Uvitex2B: a rapid and efficient stain for detection of arbuscular mycorrhizal fungi within plant roots
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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.
KeywordsArbuscular mycorrhizal fungi (AMF) Uvitex2B Confocal laser scanning microscopy (CLSM) ProMtEnod11::gus
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).
- Charron D, Pingret J, Chabaud M, Journet E, Barker DG (2004) Pharmacological evidence that multiple phospholipid signaling pathways link Rhizobium nodulation factor perception in Medicago truncatula root hairs to intracellular responses, including Ca2+ spiking and specific ENOD gene expression. Plant Physiol 136:3582–3593PubMedCrossRefGoogle Scholar
- Combes RD, Haveland-Smith RB (1982) A review of the genotoxicity of food, drug and cosmetic colours and other azo, triphenylmethane and xanthene dyes. Mutat Res Rev Genet Toxicol 98:101–243Google Scholar
- Gherbi H, Markmann K, Svistoonoff S, Estevan J, Autran D, Giczey G, Auguy F, Péret B, Laplaze L, Franche C, Parniske M, Bogusz D (2008) SymRK defines a common genetic basis for plant root endosymbioses with arbuscular mycorrhiza fungi, rhizobia, and Frankia bacteria. Proc Natl Acad Sci USA 105:4928–4932PubMedCrossRefGoogle Scholar
- Giovannetti M, Avio L, Salutini L (1991) Morphological, cytochemical, and ontogenetic characteristics of a new species of vesicular-arbuscular mycorrhizal fungus. Can J Bot 69:161–167Google Scholar
- IARC (1987) International Agency for Research on Cancer monographs on the evaluation of carcinogenic risks to humans. Overall evaluations of carcinogenicity: an updating of IARC monographs, volumes 1 to 42, supplement 7Google Scholar
- IARC (1999) International Agency for Research on Cancer monographs on the evaluation of carcinogenic risks to humans, vol. 29Google Scholar
- Smith SE, Read DJ (2008) Mycorrhizal symbiosis. Academic Press, New YorkGoogle Scholar
- Vierheilig H, Schweiger P, Brundrett M (2005) An overview of methods for the detection and observation of arbuscular mycorrhizal fungi in roots. Physiol Plant 125:393–404Google Scholar