Publications by authors named "Francisco Javier Fernández-Acero"

17 Publications

  • Page 1 of 1

Development of New Antiproliferative Compound against Human Tumor Cells from the Marine Microalgae by Applied Proteomics.

Int J Mol Sci 2020 Dec 24;22(1). Epub 2020 Dec 24.

Microbiology Laboratory, Institute of Viticulture and Agri-Food Research (IVAGRO), University of Cádiz, Pol. Río San Pedro s/n, 11510 Cádiz, Spain.

Proteomics is a crucial tool for unravelling the molecular dynamics of essential biological processes, becoming a pivotal technique for basic and applied research. Diverse bioinformatic tools are required to manage and explore the huge amount of information obtained from a single proteomics experiment. Thus, functional annotation and protein-protein interactions are evaluated in depth leading to the biological conclusions that best fit the proteomic response in the system under study. To gain insight into potential applications of the identified proteins, a novel approach named "Applied Proteomics" has been developed by comparing the obtained protein information with the existing patents database. The development of massive sequencing technology and mass spectrometry (MS/MS) improvements has allowed the application of proteomics nonmodel microorganisms, which have been deeply described as a novel source of metabolites. Between them, has been pointed out as an alternative source of biomolecules. Recently, our research group has reported the first complete proteome analysis of this microalga, which was analysed using the applied proteomics concept with the identification of 488 proteins with potential industrial applications. To validate our approach, we selected the UCA01 protein from the prohibitin family. The recombinant version of this protein showed antiproliferative activity against two tumor cell lines, Caco2 (colon adenocarcinoma) and HepG-2 (hepatocellular carcinoma), proving that proteome data have been transformed into relevant biotechnological information. From has been developed a new tool against cancer-the protein named UCA01. This protein has selective effects inhibiting the growth of tumor cells, but does not show any effect on control cells. This approach describes the first practical approach to transform proteome information in a potential industrial application, named "applied proteomics". It is based on a novel bioalgorithm, which is able to identify proteins with potential industrial applications. From hundreds of proteins described in the proteome of , the bioalgorithm identified over 400 proteins with potential uses; one of them was selected as UCA01, "in vitro" and its potential was demonstrated against cancer. This approach has great potential, but the applications are potentially numerous and undefined.
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http://dx.doi.org/10.3390/ijms22010096DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7795124PMC
December 2020

New Perspectives Related to the Bioluminescent System in Dinoflagellates: , a Case Study.

Int J Mol Sci 2020 Mar 5;21(5). Epub 2020 Mar 5.

Microbiology Laboratory, Institute of Viticulture and Agri-food Research (IVAGRO), Environmental and Marine Sciences Faculty. University of Cadiz (UCA), 11510 Puerto Real, Spain.

is considered a model organism due to its bioluminescence capacity linked to circadian rhythms. The mechanisms underlying the bioluminescent phenomenon have been well characterized in dinoflagellates; however, there are still some aspects that remain an enigma. Such is the case of the presence and diversity of the luciferin-binding protein (LBP), as well as the synthesis process of luciferin. Here we carry out a review of the literature in relation to the molecular players responsible for bioluminescence in dinoflagellates, with particular interest in . We also carried out a phylogenetic analysis of the conservation of protein sequence, structure and evolutionary pattern of these key players. The basic structure of the luciferase (LCF) is quite conserved among the sequences reported to date for dinoflagellate species, but not in the case of the LBP, which has proven to be more variable in terms of sequence and structure. In the case of luciferin, its synthesis has been shown to be complex process with more than one metabolic pathway involved. The glutathione S-transferase (GST) and the P630 or blue compound, seem to be involved in this process. In the same way, various hypotheses regarding the role of bioluminescence in dinoflagellates are exposed.
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http://dx.doi.org/10.3390/ijms21051784DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7084563PMC
March 2020

An "omic" approach to Pyrocystis lunula: New insights related with this bioluminescent dinoflagellate.

J Proteomics 2019 10 26;209:103502. Epub 2019 Aug 26.

