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Agroinfiltration

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Dominique Michaud – 1st expert on this subject based on the ideXlab platform

  • ph gradient mitigation in the leaf cell secretory pathway attenuates the defense response of nicotiana benthamiana to Agroinfiltration
    Journal of Proteome Research, 2020
    Co-Authors: Philippe V. Jutras, Marcandre Daoust, Pierreolivier Lavoie, Marieclaire Goulet, Frank Sainsbury, Rachel Tardif, Louis-philippe Hamel, Dominique Michaud

    Abstract:

    Partial neutralization of the Golgi lumen pH by the ectopic expression of influenza virus M2 proton channel is useful to stabilize acid-labile recombinant proteins in plant cells, but the impact of pH gradient mitigation on host cellular functions has not been investigated. Here, we assessed the unintended effects of M2 expression on the leaf proteome of Nicotiana benthamiana infiltrated with the bacterial gene vector Agrobacterium tumefaciens. An isobaric tags for relative and absolute quantification quantitative proteomics procedure was followed to compare the leaf proteomes of plants agroinfiltrated with either an “empty” vector or an M2-encoding vector. Leaves infiltrated with the empty vector had a low soluble protein content compared to noninfiltrated control leaves, associated with increased levels of stress-related proteins but decreased levels of photosynthesis-associated proteins. M2 expression partly compromised these effects of Agroinfiltration to restore soluble protein content in the leaf tissue, associated with restored levels of photosynthesis-associated proteins and reduced levels of stress-related proteins in the apoplast. These data illustrate the cell-wide influence of the Golgi lumen pH homeostasis on the leaf proteome of N. benthamiana responding to microbial challenge. They also underline the relevance of assessing the eventual unintended effects of accessory proteins used to modulate specific cellular or metabolic functions in plant protein biofactories.

  • pH Gradient Mitigation in the Leaf Cell Secretory Pathway Attenuates the Defense Response of Nicotiana benthamiana to Agroinfiltration
    Journal of Proteome Research, 2019
    Co-Authors: Philippe V. Jutras, Pierreolivier Lavoie, Marieclaire Goulet, Frank Sainsbury, Rachel Tardif, Louis-philippe Hamel, Marc-andre D'aoust, Dominique Michaud

    Abstract:

    Partial neutralization of the Golgi lumen pH by ectopic expression of influenza virus M2 proton channel is useful to stabilize acid-labile recombinant proteins in plant cells but the impact of pH gradient mitigation on host cellular functions has not been investigated. Here, we assessed the unintended effects of M2 expression on the leaf proteome of Nicotiana benthamiana infiltrated with the bacterial gene vector Agrobacterium tumefaciens. A iTRAQ quantitative proteomics procedure was followed to compare the leaf proteomes of plants agroinfiltrated with either an ‘empty’ vector or an M2-encoding vector. Leaves infiltrated with the empty vector had a low soluble protein content compared to non-infiltrated control leaves, associated with increased levels of stress-related proteins but decreased levels of photosynthesis-associated proteins. M2 expression partly compromised these effects of Agroinfiltration to restore soluble protein content in leaf tissue, associated with restored levels of photosynthesis-as…

  • 2 de proteome maps for the leaf apoplast of nicotiana benthamiana
    Proteomics, 2010
    Co-Authors: Charles Goulet, Marieclaire Goulet, Dominique Michaud

    Abstract:

    We provide 2-D gel reference maps for the apoplastic proteome of Nicotiana benthamiana leaves infiltrated or not with the bacterial gene vector Agrobacterium tumefaciens. About 90 proteins were analyzed by LC-MS/MS for identification and function assignment. We show, overall, an effective response of the plant to Agroinfiltration involving a specific, cell wall maintenance-independent up-regulation of defense protein secretion. The proteome maps described should be a useful tool for systemic studies on plant-pathogen interactions or cell wall metabolism. They also should prove useful for the monitoring of secreted recombinant proteins and their possible pleiotropic effects along the cell secretory pathway of N. benthamiana leaves used as an expression platform for clinically useful proteins.

