Proteomic study of three component interactions: plant, pathogens and antagonistic fungi
The molecular factors involved in the three-way interaction between plant, pathogenic fungi and antagonistic/biocontrol fungi, such as Trichoderma, are still poorly understood, even if they represent a matter of interest for improving crop management and developing new strategies for plant diseases...
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| Language: | English |
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Zhejiang University Press
2004-07-01
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| Series: | 浙江大学学报. 农业与生命科学版 |
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| Online Access: | https://www.academax.com/doi/10.3785/1008-9209.2004.04.0449 |
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| author | Marra R Ambrosino P Scala V Romano C Vinale F Ferraioli S Ruocco M Carbone V Woo S L Turrà D Scala F Lorito M |
| author_facet | Marra R Ambrosino P Scala V Romano C Vinale F Ferraioli S Ruocco M Carbone V Woo S L Turrà D Scala F Lorito M |
| author_sort | Marra R |
| collection | DOAJ |
| description | The molecular factors involved in the three-way interaction between plant, pathogenic fungi and antagonistic/biocontrol fungi, such as Trichoderma, are still poorly understood, even if they represent a matter of interest for improving crop management and developing new strategies for plant diseases control. The aim of this work is to investigate the components involved in this interaction and, for this purpose, a proteomic approach was used. 2-D maps of the protein extracts from the single components in various interactions between plants (potato, bean, tobacco or tomato), pathogens (Botrytis cinerea, Rhizoctonia solani or Pythium ultimum) and biocontrol fungi (Trichoderma atroviride strain P1 or Trichoderma harzianum strain T22) were obtained. The proteome of each partner was collected separately and extracted by acetone precipitation in presence of trichloroacetic acid and a reducing agent (DTT). The extracted proteins were separated by isoelectrofocusing (IEF), using IPG (Immobilized pH gradient) strips, followed by SDS-PAGE. In order to improve resolution the separations were performed both on wide than narrow pH range and on different gel lengths. Differential spots were noted in the proteome of the three-way interaction when compared to each single component. These were further characterized by mass spectrometry and in silico analysis with the aim of identifying and cloning the relative genes. During the in vitro interaction of T. harzianum strain T22 with tomato and the culture filtrate or cell walls of pathogens, the spot number was higher than in the presence of pathogen biomass. In terms of Trichoderma differential proteins displayed on 2D gels, the most important changes were obtained in the presence of P. ultimum. During the in vivo interaction with tomato, the antagonist proteome changed much more in presence of soilbome fungi R. solani and P. ultimum than with the foliar fungus B. cinerea, both in terms of total and increased or novel spots In silico analysis of some of those spots revealed homology with intracellular enzymes (GTPases, hydrolases) and with stress-related proteins (heat shock proteins HSP70, bacteriocin cloacin). Specific proteins in the plant proteome, i.e. pathogenesis-related proteins, have been identified during the in vivo interaction of bean with R. solani and T. atroviride strain P1. This is in agreement with the demonstrated ability of these beneficial fungi to induce plant systemic disease resistance by activating expression of defence-related genes. Proteins extracted from T. atrovride strain P1 which were analysed by mass spectrometry, revealed some interesting homologies with a fungal hydrophobin of Pleurotus ostreatus and an ABC transporter of Ralstonia metallidurans. These could represent molecular factors involved in the antagonistic mechanisms of Trichoderma and play a role in the three-way interaction with the plant and other microbes. |
| format | Article |
| id | doaj-art-b0d6f87325844eccafd4435cebfbd2fd |
| institution | Kabale University |
| issn | 1008-9209 2097-5155 |
| language | English |
| publishDate | 2004-07-01 |
| publisher | Zhejiang University Press |
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| series | 浙江大学学报. 农业与生命科学版 |