Microbiology Laboratory, Institute of Viticulture and Agri-food Research (IVAGRO), University of Cadiz, 11510 Puerto Real, Spain. Electronic address:

Pyrocystis lunula (Schutt) is a photoautotrophic dinoflagellate without armored form, frequently found in marine environments. Today, there are several biotechnological applications derived from the bioluminescent system of this species. From a post-genomic perspective, in order to have a starting point for studying the proteome of P. lunula, an "omics" approach (transcriptomics-proteomics) was assessed using fresh microalgae samples. A total of 80,874,825 raw reads were generated (11,292,087,505 bp; 55.82% GC) by mRNA sequencing. Very high-quality sequences were assembled into 414,295 contigs (219,203,407 bp; 55.38% GC) using Trinity software, generating a comprehensive reference transcriptome for this species. Then, a P. lunula proteome was inferred and further employed for its analysis on this species. A total of 17,461 peptides were identified, yielding 3182 protein identification hits, including 175 novel proteins. The identified proteins were further categorized according to functional description and gene ontology classification. SIGNIFICANCE: The major contribution of the present work is making available a reference transcriptome and proteome of P. lunula, that is now accessible for the research community, and a functional description of the 3182 proteins inferred from the transcriptome, including 175 novel proteins, which have already been deposited in the ProteomeXchange and NCBI SRA databases, respectively. In addition to this, a series of important factors related to the bioluminescent system and the regulation of gene expression, were identified and described.
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http://dx.doi.org/10.1016/j.jprot.2019.103502DOI Listing
October 2019

Proteomic study of the membrane components of signalling cascades of Botrytis cinerea controlled by phosphorylation.

Sci Rep 2019 07 8;9(1):9860. Epub 2019 Jul 8.

Andalusian Center for Grape and Grapevine Research (IVAGRO), Microbiology Lab, University of Cadiz, Puerto Real, 11510, Spain.

Protein phosphorylation and membrane proteins play an important role in the infection of plants by phytopathogenic fungi, given their involvement in signal transduction cascades. Botrytis cinerea is a well-studied necrotrophic fungus taken as a model organism in fungal plant pathology, given its broad host range and adverse economic impact. To elucidate relevant events during infection, several proteomics analyses have been performed in B. cinerea, but they cover only 10% of the total proteins predicted in the genome database of this fungus. To increase coverage, we analysed by LC-MS/MS the first-reported overlapped proteome in phytopathogenic fungi, the "phosphomembranome" of B. cinerea, combining the two most important signal transduction subproteomes. Of the 1112 membrane-associated phosphoproteins identified, 64 and 243 were classified as exclusively identified or overexpressed under glucose and deproteinized tomato cell wall conditions, respectively. Seven proteins were found under both conditions, but these presented a specific phosphorylation pattern, so they were considered as exclusively identified or overexpressed proteins. From bioinformatics analysis, those differences in the membrane-associated phosphoproteins composition were associated with various processes, including pyruvate metabolism, unfolded protein response, oxidative stress response, autophagy and cell death. Our results suggest these proteins play a significant role in the B. cinerea pathogenic cycle.
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http://dx.doi.org/10.1038/s41598-019-46270-0DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6614480PMC
July 2019

Valorisation of the microalgae Nannochloropsis gaditana biomass by proteomic approach in the context of circular economy.

J Proteomics 2019 02 29;193:239-242. Epub 2018 Oct 29.

Research Project Manager of Innovation Department, Endesa Generación, S.A., Spain.

Nannochloropsis gaditana is a non-flagellated microalgae that has been widely used for different purposes, mostly related with the industrial production of biofuels or aquiculture. However, in order to increase the economic viability of the obtained microalgae biomass from a production plant coupled to a coal power plant, a proteomic approach was initiated by using fresh and atomized microalgae samples, as the main used commercial forms. Above 51,000 high quality spectra were obtained per sample in the MS/MS analysis of whole proteome of N. gaditana, yielding above 7,500 peptides, leading the identification of 1,950 proteins, from the N. gaditana protein database, where 655 proteins were presented in all the replicates. The identified proteins were categorized according to gene ontology classification by molecular function and biological process. In this study, it has been described the first proteomic analysis of the microalgae N. gaditana under industrial conditions containing an important number of identified proteins. A significative presence of proteins with a potential role in different agri-food and biomedical applications was detected and studied being the core of future N. gaditana research to expand the current biotechnological applications of this microalga. SIGNIFICANCE OF THE STUDY: Three quarters of the planet earth correspond to seas and oceans, however its potential biotechnological use is still unknown. We described the first proteomic description of the microalgae N. gaditana under industrial conditions. Following the spirit of the EU initiatives of blue growth and the statements of circular economy, CO waste from a coal plant power has been transformed in a resource for microalgae biomass production, common product presentations were evaluated by proteomic, and its potential use of identified proteins in Agri-food and Biomedicine has been revealed.
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http://dx.doi.org/10.1016/j.jprot.2018.10.015DOI Listing
February 2019

Dataset of the phosphoproteome induced by different plant-based elicitors.

