Interspecific variation in extracellular polysaccharide content and colony formation of <Emphasis Type="Italic">Microcystis</Emphasis> spp. cultured under different light intensities and temperatures

Single cells of five different Microcystis species (M. ichthyoblabe, M. viridis, M. flos-aquae, M. wesenbergii, and M. aeruginosa) were batch-cultured at different temperatures and light intensities: (a) 25 °C and 50 μmol photons m^?2 s^?1 (control culture); (b) 25 °C and 10 μmol photons m^?2 s^?1; and (c) 15 °C and 50 μmol photons m^?2 s^?1. The extracellular polysaccharide content was significantly higher in treatments b and c than in the control treatment. All Microcystis species existed as single cells under the control treatment but formed colonies in treatments b and c. All of the colonies were irregular with indistinct margins. M. ichthyoblabe, M. viridis, M. flos-aquae, and M. wesenbergii formed colonies with similar morphologies and their cells were loosely aggregated. In contrast, M. aeruginosa formed denser colonies with no distinct holes. The colony morphologies differed from the classic morphology of M. ichthyoblabe field-grown colonies but resembled that of small colonies found in Lake Taihu (Yangtze Delta Plain, China) during early spring. This indicates that field- and laboratory-grown colonies are governed by similar formation processes. We suggest that in laboratory and field environments, M. ichthyoblabe (or M. flos-aquae) colonies are representative of small colonies formed from single Microcystis cells, whereas the morphology of older colonies evolves to resemble M. wesenbergii and M. aeruginosa colonies.     

Cyanobacterial blooms and water quality in Greek waterbodies

The cyanobacterial species composition of nine Greek waterbodies of different type and trophic status was examined during the warm period of the year (May–October). Cyanobacterial water blooms were observed in all waterbodies. Forty-six cyanobacterial taxa were identified, 11 of which are known to be toxic. Eighteen species are reported for the first time in these waterbodies, 8 of which are known to produce toxins. Toxin producing species were found in all of the waterbodies and were primarily dominant in bloom formations (e.g., Microcystis aeruginosa, Anabaena flos-aquae, Aphanizomenon flos-aquae and Cylindrospermopsis raciborskii). Cosmopolitan species (e.g., M. aeruginosa), pantropic (e.g., Anabaenopsis tanganyikae) and holarctic species (e.g., Anabaena flos-aquae) were encountered. Shallow, eutrophic waterbodies had blooms dominated by Microcystis species and were characterized by phytoplankton association M. Anabaena and Aphanizomenon species of association H were dominant in waterbodies with low dissolved inorganic nitrogen and thermal stratification in the summer. Total cyanobacterial biovolumes (CBV) ranged from 7 to 9,507 cm³ m⁻³ and were higher than Alert Level 2 and Guidance Level 2 (10 cm³ m⁻³; World Health Organization; WHO) in seven of the waterbodies. Chlorophyll a concentrations ranged from 6 to 90,000 mg m⁻³ and were higher than Alert Level 2 and Guidance Level 2 (50 mg m⁻³; WHO) in eight of the waterbodies. There is also an elevated risk of acute toxicosis (Guidance Level 3; WHO) in five waterbodies. Water of an undesirable quality, hazardous to humans and animals occurs in several Greek waterbodies.     

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.     

Diurnal changes in the vertical distribution of phytoplankton in hypertrophic Lake Kasumigaura,Japan

The daily vertical migration of five species;Microcystis aeruginosa (Kütz.) Trevis,Anabaena spiroides Klebahn f.crassa (L.) Elenkin,Aphanizomenon flos-aquae (L.) Ralfs,Melosira granulata (E). Ralfs, andCoscinodiscus lacustris Grun. was studied using a close-interval water sampler on a calm summer day in Lake Kasumigaura. Many colonies ofMicrocystis were observed at the middle of the water column (approx. 1.5 m depth) in the afternoon, and at the surface in the early morning.Anabaena occurred mostly in the upper layer whileAphanizomenon tended to be uniformly distributed. The difference in migration patterns suggests thatMicrocystis is superior toAnabaena andAphanizomenon in obtaining both light and nutrients from this lake. Among diatoms,Melosira remained at the bottom of the water column throughout day and night, but Coscinodiscus was uniformly distributed.     