R Roth – 2nd expert on this subject based on the ideXlab platform

  • Agroinfiltration is a versatile tool that facilitates comparative analyses of avr9 cf 9 induced and avr4 cf 4 induced necrosis
    Molecular Plant-microbe Interactions, 2000
    Co-Authors: R A L Van Der Hoorn, Franck Laurent, R Roth

    Abstract:

    The avirulence genes Avr9 and Avr4 from the fungal tomato pathogen Cladosporium fulvum encode extracellular proteins that elicit a hypersensitive response when injected into leaves of tomato plants carrying the matching resistance genes, Cf-9 and Cf-4, respectively. We successfully expressed both Avr9 and Avr4 genes in tobacco with the Agrobacterium tumefaciens transient transformation assay (Agroinfiltration). In addition, we expressed the matching resistance genes, Cf-9 and Cf-4, through Agroinfiltration. By combining transient Cf gene expression with either transgenic plants expressing one of the gene partners, Potato virus X (PVX)-mediated Avr gene expression, or elicitor injections, we demonstrated that Agroinfiltration is a reliable and versatile tool to study Avr/Cf-mediated recognition. Significantly, Agroinfiltration can be used to quantify and compare Avr/Cf-induced responses. Comparison of different Avr/Cf-interactions within one tobacco leaf showed that Avr9/Cf-9-induced necrosis developed slo…

  • Agroinfiltration is a versatile tool that facilitates comparative analyses of Avr9/Cf-9-induced and Avr4/Cf-4-induced necrosis.
    Molecular Plant-microbe Interactions, 2000
    Co-Authors: R A L Van Der Hoorn, Franck Laurent, R Roth

    Abstract:

    The avirulence genes Avr9 and Avr4 from the fungal tomato pathogen Cladosporium fulvum encode extracellular proteins that elicit a hypersensitive response when injected into leaves of tomato plants carrying the matching resistance genes, Cf-9 and Cf-4, respectively. We successfully expressed both Avr9 and Avr4 genes in tobacco with the Agrobacterium tumefaciens transient transformation assay (Agroinfiltration). In addition, we expressed the matching resistance genes, Cf-9 and Cf-4, through Agroinfiltration. By combining transient Cf gene expression with either transgenic plants expressing one of the gene partners, Potato virus X (PVX)-mediated Avr gene expression, or elicitor injections, we demonstrated that Agroinfiltration is a reliable and versatile tool to study Avr/Cf-mediated recognition. Significantly, Agroinfiltration can be used to quantify and compare Avr/Cf-induced responses. Comparison of different Avr/Cf-interactions within one tobacco leaf showed that Avr9/Cf-9-induced necrosis developed slo…

R A L Van Der Hoorn – 3rd expert on this subject based on the ideXlab platform

  • Agroinfiltration is a versatile tool that facilitates comparative analyses of avr9 cf 9 induced and avr4 cf 4 induced necrosis
    Molecular Plant-microbe Interactions, 2000
    Co-Authors: R A L Van Der Hoorn, Franck Laurent, R Roth

    Abstract:

    The avirulence genes Avr9 and Avr4 from the fungal tomato pathogen Cladosporium fulvum encode extracellular proteins that elicit a hypersensitive response when injected into leaves of tomato plants carrying the matching resistance genes, Cf-9 and Cf-4, respectively. We successfully expressed both Avr9 and Avr4 genes in tobacco with the Agrobacterium tumefaciens transient transformation assay (Agroinfiltration). In addition, we expressed the matching resistance genes, Cf-9 and Cf-4, through Agroinfiltration. By combining transient Cf gene expression with either transgenic plants expressing one of the gene partners, Potato virus X (PVX)-mediated Avr gene expression, or elicitor injections, we demonstrated that Agroinfiltration is a reliable and versatile tool to study Avr/Cf-mediated recognition. Significantly, Agroinfiltration can be used to quantify and compare Avr/Cf-induced responses. Comparison of different Avr/Cf-interactions within one tobacco leaf showed that Avr9/Cf-9-induced necrosis developed slo…

  • Agroinfiltration is a versatile tool that facilitates comparative analyses of Avr9/Cf-9-induced and Avr4/Cf-4-induced necrosis.
    Molecular Plant-microbe Interactions, 2000
    Co-Authors: R A L Van Der Hoorn, Franck Laurent, R Roth

    Abstract:

    The avirulence genes Avr9 and Avr4 from the fungal tomato pathogen Cladosporium fulvum encode extracellular proteins that elicit a hypersensitive response when injected into leaves of tomato plants carrying the matching resistance genes, Cf-9 and Cf-4, respectively. We successfully expressed both Avr9 and Avr4 genes in tobacco with the Agrobacterium tumefaciens transient transformation assay (Agroinfiltration). In addition, we expressed the matching resistance genes, Cf-9 and Cf-4, through Agroinfiltration. By combining transient Cf gene expression with either transgenic plants expressing one of the gene partners, Potato virus X (PVX)-mediated Avr gene expression, or elicitor injections, we demonstrated that Agroinfiltration is a reliable and versatile tool to study Avr/Cf-mediated recognition. Significantly, Agroinfiltration can be used to quantify and compare Avr/Cf-induced responses. Comparison of different Avr/Cf-interactions within one tobacco leaf showed that Avr9/Cf-9-induced necrosis developed slo…