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Publication Details
AFRICAN RESEARCH NEXUS
SHINING A SPOTLIGHT ON AFRICAN RESEARCH
agricultural and biological sciences
An immunocytochemical procedure for protein localization in various nematode life stages combined with plant tissues using methylacrylate-embedded specimens
Phytopathology, Volume 102, No. 10, Year 2012
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Description
Plant-parasitic nematodes possess a large number of proteins that are secreted in planta, allowing them to be successful parasites of plants. The majority of these proteins are synthesized mainly in the nematode subventral and dorsal glands as well as in other organs. To improve the immunovisualization of these proteins, we adapted a methacrylate embedding method for the localization of proteins inside nematode tissues, and extracellularly when secreted in planta or within plant cells. An important advantage is that the method is applicable for all nematode stages: preparasitic as well as parasitic stages, including large mature females. Herein, the method has been successfully applied for the localization of four nematode secreted proteins, such as Mi-MAP-1, Mi-CBM2-bearing proteins, Mi-PEL3, and Mi-6D4. In addition, we could also localize 14-3-3 proteins, as well as two cytoskeletal proteins, by double-immunolabeling on preparasitic juveniles. Superior preservation of nematode and plant morphology, allowed more accurate protein localization as compared with other methods. Besides excellent epitope preservation, dissolution of methacrylate from tissue sections unmasks target proteins and thereby drastically increases antibody access. © 2012 The American Phytopathological Society.
Authors & Co-Authors
Vieira, P. C.
France, Sophia Antipolis
Institut Sophia Agrobiotech - Isa
France, Paris
Cnrs Centre National de la Recherche Scientifique
France, Nice
Université Côte D'azur
Portugal, Evora
University of Évora
Youssef Banora, Mohamed Youssef
France, Sophia Antipolis
Institut Sophia Agrobiotech - Isa
France, Paris
Cnrs Centre National de la Recherche Scientifique
France, Nice
Université Côte D'azur
Egypt, Cairo
College of Agriculture
Castagnone-Sereno, Philippe
France, Sophia Antipolis
Institut Sophia Agrobiotech - Isa
France, Paris
Cnrs Centre National de la Recherche Scientifique
France, Nice
Université Côte D'azur
Rosso, Marie Noëlle
France, Sophia Antipolis
Institut Sophia Agrobiotech - Isa
France, Paris
Cnrs Centre National de la Recherche Scientifique
France, Nice
Université Côte D'azur
Engler, Gilbert J.
France, Sophia Antipolis
Institut Sophia Agrobiotech - Isa
France, Paris
Cnrs Centre National de la Recherche Scientifique
France, Nice
Université Côte D'azur
De-Almeida-Engler, Janice
France, Sophia Antipolis
Institut Sophia Agrobiotech - Isa
France, Paris
Cnrs Centre National de la Recherche Scientifique
France, Nice
Université Côte D'azur
Statistics
Citations: 11
Authors: 6
Affiliations: 5
Identifiers
Doi:
10.1094/PHYTO-02-12-0031-R
ISSN:
0031949X
Research Areas
Health System And Policy
Participants Gender
Female