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Uvitex2B: a rapid and efficient stain for detection of arbuscular mycorrhizal fungi within plant roots
Authors:Nathalie Diagne  Jacques Escoute  Marc Lartaud  Jean Luc Verdeil  Claudine Franche  Aboubacry Kane  Didier Bogusz  Diegane Diouf  Robin Duponnois  Sergio Svistoonoff
Affiliation:(1) D?partement de Biologie V?g?tale, Universit? Cheikh Anta Diop (UCAD), BP 5005, Dakar, Senegal;(2) Laboratoire Commun de Microbiologie IRD/ISRA/UCAD, Centre de Recherche de Bel Air, BP 1386, Dakar, Senegal;(3) Groupe Rhizogen?se, Unit? Mixte de Recherche Diversit? et Adaptation des Plantes Cultiv?es (DIAPC), Institut de Recherche pour le D?veloppement (IRD), 911 Avenue Agropolis, BP 64501, 34394 Montpellier Cedex 5, France;(4) Plateau d’Histocytologie et d’Imagerie V?g?tale, MRI/ CIRAD/BIOS/UMR DAP, Avenue Agropolis, 34398 Montpellier Cedex 5, France;(5) Laboratoire des Symbioses Tropicales et M?diterran?ennes (LSTM), TA10/J, IRD, UMR 113 CIRAD/INRA/IRD/SUP-AGRO/UM2, 34398, Campus International de Baillarguet, Montpellier Cedex 5, France
Abstract:The study of arbuscular mycorrhiza often requires the staining of fungal structures using specific dyes. Fluorescent dyes such as acid fuchsin and wheat germ agglutinin conjugates give excellent results, but these compounds are either hazardous or very expensive. Here, we show that a safer and inexpensive dye, Uvitex2B, can be efficiently used to stain intraradical fungal structures formed by the arbuscular mycorrhizal fungus Glomus intraradices in three plant species: carrot, Casuarina equisetifolia, and Medicago truncatula. The intensity and stability of Uvitex2B allow the acquisition of high-quality images using not only confocal laser scanning microscopy but also epifluorescence microscopy coupled with image deconvolution. Furthermore, we demonstrate that Uvitex2B and β-glucuronidase staining are compatible and can thus be used to reveal arbuscular mycorrhizal structures in the context of promoter activation analysis.
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