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Additional file 7. Differentially expressed proteins of TSH1188 and Catongo during interaction with M. perniciosa subjected PPI analysis. Networks of up regulated (A) and down regulated (B) proteins in TSH1188 at 72HAI. Networks of up... more
Additional file 7. Differentially expressed proteins of TSH1188 and Catongo during interaction with M. perniciosa subjected PPI analysis. Networks of up regulated (A) and down regulated (B) proteins in TSH1188 at 72HAI. Networks of up regulated (C) and down regulated (D) proteins in Catongo at 72HAI. Network nodes represent proteins in which each node represents all the protein by a single protein-coding gene locus. Small nodes indicate proteins of unknown 3D structure, large nodes indicate proteins which 3D structures are known or predict (can be visualized by close-up the nodes). Different line colors indicate the types of evidence for the associations. Query proteins not connected with network were removed for better visualization.
Additional file 1. Example of bidimensional gels (triplicates) highlighting the TSH1188 genotype in 45DAI infected with M. perniciosa. Total proteins extract (500 μg) were focused on IPG strips (13 cm), pH ranging from 3 to 10 NL,... more
Additional file 1. Example of bidimensional gels (triplicates) highlighting the TSH1188 genotype in 45DAI infected with M. perniciosa. Total proteins extract (500 μg) were focused on IPG strips (13 cm), pH ranging from 3 to 10 NL, separated by SDS-PAGE (12.5%) and stained with CBB G-250.
Identification of spots marked with arrows on the reference gel. (XLS 213 kb)
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Dispersion graph among the SDS-PAGE replicates. (TIF 489 kb)
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Orthologous sequences of genes from Ustilago maydis used to construct the network along with their annotations. (XLS 33 kb)
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Gene ontology for the biological processes of all proteins in the network using BiNGOA plugin, and the cluster analysis performed with the MCode plugin. (XLSX 145 kb)
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Distribution of the spots and their respective isoelectric point in the 0, 2 and 4 hag. (TIFF 741 kb)
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Sequence alignment of the target proteins identified in the western blots and their homologous in plant recognized by the antibodies. A – Sequence alignment between BiP from A. thaliana and the HSP70 from Moniliophthora perniciosa. B –... more
Sequence alignment of the target proteins identified in the western blots and their homologous in plant recognized by the antibodies. A – Sequence alignment between BiP from A. thaliana and the HSP70 from Moniliophthora perniciosa. B – Sequence alignment of the ATP synthase from A. thaliana and its homologous in Moniliophthora perniciosa. C – Sequence alignment of the catalase A from A. thaliana and its homologous in Moniliophthora perniciosa. (JPEG 3987 kb)
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Additional file 9. Detailed protein extraction method.
Additional file 5. List of complete differentially Expressed Proteins identified in Catongo.
Table S6. List of common and unique clusters and singletons. (XLSX 16 kb)
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Table S5. Annotation of the CSEPs. (XLSX 50 kb)
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Table S4. Lists of CSEPs. (XLSX 26 kb)
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Figure S1. Venn diagrams: comparison of results of the effectors: (I) Nuclear Location Signal (NLS), (II) small and cysteine rich (SCR), and (III) repeats containing protein (RCP) and effectors predicted by EffectorP. (DOCX 759 kb)
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Additional file 8. Complete list of orthologous proteins subjected to PPI analysis.
Table S1. Bacteria identified and classified according to phylum, class, order, family, and genus for in the genotypes CCN51 and Catongo, with a threshold of 99% identity against the Greengen database version 13.8 16S rRNA. (DOCX 19 kb)
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Figure S3. Quantification of libraries. (A) Electrophoresis on 1% (w / v) agarose gel with the six standards, 12 libraries (quantified in triplicates) and three negative controls – a, b and c: first biological sample - CCN51; d, e and f:... more
Figure S3. Quantification of libraries. (A) Electrophoresis on 1% (w / v) agarose gel with the six standards, 12 libraries (quantified in triplicates) and three negative controls – a, b and c: first biological sample - CCN51; d, e and f: first biological sample - Catongo; g, h and i: second biological sample - CCN51; j, k and l: second biological sample - Catongo; NC: negative control. (B) Dissociation curve – a: libraries; b: negative control. (DOCX 307 kb)
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Figure S1. Distribution of plants in the greenhouse. (A) Selected plants: green (first biological), red (second biological). (B) Panoramic photo of plants. (DOCX 2745 kb)
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Additional file 3. Venn diagrams representing the total number of spots detected in both genotypes and treatments. Spots are discriminated by their occurrence: Gray dashed circles represent non-inoculated treatments; black circles... more
Additional file 3. Venn diagrams representing the total number of spots detected in both genotypes and treatments. Spots are discriminated by their occurrence: Gray dashed circles represent non-inoculated treatments; black circles represent inoculated treatments. In the diagram's intersections the total number of common spots and the number of common significantly altered with FC ≥ 1.5 are shown.
