Pedro N. Leão

Allelopathy in freshwater cyanobacteria.

Freshwater cyanobacteria produce several bioactive secondary metabolites with diverse chemical structure, which may achieve high concentrations in the aquatic medium when cyanobacterial blooms occur. Some of the compounds released by cyanobacteria have allelopathic properties, influencing the biological processes of other phytoplankton or aquatic plants. These kinds of interactions are more easily detectable under laboratory studies; however their ecological relevance is often debated. Recent research has discovered new allelopathic properties in some cyanobacteria species, new allelochemicals and elucidated some of the allelopathic mechanisms. Ecosystem-level approaches have shed some light on the factors that influence allelopathic interactions, as well as how cyanobacteria may be able to modulate their surrounding environment by means of allelochemical release. Nevertheless, the role of allelopathy in cyanobacteria ecology is still not well understood, and its clarification should benefit from carefully designed field studies, chemical characterization of allelochemicals and new methodological approaches at the “omics” level.

Synergistic allelochemicals from a freshwater cyanobacterium

The ability of cyanobacteria to produce complex secondary metabolites with potent biological activities has gathered considerable attention due to their potential therapeutic and agrochemical applications. However, the precise physiological or ecological roles played by a majority of these metabolites have remained elusive. Several studies have shown that cyanobacteria are able to interfere with other organisms in their communities through the release of compounds into the surrounding medium, a phenomenon usually referred to as allelopathy. Exudates from the freshwater cyanobacterium Oscillatoria sp. had previously been shown to inhibit the green microalga Chlorella vulgaris. In this study, we observed that maximal allelopathic activity is highest in early growth stages of the cyanobacterium, and this provided sufficient material for isolation and chemical characterization of active compounds that inhibited the growth of C. vulgaris. Using a bioassay-guided approach, we isolated and structurally characterized these metabolites as cyclic peptides containing several unusually modified amino acids that are found both in the cells and in the spent media of Oscillatoria sp. cultures. Strikingly, only the mixture of the two most abundant metabolites in the cells was active toward C. vulgaris. Synergism was also observed in a lung cancer cell cytotoxicity assay. The binary mixture inhibited other phytoplanktonic organisms, supporting a natural function of this synergistic mixture of metabolites as allelochemicals.

The chemical ecology of cyanobacteria

This review covers the literature on the chemically mediated ecology of cyanobacteria, including ultraviolet radiation protection, feeding-deterrence, allelopathy, resource competition, and signalling. To highlight the chemical and biological diversity of this group of organisms, evolutionary and chemotaxonomical studies are presented. Several technologically relevant aspects of cyanobacterial chemical ecology are also discussed.

Cylindrospermopsis raciborskii: review of the distribution, phylogeography, and ecophysiology of a global invasive species

Cylindrospermopsis raciborskii is a cyanobacterial species extensively studied for its toxicity, bloom formation and invasiveness potential, which have consequences to public and environmental health. Its current geographical distribution, spanning different climates, suggests that C. raciborskii has acquired the status of a cosmopolitan species. From phylogeography studies, a tropical origin for this species seems convincing, with different conjectural routes of expansion toward temperate climates. This expansion may be a result of the species physiological plasticity, or of the existence of different ecotypes with distinct environmental requirements. In particular, C. raciborskii is known to tolerate wide temperature and light regimes and presents diverse nutritional strategies. This cyanobacterium is also thought to have benefited from climate change conditions, regarding its invasiveness into temperate climates. Other factors, recently put forward, such as allelopathy, may also be important to its expansion. The effect of C. raciborskii in the invaded communities is still mostly unknown but may strongly disturb species diversity at different trophic levels. In this review we present an up-to-date account of the distribution, phylogeography, ecophysiology, as well some preliminary reports of the impact of C. raciborskii in different organisms.

Differential protein expression in Corbicula fluminea upon exposure to a Microcystis aeruginosa toxic strain.

Changes in protein expression induced by a Microcystis aeruginosa toxic strain in the freshwater clam Corbicula fluminea were studied using a proteomic approach in an effort to identify new molecular biomarkers. Clams were fed with 1 x 10(6) cells mL(-1) of a M. aeruginosa toxic strain (IZANCYA 2), during 24 b. Cytosolic fractions of gills and digestive tract were analyzed by two-dimensional (2D) electrophoresis in 7 cm IPG strips (pH 4-7). On average, about 400 spots were resolved using Coomassie staining. Altered protein expression was quantitatively detected in 16-13 spots in gills and digestive tract, respectively. In 2D electrophoresis gel protein maps from gills, 10 of 16 spots were downregulated. In the digestive tract, the general tendency was an increase in the protein expression level after the exposure. The altered protein spots were excised and analyzed by MALDI-TOF-MS, with identification of 8 proteins in gills and 5 in the digestive tract. Most of the identified proteins are involved in cytoskeleton assembly. Metabolic proteins were also detected. These results are in agreement with predicted effects of PP1 and PP2A phosphatase inhibition as major effect of microcystins-related toxicity.

Allelopathic activity of cyanobacteria on green microalgae at low cell densities.

