Janaina Rigonato

The bacterial diversity in a Brazilian non-disturbed mangrove sediment

The bacterial diversity present in sediments of a well-preserved mangrove in Ilha do Cardoso, located in the extreme south of São Paulo State coastline, Brazil, was assessed using culture-independent molecular approaches (denaturing gradient gel electrophoresis (DGGE) and analysis of 166 sequences from a clone library). The data revealed a bacterial community dominated by Alphaproteobacteria (40.36% of clones), Gammaproteobacteria (19.28% of clones) and Acidobacteria (27.71% of clones), while minor components of the assemblage were affiliated to Betaproteobacteria, Deltaproteobacteria, Firmicutes, Actinobacteria and Bacteroidetes. The clustering and redundancy analysis (RDA) based on DGGE were used to determine factors that modulate the diversity of bacterial communities in mangroves, such as depth, seasonal fluctuations, and locations over a transect area from the sea to the land. Profiles of specific DGGE gels showed that both dominant (‘universal’ Bacteria and Alphaproteobacteria) and low-density bacterial communities (Betaproteobacteria and Actinobacteria) are responsive to shifts in environmental factors. The location within the mangrove was determinant for all fractions of the community studied, whereas season was significant for Bacteria, Alphaproteobacteria, and Betaproteobacteria and sample depth determined the diversity of Alphaproteobacteria and Actinobacteria.

Drivers of cyanobacterial diversity and community composition in mangrove soils in south-east Brazil

Cyanobacteria act as primary producers of carbon and nitrogen in nutrient-poor ecosystems such as mangroves. This important group of microorganisms plays a critical role in sustaining the productivity of mangrove ecosystems, but the structure and function of cyanobacteria assemblages can be perturbed by anthropogenic influences. The aim of this work was to assess the community structure and ecological drivers that influence the cyanobacterial community harboured in two Brazilian mangrove soils, and examine the long-term effects of oil contamination on these keystone species. Community fingerprinting results showed that, although cyanobacterial communities are distinct between the two mangroves, the structure and diversity of the assemblages exhibit similar responses to environmental gradients. In each ecosystem, cyanobacteria occupying near-shore areas were similar in composition, indicating importance of marine influences for structuring the community. Analysis of 16S rRNA sequences revealed the presence of diverse cyanobacterial communities in mangrove sediments, with clear differences among mangrove habitats along a transect from shore to forest. While near-shore sites in both mangroves were mainly occupied by Prochlorococcus and Synechococcus genera, sequences retrieved from other mangrove niches were mainly affiliated with uncultured cyanobacterial 16S rRNA. The most intriguing finding was the large number of potentially novel cyanobacteria 16S rRNA sequences obtained from a previously oil-contaminated site. The abundance of cyanobacterial 16S rRNA sequences observed in sites with a history of oil contamination was significantly lower than in the unimpacted areas. This study emphasized the role of environmental drivers in determining the structure of cyanobacterial communities in mangrove soils, and suggests that anthropogenic impacts may also act as ecological filters that select cyanobacterial taxa. These results are an important contribution to our understanding of the composition and relative abundance of previously poorly described cyanobacterial assemblages in mangrove ecosystems.

Draft Genome Sequence of the Brazilian Toxic Bloom-Forming Cyanobacterium Microcystis aeruginosa Strain SPC777

ABSTRACT Microcystis aeruginosa strain SPC777 is an important toxin-producing cyanobacterium, isolated from a water bloom of the Billings reservoir (São Paulo State, Brazil). Here, we report the draft genome sequence and initial findings from a preliminary analysis of strain SPC777, including several gene clusters involved in nonribosomal and ribosomal synthesis of secondary metabolites.

