Freshwater Strains Research Articles

Evaluation of Diverse Microalgal Species as Potential Biofuel Feedstocks Grown Using Municipal Wastewater

Microalgae offer great potential as a third-generation biofuel feedstock, especially when grown on wastewater, as they have the dual application for wastewater treatment and as a biomass feedstock for biofuel production. The potential for growth on wastewater centrate was evaluated for forty microalgae strains from fresh (11), brackish (11), or saltwater (18) genera. Generally, freshwater strains were able to grow at high concentrations of centrate, with two strains, Neochloris pseudostigmata and N. conjuncta, demonstrating growth at up to 40% v/v centrate. Fourteen of eighteen salt water Dunaliella strains also demonstrated growth in centrate concentrations at or above 40% v/v. Lipid profiles of freshwater strains with high-centrate tolerance were determined using gas chromatography-mass spectrometry (GC-MS) and compared against those obtained on cells grown on defined maintenance media. The major lipid compounds were found to be palmitic (16:0), oleic (18:1), and linoleic (18:2) acids for all freshwater strains grown on either centrate or their respective maintenance medium. These results demonstrate the highly concentrated wastewater can be used to grow microalgae, which limits the need to dilute wastewater prior to algal production. In addition, the algae produced generate lipids suitable for biodiesel or green diesel production.

Heterogeneity of alkane chain length in freshwater and marine cyanobacteria.

The potential utilization of cyanobacteria for the biological production of alkanes represents an exceptional system for the next generation of biofuels. Here, we analyzed a diverse group of freshwater and marine cyanobacterial isolates from Indian culture collections for their ability to produce both alkanes and alkenes. Among the 50 cyanobacterial isolates screened, 32 isolates; 14 freshwater and 18 marine isolates; produced predominantly alkanes. The GC-MS/MS profiles revealed a higher percentage of pentadecane and heptadecane production for marine and freshwater strains, respectively. Oscillatoria species were found to be the highest producers of alkanes. Among the freshwater isolates, Oscillatoria CCC305 produced the maximum alkane level with 0.43 μg/mg dry cell weight, while Oscillatoria formosa BDU30603 was the highest producer among the marine isolates with 0.13 μg/mg dry cell weight. Culturing these strains under different media compositions showed that the alkane chain length was not influenced by the growth medium but was rather an inherent property of the strains. Analysis of the cellular fatty acid content indicated the presence of predominantly C16 chain length fatty acids in marine strains, while the proportion of C18 chain length fatty acids increased in the majority of freshwater strains. These results correlated with alkane chain length specificity of marine and freshwater isolates indicating that alkane chain lengths may be primarily determined by the fatty acid synthesis pathway. Moreover, the phylogenetic analysis showed clustering of pentadecane-producing marine strains that was distinct from heptadecane-producing freshwater strains strongly suggesting a close association between alkane chain length and the cyanobacteria habitat.

Indole alkaloids from Fischerella inhibit vertebrate development in the zebrafish (Danio rerio) embryo model.

Cyanobacteria are recognized producers of toxic or otherwise bioactive metabolite associated, in particular, with so-called “harmful algal blooms” (HABs) and eutrophication of freshwater systems. In the present study, two apparently teratogenic indole alkaloids from a freshwater strain of the widespread cyanobacterial genus, Fischerella (Stigonemataceae), were isolated by bioassay-guided fractionation, specifically using the zebrafish (Danio rerio) embryo, as a model of vertebrate development. The two alkaloids include the previously known 12-epi-hapalindole H isonitrile (1), and a new nitrile-containing variant, 12-epi-ambiguine B nitrile (2). Although both compounds were toxic to developing embryos, the former compound was shown to be relatively more potent, and to correlate best with the observed embryo toxicity. Related indole alkaloids from Fischerella, and other genera in the Stigonemataceae, have been widely reported as antimicrobial compounds, specifically in association with apparent allelopathy. However, this is the first report of their vertebrate toxicity, and the observed teratogenicity of these alkaloids supports a possible contribution to the toxicity of this widespread cyanobacterial family, particularly in relation to freshwater HABs and eutrophication.

