Filamentous Cyanobacteria Research Articles

Phytoplankton settling quality has a subtle but significant effect on sediment microeukaryotic and bacterial communities

In coastal aphotic sediments, organic matter (OM) input from phytoplankton is the primary food resource for benthic organisms. Current observations from temperate ecosystems like the Baltic Sea report a decline in spring bloom diatoms, while summer cyanobacteria blooms are becoming more frequent and intense. These climate-driven changes in phytoplankton communities may in turn have important consequences for benthic biodiversity and ecosystem functions, but such questions are not yet sufficiently explored experimentally. Here, in a 4-week experiment, we investigated the response of microeukaryotic and bacterial communities to different types of OM inputs comprising five ratios of two common phytoplankton species in the Baltic Sea, the diatom Skeletonema marinoi and filamentous cyanobacterium Nodularia spumigena. Metabarcoding analyses on 16S and 18S ribosomal RNA (rRNA) at the experiment termination revealed subtle but significant changes in diversity and community composition of microeukaryotes in response to settling OM quality. Sediment bacteria were less affected, although we observed a clear effect on denitrification gene expression (nirS and nosZ), which was positively correlated with increasing proportions of cyanobacteria. Altogether, these results suggest that future changes in OM input to the seafloor may have important effects on both the composition and function of microbenthic communities.

Transcriptional responses of Trichodesmium to natural inverse gradients of Fe and P availability

The filamentous diazotrophic cyanobacterium Trichodesmium is responsible for a significant fraction of marine di-nitrogen (N2) fixation. Growth and distribution of Trichodesmium and other diazotrophs in the vast oligotrophic subtropical gyres is influenced by iron (Fe) and phosphorus (P) availability, while reciprocally influencing the biogeochemistry of these nutrients. Here we use observations across natural inverse gradients in Fe and P in the North Atlantic subtropical gyre (NASG) to demonstrate how Trichodesmium acclimates in situ to resource availability. Transcriptomic analysis identified progressive upregulation of known iron-stress biomarker genes with decreasing Fe availability, and progressive upregulation of genes involved in the acquisition of diverse P sources with decreasing P availability, while genes involved in N2 fixation were upregulated at the intersection under moderate Fe and P availability. Enhanced N2 fixation within the Fe and P co-stressed transition region was also associated with a distinct, consistent metabolic profile, including the expression of alternative photosynthetic pathways that potentially facilitate ATP generation required for N2 fixation with reduced net oxygen production. The observed response of Trichodesmium to availability of both Fe and P supports suggestions that these biogeochemically significant organisms employ unique molecular, and thus physiological responses as adaptations to specifically exploit the Fe and P co-limited niche they construct.

The primary transcriptome of hormogonia from a filamentous cyanobacterium defined by cappable-seq.

Hormogonia are motile filaments produced by many filamentous cyanobacteria that function in dispersal, phototaxis and the establishment of nitrogen-fixing symbioses. The gene regulatory network promoting hormogonium development is initiated by the hybrid histidine kinase HrmK, which in turn activates a sigma factor cascade consisting of SigJ, SigC and SigF. In this study, cappable-seq was employed to define the primary transcriptome of developing hormogonia in the model filamentous cyanobacterium Nostoc punctiforme ATCC 29133 in both the wild-type, and sigJ, sigC and sigF mutant strains 6 h post-hormogonium induction. A total of 1544 transcriptional start sites (TSSs) were identified that are associated with protein-coding genes and are expressed at levels likely to lead to biologically relevant transcripts in developing hormogonia. TSS expression among the sigma-factor deletion strains was highly consistent with previously reported gene expression levels from RNAseq experiments, and support the current working model for the role of these genes in hormogonium development. Analysis of SigJ-dependent TSSs corroborated the presence of the previously identified J-Box in the -10 region of SigJ-dependent promoters. Additionally, the data presented provides new insights on sequence conservation within the -10 regions of both SigC- and SigF-dependent promoters, and demonstrates that SigJ and SigC coordinate complex co-regulation not only of hormogonium-specific genes at different loci, but within an individual operon. As progress continues on defining the hormogonium gene regulatory network, this data set will serve as a valuable resource.