| spelling | doaj-art-b0d6f87325844eccafd4435cebfbd2fd2025-08-20T03:34:18ZengZhejiang University Press浙江大学学报. 农业与生命科学版1008-92092097-51552004-07-013044944910.3785/1008-9209.2004.04.044910089209Proteomic study of three component interactions: plant, pathogens and antagonistic fungiMarra RAmbrosino PScala VRomano CVinale FFerraioli SRuocco MCarbone VWoo S LTurrà DScala FLorito MThe molecular factors involved in the three-way interaction between plant, pathogenic fungi and antagonistic/biocontrol fungi, such as Trichoderma, are still poorly understood, even if they represent a matter of interest for improving crop management and developing new strategies for plant diseases control. The aim of this work is to investigate the components involved in this interaction and, for this purpose, a proteomic approach was used. 2-D maps of the protein extracts from the single components in various interactions between plants (potato, bean, tobacco or tomato), pathogens (Botrytis cinerea, Rhizoctonia solani or Pythium ultimum) and biocontrol fungi (Trichoderma atroviride strain P1 or Trichoderma harzianum strain T22) were obtained. The proteome of each partner was collected separately and extracted by acetone precipitation in presence of trichloroacetic acid and a reducing agent (DTT). The extracted proteins were separated by isoelectrofocusing (IEF), using IPG (Immobilized pH gradient) strips, followed by SDS-PAGE. In order to improve resolution the separations were performed both on wide than narrow pH range and on different gel lengths. Differential spots were noted in the proteome of the three-way interaction when compared to each single component. These were further characterized by mass spectrometry and in silico analysis with the aim of identifying and cloning the relative genes. During the in vitro interaction of T. harzianum strain T22 with tomato and the culture filtrate or cell walls of pathogens, the spot number was higher than in the presence of pathogen biomass. In terms of Trichoderma differential proteins displayed on 2D gels, the most important changes were obtained in the presence of P. ultimum. During the in vivo interaction with tomato, the antagonist proteome changed much more in presence of soilbome fungi R. solani and P. ultimum than with the foliar fungus B. cinerea, both in terms of total and increased or novel spots In silico analysis of some of those spots revealed homology with intracellular enzymes (GTPases, hydrolases) and with stress-related proteins (heat shock proteins HSP70, bacteriocin cloacin). Specific proteins in the plant proteome, i.e. pathogenesis-related proteins, have been identified during the in vivo interaction of bean with R. solani and T. atroviride strain P1. This is in agreement with the demonstrated ability of these beneficial fungi to induce plant systemic disease resistance by activating expression of defence-related genes. Proteins extracted from T. atrovride strain P1 which were analysed by mass spectrometry, revealed some interesting homologies with a fungal hydrophobin of Pleurotus ostreatus and an ABC transporter of Ralstonia metallidurans. These could represent molecular factors involved in the antagonistic mechanisms of Trichoderma and play a role in the three-way interaction with the plant and other microbes.https://www.academax.com/doi/10.3785/1008-9209.2004.04.0449differential proteins<italic>in vivo</italic> interactionsinduced disease resistance |
| spellingShingle | Marra R Ambrosino P Scala V Romano C Vinale F Ferraioli S Ruocco M Carbone V Woo S L Turrà D Scala F Lorito M Proteomic study of three component interactions: plant, pathogens and antagonistic fungi 浙江大学学报. 农业与生命科学版 differential proteins <italic>in vivo</italic> interactions induced disease resistance |
| title | Proteomic study of three component interactions: plant, pathogens and antagonistic fungi |
| title_full | Proteomic study of three component interactions: plant, pathogens and antagonistic fungi |
| title_fullStr | Proteomic study of three component interactions: plant, pathogens and antagonistic fungi |
| title_full_unstemmed | Proteomic study of three component interactions: plant, pathogens and antagonistic fungi |
| title_short | Proteomic study of three component interactions: plant, pathogens and antagonistic fungi |
| title_sort | proteomic study of three component interactions plant pathogens and antagonistic fungi |
| topic | differential proteins <italic>in vivo</italic> interactions induced disease resistance |
| url | https://www.academax.com/doi/10.3785/1008-9209.2004.04.0449 |
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