Heritability Estimates and Genetic Correlations Among Body Weight and Measurement Traits of Large‐size Rainbow Trout, Oncorhynchus mykiss, in China

AbstractThe variance components of additive effects and genetic correlations of body weight (BW) and body measurement traits were estimated in a freshwater strain selected for large size in a breeding program for rainbow trout, Oncorhynchus mykiss, in China. Phenotypic data were collected for BW and measurement traits from 2362 individuals with a two‐generation pedigree, and the average information‐restricted maximum likelihood method was used for the genetic analysis. The genetic parameters of BW were estimated at three time points (550, 850, and 1000 days post‐fertilization [dpf]), showing high heritability in a range from 0.33 to 0.98. However, the heritability estimates of total length (TL) showed lower values ranging from 0.05 to 0.51, with high standard errors. Genetic correlation between TL and BW was almost equal to unity at 550 dpf, and the genetic correlations between TL at 550 dpf and BW at two other time points were all positive and very high (ca. 0.95–0.99). These results indicate that BW and TL selected at 550 dpf had a direct positive effect on BW performance at 1000 dpf, which was 450 d before the date of the target trait observed.

Isolation and Characterization of New Temperature Tolerant Microalgal Strains for Biomass Production

Microalgae exhibit great potential for biomass production. Although microalgae display an enormous biodiversity, surprisingly only 15 species are used for large scale production processes worldwide. The implementation of new production strains with good process-oriented properties, especially fast growth rate and heat resistance, could improve production efficiency and reduce costs. In this study 130 environmental samples collected in Germany, Spain, Italy and Portugal were investigated for fast growing thermotolerant photosynthetic species. Isolates were characterized and identified on a molecular level. In total 21 of the isolated freshwater strains were able to grow at 40 °C. Additionally, 13 of those 21 strains are able to grow at 45 °C. The highest growth rate at room temperature was 1.16 per day (isolate T306A), compared to 0.053 per day at 45 °C (isolate Sp13). In three thermotolerant strains pigment production was induced. Molecular identification by 18S rDNA sequencing revealed that the isolates were all chlorophytes belonging to four different families.

Strain Selection, Biomass to Biofuel Conversion, and Resource Colocation have Strong Impacts on the Economic Performance of Algae Cultivation Sites

Decisions involving strain selection, biomass to biofuel technology, and the location of cultivation facilities can strongly influence the economic viability of an algae-based biofuel enterprise. We summarize our past results in a new analysis to explore the relative economic impact of these design choices. Our growth model is used to predict average biomass production for two saline strains (Nannocloropsis salina, Arthrospira sp.), one fresh to brackish strain (Chlorella sp., DOE strain 1412), and one freshwater strain (order Sphaeropleales). Biomass to biofuel conversion is compared between lipid extraction (LE) and hydrothermal liquefaction (HTL) technologies. National-scale models of water, CO2 (as flue gas), land acquisition, site leveling, construction of connecting roads, and transport of HTL oil to existing refineries are used in conjunction with estimates of fuel value (from HTL) to prioritize and select from 88,692 unit farms (UF, 405 ha in pond area), a number sufficient to produce 136E+9 L yr-1 of renewable diesel (36 billion gallons yr-1). Strain selection and choice of conversion technology have large economic impacts, with differences between combinations of strains and biomass to biofuel technologies being up to $10 million dollars yr-1 UF-1. Results based on the most productive strain, HTL-based fuel conversion, and resource costs show that the economic potential between geographic locations within the selection can differ by up to $4 million yr-1 UF-1, with 1.8 BGY of production possible from the most cost-effective sites. The local spatial variability in site rank is extreme, with very high and low sites within 10s of km of each other. Colocation with flue gas sources has a strong influence on rank, but the most costly resource component varies from site to site. The highest rank UFs are located predominantly in Florida and Texas, but most states south of 37°N latitude contain promising locations.

Growth of microalgae using CO2 enriched air for biodiesel production in supercritical CO2

The optimum conditions for lipids productivity and CO2 fixation of two freshwater strains, namely Chlorella sp. and Pseudochlorococcum sp. and a marine strain; namely Nannochlorpsis sp. have been determined in this work. The species were grown autotrophically under aeration with different CO2 concentrations, ranging from 0.04 to 2% (v/v). The growth was tested in nitrogen sufficient and deficient media at different salinities (0.49–680 mM) and temperatures of 27 and 31 °C. The optimum CO2 enrichment was found to be 1% (v/v) in both media. Nitrogen starvation resulted in an increase in lipid contents, but at lower growth rate, which resulted in a lower overall lipid productivity. The experimental data were used to determine the kinetic parameters of Haldane model. The Chlorella sp. grew well at salinity levels of up to 460 mM. The highest CO2 biofixation rate was observed when Chlorella sp. was grown at 27 °C in seawater (230 mM NaCl).Lipids were extracted from harvested marine strain, Nannochlorpsis sp., and enzymatically transesterified to produce biodiesel in supercritical CO2 (SC–CO2) medium. It was found that the conversion of biodiesel produced from microalgae lipids was 35% higher than that achieved using lamb fat in a similar system.