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.     

Comparative studies on photosynthesis and phosphate metabolism of Cylindrospermopsis raciborskii with Microcystis aeruginosa and Aphanizomenon flos-aquae

The physiological differences for three bloom-forming cyanobacteria (Cylindrospermopsis raciborskii, Microcystis aeruginosa, and Aphanizomenon flos-aquae) were investigated. In comparison with M. aeruginosa and A. flos-aquae, C. raciborskii exhibited a significantly higher concentration of carotenoids, higher values in maximum photosynthesis rate (P_m), apparent photosynthetic efficieny (a), and maximum electron transport rate (ETR_max) during the growth period. In addition, higher extracellular alkaline phosphatase activities and lower light compensation point (I_c) were also detected in C. raciborskii (p < 0.05, ANOVA). Therefore, it is suggested that the higher photosynthetic activities, more effective uptake and utilization to phosphate, and low light requirements might play important roles in the occurrence and invasive behavior of C. raciborskii.     

Molecular Response of the Bloom-Forming Cyanobacterium, Microcystis aeruginosa, to Phosphorus Limitation

Cyanobacteria blooms caused by species such as Microcystis have become commonplace in many freshwater ecosystems. Although phosphorus (P) typically limits the growth of freshwater phytoplankton populations, little is known regarding the molecular response of Microcystis to variation in P concentrations and sources. For this study, we examined genes involved in P acquisition in Microcystis including two high-affinity phosphate-binding proteins (pstS and sphX) and a putative alkaline phosphatase (phoX). Sequence analyses among ten clones of Microcystis aeruginosa and one clone of Microcystis wesenbergii indicates that these genes are present and conserved within the species, but perhaps not the genus, as phoX was not identified in M. wesenbergii. Experiments with clones of M. aeruginosa indicated that expression of these three genes was strongly upregulated (50- to 400-fold) under low inorganic P conditions and that the expression of phoX was correlated with alkaline phosphatase activity (p < 0.005). In contrast, cultures grown exclusively on high levels of organic phosphorus sources (adenosine 5′-monophosphate, β-glycerol phosphate, and d-glucose-6-phosphate) or under nitrogen-limited conditions displayed neither high levels of gene expression nor alkaline phosphatase activity. Since Microcystis dominates phytoplankton assemblages in summer when levels of inorganic P (P_i) are often low and/or dominate lakes with low P_i and high organic P, our findings suggest this cyanobacterium may rely on pstS, sphX, and phoX to efficiently transport P_i and exploit organic sources of P to form blooms.     

Colony formation in the cyanobacterium Microcystis

Morphological evolution from a unicellular to multicellular state provides greater opportunities for organisms to attain larger and more complex living forms. As the most common freshwater cyanobacterial genus, Microcystis is a unicellular microorganism, with high phenotypic plasticity, which forms colonies and blooms in lakes and reservoirs worldwide. We conducted a systematic review of field studies from the 1990s to 2017 where Microcystis was dominant. Microcystis was detected as the dominant genus in waterbodies from temperate to subtropical and tropical zones. Unicellular Microcystis spp. can be induced to form colonies by adjusting biotic and abiotic factors in laboratory. Colony formation by cell division has been induced by zooplankton filtrate, high Pb²⁺ concentration, the presence of another cyanobacterium (Cylindrospermopsis raciborskii), heterotrophic bacteria, and by low temperature and light intensity. Colony formation by cell adhesion can be induced by zooplankton grazing, high Ca²⁺ concentration, and microcystins. We hypothesise that single cells of all Microcystis morphospecies initially form colonies with a similar morphology to those found in the early spring. These colonies gradually change their morphology to that of M. ichthyoblabe, M. wesenbergii and M. aeruginosa with changing environmental conditions. Colony formation provides Microcystis with many ecological advantages, including adaption to varying light, sustained growth under poor nutrient supply, protection from chemical stressors and protection from grazing. These benefits represent passive tactics responding to environmental stress. Microcystis colonies form at the cost of decreased specific growth rates compared with a unicellular habit. Large colony size allows Microcystis to attain rapid floating velocities (maximum recorded for a single colony, ∼ 10.08 m h⁻¹) that enable them to develop and maintain a large biomass near the surface of eutrophic lakes, where they may shade and inhibit the growth of less‐buoyant species in deeper layers. Over time, accompanying species may fail to maintain viable populations, allowing Microcystis to dominate. Microcystis blooms can be controlled by artificial mixing. Microcystis colonies and non‐buoyant phytoplankton will be exposed to identical light conditions if they are evenly distributed over the water column. In that case, green algae and diatoms, which generally have a higher growth rate than Microcystis, will be more successful. Under such mixing conditions, other phytoplankton taxa could recover and the dominance of Microcystis would be reduced. This review advances our understanding of the factors and mechanisms affecting Microcystis colony formation and size in the field and laboratory through synthesis of current knowledge. The main transition pathways of morphological changes in Microcystis provide an example of the phenotypic plasticity of organisms during morphological evolution from a unicellular to multicellular state. We emphasise that the mechanisms and factors influencing competition among various close morphospecies are sometimes paradoxical because these morphospecies are potentially a single species. Further work is required to clarify the colony‐forming process in different Microcystis morphospecies and the seasonal variation in this process. This will allow researchers to grow laboratory cultures that more closely reflect field morphologies and to optimise artificial mixing to manage blooms more effectively.     