Data Brief 2016 Jun 22;7:1447-1450. Epub 2016 Apr 22.

Andalusian Center for Grape and Grapevine Research, CeiA3, Marine and Environmental Sciences Faculty, University of Cadiz, Puerto Real, 11510 Cádiz, Spain.

Phosphorylation is one of the main post-translational modification (PTM) involved in signaling network in the ascomycete , one of the most relevant phytopathogenic fungus. The data presented in this article provided a differential mass spectrometry-based analysis of the phosphoproteome of under two different phenotypical conditions induced by the use of two different elicitors: glucose and deproteinized Tomate Cell Walls (TCW). A total 1138 and 733 phosphoproteins were identified for glucose and TCW culture conditions respectively. Raw data are deposited at the ProteomeXchange Consortium via the PRIDE partner repository with the data set identifier (PRIDE: http://www.ebi.ac.uk/pride/archive/projects/PXD003099). Further interpretation and discussion of these data are provided in our research article entitled "Phosphoproteome analysis of in response to different plant-based elicitors" (Liñeiro et al., 2016) [1].
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http://dx.doi.org/10.1016/j.dib.2016.04.039DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC5063813PMC
June 2016

Modifications of fungal membrane proteins profile under pathogenicity induction: A proteomic analysis of Botrytis cinerea membranome.

Proteomics 2016 09 5;16(17):2363-76. Epub 2016 Aug 5.

Andalusian Center for Grape and Grapevine Research (IVAGRO), CeiA3, Marine and Environmental Sciences Faculty, University of Cadiz, Puerto Real, Spain.

Botrytis cinerea is a model fungus for the study of phytopathogenicity that exhibits a wide arsenal of tools to infect plant tissues. Most of these factors are related to signal transduction cascades, in which membrane proteins play a key role as a bridge between environment and intracellular molecular processes. This work describes the first description of the membranome of Botrytis under different pathogenicity conditions induced by different plant-based elicitors: glucose and tomato cell wall (TCW). A discovery proteomics analysis of membrane proteins was carried out by mass spectrometry. A total of 2794 proteins were successfully identified, 46% of them were classified as membrane proteins based on the presence of transmembrane regions and lipidation. Further analyses showed significant differences in the membranome composition depending on the available carbon source: 804 proteins were exclusively identified when the fungus was cultured with glucose as a sole carbon source, and 251 proteins were exclusively identified with TCW. Besides, among the 1737 common proteins, a subset of 898 proteins presented clear differences in their abundance. GO enrichment and clustering interaction analysis revealed changes in the composition of membranome with increase of signalling function in glucose conditions and carbohydrate degradation process in TCW conditions. All MS data have been deposited in the ProteomeXchange with identifier PXD003099 (http://proteomecentral.proteomexchange.org/dataset/PXD003099).
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http://dx.doi.org/10.1002/pmic.201500496DOI Listing
September 2016

Phosphoproteome analysis of B. cinerea in response to different plant-based elicitors.

J Proteomics 2016 Apr 18;139:84-94. Epub 2016 Mar 18.

Andalusian Center for Grape and Grapevine Research, CeiA3, Marine and Environmental Sciences Faculty, University of Cadiz, Puerto Real, 11510 Cádiz, Spain. Electronic address:

Unlabelled: The phytopathogen Botrytis cinerea is a ubiquitous fungus with a high capacity to adapt its metabolism to different hosts and environmental conditions in order to deploy a variety of virulence and pathogenicity factors and develop a successful plant infection. Here we report the first comparative phosphoproteomic study of B. cinerea, aimed to analyze the phosphoprotein composition of the fungus and its changes under different phenotypical conditions induced by two different carbon sources as plant based elicitors: glucose and deproteinized tomato cell wall (TCW). A total of 2854 and 2269 different phosphosites (2883 and 1137 phosphopeptides) were identified in glucose and TCW respectively, which map to 1338 phosphoproteins in glucose and 733 in TCW. Out of the identified phosphoproteins, 173 were exclusively found when glucose was the only carbon source and 11 when the carbon source was TCW. Differences in the pattern of phosphorylation-sites were also detected according to the carbon source. Gene ontology classification of the identified phosphoproteins showed that most of the characteristic proteins of the different carbon sources were related to signalling and transmembrane transport, thus highlighting the importance of these processes in the fungal adaptation to the surrounding conditions.

