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Development of Microscopic Techniques for the Visualization of Plant–Root-Knot Nematode Interaction

Plant-parasitic nematodes are a significant cause of yield losses and food security issues. Specifically, nematodes of the genus Meloidogyne can cause significant production losses in horticultural crops around the world. Understanding the mechanisms of the ever-changing physiology of plant roots by...

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Autores principales: Vernet, Helena, Fullana, Aïda Magdalena, Sorribas, Francisco Javier, Gualda, Emilio J.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: MDPI 2022
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9104198/
https://www.ncbi.nlm.nih.gov/pubmed/35567165
http://dx.doi.org/10.3390/plants11091165
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author Vernet, Helena
Fullana, Aïda Magdalena
Sorribas, Francisco Javier
Gualda, Emilio J.
author_facet Vernet, Helena
Fullana, Aïda Magdalena
Sorribas, Francisco Javier
Gualda, Emilio J.
author_sort Vernet, Helena
collection PubMed
description Plant-parasitic nematodes are a significant cause of yield losses and food security issues. Specifically, nematodes of the genus Meloidogyne can cause significant production losses in horticultural crops around the world. Understanding the mechanisms of the ever-changing physiology of plant roots by imaging the galls induced by nematodes could provide a great insight into their control. However, infected roots are unsuitable for light microscopy investigation due to the opacity of plant tissues. Thus, samples must be cleared to visualize the interior of whole plants in order to make them transparent using clearing agents. This work aims to identify which clearing protocol and microscopy system is the most appropriate to obtain 3D images of tomato cv. Durinta and eggplant cv. Cristal samples infected with Meloidogyne incognita to visualize and study the root–nematode interaction. To that extent, two clearing solutions (BABB and ECi), combined with three different dehydration solvents (ethanol, methanol and 1-propanol), are tested. In addition, the advantages and disadvantages of alternative imaging techniques to confocal microscopy are analyzed by employing an experimental custom-made setup that combines two microscopic techniques, light sheet fluorescence microscopy and optical projection tomography, on a single instrument.
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spelling pubmed-91041982022-05-14 Development of Microscopic Techniques for the Visualization of Plant–Root-Knot Nematode Interaction Vernet, Helena Fullana, Aïda Magdalena Sorribas, Francisco Javier Gualda, Emilio J. Plants (Basel) Article Plant-parasitic nematodes are a significant cause of yield losses and food security issues. Specifically, nematodes of the genus Meloidogyne can cause significant production losses in horticultural crops around the world. Understanding the mechanisms of the ever-changing physiology of plant roots by imaging the galls induced by nematodes could provide a great insight into their control. However, infected roots are unsuitable for light microscopy investigation due to the opacity of plant tissues. Thus, samples must be cleared to visualize the interior of whole plants in order to make them transparent using clearing agents. This work aims to identify which clearing protocol and microscopy system is the most appropriate to obtain 3D images of tomato cv. Durinta and eggplant cv. Cristal samples infected with Meloidogyne incognita to visualize and study the root–nematode interaction. To that extent, two clearing solutions (BABB and ECi), combined with three different dehydration solvents (ethanol, methanol and 1-propanol), are tested. In addition, the advantages and disadvantages of alternative imaging techniques to confocal microscopy are analyzed by employing an experimental custom-made setup that combines two microscopic techniques, light sheet fluorescence microscopy and optical projection tomography, on a single instrument. MDPI 2022-04-26 /pmc/articles/PMC9104198/ /pubmed/35567165 http://dx.doi.org/10.3390/plants11091165 Text en © 2022 by the authors. https://creativecommons.org/licenses/by/4.0/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 (https://creativecommons.org/licenses/by/4.0/).
spellingShingle Article
Vernet, Helena
Fullana, Aïda Magdalena
Sorribas, Francisco Javier
Gualda, Emilio J.
Development of Microscopic Techniques for the Visualization of Plant–Root-Knot Nematode Interaction
title Development of Microscopic Techniques for the Visualization of Plant–Root-Knot Nematode Interaction
title_full Development of Microscopic Techniques for the Visualization of Plant–Root-Knot Nematode Interaction
title_fullStr Development of Microscopic Techniques for the Visualization of Plant–Root-Knot Nematode Interaction
title_full_unstemmed Development of Microscopic Techniques for the Visualization of Plant–Root-Knot Nematode Interaction
title_short Development of Microscopic Techniques for the Visualization of Plant–Root-Knot Nematode Interaction
title_sort development of microscopic techniques for the visualization of plant–root-knot nematode interaction
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9104198/
https://www.ncbi.nlm.nih.gov/pubmed/35567165
http://dx.doi.org/10.3390/plants11091165
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