Chemotaxonomy of planktonic cyanobacteria based on non-polar and 3-hydroxy fatty acid composition

Twenty-eight axenio planktonic cyanobacterial strains (10 Microcystis, three Oscillatoria, one Spirulina, one Aphanizomenon, 13 Anabaena) were investigated for their fatty acid composition by measurement of non-polar and hydroxy fatty acids. No 2-hydroxy fatty acids were detected in any strain, but 3-hydroxy fatty acids were detected in minor quantities in 24 strains. The highest portion of total fatty acids were non-polar fatty acids. Qualitative and quantitative analyses of 3-hydroxy fatty acids showed no taxonomic value in these strains, while the type of non-polar fatty acid composition was shown to be consistent within Microcystis and Anabaena strains, distinguishing them as type 4, characterized by the presence of 18:4, and type 2, characterized by 18:3 (α) of the Kenyon-Murata system. Two Oscillatoria agardhii Gomont strains were also included in the type 2 group due to the presence of 18: 3 (α), but the difference in characteristics of 16:2 and 16:3 between O. agardhii and Anabaena further divided type 2 into two subgroups: type 2A for Anabaena and type 2B for O. agardhii. A simplified unweighted pair group method with arithmetic averages (UPGMA) dendrogram demonstrated that the classification of 28 strains (Microcystis spp., Anabaena spp., Aphanizomenon flos-aquae (Lemmermann) Ralfs f. gracile (Lemmermann) Elenkin, O. agardhii and Spirullnasubsalsa Oersted ex Gomont based on numerical analysis of non-polar fatty acids corresponded to morphological species criteria, suggesting that non-polar fatty acid composition is a valuable chemical marker in the taxonomy of planktonic cyanobacteria. However, the fatty acid composition in Oscillatoria raciborskii is similar to that of Microcystis and very different from that of O. agardhii, suggesting its special position in Oscillatoria and the chemical diversity in the genus Oscillatoria.     

Strong effects of amoebae grazing on the biomass and genetic structure of a Microcystis bloom (Cyanobacteria)

Despite its importance for bloom toxicity, the factors determining the population structure of cyanobacterial blooms are poorly understood. Here, we report the results of a two‐year field survey of the population dynamics of Microcystis blooms in a small hypertrophic urban pond. Microscopic enumeration of Microcystis and its predators and parasites was combined with pigment and microcystin analysis and denaturing gradient gel electrophoresis of the ITS rDNA region to assess population dynamics and structure. Two main Microcystis morpho‐ and ITS types were revealed, corresponding to M. aeruginosa and M. viridis. In both years, high population densities of naked amoebae grazing on Microcystis coincided with rapid decreases in Microcystis biomass. In one year, there was a shift from heavily infested M. aeruginosa to the less‐infested M. viridis, allowing the bloom to rapidly recover. The preference of amoebae for M. aeruginosa was confirmed by grazing experiments, in which several amoeba strains were capable of grazing down a strain of M. aeruginosa, but not of M. viridis. Zooplankton and chytrid parasites appeared to be of minor importance for these strong and fast reductions in Microcystis biomass. These findings demonstrate a strong impact of small protozoan grazers on the biomass and genetic structure of Microcystis blooms.     

