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Monitoring the persistence of Laccaria bicolor as an ectomycorrhizal symbiont of nursery-grown Douglas fir by PCR of the rDNA intergenic spacer

Authors

  • B. HENRION,

    1. Equipe de Microbiologic Forestière, Centre de Recherches de Nancy, Institut National de la Recherche Agronomique, 54280 Champenoux, France
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  • C. DI BATTISTA,

    1. Equipe de Microbiologic Forestière, Centre de Recherches de Nancy, Institut National de la Recherche Agronomique, 54280 Champenoux, France
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  • D. BOUCHARD,

    1. Equipe de Microbiologic Forestière, Centre de Recherches de Nancy, Institut National de la Recherche Agronomique, 54280 Champenoux, France
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  • D. VAIRELLES,

    1. Equipe de Microbiologic Forestière, Centre de Recherches de Nancy, Institut National de la Recherche Agronomique, 54280 Champenoux, France
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  • B. D. THOMPSON,

    1. *Arborgen Europe, Poelweg 46, 1424 PB de Kwakel, the Netherlands
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    • †Institute of Ecology and Resource Management, Edinburgh University, Darwin Building, Mayfield Road, Edinburgh, Scotland EH9 3JU, UK.

  • F. LE TACON,

    1. Equipe de Microbiologic Forestière, Centre de Recherches de Nancy, Institut National de la Recherche Agronomique, 54280 Champenoux, France
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  • F. MARTIN

    Corresponding author
    1. Equipe de Microbiologic Forestière, Centre de Recherches de Nancy, Institut National de la Recherche Agronomique, 54280 Champenoux, France
      Equipe de Microbiologic Forestière, INRA Centre de Recherches de Nancy, 54280 Champenoux, France. Tel. 33–83 39 40 80; fax 33–83 39 40 69; e-mail: fmartin@nancy.inra.fr
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  • This paper results from collaborative research between the groups of F. Martin and F. Le Tacon into development of PCR-based marker systems for detection of ectomycorrhizal fungi in the environment. Experimental work was carried out by B. Henrion and C. Di Battista. D. Bouchard is technical officer and responsible for setting up and maintaining the field sites. D. Vairelles is technical officer and works on the capillary electrophoresis analysis of PCR products. B. Thompson is at Arbrogen Europe and provided the seedlings.

Equipe de Microbiologic Forestière, INRA Centre de Recherches de Nancy, 54280 Champenoux, France. Tel. 33–83 39 40 80; fax 33–83 39 40 69; e-mail: fmartin@nancy.inra.fr

Abstract

The large-scale inoculation of selected beneficial ectomycorrhizal fungi in forest nurseries has generated renewed interest in the ecology of these symbiotic fungi. However, information on the dissemination and persistence of introduced symbionts is scarce due to the limitation of the current identification methods. To identify ectomycorrhizal fungi on single root tips, we investigated the polymorphism of the PCR-amplified ribosomal DNA intergenic spacer (IGS) from a wide range of ectomycorrhizal fungi. To investigate the reliability of this molecular approach in large-scale surveys, the dissemination and persistence on Douglas fir seedlings of the introduced Laccaria bicolor S238N were assessed in a forest nursery in the Massif Central (France). Several hundred ectomycorrhizas and fruiting bodies were sampled from plots where control and L. bicolor inoculated-Douglas fir seedlings were grown for 1.5 years. PCR typing of mycorrhizas indicated that trees inoculated with L. bicolor S238N remained exclusively colonized by that isolate (or sexually derived isolates) for the entire test period. In contrast, control seedlings were infected by indigenous isolates of Laccaria laccata and Thelephora terrestris. The molecular evidence for the persistence of the introduced mycobiont despite the competition from indigenous isolates of the same species provides further illustration of the potential of exotic species for large-scale microbial application.

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