Biological Significance: The characterization of the B. cinerea phosphoproteome under different induction conditions reported here is the first comparative phosphoproteomic approach in this model phytopathogenic fungus. The identified phosphopeptides contribute to expand the map of known phosphoproteins in this pathogen and the observed changes according to the used carbon source contribute to understand the adaptation of the fungus to the environment changes. This knowledge improves the understanding of the adaptation mechanism, defines the role of the phosphoproteins involved in this process, and enables the advance in the design of novel strategies against the fungi.
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http://dx.doi.org/10.1016/j.jprot.2016.03.019DOI Listing
April 2016

Analysis of temperature-mediated changes in the wine yeast Saccharomyces bayanus var uvarum. An oenological study of how the protein content influences wine quality.

Proteomics 2016 Feb 25;16(4):576-92. Epub 2016 Jan 25.

Andalusian Center for Grape and Grapevine Research, CeIA3, Marine and Environmental Sciences Faculty, University of Cadiz, Cádiz, Spain.

Saccharomyces bayanus var. uvarum plays an important role in the fermentation of red wine from the D.O. Ribera del Duero. This is due to the special organoleptic taste that this yeast gives the wines and their ability to ferment at low temperature. To determine the molecular factors involved in the fermentation process at low temperature, a differential proteomic approach was performed by using 2D-DIGE, comparing, qualitatively and quantitatively, the profiles obtained at 13 and 25°C. A total of 152 protein spots were identified. We detected proteins upregulated at 13°C that were shown to be related to temperature stress, the production of aromatic compounds involved in the metabolism of amino acids, and the production of fusel alcohols and their derivatives, each of which is directly related to the quality of the wines. To check the temperature effects, an aromatic analysis by GC-MS was performed. The proteomic and "aromatomic" results are discussed in relation to the oenological properties of S. bayanus var. uvarum.
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http://dx.doi.org/10.1002/pmic.201500137DOI Listing
February 2016

Proteomic profiling of Botrytis cinerea conidial germination.

Arch Microbiol 2015 Mar 21;197(2):117-33. Epub 2014 Aug 21.

Laboratory of Microbiology, Marine and Environmental Sciences Faculty, Andalusian Center for Grape and Grapevine Research, CeiA3 International Campus of Excellence in Agrifood, University of Cádiz, Pol. Río San Pedro s/n, 11510, Puerto Real, Cádiz, Spain.

Botrytis cinerea is one of the most relevant plant pathogenic fungi. The first step during its infection process is the germination of the conidia. Here, we report on the first proteome analysis during the germination of B. cinerea conidia, where 204 spots showed significant differences in their accumulation between ungerminated and germinated conidia by two-dimensional polyacrylamide gel electrophoresis and qPCR. The identified proteins were grouped by gene ontology revealing that the infective tools are mainly preformed inside the ungerminated conidia allowing a quick fungal development at the early stages of conidial germination. From 118 identified spots, several virulence factors have been identified while proteins, such as mannitol-1-phosphate dehydrogenase, 6,7-dimethyl-8-ribityllumazine synthase or uracil phosphoribosyltransferase, have been disclosed as a new potential virulence factors in botrytis whose role in pathogenicity needs to be studied to gain new insights about the role of these proteins as therapeutic targets and virulence factors.
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http://dx.doi.org/10.1007/s00203-014-1029-4DOI Listing
March 2015

Molecular analysis of red wine yeast diversity in the Ribera del Duero D.O. (Spain) area.

Arch Microbiol 2013 May 9;195(5):297-302. Epub 2013 Feb 9.

Laboratorio de Microbiología y Genética, Departamento de Biomedicina, Biotecnología y Salud Pública, Facultad de Ciencias del Mar y Ambientales, Universidad de Cádiz, Polígono del Río San Pedro s/n, 11510 Puerto Real, Cádiz, Spain.

Molecular characterization of wine yeast population during spontaneous fermentation in biodynamic wines from Ribera del Duero D.O. located at northern plateau of Spain has been carried out during two consecutive years. A total of 829 yeast strains were isolated from the samples and characterized by electrophoretic karyotype. The results show the presence of three population of yeast differentiated by their electrophoretic karyotypes, (1) non-Saccharomyces yeast dominant in the initial phase of the fermentations (NS); (2) Saccharomyces bayanus var uvarum detected mainly mid-way through the fermentation process at 20-25 °C; and (3) Saccharomyces cerevisiae which remained dominant until the end of the fermentation. This is the first study showing the population dynamic of S. bayanus var. uvarum in red wines produced in Ribera del Duero that could represent an important source of autochthonous wine yeasts with novel oenological properties.
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http://dx.doi.org/10.1007/s00203-013-0872-zDOI Listing
May 2013

Proteomic analysis of conidia germination in Colletotrichum acutatum.