Additional file 4. List of complete differentially Expressed Proteins identified in TSH1188.
Additional file 2. Principal Component Analysis showing the grouping of samples regarding different treatments. In A, biplot for all treatments of the Catongo genotype. B, biplot for all treatments of the TSH1188 genotype. C, biplot for... more
Additional file 2. Principal Component Analysis showing the grouping of samples regarding different treatments. In A, biplot for all treatments of the Catongo genotype. B, biplot for all treatments of the TSH1188 genotype. C, biplot for all treatments of the two genotypes analyzed together. Each dot represents a triplicate, named as follows: Initial sequence letters representing the genotypes, followed by the numbers represented by the treatment period, 72HAI and 45DAI and the final letters representing the inoculated (I) and not inoculated (N) treatment.
Gene ontology for the biological processes of all proteins in the network using BiNGOA plugin, and the cluster analysis performed with the MCode plugin. (XLSX 145 kb)
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Table S3. Repeat elements in the genomes. (DOCX 14 kb)
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Cocoa is a fruit of great economic importance, being the main raw material in the manufacture of chocolate. Among the stages of pre-processing, the main and most important is the spontaneous fermentation of the cocoa pulp by... more
Cocoa is a fruit of great economic importance, being the main raw material in the manufacture of chocolate. Among the stages of pre-processing, the main and most important is the spontaneous fermentation of the cocoa pulp by microorganisms, especially the yeasts, which initiate the process and contribute to the death of the germ of the seed, releasing compounds that directly influence the quality of the final product (flavor and aroma). Poorly fermented almonds confer bitter and astringent taste on chocolate, so it is advantageous to select autochthonous yeasts with better performance in the fermentation (producing enzymes of interest in the process) to be used as inoculum starter when added in the spontaneous fermentation, where they can accelerate the fermentation and contribute to raising the quality of the product. Therefore, the objective of this work was to qualitatively determine the production of enzymes of biotechnological interest by yeasts for the fermentation of cocoa th...
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The hemibiotrophic pathogens Moniliophthora perniciosa (witches' broom disease) and Moniliophthora roreri (frosty pod rot disease) are among the most important pathogens of cacao. Moniliophthora perniciosa has a broad host range and... more
The hemibiotrophic pathogens Moniliophthora perniciosa (witches' broom disease) and Moniliophthora roreri (frosty pod rot disease) are among the most important pathogens of cacao. Moniliophthora perniciosa has a broad host range and infects a variety of meristematic tissues in cacao plants, whereas M. roreri infects only pods of Theobroma and Herrania genera. Comparative pathogenomics of these fungi is essential to understand Moniliophthora infection strategies, therefore the detection and in silico functional characterization of effector candidates are important steps to gain insight on their pathogenicity. Candidate secreted effector proteins repertoire were predicted using the genomes of five representative isolates of M. perniciosa subpopulations (three from cacao and two from solanaceous hosts), and one representative isolate of M. roreri from Peru. Many putative effectors candidates were identified in M. perniciosa: 157 and 134 in cacao isolates from Bahia, Brazil; 109 in ...
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Water scarcity can elicit drastic changes in plant metabolic and hormonal regulation, which may be of fundamental importance to stress tolerance. The study of plant the metabolic alterations in response to water deficit, especially the... more
Water scarcity can elicit drastic changes in plant metabolic and hormonal regulation, which may be of fundamental importance to stress tolerance. The study of plant the metabolic alterations in response to water deficit, especially the effects of the rootstocks level, is important to elucidate the mechanisms associated to drought tolerance. To verify the influence of rootstock and grafting on the tolerance to drought in citrus plants, we analyzed the growth, phytohormone levels and flavonoid profiles in grafted and ungrafted citrus plants subjected to different soil water regimes on plant status (well-watered, moderate drought and severe drought and rehydrated) under field conditions. The experiments were conducted under field conditions in the Brazilian Agricultural Research Corporation (EMBRAPA), Cruz das Almas, BA, Brazil. Water deficit reduced the total leaf area per plant in all canopy/rootstock combinations. Self-grafting reduce root volume, area and length when compared to un...