Composition and Dominance of Edible and Inedible Phytoplankton Predict Responses of Baltic Sea Summer Communities to Elevated Temperature and CO2.

Previous studies with Baltic Sea phytoplankton combining elevated seawater temperature with CO2 revealed the importance of size trait-based analyses, in particular dividing the plankton into edible (>5 and <100 µm) and inedible (<5 and >100 µm) size classes for mesozoopankton grazers. While the edible phytoplankton responded predominantly negative to warming and the inedible group stayed unaffected or increased, independent from edibility most phytoplankton groups gained from CO2. Because the ratio between edible and inedible taxa changes profoundly over seasons, we investigated if community responses can be predicted according to the prevailing composition of edible and inedible groups. We experimentally explored the combined effects of elevated temperatures and CO2 concentrations on a late-summer Baltic Sea community. Total phytoplankton significantly increased in response to elevated CO2 in particular in combination with temperature, driven by a significant gain of the inedible <5 µm fraction and large filamentous cyanobacteria. Large flagellates disappeared. The edible group was low as usual in summer and decreased with both factors due to enhanced copepod grazing and overall decline of small flagellates. Our results emphasize that the responses of summer communities are complex, but can be predicted by the composition and dominance of size classes and groups.

Non-conventional octameric structure of C-phycocyanin

C-phycocyanin (CPC), a blue pigment protein, is an indispensable component of giant phycobilisomes, which are light-harvesting antenna complexes in cyanobacteria that transfer energy efficiently to photosystems I and II. X-ray crystallographic and electron microscopy (EM) analyses have revealed the structure of CPC to be a closed toroidal hexamer by assembling two trimers. In this study, the structural characterization of non-conventional octameric CPC is reported for the first time. Analyses of the crystal and cryogenic EM structures of the native CPC from filamentous thermophilic cyanobacterium Thermoleptolyngbya sp. O–77 unexpectedly illustrated the coexistence of conventional hexamer and novel octamer. In addition, an unusual dimeric state, observed via analytical ultracentrifugation, was postulated to be a key intermediate structure in the assemble of the previously unobserved octamer. These observations provide new insights into the assembly processes of CPCs and the mechanism of energy transfer in the light-harvesting complexes.

Geobiology of Andean Microbial Ecosystems Discovered in Salar de Atacama, Chile.

The Salar de Atacama in the Chilean Central Andes harbors unique microbial ecosystems due to extreme environmental conditions, such as high altitude, low oxygen pressure, high solar radiation, and high salinity. Combining X-ray diffraction analyses, scanning electron microscopy and molecular diversity studies, we have characterized twenty previously unexplored Andean microbial ecosystems in eight different lakes and wetlands from the middle-east and south-east regions of this salt flat. The mats and microbialites studied are mainly formed by calcium carbonate (aragonite and calcite) and halite, whereas the endoevaporites are composed predominantly of gypsum and halite. The carbonate-rich mats and microbialites are dominated by Bacteroidetes and Proteobacteria phyla. Within the phylum Proteobacteria, the most abundant classes are Alphaproteobacteria, Gammaproteobacteria and Deltaproteobacteria. While in the phylum Bacteroidetes, the most abundant classes are Bacteroidia and Rhodothermia. Cyanobacteria, Chloroflexi, Planctomycetes, and Verrucomicrobia phyla are also well-represented in the majority of these systems. Gypsum endoevaporites, on the contrary, are dominated by Proteobacteria, Bacteroidetes, and Euryarchaeota phyla. The Cyanobacteria phylum is also abundant in these systems, but it is less represented in comparison to mats and microbialites. Regarding the eukaryotic taxa, diatoms are key structural components in most of the microbial ecosystems studied. The genera of diatoms identified were Achnanthes, Fallacia, Halamphora, Mastogloia, Navicula, Nitzschia, and Surirella. Normally, in the mats and microbialites, diatoms form nano-globular carbonate aggregates with filamentous cyanobacteria and other prokaryotic cells, suggesting their participation in the mineral precipitation process. This work expands our knowledge of the microbial ecosystems inhabiting the extreme environments from the Central Andes region, which is important to ensure their protection and conservation.

