Carolina Weigert Galvão

Prokaryotic communities of acidic peatlands from the southern Brazilian Atlantic Forest

The acidic peatlands of southern Brazil are ecosystems essential for the maintenance of the Atlantic Forest, one of the 25 hot-spots of biodiversity in the world. In this work, we investigated the composition of prokaryotic communities in four histosols of three acidic peatland regions by constructing small-subunit (SSU) rRNA gene libraries and sequencing. SSU rRNA gene sequence analysis showed the prevalence of Acidobacteria (38.8%) and Proteobacteria (27.4%) of the Bacteria domain and Miscellaneous (58%) and Terrestrial (24%) groups of Crenarchaeota of the Archaea domain. As observed in other ecosystems, archaeal communities showed lower richness than bacterial communities. We also found a limited number of Euryarchaeota and of known methanotrophic bacteria in the clone libraries.

Microbial community and performance of slaughterhouse wastewater treatment filters.

The performance of anaerobic filter bioreactors (AFs) is influenced by the composition of the substrate, support medium, and the microbial species present in the sludge. In this study, the efficiency of a slaughterhouse effluent treatment using three AFs containing different support media was tested, and the microbial diversity was investigated by amplified ribosomal DNA restriction analysis and 16S rRNA gene sequencing. The physicochemical analysis of the AF systems tested suggested their feasibility, with rates of chemical oxygen demand removal of 72±8% in hydraulic retention times of 1 day. Analysis of pH, alkalinity, volatile acidity, total solids, total volatile solids, total Kjeldahl nitrogen, and the microbial community structures indicated high similarity among the three AFs. The composition of prokaryotic communities showed a prevalence of Proteobacteria (27.3%) and Bacteroidetes (18.4%) of the Bacteria domain and Methanomicrobiales (36.4%) and Methanosarcinales (35.3%) of the Archaea domain. Despite the high similarity of the microbial communities among the AFs, the reactor containing pieces of clay brick as a support medium presented the highest richness and diversity of bacterial and archaeal operational taxonomic units.

Effect of ethylene treatment on phytochemical and ethylene-related gene expression during ripening in strawberry fruit Fragaria x ananassa cv. Camino Real.

In contrast to climacteric fruits, in which ethylene is known to be pivotal, the regulation of ripening in non-climacteric fruits is not well understood. The strawberry is a typical example of a non-climacteric fruit, which has been used as a model system of these types of fruit. In this study, the effect of exogenous ethephon on the expression of ethylene biosynthesis and signaling genes, FaERF2 and FaACO1, was analyzed in the Fragaria ananassa cultivar Camino Real by quantitative real-time polymerase chain reaction, and the physicochemical and phytochemical characteristics of fruits were evaluated in field trials and postharvest tests. Transcript accumulation was influenced by exogenous treatment with ethephon, which affected the pattern of gene expression during different stages of growth and fruit development, with the highest expression occurring during postharvest tests. In addition, ethephon significantly influenced the phytochemical profile of sugars, anthocyanins, phenolic compounds, and vitamin C contents both in the field- and postharvest-treated fruits at different stages. These results indicate that ethylene regulates the phenylpropanoid maturation pathway in strawberry fruit.

The RecX protein interacts with the RecA protein and modulates its activity in Herbaspirillum seropedicae

DNA repair is crucial to the survival of all organisms. The bacterial RecA protein is a central component in the SOS response and in recombinational and SOS DNA repairs. The RecX protein has been characterized as a negative modulator of RecA activity in many bacteria. The recA and recX genes of Herbaspirillum seropedicae constitute a single operon, and evidence suggests that RecX participates in SOS repair. In the present study, we show that the H. seropedicae RecX protein (RecXHs) can interact with the H. seropedicae RecA protein (RecAHs) and that RecAHs possesses ATP binding, ATP hydrolyzing and DNA strand exchange activities. RecXHs inhibited 90% of the RecAHs DNA strand exchange activity even when present in a 50-fold lower molar concentration than RecAHs. RecAHs ATP binding was not affected by the addition of RecX, but the ATPase activity was reduced. When RecXHs was present before the formation of RecA filaments (RecA-ssDNA), inhibition of ATPase activity was substantially reduced and excess ssDNA also partially suppressed this inhibition. The results suggest that the RecXHs protein negatively modulates the RecAHs activities by protein-protein interactions and also by DNA-protein interactions.