Inducible expression system for the marine cyanobacterium Synechococcus sp. strain NKBG 15041c

Cyanobacteria have been recognized as promising host organisms to produce biofuel materials including hydrogen and hydrocarbons due to high biomass productivity and capability of transformation. Metabolic engineering has recently been employed for further enhancement of bioenergy production, while excess fuel materials could give rise to cytotoxicity. Therefore genetic tools are required in order to precisely control the gene expression involved in the fuel production metabolism. Thus far, inducible-expression systems in cyanobacteria have been established mainly in freshwater strains. However there are a few reports with regard to marine strains, which can use vast of ocean for cultivation. In this study, we established a tetracycline-inducible gene expression system in the marine cyanobacterium Synechococcus sp. strain NKBG 15041c. Since tetracycline-inducible system is a non-native regulation, it is expected for inherent metabolisms not to be disrupted. The de novo designed promoter including tetracycline operator elements was inactivated by the repressor protein, and inducer addition successfully initiated target protein expression. This is the first report to demonstrate tetracycline-induction system with strict on/off switching in marine cyanobacteria, and it should be useful for future bioenergy production.

Salt tolerance and polyphyly in the cyanobacterium Chroococcidiopsis (Pleurocapsales).

Chroococcidiopsis Geitler (Geitler 1933) is a genus of cyanobacteria containing desiccation and radiation resistant strains. Members of the genus live in habitats ranging from hot and cold deserts to fresh and saltwater environments. Morphology and cell division pattern have historically been used to define the genus. To better understand the evolution and ability of the Chroococcidiopsis genus to survive in diverse environments we investigated how salt tolerance varies among 15 strains previously isolated from different locations, and if salt tolerant strains are monophyletic to those isolated from freshwater and land environments. Four markers were sequenced from these 15 strains, the 16S rRNA, rbcL, desC1, and gltX genes. Phylogenetic trees were generated which identified a distinct clade of salt-tolerant strains. This study demonstrates that the genus is polyphyletic based on saltwater and freshwater phenotypes. To understand the resistance to salt in more details, the strains were grown on a range of sea salt concentrations which demonstrated that the freshwater strains were salt-intolerant whilst the saltwater strains required salt for growth. This study shows an increased resolution of the phylogeny of Chroococcidiopsis and provides further evidence that the genus is polyphyletic and should be reclassified to improve clarity in the literature.

Comprehensive modeling and investigation of the effect of iron on the growth rate and lipid accumulation of Chlorella vulgaris cultured in batch photobioreactors

Recent works have shown that specific strains of microalgae are capable to simultaneously increase their growth rate and lipid content when cultured under suitable concentrations of iron. While these results are promising in view of the exploitation of microalgae for producing biofuels, to the best of our knowledge, no mathematical model capable to describe the effect of iron on lipid accumulation in microalgae, has been so far proposed. A comprehensive mathematical model describing the effect of iron on chlorophyll synthesis, nitrogen assimilation, growth rate and lipid accumulation in a freshwater strain of Chlorella vulgaris is then proposed in this work. Model results are successfully compared with experimental data which confirm the positive effect of growing iron concentrations on lipid productivity of C. vulgaris. Thus, the proposed model might represent a useful tool to optimize iron-based strategies to improve the lipid productivity of microalgal cultures.