Harnessing Nutritional Benefits of Spirulina platensis: Standardization of Cultivating Conditions of Spirulina in Kilimanjaro

Malnutrition remains a challenge in Tanzania, notwithstanding government initiatives and health education geared towards assuaging the problem. According to the World Health Organization (WHO), there will be more than 600,000 severely malnourished children in Tanzania by the year 2030. In particular, protein based malnutrition remains the greatest challenge due to the irreplaceable nature of its essential amino acids. Macronutrients and micronutrients which are found in Spirulina platensis have been recommended by WHO to address malnutrition in developing countries. Spirulina platensis is a filamentous Cyanobacterium microalgae with the highest recorded protein content of plant origin with several immune boosting nutrients. Spirulina cultivation requires sufficient aeration, proper light intensity and salinity for maximum biomass yield, cell productivity, specific growth rate, and protein content. This paper presents the findings of a study carried out in Kilimanjaro on the optimized conditions, locally and economically tailored approach system required to grow spirulina in the region. The study established the use of food grade organic media with low-cost urea as nitrogen source, a greenhouse average temperature of 30–32 °C in the months between December to March, alkalinity of 5 g/L, mixing frequency of 3 times per day/100 L and partial shading, as the optimum conditions for outdoor cultivation of spirulina.&#x0D; Keywords: Malnutrition, Spirulina, Protein, growth condition

Functions of the Essential Gene mraY in Cellular Morphogenesis and Development of the Filamentous Cyanobacterium Anabaena PCC 7120.

Bacterial cell shape is determined by the peptidoglycan (PG) layer. The cyanobacterium Anabaena sp. PCC 7120 (Anabaena) is a filamentous strain with ovoid-shaped cells connected together with incomplete cell constriction. When deprived of combined nitrogen in the growth medium, about 5–10% of the cells differentiate into heterocysts, cells devoted to nitrogen fixation. It has been shown that PG synthesis is modulated during heterocyst development and some penicillin-binding proteins (PBPs) participating in PG synthesis are required for heterocyst morphogenesis or functioning. Anabaena has multiple PBPs with functional redundancy. In this study, in order to examine the function of PG synthesis and its relationship with heterocyst development, we created a conditional mutant of mraY, a gene necessary for the synthesis of the PG precursor, lipid I. We show that mraY is required for cell and filament integrity. Furthermore, when mraY expression was being limited, persistent septal PG synthetic activity was observed, resulting in increase in cell width. Under non-permissive conditions, filaments and cells were rapidly lysed, and no sign of heterocyst development within the time window allowed was detected after nitrogen starvation. When mraY expression was being limited, a high percentage of heterocyst doublets were found. These doublets are formed likely as a consequence of delayed cell division and persistent septal PG synthesis. MraY interacts with components of both the elongasome and the divisome, in particular those directly involved in PG synthesis, including HetF, which is required for both cell division and heterocyst formation.

Cryoconite – From minerals and organic matter to bioengineered sediments on glacier's surfaces

Cryoconite is a mixture of mineral and organic material covering glacial ice, playing important roles in biogeochemical cycles and lowering the albedo of a glacier surface. Understanding the differences in structure of cryoconite across the globe can be important in recognizing past and future changes in supraglacial environments and ice-organisms-minerals interactions. Despite the worldwide distribution and over a century of studies, the basic characteristics of cryoconite, including its forms and geochemistry, remain poorly studied. The major purpose of our study is the presentation and description of morphological diversity, chemical and photoautotrophs composition, and organic matter content of cryoconite sampled from 33 polar and mountain glaciers around the globe. Observations revealed that cryoconite is represented by various morphologies including loose and granular forms. Granular cryoconite includes smooth, rounded, or irregularly shaped forms; with some having their surfaces covered by cyanobacteria filaments. The occurrence of granules increased with the organic matter content in cryoconite. Moreover, a major driver of cryoconite colouring was the concentration of organic matter and its interplay with minerals. The structure of cyanobacteria and algae communities in cryoconite differs between glaciers, but representatives of cyanobacteria families Pseudanabaenaceae and Phormidiaceae, and algae families Mesotaeniaceae and Ulotrichaceae were the most common. The most of detected cyanobacterial taxa are known to produce polymeric substances (EPS) that may cement granules. Organic matter content in cryoconite varied between glaciers, ranging from 1% to 38%. The geochemistry of all the investigated samples reflected local sediment sources, except of highly concentrated Pb and Hg in cryoconite collected from European glaciers near industrialized regions, corroborating cryoconite as element-specific collector and potential environmental indicator of anthropogenic activity. Our work supports a notion that cryoconite may be more than just simple sediment and instead exhibits complex structure with relevance for biodiversity and the functioning of glacial ecosystems.