Trophic coupling of a testate amoeba and <Emphasis Type="Italic">Microcystis</Emphasis> species in a hypertrophic pond

Seasonal changes in abundance of the testate amoeba Penardochlamys sp. and its food vacuole contents were investigated in relation to blooms of the cyanobacteria Microcystis spp. in a hypertrophic pond from April 1999 to March 2000. The behavior of the amoeba feeding on M. aeruginosa and M. wesenbergii was also observed in the laboratory. The amoeba was detectable from late May to November 1999 during the blooms of Microcystis spp. Cell densities of the amoeba fluctuated between 1.4 and 350 cells ml^–1 with some sporadic peaks, which did not coincide with rapid decreases in the abundance of Microcystis spp. Food vacuoles contained only Microcystis cells; other prey items were not found, suggesting that this amoeba utilized only the cyanobacteria as food. The amoeba was frequently found attached to Microcystis colonies, but was not associated with other suspended particles. Observation of the amoeba feeding revealed the feeding mechanism and that the amoeba was able to graze on both species of Microcystis. These results suggest that the trophic coupling of these organisms is substantial, although grazing by the amoeba is not sufficient to regulate the dynamics of Microcystis populations in this hypertrophic pond.     

Allelopathic control of freshwater phytoplankton by the submerged macrophyte Najas minor All

Water blooms in eutrophic waters have been serious environmental problems in recent years. To explore effective measures to control this issue has been an interest of research. Our current study was designed to investigate the effects of submerged macrophyte Najas minor All. exudates on the growth of four freshwater phytoplankton species, toxic Microcystis aeruginosa, toxic Anabaena flos-aquae, Chlorella pyrenoidosa and Scenedesmus obliquus as well as natural phytoplankton assemblages of pond water. We also conducted a reciprocal response between N. minor and toxic M. aeruginosa using coexistence experiments. Our results showed that: (1) N. minor exudates significantly inhibited the growth of toxic M. aeruginosa, toxic A. flos-aquae and S. obliquus, with M. aeruginosa being the most sensitive, followed by toxic A. flos-aquae, and S. obliquus the least. N. minor exudates did not show inhibitory effect on C. pyrenoidosa; (2) N. minor and toxic M. aeruginosa have reciprocal inhibitory effect, and the allelopathic interactions between the two different organisms are density dependent and affect their mutual growth; (3) N. minor exudates also can induce a decrease in chlorophyll a content and an inhibition in total dehydrogenase activity of the phytoplankton assemblages. Our present studies indicated the submerged macrophyte N. minor might be a potential useful tool to control phytoplankton blooms.     

The importance of morphological versus chemical defences for the bloom-forming cyanobacterium Microcystis against amoebae grazing

Amoebae grazing can be an important loss factor for blooms of the common cyanobacterium Microcystis. Some Microcystis strains seem to be protected against amoebae grazing, but it is unclear whether this is achieved by their colony morphology or biochemically. These factors were investigated in grazing experiments using two Microcystis-grazing amoebae (Korotnevella sp. and Vannella sp.) and two Microcystis strains with differing colony morphology (aeruginosa and viridis morphotype) and different sensitivity to amoebae grazing. Amoebae did not increase in density and failed to reduce the growth rate of cultures of the amoebae insensitive viridis strain, irrespective of whether the Microcystis strain was colonial or unicellular. This suggests that the extended mucilage matrix surrounding viridis colonies is not the main defence mechanism against amoebae grazing. At the same time, the growth rate of both unicellular and colonial cultures of the amoebae-sensitive aeruginosa strain was heavily reduced by the growing amoebae. The addition of filtered viridis-conditioned medium to aeruginosa cultures significantly decreased both amoebae growth and its effect on aeruginosa growth rates, which indicates that extracellular compounds constitutively produced by viridis are at least partially responsible for their insensitivity to amoebae grazing. These results demonstrate the potential importance of chemical interactions between lower trophic levels (protists) for Microcystis bloom dynamics.     

