Secretome analysis identifies potential virulence factors of Diplodia corticola, a fungal pathogen involved in cork oak (Quercus suber) decline.
Identifieur interne : 002002 ( Main/Exploration ); précédent : 002001; suivant : 002003Secretome analysis identifies potential virulence factors of Diplodia corticola, a fungal pathogen involved in cork oak (Quercus suber) decline.
Auteurs : Isabel Fernandes [Belgique] ; Artur Alves [Portugal] ; Ant Nio Correia [Portugal] ; Bart Devreese [Belgique] ; Ana Cristina Esteves [Portugal]Source :
- Fungal biology [ 1878-6146 ]
English descriptors
- KwdEn :
- Ascomycota (chemistry), Ascomycota (genetics), Ascomycota (isolation & purification), Ascomycota (metabolism), Electrophoresis, Gel, Two-Dimensional, Fungal Proteins (chemistry), Fungal Proteins (genetics), Fungal Proteins (metabolism), Molecular Sequence Data, Plant Diseases (microbiology), Protein Transport, Proteomics, Quercus (microbiology), Virulence Factors (chemistry), Virulence Factors (genetics), Virulence Factors (metabolism).
- MESH :
- chemical , chemistry : Fungal Proteins, Virulence Factors.
- chemistry : Ascomycota.
- genetics : Ascomycota, Fungal Proteins, Virulence Factors.
- isolation & purification : Ascomycota.
- metabolism : Ascomycota, Fungal Proteins, Virulence Factors.
- microbiology : Plant Diseases, Quercus.
- Electrophoresis, Gel, Two-Dimensional, Molecular Sequence Data, Protein Transport, Proteomics.
Abstract
The characterisation of the secretome of phytopathogenic fungi may contribute to elucidate the molecular mechanisms of pathogenesis. This is particularly relevant for Diplodia corticola, a fungal plant pathogen belonging to the family Botryosphaeriaceae, whose genome remains unsequenced. This phytopathogenic fungus is recognised as one of the most important pathogens of cork oak, being related to the decline of cork oak forests in the Iberian Peninsula. Unfortunately, secretome analysis of filamentous fungi is limited by the low protein concentration and by the presence of many interfering substances, such as polysaccharides, which affect the separation and analysis by 1D and 2D gel electrophoresis. We compared six protein extraction protocols concerning their suitability for further application with proteomic workflows. The protocols involving protein precipitation were the most efficient, with emphasis on TCA-acetone protocol, allowing us to identify the most abundant proteins on the secretome of this plant pathogen. Approximately 60% of the spots detected were identified, all corresponding to extracellular proteins. Most proteins identified were carbohydrate degrading enzymes and proteases that may be related to D. corticola pathogenicity. Although the secretome was assessed in a noninfection environment, potential virulence factors such as the putative glucan-β-glucosidase, neuraminidase, and the putative ferulic acid esterase were identified. The data obtained forms a useful basis for a deeper understanding of the pathogenicity and infection biology of D. corticola. Moreover, it will contribute to the development of proteomics studies on other members of the Botryosphaeriaceae.
