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Fluorescently-primed in situ PCR in arbuscular mycorrhizas

Published online by Cambridge University Press:  01 December 1998

B. BAGO
Affiliation:
Centre de recherche en biologie forestière (CRBF), Pavillon Charles-Eugène-Marchand, Université Laval, Québec, G1K 7P4 Canada Present address: Plant and Soil Biophysics Lab, Eastern Regional Research Center, USDA/ARS, 600 E. Mermaid Lane, Wyndmoor, PA 19038, U.S.A.
Y. PICHÉ
Affiliation:
Centre de recherche en biologie forestière (CRBF), Pavillon Charles-Eugène-Marchand, Université Laval, Québec, G1K 7P4 Canada
L. SIMON
Affiliation:
Vice-rectorat à la recherche, Pavillon des sciences de l'éducation, Université Laval, Québec, G1K 7P4 Canada
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Abstract

In situ gene amplification (in situ PCR) is a recent, powerful molecular technique which allows the localization of low abundance nucleic acids targets directly within tissue sections. The work presented here is, to our knowledge, the first report of successful direct detection of in situ PCR amplification with fluorescently-labelled primers, and the first successful in situ PCR performed on arbuscular mycorrhizal (AM) fungi. Ribosomal SSU genes within AM fungal spore sections were amplified by using fluorescent, glomalean-specific primers, then directly detected by means of epifluorescence microscopy. Different controls confirmed the successfulness of the in situ amplification. These results open new avenues in the study of arbuscular mycorrhizas, where genetic processes seem to be transient and very localized.

Type
Research Article
Copyright
© The British Mycological Society 1998

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