Contribution of benthic blue-green algal recruitment to lake populations and phosphorus translocation

1. A two-season investigation was undertaken to determine the contribution of benthic recruitment to the population development of several species of blue-green algae. 2. Gloevtrichia echinulata populations were shown to be heavily subsidized by benthic recruitment, deriving on average 40% of their planktonic colonies from the benthos. 3. Benthic recruitment of Aphanizomenon flos-aquae, Anabaena flos-aquae and a second Anabaena species contributed less than 2% to planktonic increases, while Microcystis aeruginosa recruitment was negligible. 4. Phosphorus translocation via migrating G. echinulata accounted for most of the phosphorus (P) in the planktonic colonies, and constituted a significant portion of the internal loading of the lake. 5. Estimated P translocation via Aph. flos-aquae was relatively minor, although there was evidence of luxury uptake in the benthos.     

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.     

The Influence of Toxic and Filamentous Blue-green Algae on Feeding and Population Growth of the Rotifer Brachionus rubens

The effect of toxic and of filamentous blue-green algae on feeding and population growth of the rotifer Brachionus rubens was investigated in laboratory experiments. A toxic strain of Microcystis aeruginosa was ingested, but rotifers cultured with Microcystis died faster than nonfed controls. The rigid filaments of Cylindrospermopsis raciborskii were not ingested, reduced the ingestion of simultaneously offered algae, and depressed population growth rates. The soft filaments of Anabaena flos-aquae were ingested at a moderate rate, did not reduce ingestion of other algae, and were used as an additional food in population growth experiments.     

Rising temperatures may increase growth rates and microcystin production in tropical Microcystis species

Rising temperatures (1.4–6 °C) due to climate change have been predicted to increase cyanobacterial bloom occurrences in temperate water bodies; however, the impacts of warming on tropical cyanobacterial blooms are unknown. We examined the effects of four different temperatures on the growth rates and microcystin (MC) production of five tropical Microcystis isolates (M. ichthyoblabe (two strains), M. viridis, M. flos-aquae, and M. aeruginosa). The temperature treatments are based on current temperature range in Singapore's reservoirs (27 °C and 30 °C), as well as projected mean (33 °C) and maximum temperatures (36 °C) based on tropical climate change estimates of +6 °C in air temperature. Increasing temperatures did not significantly affect the maximum growth rates of most Microcystis strains. Higher growth rates were only observed in one M. ichthyoblabe strain at 33 °C and M. flos-aquae at 30 °C where both were isolated from the same reservoir. MC-RR and MC-LR were produced in varying amounts by all four species of Microcystis. Raised temperatures of 33 °C were found to boost total MC cell quota for three Microcystis strains although further increase to 36 °C led to a sharp decrease in total MC cell quota for all five Microcystis strains. Increasing temperature also led to higher MC-LR:MC-RR cell quota ratios in M. ichthyoblabe. Our study suggests that higher mean water temperatures resulting from climate change will generally not influence growth rates of Microcystis spp. in Singapore except for increases in M. ichthyoblabe strains. However, toxin cell quota may increase under moderate warming scenarios depending on the species.     

Effects of Iron on Growth,Pigment Content,Photosystem II Efficiency,and Siderophores Production of <Emphasis Type="Italic">Microcystis aeruginosa</Emphasis> and <Emphasis Type="Italic">Microcystis wesenbergii</Emphasis>

Changes in growth, photosynthetic pigments, and photosystem II (PS II) photochemical efficiency as well as production of siderophores of Microcystis aeruginosa and Microcystis wesenbergii were determined in this experiment. Results showed growths of M. aeruginosa and M. wesenbergii, measured by means of optical density at 665 nm, were severely inhibited under an iron-limited condition, whereas they thrived under an iron-replete condition. The contents of chlorophyll-a, carotenoid, phycocyanin, and allophycocyanin under an iron-limited condition were lower than those under an iron-replete condition, and they all reached maximal contents on day 4 under the iron-limited condition. PS II photochemical efficiencies (maximal PS II quantum yield), saturating light levels (I _k ) and maximal electron transport rates (ETR_max) of M. aeruginosa and M. wesenbergii declined sharply under the iron-limited condition. The PS II photochemical efficiency and ETR_max of M. aeruginosa rose , whereas in the strain of M. wesenbergii, they declined gradually under the iron-replete condition. In addition, I _k of M. aeruginosa and M. wesenbergii under the iron-replete condition did not change obviously. Siderophore production of M. aeruginosa was higher than that of M. wesenbergii under the iron-limited condition. It was concluded that M. aeruginosa requires higher iron concentration for physiological and biochemical processes compared with M. wesenbergii, but its tolerance against too high a concentration of iron is weaker than M. wesenbergii.     