Arch Microbiol 2013 Apr 1;195(4):227-46. Epub 2013 Feb 1.

Microbiology Laboratory, Wine and Grapewine Research Institute, Faculty of Marine and Environmental Sciences, University of Cádiz, Pol. Río San Pedro s/n, Puerto Real, 11510 Cádiz, Spain.

Colletotrichum acutatum is an important phytopathogenic fungus causing anthracnose in commercially important fruit crops, such as strawberry. The conidia produced by the fungus are survival structures which play a key role in host infection and fungal propagation. Despite its relevance to the fungal life cycle, conidial biology has not been extensively investigated. Here, we provide the first proteomic description of the conidial germination in C. acutatum by comparing the proteomic profiles of ungerminated and germinated conidia. Using two-dimensional electrophoresis combined with MALDI-TOF/TOF mass spectrometry, we have identified 365 proteins in 354 spots, which represent 245 unique proteins, including some proteins with key functions in pathogenesis. All these proteins have been classified according to their molecular function and their involvement in biological processes, including cellular energy production, oxidative metabolism, stress, fatty acid synthesis, protein synthesis, and folding. This report constitutes the first comprehensive study of protein expression during the early stage of the C. acutatum conidial germination. It advances our understanding of the molecular mechanisms involved in the conidial germination process, and provides a useful basis for the further characterization of proteins involved in fungal biology and fungus life cycles.
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http://dx.doi.org/10.1007/s00203-013-0871-0DOI Listing
April 2013

Development of proteomics-based fungicides: new strategies for environmentally friendly control of fungal plant diseases.

Int J Mol Sci 2011 Jan 21;12(1):795-816. Epub 2011 Jan 21.

Laboratory of Microbiology, Faculty of Marine and Environmental Sciences, University of Cádiz, Pol. Río San Pedro s/n, 11510 Puerto Real, Spain.

Proteomics has become one of the most relevant high-throughput technologies. Several approaches have been used for studying, for example, tumor development, biomarker discovery, or microbiology. In this "post-genomic" era, the relevance of these studies has been highlighted as the phenotypes determined by the proteins and not by the genotypes encoding them that is responsible for the final phenotypes. One of the most interesting outcomes of these technologies is the design of new drugs, due to the discovery of new disease factors that may be candidates for new therapeutic targets. To our knowledge, no commercial fungicides have been developed from targeted molecular research, this review will shed some light on future prospects. We will summarize previous research efforts and discuss future innovations, focused on the fight against one of the main agents causing a devastating crops disease, fungal phytopathogens.
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http://dx.doi.org/10.3390/ijms12010795DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC3039980PMC
January 2011

2-DE proteomic approach to the Botrytis cinerea secretome induced with different carbon sources and plant-based elicitors.

Proteomics 2010 Jun;10(12):2270-80

Marine and Environmental Sciences Faculty, University of Cádiz, Puerto Real, Cádiz, Spain.

Botrytis cinerea is a phytopathogenic fungus infecting a number of crops (tomatoes, grapes and strawberries), which has been adopted as a model system in molecular phytopathology. B. cinerea uses a wide variety of infection strategies, which are mediated by a set of genes/proteins called pathogenicity/virulence factors. Many of these factors have been described as secreted proteins, and thus the study of this sub-proteome, the secretome, under changing circumstances can help us to understand the roles of these factors, possibly revealing new loci for the fight against the pathogen. A 2-DE, MALDI TOF/TOF-based approach has been developed to establish the proteins secreted to culture media supplemented with different carbon sources and plant-based elicitors (in this study: glucose, cellulose, starch, pectin and tomato cell walls). Secreted proteins were obtained from the culture media by deoxycholate-trichloroacetic acid/phenol extraction, and 76 spots were identified, yielding 95 positive hits that correspond to 56 unique proteins, including several known virulence factors (i.e. pectin methyl esterases, xylanases and proteases). The observed increases in secretion of proteins with established virulence-related functions indicate that this in vitro-induction/proteome-mining approach is a promising strategy for discovering new pathogenicity factors and dissecting infection mechanisms in a discrete fashion.
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http://dx.doi.org/10.1002/pmic.200900408DOI Listing
June 2010

Proteomic analysis of the phytopathogenic fungus Botrytis cinerea during cellulose degradation.