DOI: 10.1016/j.funbio.2014.04.006
PubMed: 24863480
Affiliations:
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Electronic address: acesteves@ua.pt.</nlm:affiliation><country xml:lang="fr">Portugal</country><wicri:regionArea>Department of Biology, CESAM, Aveiro University, Campus Universitário de Santiago, 3810-193 Aveiro</wicri:regionArea><wicri:noRegion>3810-193 Aveiro</wicri:noRegion></affiliation></author></analytic><series><title level="j">Fungal biology</title><idno type="ISSN">1878-6146</idno></series></biblStruct></sourceDesc></fileDesc><profileDesc><textClass><keywords scheme="KwdEn" xml:lang="en"><term>Ascomycota (chemistry)</term><term>Ascomycota (genetics)</term><term>Ascomycota (isolation & purification)</term><term>Ascomycota (metabolism)</term><term>Electrophoresis, Gel, Two-Dimensional</term><term>Fungal Proteins (chemistry)</term><term>Fungal Proteins (genetics)</term><term>Fungal Proteins (metabolism)</term><term>Molecular Sequence Data</term><term>Plant Diseases (microbiology)</term><term>Protein Transport</term><term>Proteomics</term><term>Quercus (microbiology)</term><term>Virulence Factors (chemistry)</term><term>Virulence Factors (genetics)</term><term>Virulence Factors (metabolism)</term></keywords><keywords scheme="MESH" type="chemical" qualifier="chemistry" xml:lang="en"><term>Fungal Proteins</term><term>Virulence Factors</term></keywords><keywords scheme="MESH" qualifier="chemistry" xml:lang="en"><term>Ascomycota</term></keywords><keywords scheme="MESH" qualifier="genetics" xml:lang="en"><term>Ascomycota</term><term>Fungal Proteins</term><term>Virulence Factors</term></keywords><keywords scheme="MESH" qualifier="isolation & purification" xml:lang="en"><term>Ascomycota</term></keywords><keywords scheme="MESH" qualifier="metabolism" xml:lang="en"><term>Ascomycota</term><term>Fungal Proteins</term><term>Virulence Factors</term></keywords><keywords scheme="MESH" qualifier="microbiology" xml:lang="en"><term>Plant Diseases</term><term>Quercus</term></keywords><keywords scheme="MESH" xml:lang="en"><term>Electrophoresis, Gel, Two-Dimensional</term><term>Molecular Sequence Data</term><term>Protein Transport</term><term>Proteomics</term></keywords></textClass></profileDesc></teiHeader><front><div type="abstract" xml:lang="en">The characterisation of the secretome of phytopathogenic fungi may contribute to elucidate the molecular mechanisms of pathogenesis. This is particularly relevant for Diplodia corticola, a fungal plant pathogen belonging to the family Botryosphaeriaceae, whose genome remains unsequenced. This phytopathogenic fungus is recognised as one of the most important pathogens of cork oak, being related to the decline of cork oak forests in the Iberian Peninsula. Unfortunately, secretome analysis of filamentous fungi is limited by the low protein concentration and by the presence of many interfering substances, such as polysaccharides, which affect the separation and analysis by 1D and 2D gel electrophoresis. We compared six protein extraction protocols concerning their suitability for further application with proteomic workflows. The protocols involving protein precipitation were the most efficient, with emphasis on TCA-acetone protocol, allowing us to identify the most abundant proteins on the secretome of this plant pathogen. Approximately 60% of the spots detected were identified, all corresponding to extracellular proteins. Most proteins identified were carbohydrate degrading enzymes and proteases that may be related to D. corticola pathogenicity. Although the secretome was assessed in a noninfection environment, potential virulence factors such as the putative glucan-β-glucosidase, neuraminidase, and the putative ferulic acid esterase were identified. The data obtained forms a useful basis for a deeper understanding of the pathogenicity and infection biology of D. corticola. Moreover, it will contribute to the development of proteomics studies on other members of the Botryosphaeriaceae.</div></front></TEI><affiliations><list><country><li>Belgique</li><li>Portugal</li></country><region><li>Province de Flandre-Orientale</li></region><settlement><li>Gand</li></settlement><orgName><li>Université de Gand</li></orgName></list><tree><country name="Belgique"><region name="Province de Flandre-Orientale"><name sortKey="Fernandes, Isabel" sort="Fernandes, Isabel" uniqKey="Fernandes I" first="Isabel" last="Fernandes">Isabel Fernandes</name></region><name sortKey="Devreese, Bart" sort="Devreese, Bart" uniqKey="Devreese B" first="Bart" last="Devreese">Bart Devreese</name></country><country name="Portugal"><noRegion><name sortKey="Alves, Artur" sort="Alves, Artur" uniqKey="Alves A" first="Artur" last="Alves">Artur Alves</name></noRegion><name sortKey="Correia, Ant Nio" sort="Correia, Ant Nio" uniqKey="Correia A" first="Ant Nio" last="Correia">Ant Nio Correia</name><name sortKey="Esteves, Ana Cristina" sort="Esteves, Ana Cristina" uniqKey="Esteves A" first="Ana Cristina" last="Esteves">Ana Cristina Esteves</name></country></tree></affiliations></record>Pour manipuler ce document sous Unix (Dilib)
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