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A Phenotyping Method of Giant Cells from Root-Knot Nematode Feeding Sites by Confocal Microscopy Highlights a Role for CHITINASE-LIKE 1 in Arabidopsis

Most effective nematicides for the control of root-knot nematodes are banned, which demands a better understanding of the plant-nematode interaction. Understanding how gene expression in the nematode-feeding sites relates to morphological features may assist a better characterization of the interact...

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Autores principales: Cabrera, Javier, Olmo, Rocio, Ruiz-Ferrer, Virginia, Abreu, Isidro, Hermans, Christian, Martinez-Argudo, Isabel, Fenoll, Carmen, Escobar, Carolina
Formato: Online Artículo Texto
Lenguaje:English
Publicado: MDPI 2018
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5855651/
https://www.ncbi.nlm.nih.gov/pubmed/29389847
http://dx.doi.org/10.3390/ijms19020429
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author Cabrera, Javier
Olmo, Rocio
Ruiz-Ferrer, Virginia
Abreu, Isidro
Hermans, Christian
Martinez-Argudo, Isabel
Fenoll, Carmen
Escobar, Carolina
author_facet Cabrera, Javier
Olmo, Rocio
Ruiz-Ferrer, Virginia
Abreu, Isidro
Hermans, Christian
Martinez-Argudo, Isabel
Fenoll, Carmen
Escobar, Carolina
author_sort Cabrera, Javier
collection PubMed
description Most effective nematicides for the control of root-knot nematodes are banned, which demands a better understanding of the plant-nematode interaction. Understanding how gene expression in the nematode-feeding sites relates to morphological features may assist a better characterization of the interaction. However, nematode-induced galls resulting from cell-proliferation and hypertrophy hinders such observation, which would require tissue sectioning or clearing. We demonstrate that a method based on the green auto-fluorescence produced by glutaraldehyde and the tissue-clearing properties of benzyl-alcohol/benzyl-benzoate preserves the structure of the nematode-feeding sites and the plant-nematode interface with unprecedented resolution quality. This allowed us to obtain detailed measurements of the giant cells’ area in an Arabidopsis line overexpressing CHITINASE-LIKE-1 (CTL1) from optical sections by confocal microscopy, assigning a role for CTL1 and adding essential data to the scarce information of the role of gene repression in giant cells. Furthermore, subcellular structures and features of the nematodes body and tissues from thick organs formed after different biotic interactions, i.e., galls, syncytia, and nodules, were clearly distinguished without embedding or sectioning in different plant species (Arabidopsis, cucumber or Medicago). The combination of this method with molecular studies will be valuable for a better understanding of the plant-biotic interactions.
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spelling pubmed-58556512018-03-20 A Phenotyping Method of Giant Cells from Root-Knot Nematode Feeding Sites by Confocal Microscopy Highlights a Role for CHITINASE-LIKE 1 in Arabidopsis Cabrera, Javier Olmo, Rocio Ruiz-Ferrer, Virginia Abreu, Isidro Hermans, Christian Martinez-Argudo, Isabel Fenoll, Carmen Escobar, Carolina Int J Mol Sci Article Most effective nematicides for the control of root-knot nematodes are banned, which demands a better understanding of the plant-nematode interaction. Understanding how gene expression in the nematode-feeding sites relates to morphological features may assist a better characterization of the interaction. However, nematode-induced galls resulting from cell-proliferation and hypertrophy hinders such observation, which would require tissue sectioning or clearing. We demonstrate that a method based on the green auto-fluorescence produced by glutaraldehyde and the tissue-clearing properties of benzyl-alcohol/benzyl-benzoate preserves the structure of the nematode-feeding sites and the plant-nematode interface with unprecedented resolution quality. This allowed us to obtain detailed measurements of the giant cells’ area in an Arabidopsis line overexpressing CHITINASE-LIKE-1 (CTL1) from optical sections by confocal microscopy, assigning a role for CTL1 and adding essential data to the scarce information of the role of gene repression in giant cells. Furthermore, subcellular structures and features of the nematodes body and tissues from thick organs formed after different biotic interactions, i.e., galls, syncytia, and nodules, were clearly distinguished without embedding or sectioning in different plant species (Arabidopsis, cucumber or Medicago). The combination of this method with molecular studies will be valuable for a better understanding of the plant-biotic interactions. MDPI 2018-01-31 /pmc/articles/PMC5855651/ /pubmed/29389847 http://dx.doi.org/10.3390/ijms19020429 Text en © 2018 by the authors. Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (http://creativecommons.org/licenses/by/4.0/).
spellingShingle Article
Cabrera, Javier
Olmo, Rocio
Ruiz-Ferrer, Virginia
Abreu, Isidro
Hermans, Christian
Martinez-Argudo, Isabel
Fenoll, Carmen
Escobar, Carolina
A Phenotyping Method of Giant Cells from Root-Knot Nematode Feeding Sites by Confocal Microscopy Highlights a Role for CHITINASE-LIKE 1 in Arabidopsis
title A Phenotyping Method of Giant Cells from Root-Knot Nematode Feeding Sites by Confocal Microscopy Highlights a Role for CHITINASE-LIKE 1 in Arabidopsis
title_full A Phenotyping Method of Giant Cells from Root-Knot Nematode Feeding Sites by Confocal Microscopy Highlights a Role for CHITINASE-LIKE 1 in Arabidopsis
title_fullStr A Phenotyping Method of Giant Cells from Root-Knot Nematode Feeding Sites by Confocal Microscopy Highlights a Role for CHITINASE-LIKE 1 in Arabidopsis
title_full_unstemmed A Phenotyping Method of Giant Cells from Root-Knot Nematode Feeding Sites by Confocal Microscopy Highlights a Role for CHITINASE-LIKE 1 in Arabidopsis
title_short A Phenotyping Method of Giant Cells from Root-Knot Nematode Feeding Sites by Confocal Microscopy Highlights a Role for CHITINASE-LIKE 1 in Arabidopsis
title_sort phenotyping method of giant cells from root-knot nematode feeding sites by confocal microscopy highlights a role for chitinase-like 1 in arabidopsis
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5855651/
https://www.ncbi.nlm.nih.gov/pubmed/29389847
http://dx.doi.org/10.3390/ijms19020429
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