The Selection Exerted by Oil Contamination on Mangrove Fungal Communities

Mangrove ecosystems are tropical environments that are characterized by the interaction between the land and the sea. As such, this ecosystem is vulnerable to oil spills. Here, we show a culture-independent survey of fungal communities that are found in the sediments of the following two mangroves that are located on the coast of Sao Paulo State (Brazil): (1) an oil-spill-affected mangrove and (2) a nearby unaffected mangrove. Samples were collected from each mangrove forest at three distinct locations (transect from sea to land), and the samples were analyzed by quantitative PCR and internal transcribed spacer (ITS)-based PCR-DGGE analysis. The abundance of fungi was found to be higher in the oil-affected mangrove. Visual observation and correspondence analysis (CA) of the ITS-based PCR-DGGE profiles revealed differences in the fungal communities between the sampled areas. Remarkably, the oil-spilled area was quite distinct from the unaffected sampling areas. On the basis of the ITS sequences, fungi that are associated with the Basidiomycota and Ascomycota taxa were most common and belonged primarily to the genera Epicoccum, Nigrospora, and Cladosporium. Moreover, the Nigrospora fungal species were shown to be sensitive to oil, whereas a group that was described as “uncultured Basidiomycota” was found more frequently in oil-contaminated areas. Our results showed an increase in fungal abundance in the oil-polluted mangrove regions, and these data indicated potential fungal candidates for remediation of the oil-affected mangroves.

Cyanobacteria in mangrove ecosystems

Mangroves are subject to the effects of tides and fluctuations in environmental conditions, which may reach extreme conditions. These ecosystems are severely threatened by human activities despite their ecological importance. Although mangroves are characterized by a highly specialized but low plant diversity in comparison to most other tropical ecosystems, they support a diverse microbial community. Adapted microorganisms in soil, water, and on plant surfaces perform fundamental roles in nutrient cycling, especially nitrogen and phosphorus. Cyanobacteria contribute to carbon and nitrogen fixation and their cells act as phosphorus storages in ecosystems with extreme or oligotrophic environmental conditions such as those found in mangroves. As the high plant productivity in mangroves is only possible due to interactions with microorganisms, cyanobacteria may contribute to these ecosystems by providing fixed nitrogen, carbon, and herbivory-defense molecules, xenobiotic biosorption and bioremediation, and secreting plant growth-promoting substances. In addition to water, cyanobacterial colonies have been detected on sediments, rocks, decaying wood, underground and aerial roots, trunks, and leaves. Some mangrove cyanobacteria were also found in association to algae or seagrasses. Few studies on mangrove cyanobacteria are available, but together they have reported a substantial number of species in these ecosystems. However, the cyanobacterial diversity in this biome has been traditionally underestimated. Though mangrove communities generally host cyanobacterial taxa commonly found in marine environments, unique microhabitats found in mangroves potentially harbor several undescribed cyanobacterial taxa. The relevance of cyanobacteria for mangrove conservation is highlighted in their use for the recovery of degraded mangroves as biostimulants or in bioremediation.

Phyllonema aviceniicola gen. nov., sp nov and Foliisarcina bertiogensis gen. nov., sp nov., epiphyllic cyanobacteria associated with Avicennia schaueriana leaves

Cyanobacteria dwelling on the salt-excreting leaves of the mangrove tree Avicennia schaueriana were isolated and characterized by ecological, morphological and genetic approaches. Leaves were collected in a mangrove with a history of oil contamination on the coastline of São Paulo state, Brazil, and isolation was achieved by smearing leaves on the surface of solid media or by submerging leaves in liquid media. Twenty-nine isolated strains were shown to belong to five cyanobacterial orders (thirteen to Synechococcales, seven to Nostocales, seven to Pleurocapsales, one to Chroococcales, and one to Oscillatoriales) according to morphological and 16S rRNA gene sequence evaluations. More detailed investigations pointed six Rivulariacean and four Xenococcacean strains as novel taxa. These strains were classified as Phyllonema gen. nov. (type species Phyllonema aviceniicola sp. nov. with type strain CENA341T) and Foliisarcina gen. nov. (type species Foliisarcina bertiogensis sp. nov. with type strain CENA333T), according to the International Code of Nomenclature for Algae, Fungi, and Plants. This investigation shows some of the unique cyanobacteria inhabiting the phyllosphere of Avicennia schaueriana can be retrieved by culturing techniques, improving current taxonomy and providing new insights into the evolution, ecology, and biogeography of this phylum.