Ardra profiles of bacteria and archaea in mangrove sediments with different levels of contamination in the estuarine complex of Paranaguá, Brazil

The mangroves sediments from the coastal areas under human activities may contain significant contaminations by hydrocarbons, even when there are no visual evidences of it. The microorganisms are essential to these ecosystems, especially in the control of their chemical environment. Sediment samples were collected in two regions under different environment conditions (pristine and contaminated) of the Paranagua Estuarine Complex (Paranagua Bay and Laranjeiras Bay), Brazil. Aliphatic hydrocarbons were determined by the GC-FID to assess the status of contamination of the studied areas. The total DNA was extracted from these samples. The 16S rRNA gene was amplified by the PCR reactions with the pair of primers 21F and 958R for the archaeal domain, and 27F and 1492R for the bacterial domain. Comparisons of communities were made by the ARDRA technique, using the HinfI restriction enzyme. The phosphate concentration showed significant differences between the two regions. The aliphatic hydrocarbons analysis showed the presence of unresolved complex mixture (UCM), an indicator of oil contamination, in the samples from the Paranagua Bay, which was corroborated by the concentration of total aliphatic hydrocarbons. The ARDRA profile indicated that the structure of archaeal and bacterial communities of the sampled areas was very similar. Therefore, the anthropogenic influences in the Paranagua Bay showed to be not sufficient to produce disturbances in the prokaryotic dominant groups.

Seasonal changes in dominant bacterial taxa from acidic peatlands of the Atlantic Rain Forest.

The acidic peatlands of southern Brazil are essential for maintenance of the Atlantic Rain Forest, one of the 25 hot-spots of biodiversity in the world. While these ecosystems are closely linked to conservation issues, their microbial community ecology and composition remain unknown. In this work, histosol samples were collected from three acidic peatland regions during dry and rainy seasons and their chemical and microbial characteristics were evaluated. Culturing and culture-independent approaches based on SSU rRNA gene pyrosequencing were used to survey the bacterial community and to identify environmental factors affecting the biodiversity and microbial metabolic potential of the Brazilian peatlands. All acidic peatlands were dominated by the Acidobacteria phylum (56-22%) followed by Proteobacteria (28-12%). The OTU richness of these phyla and the abundance of their Gp1, Gp2, Gp3, Gp13, Rhodospirillales and Caulobacteriales members varied according to the period of collection and significantly correlated with the rainy season. However, despite changes in acidobacterial and proteobacterial communities, rainfall did not affect the microbial metabolic potential of the southern Brazilian Atlantic Rain Forest peatlands, as judged by the metabolic capabilities of the microbial community.

Ribopeaks: a web tool for bacterial classification through m/z data from ribosomal proteins

Summary MALDI-TOF MS is a rapid, sensitive and economic tool for bacterial identification. Highly abundant bacterial proteins are detected by this technique, including ribosomal proteins (r-protein), and the generated mass spectra are compared with a MALDI-TOF MS spectra database. Currently, it allows mainly the classification of clinical bacteria due to the limited number of environmental bacteria included in the spectra database. We present a wide-ranging bacterium classifier tool, called Ribopeaks, which was created based on r-protein data from the Genbank. The Ribopeaks database has more than 28 500 bacterial taxonomic records. It compares the incoming m/z data from MALDI-TOF MS analysis with models stored in the Ribopeaks database created by machine learning and then taxonomically classifies the bacteria. Availability and implementation The software is available at http://www.ribopeaks.com. Supplementary information Supplementary data are available at Bioinformatics online.

