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Applied and Environmental Microbiology, May 2004, p. 3073-3081, Vol. 70, No. 5
0099-2240/04/$08.00+0     DOI: 10.1128/AEM.70.5.3073-3081.2004
Copyright © 2004, American Society for Microbiology. All Rights Reserved.

In Vivo Study of Trichoderma-Pathogen-Plant Interactions, Using Constitutive and Inducible Green Fluorescent Protein Reporter Systems

Zexun Lu,1 Riccardo Tombolini,2 Sheridan Woo,3 Susanne Zeilinger,4 Matteo Lorito,3 and Janet K. Jansson1,5*

Section for Natural Sciences, Södertörn University College, 14189 Huddinge,1 Plant Pathology and Biocontrol Unit and,2 Department of Microbiology, Swedish University of Agricultural Sciences, 75007 Uppsala, Sweden,5 Universitá degli Studi di Napoli Federico II and Centro di Studio CNR per le Tecniche di Lotta Biologica, 80050 Portici, Italy,3 Abteilung für Mikrobielle Biochemie, Institut für Biochemische Technologie und Mikrobiologie, Technische Universität Wien, A-1060 Vienna, Austria4

Received 14 July 2003/ Accepted 27 January 2004

Plant tissue colonization by Trichoderma atroviride plays a critical role in the reduction of diseases caused by phytopathogenic fungi, but this process has not been thoroughly studied in situ. We monitored in situ interactions between gfp-tagged biocontrol strains of T. atroviride and soilborne plant pathogens that were grown in cocultures and on cucumber seeds by confocal scanning laser microscopy and fluorescence stereomicroscopy. Spores of T. atroviride adhered to Pythium ultimum mycelia in coculture experiments. In mycoparasitic interactions of T. atroviride with P. ultimum or Rhizoctonia solani, the mycoparasitic hyphae grew alongside the pathogen mycelia, and this was followed by coiling and formation of specialized structures similar to hooks, appressoria, and papillae. The morphological changes observed depended on the pathogen tested. Branching of T. atroviride mycelium appeared to be an active response to the presence of the pathogenic host. Mycoparasitism of P. ultimum by T. atroviride occurred on cucumber seed surfaces while the seeds were germinating. The interaction of these fungi on the cucumber seeds was similar to the interaction observed in coculture experiments. Green fluorescent protein expression under the control of host-inducible promoters was also studied. The induction of specific Trichoderma genes was monitored visually in cocultures, on plant surfaces, and in soil in the presence of colloidal chitin or Rhizoctonia by confocal microscopy and fluorescence stereomicroscopy. These tools allowed initiation of the mycoparasitic gene expression cascade to be monitored in vivo.

* Corresponding author. Mailing address: Department of Microbiology, Swedish University of Agricultural Sciences, 75007 Uppsala, Sweden. Phone: 46-18-673201. Fax: 46-18-673393. E-mail: janet.jansson{at}mikrob.slu.se.

Applied and Environmental Microbiology, May 2004, p. 3073-3081, Vol. 70, No. 5
0099-2240/04/$08.00+0     DOI: 10.1128/AEM.70.5.3073-3081.2004
Copyright © 2004, American Society for Microbiology. All Rights Reserved.



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  • Zeilinger, S., Reithner, B., Scala, V., Peissl, I., Lorito, M., Mach, R. L. (2005). Signal Transduction by Tga3, a Novel G Protein {alpha} Subunit of Trichoderma atroviride. Appl. Environ. Microbiol. 71: 1591-1597 [Abstract] [Full Text]  


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