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Applied and Environmental Microbiology, October 2000, p. 4305-4314, Vol. 66, No. 10
0099-2240/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.

Cytological Effects of Cellulases in the Parasitism of Phytophthora parasitica by Pythium oligandrum

Karine Picard,1 Yves Tirilly,1 and Nicole Benhamou2,*

Laboratoire de Microbiologie et Sécurité Alimentaire, Université de Brest, Technopôle Brest-Iroise, 29200 Plouzané, France,1 and Recherche en Sciences de la vie et de la santé, Pavillon Charles-Eugène Marchand, Université Laval, Sainte-Foy, Québec, Canada, G1K 7P42

Received 19 January 2000/Accepted 27 July 2000

The ubiquitous oomycete Pythium oligandrum is a potential biocontrol agent for use against a wide range of pathogenic fungi and an inducer of plant disease resistance. The ability of P. oligandrum to compete with root pathogens for saprophytic colonization of substrates may be critical for pathogen increase in soil, but other mechanisms, including antibiosis and enzyme production, also may play a role in the antagonistic process. We used transmission electron microscopy and gold cytochemistry to analyze the intercellular interaction between P. oligandrum and Phytophthora parasitica. Growth of P. oligandrum towards Phytophthora cells correlated with changes in the host, including retraction of the plasma membrane and cytoplasmic disorganization. These changes were associated with the deposition onto the inner host cell surface of a cellulose-enriched material. P. oligandrum hyphae could penetrate the thickened host cell wall and the cellulose-enriched material, suggesting that large amounts of cellulolytic enzymes were produced. Labeling of cellulose with gold-complexed exoglucanase showed that the integrity of the cellulose was greatly affected both along the channel of fungal penetration and also at a distance from it. We measured cellulolytic activity of P. oligandrum in substrate-free liquid medium. The enzymes present were almost as effective as those from Trichoderma viride in degrading both carboxymethyl cellulose and Phytophthora wall-bound cellulose. P. oligandrum and its cellulolytic enzymes may be useful for biological control of oomycete pathogens, including Phytophthora and Pythium spp., which are frequently encountered in field and greenhouse production.


* Corresponding author. Mailing address: Recherche en sciences de la vie et de la santé, Pavillon C.E. Marchand, Université Laval, Sainte-Foy, Québec, Canada G1K 7P4. Phone: (418) 656-7517. Fax: (418) 656-7176. E-mail: nben{at}rsvs.ulaval.ca.


Applied and Environmental Microbiology, October 2000, p. 4305-4314, Vol. 66, No. 10
0099-2240/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.



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