Draft Genome Sequence of the Phototropic Cyanobacterium Rivularia sp. Strain IAM M-261

ABSTRACTRivularia sp. strain IAM M-261 is a filamentous cyanobacterium with tapering morphology and basal-apical polarity. The apical filament of this cyanobacterium exhibits positive phototropism toward visible light. To elucidate the molecular basis for this phototropism, we determined the draft genome sequence of this strain.

Morphogenesis of digitate structures in hot spring silica sinters of the El Tatio geothermal field, Chile

In silica‐rich hot spring environments, internally laminated, digitate sinter deposits are often interpreted as bio‐mediated structures. The organic components of microbial communities (cell surfaces, sheaths and extracellular polymeric substances) can act as templates for silica precipitation, therefore influencing digitate sinter morphogenesis. In addition to biologic surface‐templating effects, various microenvironmental factors (hydrodynamics, local pH and fluctuating wind patterns) can also influence silica precipitation, and therefore the morphology of resulting digitate sinters. Digitate sinter morphology thus depends on the dynamic interplay between microenvironmentally driven silica precipitation and microbial growth, but the relative contributions of both factors are a topic of continuing research. Here we present a detailed study of digitate silica sinters in distal, low‐temperature regimes of the El Tatio geothermal field, Chile. This high‐altitude geothermal field is extremely arid and windy, and has one of the highest silica precipitation rates found in the world. We find that digitate silica sinters at El Tatio always accrete into the prevailing eastward wind direction and exhibit laminar growth patterns coinciding with day–night cycles of wind‐ and thermally driven evaporation and rewetting. Subaerial parts of digitate sinters lack preserved organics and sinter textures that would indicate past microbial colonization, while filamentous cyanobacteria with resistant, silicified sheaths only inhabit subaqueous cavities that crosscut the primary laminations. We conclude that, although fragile biofilms of extremophile micro‐organisms may have initially been present and templated silica precipitation at the tips of these digitate sinters, the saltation of sand grains and precipitation of silica by recurrent wind‐ and thermally driven environmental forcing at El Tatio are important, if not dominant factors shaping the morphology of these digitate structures. Our study sheds light on the relative contributions of biogenic and abiogenic factors in sinter formation in geothermal systems, with geobiological implications for the cautious interpretation of stromatolite‐like features in ancient silica deposits on Earth and Mars.

Phenotypic plasticity and developmental noise in hybrid and parental clones of Daphnia longispina complex

According to the “temporal hybrid superiority hypothesis”, seasonal variability in environmental factors in temperate lakes gives hybrid clones within the D. longispina complex a temporary fitness advantage, thus allowing long-term, dynamic coexistence of hybrids and maternal taxa. However, the maintenance of hybrids would not require their superiority under any given set of environmental conditions if their average fitness over longer periods surpassed that of more specialized and less flexible parental clones. Phenotypic plasticity and developmental noise of several hybrid and maternal clones of Daphnia (Daphnia galeata, Daphnia hyalina, their hybrids and backcrosses) were compared in a series of laboratory experiments. Changes in depth selection and body size at first reproduction were scored in Daphnia exposed to predator (planktivorous fish) threat, to the presence of filamentous cyanobacteria (Cylindrospermopsis raciborskii), and to the presence of toxic compounds (PCB52 and PCB153). The hybrid clones were found to exhibit the broadest phenotypic plasticity of the studied traits in response to the different stress factors. Developmental noise in depth selection behaviour was the lowest in Daphnia galeata, the highest in Daphnia hyalina, and intermediate in hybrid and backcross clones. This diversity of reaction norms might permit the coexistence of closely related Daphnia clones in the variable and often unpredictable lake environment.