Allelopathic interactions are thought to play an important role in phytoplankton ecology. Many cyanobacterial genera have been shown to produce compounds with allelopathic activity; however, most studies have used high cyanobacterial cell densities, typical of bloom situations. We investigated whether low cell densities of cyanobacteria exhibit allelopathic activity. Twenty-two cyanobacterial strains were cultured in laboratory conditions, and the effects of their exudates were tested on the microalgae Ankistrodesmus falcatus and Chlorella vulgaris. We found that the exudates from one strain of Cylindrospermopsis raciborskii caused an enhancement of A. falcatus growth, followed by a marked decrease in growth rates, and that exudates from one strain of Oscillatoria sp. strongly inhibited C. vulgaris, as confirmed by dose–response assays. After the removal of the stressors, the microalgae grew at normal rates. The responsible compounds showed low sensitivity to heat and proteinase treatments. However, the majority of the strains did not exhibit appreciable allelopathic activity. Allelopathy may thus be a less frequent process in cyanobacteria at low cell-densities.

Antitumor Activity of Hierridin B, a Cyanobacterial Secondary Metabolite Found in both Filamentous and Unicellular Marine Strains

Cyanobacteria are widely recognized as a valuable source of bioactive metabolites. The majority of such compounds have been isolated from so-called complex cyanobacteria, such as filamentous or colonial forms, which usually display a larger number of biosynthetic gene clusters in their genomes, when compared to free-living unicellular forms. Nevertheless, picocyanobacteria are also known to have potential to produce bioactive natural products. Here, we report the isolation of hierridin B from the marine picocyanobacterium Cyanobium sp. LEGE 06113. This compound had previously been isolated from the filamentous epiphytic cyanobacterium Phormidium ectocarpi SAG 60.90, and had been shown to possess antiplasmodial activity. A phylogenetic analysis of the 16S rRNA gene from both strains confirmed that these cyanobacteria derive from different evolutionary lineages. We further investigated the biological activity of hierridin B, and tested its cytotoxicity towards a panel of human cancer cell lines; it showed selective cytotoxicity towards HT-29 colon adenocarcinoma cells.

Biosynthesis‐Assisted Structural Elucidation of the Bartolosides, Chlorinated Aromatic Glycolipids from Cyanobacteria

The isolation of the bartolosides, unprecedented cyanobacterial glycolipids featuring aliphatic chains with chlorine substituents and C-glycosyl moieties, is reported. Their chlorinated dialkylresorcinol (DAR) core presented a major structural-elucidation challenge. To overcome this, we discovered the bartoloside (brt) biosynthetic gene cluster and linked it to the natural products through in vitro characterization of the DAR-forming ketosynthase and aromatase. Bioinformatic analysis also revealed a novel potential halogenase. Knowledge of the bartoloside biosynthesis constrained the DAR core structure by defining key pathway intermediates, ultimately allowing us to determine the full structures of the bartolosides. This work illustrates the power of genomics to enable the use of biosynthetic information for structure elucidation.

Role of marine cyanobacteria in trace metal bioavailability in seawater.

In seawater, several trace metals with biological significance are highly complexed with organic matter. Marine cyanobacteria are an important phytoplanktonic group, with the ability to release trace metal-binding compounds to the seawater medium, which in turn modulates their bioavailability and influences their biogeochemical cycles. Such interactions may allow cyanobacteria to more easily access less available trace metals essential for their metabolic processes, or, conversely, keep the toxic forms of the trace metals from reaching intolerable levels. In this minireview, Cu and Fe interactions with cyanobacteria received special attention, although other trace metals (Co, Pb, Zn, and Cd) are also covered. Recent research has shed light on many aspects of trace metal–cyanobacteria ecology in seawater; nevertheless, the biochemical processes behind this dynamics and the structure of the vast majority of the metal binding compounds remain unclear.

Picocyanobacteria From a Clade of Marine Cyanobium Revealed Bioactive Potential Against Microalgae, Bacteria, and Marine Invertebrates

The production of bioactive compounds either toxic or with pharmacological applications by cyanobacteria is well established. However, picoplanktonic forms within this group of organisms have rarely been studied in this context. In this study, the toxicological potential of picocyanobacteria from a clade of marine Cyanobium strains isolated from the Portuguese coast was examined using different biological models. First, strains were identified by applying morphological and molecular approaches and cultured under lab conditions. A crude extract and three fractions reflecting a preliminary segregation of lipophilic metabolites were tested for toxicity with the marine microalga Nannochloropsis sp., the bacteria Pseudomonas sp., the brine shrimp Artemia salina, and fertilized eggs of the sea urchin Paracentrotus lividus. No significant apparent adverse effects were noted against Artemia salina. However, significant adverse effects were found in all other assays, with an inhibition of Nannochloropsis sp. and Pseudomonas sp. growth and marked reduction in Paracentrotus lividus larvae length. The results obtained indicated that Cyanobium genus may serve as a potential source of interesting bioactive compounds and emphasize the importance of also studying smaller picoplanktonic fractions of marine cyanobacteria.