Estimating genetic structure and diversity of cyanobacterial communities in Atlantic forest phyllosphere

Cyanobacterial communities on the phyllosphere of 4 plant species inhabiting the endangered Brazilian Atlantic Forest biome were evaluated using cultivation-independent molecular approaches. Total genomic DNA was extracted from cells detached from the surface of leaves of Euterpe edulis, Guapira opposita, Garcinia gardneriana, and Merostachys neesii sampled in 2 Brazilian Atlantic Forest locations along an elevational gradient, i.e., lowland and montane forest. The DNA fingerprinting method PCR-DGGE revealed that the cyanobacterial phyllosphere community structures were mainly influenced by the plant species; geographical location of the plant had little effect. The 16S rRNA gene sequences obtained by clone libraries showed a predominance of nitrogen-fixing cyanobacteria of the order Nostocales, even though the majority of retrieved operational taxonomic units (∼60% of the sequences) showed similarity only to uncultured cyanobacteria phylotypes. The leaf surface of Guapira opposita had the highest richness and diversity of cyanobacteria, whereas the M. neesii (bamboo) had the largest number of copies of cyanobacterial 16S rRNA gene per cm2 of leaf. This study investigated cyanobacteria diversity and its distribution pattern in Atlantic forest phyllosphere. The results indicated that plant species is the main driver of cyanobacteria community assemblage in the phyllosphere and that these communities are made up of a high diversity of cyanobacterial taxa that need to be discovered.

Sphaerocavum : a coccoid morphogenus identical to Microcystis in terms of 16S rDNA and ITS sequence phylogenies

Sphaerocavum is a typical planktonic coccoid cyanobacterium that co-occurs with Microcystis species, and it is commonly found in blooms. Sphaerocavum is a morphogenus distinguishable from Microcystis, as it presents hollow colonies and two-plane cell division. However, Sphaerocavum phylogenetic position based on marker gene(s) has not yet been established. Here, we combined morphological and molecular analyses to address the taxonomic and phylogenetic placement of Sphaerocavum isolated from Brazilian eutrophic lakes. Phylogenetic analysis revealed that Sphaerocavum and Microcystis clustered together in a well-established clade, thus not supporting the Sphaerocavum designation as a divergent genus. The type strain and isolated lineages from Brazilian tropical regions were grouped in the ITS phylogenetic analysis apart from other Microcystis strains. As such, although the infrageneric recognition of Microcystis by molecular markers is still unclear, our analyses strongly indicate that Sphaerocavum must be considered as a morphotype of Microcystis. Thus, taking together our morphological and molecular analyses, we propose the inclusion of Sphaerocavum within the genus Microcystis. In addition, we suggest that the strains used in this study should be named as Microcystis brasiliensis (Azevedo & Sant’Anna) Rigonato et al. comb. nov., with CCIBt3094 as the type strain.

Draft Genome Assembly of the Bloom-Forming Cyanobacterium Nodularia spumigena Strain CENA596 in Shrimp Production Ponds

ABSTRACT We report here the draft genome assembly of the brackish cyanobacterium Nodularia spumigena strain CENA596 isolated from a shrimp production pond in Rio Grande do Sul, Brazil. The draft genome consists of 291 contigs with a total size of 5,189,679 bp. Secondary metabolite annotations resulted in several predicted gene clusters, including those responsible for encoding the hepatotoxin nodularin.

Temporal assessment of microbial communities in soils of two contrasting mangroves

Variations in microbial communities promoted by alterations in environmental conditions are reflected in similarities/differences both at taxonomic and functional levels. Here we used a natural gradient within mangroves from seashore to upland, to contrast the natural variability in bacteria, cyanobacteria and diazotroph assemblages in a pristine area compared to an oil polluted area along a timespan of three years, based on ARISA (bacteria and cyanobacteria) and nifH T-RFLP (diazotrophs) fingerprinting. The data presented herein indicated that changes in all the communities evaluated were mainly driven by the temporal effect in the contaminated area, while local effects were dominant on the pristine mangrove. A positive correlation of community structure between diazotrophs and cyanobacteria was observed, suggesting the functional importance of this phylum as nitrogen fixers in mangroves soils. Different ecological patterns explained the microbial behavior in the pristine and polluted mangroves. Stochastic models in the pristine mangrove indicate that there is not a specific environmental factor that determines the bacterial distribution, while cyanobacteria and diazotrophs better fitted in deterministic model in the same area. For the contaminated mangrove site, deterministic models better represented the variations in the communities, suggesting that the presence of oil might change the microbial ecological structures over time. Mangroves represent a unique environment threatened by global change, and this study contributed to the knowledge of the microbial distribution in such areas and its response on persistent contamination historic events.