Selection of native freshwater microalgae and cyanobacteria for CO2 biofixation

One of the technologies available for coping with problems related to the rise in atmospheric concentrations of carbon dioxide is CO2 biofixation with microalgae or cyanobacteria. The selection of native strains that grow well at the specific site where the technology is to be used will increase the success possibilities of such a technology. Thus, with the aim of finding a suitable local variety for use in a CO2 biofixation system, three recently isolated freshwater strains, Scenedesmus sp., Chlorogonium sp. and Synechocystis sp. were studied. Chlorella sorokiniana was used as a control strain. All the strains were grown under the same culture conditions for seven days of batch culture, and various growth and CO2 biofixation parameters were determined. Synechocystis sp. showed the highest specific growth rate at 1.75 per day (l/d). Results for CO2 biofixation ranged between 0.650 and 0.953 g of carbon dioxide per litre per day (g CO2/l/d), but differences among native strains were noted, although they were not statistically significant. However, Synechocystis sp. was selected as the most suitable strain for CO2 biofixation, owing to its good capacity to use light in dense cultures, an essential requirement for sustainable commercial systems.

Co‐occurrence of phycocyanin‐ and phycoerythrin‐rich Synechococcus in subtropical estuarine and coastal waters of Hong Kong

Phylogenetic diversity of Synechococcus with different pigmentation in subtropical estuarine and coastal waters of Hong Kong was revealed by the phylogeny of cpcBA and cpeBA operons encoding for phycocyanin (PC) and phycoerythrin (PE). Synechococcus containing only PC (PC-rich Synechococcus) dominated at the estuarine station in summer, whereas PE-rich marine Synechococcus containing both PC and PE (PE-rich Synechococcus) dominated in the coastal waters. Our PC sequences are closely related to freshwater strains but differed from Baltic Sea strains, implying that they were from river discharge. Among PE-rich Synechococcus, clones grouping with strains containing only phycoerythrobilin (PEB-only) were abundant in July, while clones grouping with strains possessing a low content of phycourobilin (PUB) in addition to PEB (low PUB/PEB) were more abundant in January at both stations. Clones of high PUB/PEB types were only presented at the coastal station, but were not detected at the estuarine station. The much higher diversity of both PC-rich and PE-rich Synechococcus, as compared with the Baltic Sea, and the occurrence of the high PUB/PEB strains indicate the high dynamic nature of this subtropical estuarine-coastal environment with strong mixing of water masses ranging from Pearl River plume to oceanic South China Sea water. Our results of phylogenetic study agreed well with flow cytometric counts, which revealed the coexistence of PC-rich and PE-rich Synechococcus in the subtropical coastal waters and the dominance of the former type in the estuarine waters during summer high freshwater discharge. These results indicate that picocyanobacteria, particularly PC-rich Synechococcus, which has long been overlooked, are an important part of the primary production, and they could play an important role in the microbial food web in estuarine ecosystems.

Detection and expression of genes for phosphorus metabolism in picocyanobacteria from the Laurentian Great Lakes

The pelagic regions of Lake Superior and eastern Lake Erie (Laurentian Great Lakes) are typically phosphorus (P)-limited environments, and picocyanobacteria of the genus Synechococcus spp. are prominent primary producers during the summer. As a proxy for their utilization of organic P, the expression of two genes, phnD and phoX, was monitored. The phnD gene encodes the phosphonate binding protein of the ABC-type phosphonate transporter, whereas the phoX gene encodes a calcium-dependent alkaline phosphatase. Furthermore, to assess the ability of freshwater Synechococcus spp. to substitute sulfolipids for phospholipids, sqdX gene (cyanobacterial sulfolipid synthase) expression was examined. We employed PCR primers to detect the presence of all three genes in the endemic Synechococcus spp., and RT-PCR assays of cultured freshwater strains and environmental samples to assess the degree of P-stress in the phytoplankton. We show that the phnD gene was constitutively expressed, suggesting that freshwater picocyanobacteria were metabolizing exogenous phosphonate compounds in Lakes Erie and Superior. By contrast, phoX was regulated by P bioavailability. We also provide evidence that sqdX is expressed during increased growth rates in phosphorus-replete conditions, suggesting that sulfolipid synthesis is not a P conservation mechanism for freshwater Synechococcus spp.

