studied freshwater phytoplankton and suggests that these species constitute a potential source for producing natural BHT.     

ALLELOPATHIC GROWTH INHIBITION OF SELECTED PHYTOPLANKTON SPECIES BY SUBMERGED MACROPHYTES1

Allelopathic effects of submerged macrophytes on the growth and photosynthesis of different unialgal cultures of planktonic cyanobacteria, a diatom, and a green alga were tested in coexistence experiments using dialysis cultures. The method applied allowed measurements under conditions similar to that in lakes but without nutrient and light limitation. Growth and photosynthesis were measured with a pulse amplitude modulated fluorometer as an increase of chl a fluorescence and activity of PSII, respectively. Eurasian water milfoil Myriophyllum spicatum L. and rigid hornwort Ceratophyllum demersum L. proved to inhibit the PSII activity and then growth of the investigated phytoplankton species, whereas sago pondweed Potamogeton pectinatus L. showed no effect. Growth inhibition was dependent on biomass of M. spicatum. Considerable differences between phytoplankton groups and among species of cyanobacteria were found regarding their response to M. spicatum. Members of the Oscillatoriales and Microcystis aeruginosa Kütz. emend. Elenkin were more sensitive than the cyanobacterium Aphanizomenon flos‐aquae Ralfs ex Born. et Flah., the diatom Stephanodiscus minutulus (Kütz) Cleve et Möller, and the green alga Scenedesmus armatus Chodat. A possible contribution of this result to changes in the phytoplankton succession of lakes after loss of macrophytes is discussed.     

AN ECOLOGICAL REVIEW OF CLADOPHORA GLOMERATA (CHLOROPHYTA) IN THE LAURENTIAN GREAT LAKES1

Cladophora glomerata (L.) Kütz. is, potentially, the most widely distributed macroalga throughout the world’s freshwater ecosystems. C. glomerata has been described throughout North America, Europe, the Atlantic Islands, the Caribbean Islands, Asia, Africa, Australia and New Zealand, and the Pacific Islands. Cladophora blooms were a common feature of the lower North American Great Lakes (Erie, Michigan, Ontario) from the 1950s through the early 1980s and were largely eradicated through the implementation of a multibillion‐dollar phosphorus (P) abatement program. The return of widespread blooms in these lakes since the mid‐1990s, however, was not associated with increases in P loading. Instead, current evidence indicates that the resurgence in blooms was directly related to ecosystem level changes in substratum availability, water clarity, and P recycling associated with the establishment of dense colonies of invasive dreissenid mussels. These results support the hypothesis that dreissenid mussel invasions may induce dramatic shifts in energy and nutrient flow from pelagic zones to the benthic zone.     

SEDIMENTARY IMPRINT OF MICROCYSTIS AERUGINOSA (CYANOBACTERIA) BLOOMS IN GRANGENT RESERVOIR (LOIRE,FRANCE)1

Analysis of a sediment core taken from the Grangent reservoir in 2004 showed the presence of high concentrations of Microcystis aeruginosa Kütz. colonies at the sediment surface (250 colonies · mL sediment^?1) and also at depths of 25–35 cm (2300 colonies·mL sediment^?1) and 70 cm (600 colonies · mL sediment^?1). Measurements of radioactive isotopes (⁷Be, ¹³⁷Cs, and ²⁴¹Am) along with photographic analysis of the core were used to date the deep layers: the layer located at ?30 cm dates from summer 2003, and that located at ?70 cm from 1990 to 1991. The physiological and morphological conditions of those benthic colonies were compared with those of planktonic colonies using several techniques (environmental scanning electron microscopy [ESEM], TEM, DNA markers, cellular esterases, and toxins). The ESEM observations showed that, as these colonies age, peripheral cells disappear, with no cells remaining in the mucilage of the deepest colonies (70 cm), an indication of the survival thresholds of these organisms. In the benthic phase, the physiological conditions (enzyme activity, cell division, and intracellular toxins) and ultrastructure (particularly the gas vesicles) of the cells surviving in the heart of the colony are comparable to those of the planktonic form, with all the potential needed for growth. Maintaining cellular integrity requires a process that can provide sufficient energy and is expressed in the reduced, but still existing, enzymatic activity that we measured, which is equivalent to a quiescent state.     

GENETIC ADAPTATION AND ACCLIMATION OF PHYTOPLANKTON ALONG A STRESS GRADIENT IN THE EXTREME WATERS OF THE AGRIO RIVER–CAVIAHUE LAKE (ARGENTINA)1

We tested if different adaptation strategies were linked to a stress gradient in phytoplankton cells. For this purpose, we studied the adaptation and acclimation of Dictyosphaerium chlorelloides (Naumann) Komárek et Perman (Chlorophyta) and Microcystis aeruginosa (Kütz.) Kütz. (Cyanobacteria) to different water samples (from extremely acid, metal‐rich water to moderate stressful conditions) of the Agrio River–Caviahue Lake system (Neuquén, Argentina). Both experimental strains were isolated from pristine, slightly alkaline waters. To distinguish between physiological acclimation and genetic adaptation (an adaptive evolution event), a modified Luria‐Delbrück fluctuation analysis was carried out with both species by using as selective agent sample waters from different points along the stress gradient. M. aeruginosa did not acclimate to any of the waters tested from different points along the stress gradient nor did D. chlorelloides to the two most acidic and metal‐rich waters. However, D. chlorelloides proliferated by rapid genetic adaptation, as the consequence of a single mutation (5.4 × 10^?7 resistant mutants per cell per division) at one locus, in less extreme water and also by acclimation in the least extreme water. It is hypothesized that the stress gradient resulted in different strategies of adaptation in phytoplankton cells from nonextreme waters. Thus, very extreme conditions were lethal for both organisms, but as stressful conditions decreased, adaptation of D. chlorelloides cells was possible by the selection of resistant mutants, and in less extreme conditions, by acclimation.