Effects of temperature, and availability of nitrogen and phosphorus on the abundance of Anabaena and Microcystis in Lake Biwa, Japan: an experimental approach

Under optimal nutrient conditions, both Microcystis sp. and Anabaena sp. isolated from Lake Biwa grew optimally at 28–32°C but differed in maximal growth rates, phosphate uptake kinetics, maximal phosphorus quotas, and growth responses to nitrogen and phosphorus limitation. The maximal growth rates of Microcystis and Anabaena were 1.6 and 1.25 divisions day⁻¹, respectively. With phosphate and nitrate in the growth-limiting range, the growth of Microcystis was optimal at an N : P ratio of 100 : 1 (by weight) and declined at lower (nitrogen limitation) and higher (phosphorus limitation) ratios. In contrast, Anabaena growth rates did not change at N : P ratios from 1000 : 1 to 10 : 1. Starting with cells containing the maximal phosphorus quota, Microcystis growth in minus-phosphorus medium ceased in 7–9 days, compared with 12–13 days for Anabaena. The phosphate turnover time in cultures starved to their minimum cell quotas was 7.9 min for Microcystis and 0.6 min for Anabaena. Microcystis had a higher K _s (0.12 μg P l⁻¹ 10⁻⁶ cells) and lower V _max (9.63 μg P l⁻¹ h⁻¹ 10⁻⁶ cells), than Anabaena (K _s 0.02 μg P l⁻¹ h⁻¹ 10⁻⁶ cells; V _max 46.25 63 μg P l⁻¹ h⁻¹ 10⁻⁶ cells), suggesting that Microcystis would not be able to grow well in phosphorus-limited waters. We conclude that in spite of the higher growth rate under ideal conditions, Microcystis does not usually bloom in the North Basin because of low availability of phosphorus and nitrogen. Although Anabaena has an efficient phosphorus-uptake system, its main strategy for growth in low-phosphorus environments may depend on storage of phosphorus during periods of abundant phosphorus supply, which are rare in the North Basin. Received: July 31, 2000 / Accepted: October 18, 2000     

POLYPHASIC CHARACTERIZATION OF THREE STRAINS OF ANABAENA RENIFORMIS AND APHANIZOMENON APHANIZOMENOIDES (CYANOBACTERIA) AND THEIR RECLASSIFICATION TO SPHAEROSPERMUM GEN. NOV. (INCL. ANABAENA KISSELEVIANA)1

Occurrences of rare cyanobacteria Anabaena reniformis Lemmerm. and Aphanizomenon aphanizomenoides (Forti) Horecká et Komárek were recently detected at several localities in the Czech Republic. Two monoclonal strains of An. reniformis and one strain of Aph. aphanizomenoides were isolated from distant localities and different sampling years. They were characterized by a combination of morphological, genetic, and biochemical approaches. For the first time, partial 16S rRNA gene sequences were obtained for these morphospecies. Based on this gene, all of these strains clustered separately from other planktonic Anabaena and Aphanizomenon strains. They appeared in a cluster with Cylindrospermopsis Seenaya et Subba Raju and Raphidiopsis F. E. Fritsch et M. F. Rich, clustered closely together with two An. kisseleviana Elenkin strains available from GenBank. A new generic entity was defined (Sphaerospermum gen. nov., with the type species S. reniforme, based on the traditional species An. reniformis). These results contribute significantly to the knowledge base about genetic heterogeneity among planktonic Anabaena–like and Aphanizomenon–like morphospecies. Accordingly, the subgenus Dolichospermum, previously proposed for the group of planktonic Anabaena, should be revaluated. Secondary metabolite profiles of the An. reniformis and Aph. aphanizomenoides strains differed considerably from 17 other planktonic Anabaena strains of eight morphospecies isolated from Czech water bodies. Production of puwainaphycin A was found in both of the An. reniformis strains. Despite the relatively short phylogenetic distance from Cylidrospermopsis, the production of cylindrospermopsin was not detected in any of our strains.