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Uvitex2B: a rapid and efficient stain for detection of arbuscular mycorrhizal fungi within plant roots

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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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Acknowledgments

We are grateful to G. Bécard for providing G. intraradices starting cultures and C. Bournaud for providing carrot roots colonized with G. clarum. We would like to thank D. Moukouanga, V. Vaissayre, and J. Bonneau for their help with plant care and histochemical staining. We also thank E. Duhoux and Y. Prin for their critical reading of the manuscript. This work was funded by the IRD and CIRAD and grants from the Département Soutien et Formation des Communautés Scientifiques du Sud (DSF/IRD).

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Authors and Affiliations

  1. Département de Biologie Végétale, Université Cheikh Anta Diop (UCAD), BP 5005, Dakar, Senegal

    Nathalie Diagne, Aboubacry Kane & Diegane Diouf

  2. Laboratoire Commun de Microbiologie IRD/ISRA/UCAD, Centre de Recherche de Bel Air, BP 1386, Dakar, Senegal

    Nathalie Diagne, Aboubacry Kane, Diegane Diouf & Robin Duponnois

  3. Groupe Rhizogenèse, Unité Mixte de Recherche Diversité et Adaptation des Plantes Cultivées (DIAPC), Institut de Recherche pour le Développement (IRD), 911 Avenue Agropolis, BP 64501, 34394, Montpellier Cedex 5, France

    Nathalie Diagne, Claudine Franche, Didier Bogusz & Sergio Svistoonoff

  4. Plateau d’Histocytologie et d’Imagerie Végétale, MRI/ CIRAD/BIOS/UMR DAP, Avenue Agropolis, 34398, Montpellier Cedex 5, France

    Jacques Escoute, Marc Lartaud & Jean Luc Verdeil

  5. Laboratoire des Symbioses Tropicales et Méditerranéennes (LSTM), TA10/J, IRD, UMR 113 CIRAD/INRA/IRD/SUP-AGRO/UM2, 34398, Campus International de Baillarguet, Montpellier Cedex 5, France

    Robin Duponnois

Authors
  1. Nathalie Diagne
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  2. Jacques Escoute
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  3. Marc Lartaud
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  4. Jean Luc Verdeil
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  5. Claudine Franche
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  6. Aboubacry Kane
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  7. Didier Bogusz
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  8. Diegane Diouf
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  9. Robin Duponnois
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  10. Sergio Svistoonoff
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Corresponding author

Correspondence to Sergio Svistoonoff.

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Fig. S1a

Carrot and C. equisetifolia roots (a and b, respectively) colonized by G. intraradices stained with Uvitex2B and acquired using CLSM. The animation, assembled using a z stack, allows observation of the infection pathway: intracellular filaments give rise to several small arbuscules in the case of the carrot (=Paris-type), and intercellular filaments give rise to a single intracellular arbuscule in the case of C. equisetifolia (=Arum-type; MP4 499 kb)

Fig. S1b

(MP4 354 kb)

Fig. S2

Carrot (a), C. equisetifolia (b), and M. truncatula (c) roots colonized by G. intraradices stained with Uvitex2B, acquired using epifluorescence microscopy. These are the raw images corresponding to Fig. 1d–f before deconvolution. Bar = 10 μm (JPEG 23 kb)

High-Resolution (EPS, 297 kb)

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Diagne, N., Escoute, J., Lartaud, M. et al. Uvitex2B: a rapid and efficient stain for detection of arbuscular mycorrhizal fungi within plant roots. Mycorrhiza 21, 315–321 (2011). https://doi.org/10.1007/s00572-010-0357-8

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  • DOI: https://doi.org/10.1007/s00572-010-0357-8

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