Sulfide oxidation, nitrate respiration, carbon acquisition, and electron transport pathways suggested by the draft genome of a single orange Guaymas Basin Beggiatoa (Cand. Maribeggiatoa) sp. filament

A near-complete draft genome has been obtained for a single vacuolated orange Beggiatoa (Cand. Maribeggiatoa) filament from a Guaymas Basin seafloor microbial mat, the third relatively complete sequence for the Beggiatoaceae. Possible pathways for sulfide oxidation; nitrate respiration; inorganic carbon fixation by both Type II RuBisCO and the reductive tricarboxylic acid cycle; acetate and possibly formate uptake; and energy-generating electron transport via both oxidative phosphorylation and the Rnf complex are discussed here. A role in nitrite reduction is suggested for an abundant orange cytochrome produced by the Guaymas strain; this has a possible homolog in Beggiatoa (Cand. Isobeggiatoa) sp. PS, isolated from marine harbor sediment, but not Beggiatoa alba B18LD, isolated from a freshwater rice field ditch. Inferred phylogenies for the Calvin–Benson–Bassham (CBB) cycle and the reductive (rTCA) and oxidative (TCA) tricarboxylic acid cycles suggest that genes encoding succinate dehydrogenase and enzymes for carboxylation and/or decarboxylation steps (including RuBisCO) may have been introduced to (or exported from) one or more of the three genomes by horizontal transfer, sometimes by different routes. Sequences from the two marine strains are generally more similar to each other than to sequences from the freshwater strain, except in the case of RuBisCO: only the Guaymas strain encodes a Type II enzyme, which (where studied) discriminates less against oxygen than do Type I RuBisCOs. Genes subject to horizontal transfer may represent key steps for adaptation to factors such as oxygen and carbon dioxide concentration, organic carbon availability, and environmental variability.

Selection of microalgae for wastewater treatment and potential lipids production

In the present study, ten microalgal strains found in fresh and saline waters were cultured, and used to conduct batch experiments in order to evaluate their potential contribution to nutrient removal and biofuel production. The growth rate of microalgae was inversely analogous to their initial concentration. Three freshwater strains were selected, based on their growth rate, and their behavior with synthetic wastewater was further investigated. The strains studied were the Scenedesmus rubescens (SAG 5.95), the Neochloris vigensis (SAG 80.80), and the Chlorococcum spec. (SAG 22.83), and higher growth rate was observed with S. rubescens. Total phosphorus removal at an initial phosphate concentration of 6-7 mg P/L in the synthetic wastewater, was 53%, 25% and 11% for N. vigensis, Chlorococcum spec., and S. rubescens, respectively. Finally, the lipid content was determined at 20th and 30th day of cultivation, and the highest amount was observed at the 20th day.

Comparative Salt Stress Study on Intracellular Ion Concentration in Marine and Salt-adapted Freshwater Strains of Microalgae

Salinity imposes significant stresses in various living organisms including microalgae. High extracellular concentration of Na+ directly influences ionic balance inside the cell and subsequently the cellular activities. In the present study, the effect of such stress on growth and intracellular ions concentration (IIC) of Dunaliella salina and Chlorella Spp. was investigated. IIC was analyzed using Ion chromatography technique. D. salina showed the highest degree of resistance to increase in salinity as little changes occurred both in IIC and in growth parameters. D. salina could maintain the balance of K+ inside the cell and eject the excess Na+ even at NaCl concentrations above 1M. Moreover, D. salina accumulated β-carotene in order to protect its photosynthetic apparatus. Among Chlorella species, C. vulgaris showed signs of adaptation to high content of salinity, though it is a fresh water species by nature. Moreover, the response shown by C. vulgaris to rise in salinity was even stronger than that of C. salina, which is presumably a salt-water resistant species. In fact, C. vulgaris could maintain intracellular K+ better than C. salina in response to increasing salinity, and as a result, it could survive at NaCl concentrations as high as 0.75 M. Marine strains such as D. salina well cope with the fluctuations in salinity through the existing adaptation mechanisms i.e. maintaining the K+/N+ balance inside the cell, K+ accumulation and Na+ ejection, accumulation of photosynthetic pigments like β-carotene.

CaCO3 biomineralization on cyanobacterial surfaces: Insights from experiments with three Synechococcus strains