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Eutirucallin is a lectin isolated from the latex of Euphorbia tirucalli, a plant known for its medical properties. The present study explores various characteristics of Eutirucallin including stability, cytotoxicity against tumor cells,... more
Eutirucallin is a lectin isolated from the latex of Euphorbia tirucalli, a plant known for its medical properties. The present study explores various characteristics of Eutirucallin including stability, cytotoxicity against tumor cells, antimicrobial and antiparasitic activities. Eutirucallin was stable from 2 to 40 days at 4°C, maintained hemagglutinating activity within a restricted range, and showed optimal activity at pH 7.0-8.0. Eutirucallin presented antiproliferative activity for HeLa, PC3, MDA-MB-231, and MCF-7 tumor cells but was not cytotoxic for non-tumorigenic cells such as macrophages and fibroblasts. Eutirucallin inhibited the Ehrlich ascites carcinoma in vivo and it was also observed that Eutirucallin inhibited 62.5% of Escherichia coli growth. Also, Eutirucallin showed to be effective when tested directly against Toxoplasma gondii infection in vitro. Therefore, this study sheds perspectives for pharmacological applications of Eutirucallin.
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Oxalic acid (OA) and Nep1-like proteins (NLP) are recognized as elicitors of programmed cell death (PCD) in plants, which is crucial for the pathogenic success of necrotrophic plant pathogens and involves reactive oxygen species (ROS). To... more
Oxalic acid (OA) and Nep1-like proteins (NLP) are recognized as elicitors of programmed cell death (PCD) in plants, which is crucial for the pathogenic success of necrotrophic plant pathogens and involves reactive oxygen species (ROS). To determine the importance of oxalate as a source of ROS for OA- and NLP-induced cell death, a full-length cDNA coding for an oxalate decarboxylase (FvOXDC) from the basidiomycete Flammulina velutipes, which converts OA into CO2 and formate, was overexpressed in tobacco plants. The transgenic plants contained less OA and more formic acid compared with the control plants and showed enhanced resistance to cell death induced by exogenous OA and MpNEP2, an NLP of the hemibiotrophic fungus Moniliophthora perniciosa. This resistance was correlated with the inhibition of ROS formation in the transgenic plants inoculated with OA, MpNEP2, or a combination of both PCD elicitors. Taken together, these results have established a pivotal function for oxalate as a...
Research Interests: Genetics, Microbiology, Plant Biology, Biology, Plant Molecular Biology, and 15 moreMedicine, Genetically Modified Crops, Plant diseases, Reactive Oxygen Species, Cell Death, Programmed cell death, Molecular Plant Microbe Interactions, Fusarium oxysporum, Necrosis, Defense Responses, Calcium Oxalate, Oxalic acid, Plant Leaves, Agaricales, and reverse transcriptase polymerase chain reaction
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Research Interests: Genetics, Biology, Oxidative Stress, Molecular Genetics, Medicine, and 10 moreWood, Industry, Xylem, Eucalyptus, High Performance Liquid Chromatography, Liquid Chromatography / Electrospray Ionization Mass Spectrometry, Two-Dimensional Gel Electrophoresis, Two dimensional Gel Electrophoresis, PLANT PROTEINS, and Paper
Altered physiology, cell structure, and gene expression of Theobroma cacao seedlings subjected to Cu toxicity
Distribution of the spots and their respective molecular weight in the 0, 2 and 4 hag. (TIFF 2477 kb)
Table S2. Bacterial genera exclusive to the phylloplane of the genotypes CCN51 and Catongo. (DOCX 17 kb)
Theobroma cacao is cultivated in the shade, in a so-called 'Cabruca' system, in intercropped with Erithryna or other tree species of economic value, and in full sun as a monoculture in irrigated or chemically-irrigated systems.... more
Theobroma cacao is cultivated in the shade, in a so-called 'Cabruca' system, in intercropped with Erithryna or other tree species of economic value, and in full sun as a monoculture in irrigated or chemically-irrigated systems. Since it is a species quite intolerant to wind, it is practically impossible to implant cacao crops under full exposure to the sun, or in areas of frequent winds, without the protection of windbreaks, using arboreal species around the area of culture in the form of box. Wind can cause mechanical stimuli in plants, affecting their growth and development. The objective of this work was to evaluate the photosynthetic changes in mature leaves and the molecular, biochemical and ultrastructural changes in young and mature leaves of the CCN 51 cloned genotype of T. cacao subjected to intermittent (IW) and constant (CW) wind, with velocities of 2.5, 3.5 and 4.5 m s-1, during 3, 6 and 12 h of exposure. It was verified that CW and IW, considering different expo...
