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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Detalles Bibliográficos
Autores: Vernet Pérez, Helena, Fullana Pons, Aïda Magdalena, Sorribas Royo, Francisco Javier|||0000-0001-7465-7353, Gualda Manzano, Emilio José|||0000-0001-5948-5493
Tipo de recurso: artículo
Fecha de publicación:2022
País:España
Institución:Universitat Politècnica de Catalunya (UPC)
Repositorio:UPCommons. Portal del coneixement obert de la UPC
Idioma:inglés
OAI Identifier:oai:upcommons.upc.edu:2117/367483
Acceso en línea:https://hdl.handle.net/2117/367483
https://dx.doi.org/10.3390/plants11091165
Access Level:acceso abierto
Palabra clave:Plant nematodes
Plant 3D imaging
Root-knot nematode
Tissue clearing
Light sheet fluorescence microscopy
Optical projection tomography
Nematodes fitoparàsits
Àrees temàtiques de la UPC::Enginyeria agroalimentària::Agricultura::Fitopatologia
Descripción
Sumario: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.