In the present paper, the impact of freshwater (ARC21 and LS0519) and marine (PCC8806) Synechococcus cyanobacteria on calcium carbonate (CaCO3) precipitation has been examined in respect of the formation rates and morphology of crystals. Acid–base potentiometric titrations were employed to study surface functional groups, while CaCO3 experiments have been carried out in presence and absence of cells at low to near-equilibrium conditions in respect to CaCO3. During these experiments, the pH values have been monitored, Ca and alkalinity were measured and precipitates have been investigated by Raman spectroscopy and Atomic Force and Scanning Electron microscopy.Our results showed that the Synechococcus strains exhibited different surface reactivity with total concentration of surface functional groups of 0.342 and 0.350mMg−1 of dry bact. for freshwater strains, and 0.662mMg−1 of dry bact. for the marine strain, which are on the same order of magnitude as that reported for bacterial cell surfaces. The marine strain showed the highest CaCO3 formation rate with Ca2+ removal of 18mMg−1 dry bact. compared to 6–7mMg−1 dry bact. for freshwater strains. The morphological diversity in crystals has been linked to presence of specific functional groups. The linking cell surface properties to crystal morphologies and precipitation rates propose that bacterial surfaces may modulate CaCO3 formation. Results of this work should allow better understanding of biominiralization in marine and freshwater systems as they define the precipiatation rates in typical range of pH necessary for estimation of CaCO3 formation by cyanobacterial communities.

Identification of Monoraphidium contortum as a promising species for liquid biofuel production

In this work, 30 microalgae strains from 17 genera were investigated in regard to biomass productivity in photoautotrophic growth conditions, lipid amount, lipid quality and biomass degradability. Six strains could be identified with robust phototrophic growth properties and high biomass productivities equal or above 300mgl−1day−1. Anaerobic fermentation of the algal biomass was most efficient for the marine members of the genera Dunaliella and Navicula, while biogas production with the freshwater strains generally resulted in lower methane yields. Monoraphidium contortum was identified as promising candidate for liquid biofuel production, characterized by high biomass productivity during maximum growth (maximum increase of 896mg dry biomass weight (DW) l−1day−1) and a promising lipid profile. Neutral lipid production was strongly induced in M. contortum by nitrogen deficient conditions and accumulated to up to 20.4±2.2% of DW.

Two-stage cultivation of two Chlorella sp. strains by simultaneous treatment of brewery wastewater and maximizing lipid productivity

A cultivation system in the two-stage photoautotrophic–photoheterotrophic/mixotrophic mode was adapted to maximize lipid productivity of two freshwater strains of Chlorella sp. grown in brewery wastewater (BWW). The endogenous Chlorella sp. isolated from BWW had a higher growth rate than wild-type Chlorella vulgaris (UTEX-265) while C. vulgaris (UTEX-265) had a higher maximal biomass and lipid contents than that of endogenous Chlorella sp., resulting in more than 90% of the inorganic nutrients in both total nitrogen (TN) and phosphorus (TP) was removed during the first stage in the two-stage photoautotrophic–photoheterotrophic mode in each Chlorella sp. The maximal biomass and lipid contents of C. vulgaris (UTEX-265) for single stage photoautotrophic cultivation were 1.5g/L and 18%, respectively. Importantly, during two-stage photoautotrophic–photoheterotrophic cultivation for C. vulgaris (UTEX-265), the biomass was increased to 3.5g/L, and the lipid productivity was increased from 31.1 to 108.0mg/Lday.

Gene Flow, Recombination, and Selection in Cyanobacteria: Population Structure of Geographically Related Planktothrix Freshwater Strains

Several Planktothrix strains, each producing a distinct oligopeptide profile, have been shown to coexist within Lake Steinsfjorden (Norway). Using nonribosomal peptide synthetase (NRPS) genes as markers, it has been shown that the Planktothrix community comprises distinct genetic variants displaying differences in bloom dynamics, suggesting a Planktothrix subpopulation structure. Here, we investigate the Planktothrix variants inhabiting four lakes in southeast of Norway utilizing both NRPS and non-NRPS genes. Phylogenetic analyses showed similar topologies for both NRPS and non-NRPS genes, and the lakes appear to have similar structuring of Planktothrix genetic variants. The structure of distinct variants was also supported by very low genetic diversity within variants compared to the between-variant diversity. Incongruent topologies and split decomposition revealed recombination events between Planktothrix variants. In several strains the gene variants seem to be a result of recombination. Both NRPS and non-NRPS genes are dominated by purifying selection; however, sites subjected to positive selection were also detected. The presence of similar and well-separated Planktothrix variants with low internal genetic diversity indicates gene flow within Planktothrix populations. Further, the low genetic diversity found between lakes (similar range as within lakes) indicates gene flow also between Planktothrix populations and suggests recent, or recurrent, dispersals. Our data also indicate that recombination has resulted in new genetic variants. Stability within variants and the development of new variants are likely to be influenced by selection patterns and within-variant homologous recombination.