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The cell surface of Toxoplasma gondii is covered by antigens (SAGs) from the SRS family anchored by glycosylphosphatidylinositol (GPI) and includes antigens from the SAG2 family. Among these, the SAG2A surface antigen shows great... more
The cell surface of Toxoplasma gondii is covered by antigens (SAGs) from the SRS family anchored by glycosylphosphatidylinositol (GPI) and includes antigens from the SAG2 family. Among these, the SAG2A surface antigen shows great potential in activating humoral responses and has been used in characterizing the acute phase of infection and in the serological diagnosis of toxoplasmosis. In this study, we aimed to evaluate rSAG2A-induced proteins in BALB/c and C57BL/c mice macrophages and to evaluate the phenotypic polarization induced in the process. We treated the peritoneal macrophages from mouse strains that were resistant or susceptible to T. gondii with rSAG2A to analyze their proteomic profile by mass spectrometry and systems biology. We also examined the gene expression of these cells by RT-qPCR using the phenotypic markers of M1 and M2 macrophages. Differences were observed in the expression of proteins involved in the inflammatory process in both resistant and susceptible cel...
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Functional screening of metagenomic libraries is an important tool for the discovery of new molecules. The metabolic diversity of microorganisms enables survival in harsh environments and is related to the production of enzymes. In this... more
Functional screening of metagenomic libraries is an important tool for the discovery of new molecules. The metabolic diversity of microorganisms enables survival in harsh environments and is related to the production of enzymes. In this study, we identified a protease-producing clone from a metagenomic library derived from mangrove sediment. The protease was purified by ammonium sulphate precipitation and gel filtration chromatography, with a yield of 77.27% and a specific activity of 8.57 U μg(-1) . It had a molecular weight of approximately 70 kDa. MS/MS in ESI-Q-TOF revealed nine peptides similar to a peptidase of Bacillus safensis. The aligned partial sequence showed 47.48% identity and 82.74% similarity to the conserved domains of a glutamyl aminopeptidase from the human gut metagenome and 32.12% total coverage. The protease had an optimal pH of 8.5 and optimal activity at 60°C. At pH 9-12, its activity was greater than 80%. It had moderate thermotolerance and thermostability a...
Research Interests: Microbiology, Wetlands, Kinetics, Biotechnology, Medical Microbiology, and 15 moreBiology, Brazil, Metagenomics, Mangrove, Medicine, Sequence alignment, Bacillus, Temperature, Tandem Mass Spectrometry, Substrate Specificity, Molecular weight, Amino Acid Sequence, Salts, Geologic Sediments, and Metagenome
Cadmium (Cd) is a highly toxic metal for plants, even at low concentrations in the soil. The annual production of world cocoa beans is approximately 4 million tons. Most of these fermented and dried beans are used in the manufacture of... more
Cadmium (Cd) is a highly toxic metal for plants, even at low concentrations in the soil. The annual production of world cocoa beans is approximately 4 million tons. Most of these fermented and dried beans are used in the manufacture of chocolate. Recent work has shown that the concentration of Cd in these beans has exceeded the critical level (0.6mgkg(-1) DM). The objective of this study was to evaluate the toxicity of Cd in young plants of CCN 51 cacao genotype grown in soil with different concentrations of Cd (0, 0.05 and 0.1gkg(-1) soil) through photosynthetic, antioxidative, molecular and ultrastructural changes. The increase of Cd concentration in the soil altered mineral nutrient absorption by competition or synergism, changed photosynthetic activity caused by reduction in chloroplastidic pigment content and damage to the photosynthetic machinery evidenced by the Fv/Fm ratio and expression of the psbA gene and increased GPX activity in the root and SOD in leaves. Additionally,...
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White-rot basidiomycetes are the organisms that decompose lignin most efficiently and Trametes villosa is a promising species for the ligninolytic enzymes production. There are several publications on Trametes villosa applications for... more
White-rot basidiomycetes are the organisms that decompose lignin most efficiently and Trametes villosa is a promising species for the ligninolytic enzymes production. There are several publications on Trametes villosa applications for lignin degradation regarding the expression and secretion of laccase (Lac) and manganese peroxidase (MnP) but no reports on the identification and characterization of lignin peroxidase (LiP), a relevant enzyme for the efficient breaking down of lignin. The object of this study was to identify and partially characterize, for the first time, gDNA, mRNA and the corresponding lignin peroxidase (TvLiP) protein from a strain of Trametes villosa CCMB561 from the Brazilian semi-arid region. The presence of ligninolytic enzymes produced by this strain grown in inducer media was qualitatively and quantitatively analyzed by spectrophotometry, qPCR, and dye fading using RBBR. The spectrophotometric analysis showed that LiP activity was higher than that of MnP. The...