Proteomics 2009 May;9(10):2892-902

Laboratory of Microbiology, Marine and Environmental Sciences Faculty, University of Cádiz, Pol. Río San Pedro s/n, Puerto Real, Cádiz, Spain.

The ascomycete Botrytis cinerea is a phytopathogenic fungus infecting and causing significant yield losses in a number of crops. Moreover, in the last few years, B. cinerea has been adopted as an important model system in molecular phytopathology. In spite of these contributions, the molecular basis of the infection cycle remains unclear. Proteomic approaches have revealed significant information about the infective cycle of several pathogens, including B. cinerea. The main aim of this study is to make available a proteomic database containing a significant number of identified proteins from B. cinerea. In brief, three independent B. cinerea cultures supplemented with carboxymethylcellulose were used, and the extracted proteins were independently separated by 2-D PAGE to obtain the proteome map from B. cinerea. Two hundred and sixty-seven spots were selected for MALDI TOF/TOF MS analysis, resulting in 303 positive identifications, mostly representing unannotated proteins. Identified proteins were then classified into categories using the PANTHER classification system (www.pantherdb.org), showing the relevance of protein metabolism and modification process and oxidoreductase activity. Since cellulose is one of the major components of the plant cell wall, many of the identified proteins may have a crucial role in the pathogenicity process. In brief, this proteomic map of B. cinerea will be a useful basis for exploring the proteins involved in the infection cycle, which will in turn provide new targets for crop diagnosis and focused fungicide design.
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http://dx.doi.org/10.1002/pmic.200800540DOI Listing
May 2009

Proteomic analysis of phytopathogenic fungus Botrytis cinerea as a potential tool for identifying pathogenicity factors, therapeutic targets and for basic research.

Arch Microbiol 2007 Mar 24;187(3):207-15. Epub 2006 Nov 24.

Laboratory of Microbiology, Marine and Environmental Sciences Faculty, University of Cádiz, Pol. Río San Pedro, 11510 Puerto Real, Cádiz, Spain.

Botrytis cinerea is a phytopathogenic fungus causing disease in a substantial number of economically important crops. In an attempt to identify putative fungal virulence factors, the two-dimensional gel electrophoresis (2-DE) protein profile from two B. cinerea strains differing in virulence and toxin production were compared. Protein extracts from fungal mycelium obtained by tissue homogenization were analyzed. The mycelial 2-DE protein profile revealed the existence of qualitative and quantitative differences between the analyzed strains. The lack of genomic data from B. cinerea required the use of peptide fragmentation data from MALDI-TOF/TOF and ESI ion trap for protein identification, resulting in the identification of 27 protein spots. A significant number of spots were identified as malate dehydrogenase (MDH) and glyceraldehyde-3-phosphate dehydrogenase (GAPDH). The different expression patterns revealed by some of the identified proteins could be ascribed to differences in virulence between strains. Our results indicate that proteomic analysis are becoming an important tool to be used as a starting point for identifying new pathogenicity factors, therapeutic targets and for basic research on this plant pathogen in the postgenomic era.
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http://dx.doi.org/10.1007/s00203-006-0188-3DOI Listing
March 2007

Two-dimensional electrophoresis protein profile of the phytopathogenic fungus Botrytis cinerea.

Proteomics 2006 Apr;6 Suppl 1:S88-96

Laboratory of Microbiology, Marine and Environmental Sciences Faculty, University of Cádiz, Cádiz, Spain.

Botrytis cinerea is a phytopathogenic fungi causing disease in a number of important crops. It is considered a very complex species in which different populations seem to be adapted to different hosts. In order to characterize fungal virulence factors, a proteomic research was started. A protocol for protein extraction from mycelium tissue, with protein separation by 2-DE and MS analysis, was optimised as a first approach to defining the B. cinerea proteome. Around 400 spots were detected in 2-DE CBB-stained gels, covering the 5.4-7.7 pH and 14-85 kDa ranges. The averages of analytical and biological coefficients of variance for 64 independent spots were 16.1% and 37.5%, respectively. Twenty-two protein spots were identified by MALDI-TOF or ESI IT MS/MS, with some of them corresponding to forms of malate dehydrogenase and glyceraldehyde-3-phosphate dehydrogenase. Two more spots matched a cyclophilin and a protein with an unknown function.
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http://dx.doi.org/10.1002/pmic.200500436DOI Listing
April 2006