Phylogenetic, morphological and biochemical studies on Thermospirulina andreolii gen. & sp. nov. (Cyanophyta) from the Euganean Thermal District (Italy)

ABSTRACT The therapeutic muds of the Euganean Thermal District, the oldest and largest thermal site in Europe, are recognized by the Italian Health System as an excellent treatment for osteoarticular diseases of inflammatory origin. The beneficial effects that have been demonstrated for these muds are due not only to heat and electrolytes, but also to bioactive compounds released from the microbial community, mainly composed of cyanobacteria, living in the thermal mud surface. We isolated and characterized a new coil-shaped filamentous cyanobacterium through a polyphasic approach. Molecular, morphological and biochemical data supported the description of the microorganism as a new species, Thermospirulina andreolii, of the new genus Thermospirulina, which includes other Spirulina-like cyanobacteria living in different thermal springs around the world. Biochemical analyses highlighted the capability of the strain to synthesize high amounts of phycocyanin and exopolysaccharides, with possible antioxidant and anti-inflammatory activities, suggesting its potential for contributing to the healing properties of the Euganean muds.

Evaluation of inducible promoter-riboswitch constructs for heterologous protein expression in the cyanobacterial species Anabaena sp. PCC 7120.

Cyanobacteria are promising chassis for synthetic biology applications due to the fact that they are photosynthetic organisms capable of growing in simple, inexpensive media. Given their slower growth rate than other model organisms such as Escherichia coli and Saccharomyces cerevisiae, there are fewer synthetic biology tools and promoters available for use in model cyanobacteria. Here, we compared a small library of promoter–riboswitch constructs for synthetic biology applications in Anabaena sp. PCC 7120, a model filamentous cyanobacterium. These constructs were designed from six cyanobacterial promoters of various strengths, each paired with one of two theophylline-responsive riboswitches. The promoter–riboswitch pairs were cloned upstream of a chloramphenicol acetyltransferase (cat) gene, and CAT activity was quantified using an in vitro assay. Addition of theophylline to cultures increased the CAT activity in almost all cases, allowing inducible protein production with natively constitutive promoters. We found that riboswitch F tended to have a lower induced and uninduced production compared to riboswitch E for the weak and medium promoters, although the difference was larger for the uninduced production, in accord with previous research. The strong promoters yielded a higher baseline CAT activity than medium strength and weak promoters. In addition, we observed no appreciable difference between CAT activity measured from strong promoters cultured in uninduced and induced conditions. The results of this study add to the genetic toolbox for cyanobacteria and allow future natural product and synthetic biology researchers to choose a construct that fits their needs.

Environmental factors affecting chytrid (Chytridiomycota) infection rates on Planktothrix agardhii.

Planktothrix agardhii dominates the cyanobacterial harmful algal bloom biomass in Sandusky Bay, Lake Erie (USA) from May until September. This filamentous cyanobacterium known parasites including the chytrid fungal species Rhizophydium sp. C02, which was previously isolated from this region. The purpose of our work has been to establish how parasitic interactions affect Planktothrix population dynamics during a bloom event. Samples analyzed from the 2015 to 2019 bloom seasons using quantitative PCR investigate the spatial and temporal prevalence of chytrid infections. Abiotic factors examined in lab include manipulating temperature (17-31°C), conductivity (0.226-1.225 mS/cm) and turbulence. Planktothrix-specific chytrids are present throughout the bloom period and are occasionally at high enough densities to exert parasitic pressure on their hosts. Temperatures above 27.1°C in lab can inhibit chytrid infection, indicating the presence of a possible upper thermal refuge for the host. Data suggest that chytrids can survive conductivity spikes in lab at levels three-fold above Sandusky Bay waters if given sufficient time (7-12days), whereas increased turbulence in lab severely inhibits chytrid infections, perhaps due to disruption of chemical signaling. Overall, these data provide insights into the environmental conditions that inhibit chytrid infections during Planktothrix-dominated blooms in temperate waters.