Wheat colonization by an Azospirillum brasilense ammonium-excreting strain reveals upregulation of nitrogenase and superior plant growth promotion

AimsIn this work, an ammonium-excreting strain (HM053) of A. brasilense was further characterized genetically and biochemically, and its abilities to colonize and promote wheat growth were determined.MethodsImmunoblot, reverse transcription-qPCR, and DNA sequencing were used for HM053 characterization. To analyze wheat-A. brasilense interaction nifH::gusA fusions in the wild-type FP2 (FP2-7) and HM053 (HM053-36) backgrounds were employed.ResultsHM053 glutamine synthetase (GS) was not adenylylated in response to an ammonium shock or under any condition tested. Sequencing of the glnA gene revealed a substitution of a proline residue by a leucine at position 347 of the GS. Under axenic growth condition, HM053 was capable of colonizing the surface of wheat roots and increased by 30 and 49% the shoot and root dry weight, respectively, when compared with uninoculated plants, and by 30 and 31% when compared with the parental strain FP2. Although HM053-36 and FP2-7 showed GUS activity located mainly at lateral root emergence points, HM053-36 consistently showed stronger signals and expressed the nifH gene at a level 278 fold higher than strain FP2 in planta, according to qPCR data.ConclusionsHM053, a spontaneous mutant in GS, increased wheat root and shoot dry weight when compared to the wild-type FP2. HM053 ability to excrete ammonium and fix nitrogen constitutively, even in the presence of high NH4+ concentration, could explain why this mutant has a higher potential to promote plant growth than FP2 and suggests HM053 as a potential nitrogen biofertilizer. However, HM053 should be tested under field conditions to evaluate its abilities to compete with indigenous microflora.

Comparative proteomic analysis between early developmental stages of the Coffea arabica fruits.

Coffee is one of the most valuable agricultural commodities. There is much agronomic research on coffee, but molecular knowledge of its fruit development and ripening is limited. This study reports a comparative proteomic investigation of immature coffee fruits in two early developmental stages: stage 1, cell division and elongation of the perisperm; and stage 2, early growth of the endosperm progressively replacing the perisperm. Proteins were extracted using a modified SDS-phenol method and two-dimensional electrophoresis gels stained with Coomassie blue revealed about 300 well-resolved polypeptide spots in the pH range of 3 to 10. The differentially expressed polypeptides spots were excised, trypsin-digested, and analyzed by MALDI-TOF mass spectrometry. Peptide MS data were searched against the coffee EST database. Most of the identified protein spots are involved in the glycolytic pathway and energy reserve, and are more highly expressed at stage 2.

Invasion ecology applied to inoculation of plant growth promoting bacteria through a novel SIMPER-PCA approach

AimsPlant growth promoting bacteria (PGPB) have been used on crops for years, but inoculants that are efficient in some locations may not be efficient in others. Here, we applied classical invasion ecology theory to PGPB inoculation in order to identify patterns that can be used to predict plant growth promoting (PGP) efficiency. The hypotheses that the inoculant that causes most impact will be the most efficient PGPB, and that the most invasible locations would have higher PGP efficiency, were tested. We also aim to present our statistical approach to analyze SIMPER results.MethodsUsing next generation sequencing targeting the 16S rDNA gene in metagenomics samples, we analyzed samples of pre-planting bulk soil and rhizosphere of inoculated maize plants. Bacterial communities of inoculated plants were compared to the non-inoculated controls, in order to estimate the inoculant invasion impact. Crop yield was compared to different indexes, and a novel data exploration approach was employed.ResultsThe most efficient inoculant was not the most invasive, and a nutrient per diversity ratio was unable to predict inoculant efficiency or invasion impact. However, the efficient inoculation treatment presented an enrichment of specific pre-planting taxa.ConclusionsInvasion ecology frameworks could not anticipate field results of inoculated plants. Nonetheless, our data exploration approach, which is explained in detail, can be useful to raise new hypothesis and improve the visualization of dissimilarity data.