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An immunocytochemical procedure for protein localization in various nematode life stages combined with plant tissues using methylacrylate-embedded specimens
- Source :
- Phytopathology, Phytopathology, American Phytopathological Society, 2012, 102 (10), pp.990-996. ⟨10.1094/PHYTO-02-12-0031-R⟩, Phytopathology 10 (102), 990-996. (2012)
- Publication Year :
- 2012
- Publisher :
- HAL CCSD, 2012.
-
Abstract
- 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.
- Subjects :
- 0106 biological sciences
[SDV.SA]Life Sciences [q-bio]/Agricultural sciences
Nematoda
nématode des plantes
Plant Science
Biology
01 natural sciences
Epitope
nématode parasite
03 medical and health sciences
tissu végétal
protéine végétale
épitope
Animals
méthacrylate
[SDV.BV]Life Sciences [q-bio]/Vegetal Biology
Cytoskeleton
030304 developmental biology
nématode
0303 health sciences
Vegetal Biology
cellule végétale
Plants
biology.organism_classification
Plant cell
localisation de protéine
Protein subcellular localization prediction
Immunohistochemistry
Cell biology
Agricultural sciences
Secretory protein
Nematode
analyse immunohistochimique
Plant morphology
biology.protein
anticorps
Methacrylates
morphologie de la plante
Antibody
Agronomy and Crop Science
Sciences agricoles
Biologie végétale
010606 plant biology & botany
Subjects
Details
- Language :
- English
- ISSN :
- 0031949X
- Database :
- OpenAIRE
- Journal :
- Phytopathology, Phytopathology, American Phytopathological Society, 2012, 102 (10), pp.990-996. ⟨10.1094/PHYTO-02-12-0031-R⟩, Phytopathology 10 (102), 990-996. (2012)
- Accession number :
- edsair.doi.dedup.....d7ee6808dac5e00dc251f17fbfd74b3d