A Review of Green Synthesis of Metal Nanoparticles Using Algae.

The ability of algae to accumulate metals and reduce metal ions make them a superior contender for the biosynthesis of nanoparticles and hence they are called bio-nano factories as both the live and dead dried biomass are used for the synthesis of metallic nanoparticles. Microalgae, forming a substantial part of the planet’s biodiversity, are usually single-celled colony-forming or filamentous photosynthetic microorganisms, including several legal divisions like Chlorophyta, Charophyta, and Bacillariophyta. Whole cells of Plectonema boryanum (filamentous cyanobacteria) proved efficient in promoting the production of Au, Ag, and Pt nanoparticles. The cyanobacterial strains of Anabaena flos-aquae and Calothrix pulvinate were used to implement the biosynthesis of Au, Ag, and Pt nanoparticles. Once synthesized within the cells, the nanoparticles were released into the culture media where they formed stable colloids easing their recovery. Lyngbya majuscule and Chlorella vulgaris have been reported to be used as a cost-effective method for Ag nanoparticle synthesis. Dried edible algae (Spirulina platensis) was reported to be used for the extracellular synthesis of Au, Ag, and Au/Ag bimetallic nanoparticles. Synthesis of extracellular metal bio-nanoparticles using Sargassum wightii and Kappaphycus alvarezi has also been reported. Bioreduction of Au (III)-Au (0) using the biomass of brown alga, Fucus vesiculosus, and biosynthesis of Au nanoparticles using red algal (Chondrus crispus) and green algal (Spyrogira insignis) biomass have also been reported. Algae are relatively convenient to handle, less toxic, and less harmful to the environment; synthesis can be carried out at ambient temperature and pressure and in simple aqueous media at a normal pH value. Therefore, the study of algae-mediated biosynthesis of metallic nanoparticles can be taken toward a new branch, termed phyco-nanotechnology.

Scattered migrating colony formation in the filamentous cyanobacterium, Pseudanabaena sp. NIES-4403

BackgroundBacteria have been reported to exhibit complicated morphological colony patterns on solid media, depending on intracellular, and extracellular factors such as motility, cell propagation, and cell-cell interaction. We isolated the filamentous cyanobacterium, Pseudanabaena sp. NIES-4403 (Pseudanabaena, hereafter), that forms scattered (discrete) migrating colonies on solid media. While the scattered colony pattern has been observed in some bacterial species, the mechanism underlying such a pattern still remains obscure.ResultsWe studied the morphology of Pseudanabaena migrating collectively and found that this species forms randomly scattered clusters varying in size and further consists of a mixture of comet-like wandering clusters and disk-like rotating clusters. Quantitative analysis of the formation of these wandering and rotating clusters showed that bacterial filaments tend to follow trajectories of previously migrating filaments at velocities that are dependent on filament length. Collisions between filaments occurred without crossing paths, which enhanced their nematic alignments, giving rise to bundle-like colonies. As cells increased and bundles aggregated, comet-like wandering clusters developed. The direction and velocity of the movement of cells in comet-like wandering clusters were highly coordinated. When the wandering clusters entered into a circular orbit, they turned into rotating clusters, maintaining a more stable location. Disk-like rotating clusters may rotate for days, and the speed of cells within a rotating cluster increases from the center to the outmost part of the cluster. Using a mathematical modeling with simplified assumption we reproduced some features of the scattered pattern including migrating clusters.ConclusionBased on these observations, we propose that Pseudanabaena forms scattered migrating colonies that undergo a series of transitions involving several morphological patterns. A simplified model is able to reproduce some features of the observed migrating clusters.

