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Applied and Environmental Microbiology, November 2008, p. 6598-6605, Vol. 74, No. 21
0099-2240/08/$08.00+0     doi:10.1128/AEM.00584-08
Copyright © 2008, American Society for Microbiology. All Rights Reserved.

Gene Transcription in Lactarius quietus-Quercus petraea Ectomycorrhizas from a Forest Soil{triangledown}

P. E. Courty,* M. Poletto, F. Duchaussoy, M. Buée, J. Garbaye, and F. Martin

UMR 1136, INRA-Nancy Université, Interactions Arbres/Microorganismes, INRA-Nancy, 54280 Champenoux, France

Received 11 March 2008/ Accepted 29 August 2008

Extracting fungal mRNA from ectomycorrhizas (ECMs) and forest soil samples for monitoring in situ metabolic activities is a significant challenge when studying the role of ECMs in biogeochemical cycles. A robust, simple, rapid, and effective method was developed for extracting RNA from rhizospheric soil and ECMs by adapting previous grinding and lysis methods. The quality and yield of the extracted RNA were sufficient to be used for reverse transcription. RNA extracted from ECMs of Lactarius quietus in a 100-year-old oak stand was used to construct a cDNA library and sequence expressed sequence tags. The transcripts of many genes involved in primary metabolism and in the degradation of organic matter were found. The transcription levels of four targeted fungal genes (glutamine synthase, a general amino acid transporter, a tyrosinase, and N-acetylhexosaminidase) were measured by quantitative reverse transcription-PCR in ECMs and in the ectomycorrhizospheric soil (the soil surrounding the ECMs containing the extraradical mycelium) in forest samples. On average, levels of gene expression for the L. quietus ECM root tips were similar to those for the extraradical mycelium, although gene expression varied up to 10-fold among the samples. This study demonstrates that gene expression from ECMs and soil can be analyzed. These results provide new perspectives for investigating the role of ectomycorrhizal fungi in the functioning of forest ecosystems.

* Corresponding author. Mailing address: UMR 1136, INRA-Nancy Université, Interactions Arbres/Microorganismes, INRA-Nancy, 54280 Champenoux, France. Phone: 33 3 83 39 40 41. Fax: 33 3 83 39 40 69. E-mail: courty{at}nancy.inra.fr

{triangledown} Published ahead of print on 12 September 2008.

Applied and Environmental Microbiology, November 2008, p. 6598-6605, Vol. 74, No. 21
0099-2240/08/$08.00+0     doi:10.1128/AEM.00584-08
Copyright © 2008, American Society for Microbiology. All Rights Reserved.





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