Stigonema (Nostocales, Cyanobacteria) in the Rhynie chert (Lower Devonian, Scotland)

Filamentous cyanobacteria are morphologically diverse. The most complex morphologies occur in the genus Stigonema (Stigonemataceae, Nostocales), which is heterocystous and characterized by true-branched, multiseriate or uniseriate trichomes surrounded by a sheath. There is virtually no documentation of fossil equivalents to Stigonema. This study describes a specimen of a new filamentous cyanobacterium, Rhystigonema obscurum nov. gen. et sp., from the Lower Devonian Rhynie chert that appears to have been preserved due to the fortuitous circumstance that it was located in a microbial mat at the time of silicification. The specimen consists of a large (>50 μm wide), true-branched, multiseriate trichome enveloped in a prominent sheath. It can satisfactorily be placed in Stigonemataceae based on morphology; were it not for the lack of molecular evidence, it could also be assigned to Stigonema. Rhystigonema obscurum broadens our knowledge of cyanobacterial biodiversity in early non-marine ecosystems.

Phytoplankton Drivers of Dissolved Organic Material Production in Colorado Reservoirs and the Formation of Disinfection By-Products

Previous studies have shown that algal-derived dissolved organic matter (DOM) has a strong influence on the formation of disinfection byproducts (DBPs) during the treatment of drinking water. In the summer of 2010, we evaluated the role of nitrogen and phosphorus loading and phytoplankton abundance as drivers of the concentrations and quality of DOM and the associated DBP formation in 30 reservoirs in the mountains and plains of the State of Colorado. Optical properties such as Specific Ultraviolet Absorbance at 254 nm (SUVA254) and fluorescence spectroscopy were used to characterize DOM quality. Nutrient concentrations such as total nitrogen were also assessed and were associated with high concentrations of chlorophyll a (Chl-a). In turn, high total organic carbon (TOC) concentrations were associated with high concentrations of Chl-a, and the DOM in these reservoirs had a fluorescence signature indicative of contributions from phytoplankton growth. The reservoirs with TOC concentrations above 4 mgC/L were predominantly located in the plains and many are impacted by agricultural runoff and wastewater discharges, rather than in the mountains and are characterized by warm water conditions and shallow depths. For a subset of fourteen reservoirs, we characterized the composition of the phytoplankton using a rapid imaging microscopy technique and observed a dominance by filamentous Cyanobacteria in reservoirs with TOC concentrations above 4 mgC/L. The combination of high TOC concentrations with microbial characteristics resulted in high potential for production of two major classes of regulated DBPs, trihalomethanes and haloacetic acids. While fluorescence spectroscopy was useful in confirming the contribution of phytoplankton growth to high TOC concentrations, evaluation of predictive models for DBP yields found that all equally predictive models included SUVA254 and some of these models also included fluorescence indices or logTOC. These findings provide a limnological context in support of the recent guidelines that have been implemented for protection of high-quality drinking water supplies in the State of Colorado.

Identification of Putative Biosynthetic Gene Clusters for Tolyporphins in Multiple Filamentous Cyanobacteria.

Tolyporphins A–R are unusual tetrapyrrole macrocycles produced by the non-axenic filamentous cyanobacterium HT-58-2. A putative biosynthetic gene cluster for biosynthesis of tolyporphins (here termed BGC-1) was previously identified in the genome of HT-58-2. Here, homology searching of BGC-1 in HT-58-2 led to identification of similar BGCs in seven other filamentous cyanobacteria, including strains Nostoc sp. 106C, Nostoc sp. RF31YmG, Nostoc sp. FACHB-892, Brasilonema octagenarum UFV-OR1, Brasilonema octagenarum UFV-E1, Brasilonema sennae CENA114 and Oculatella sp. LEGE 06141, suggesting their potential for tolyporphins production. A similar gene cluster (BGC-2) also was identified unexpectedly in HT-58-2. Tolyporphins BGCs were not identified in unicellular cyanobacteria. Phylogenetic analysis based on 16S rRNA and a common component of the BGCs, TolD, points to a close evolutionary history between each strain and their respective tolyporphins BGC. Though identified with putative tolyporphins BGCs, examination of pigments extracted from three cyanobacteria has not revealed the presence of tolyporphins. Overall, the identification of BGCs and potential producers of tolyporphins presents a collection of candidate cyanobacteria for genetic and biochemical analysis pertaining to these unusual tetrapyrrole macrocycles.