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Eukaryotic Cell, November 2008, p. 1980-1993, Vol. 7, No. 11
1535-9778/08/$08.00+0     doi:10.1128/EC.00091-08
Copyright © 2008, American Society for Microbiology. All Rights Reserved.

Cell Wall Chitosaccharides Are Essential Components and Exposed Patterns of the Phytopathogenic Oomycete Aphanomyces euteiches{triangledown}

Ilham Badreddine,1 Claude Lafitte,1 Laurent Heux,2 Nicholas Skandalis,3 Zacharoula Spanou,3 Yves Martinez,1 Marie-Thérèse Esquerré-Tugayé,1 Vincent Bulone,4 Bernard Dumas,1 and Arnaud Bottin1*

Université de Toulouse, UMR 5546 CNRS-UPS, 24 Chemin de Borde-Rouge, BP 42617, Auzeville, F-31326 Castanet-Tolosan, France,1 CERMAV-CNRS and Joseph Fourier Université, BP53, F-38041 Grenoble Cedex 9, France,2 Institute of Molecular Biology and Biotechnology, University of Crete, FORTH, Vassilika Vouton, P.O. Box 1385, GR 711 10 Heraklion, Crete, Greece,3 School of Biotechnology, Royal Institute of Technology (KTH), AlbaNova University Center, 106 91 Stockholm, Sweden4

Received 12 March 2008/ Accepted 19 August 2008

Chitin is an essential component of fungal cell walls, where it forms a crystalline scaffold, and chitooligosaccharides derived from it are signaling molecules recognized by the hosts of pathogenic fungi. Oomycetes are cellulosic fungus-like microorganisms which most often lack chitin in their cell walls. Here we present the first study of the cell wall of the oomycete Aphanomyces euteiches, a major parasite of legume plants. Biochemical analyses demonstrated the presence of ca. 10% N-acetyl-D-glucosamine (GlcNAc) in the cell wall. Further characterization of the GlcNAc-containing material revealed that it corresponds to noncrystalline chitosaccharides associated with glucans, rather than to chitin per se. Two putative chitin synthase (CHS) genes were identified by data mining of an A. euteiches expressed sequence tag collection and Southern blot analysis, and full-length cDNA sequences of both genes were obtained. Phylogeny analysis indicated that oomycete CHS diversification occurred before the divergence of the major oomycete lineages. Remarkably, lectin labeling showed that the Aphanomyces euteiches chitosaccharides are exposed at the cell wall surface, and study of the effect of the CHS inhibitor nikkomycin Z demonstrated that they are involved in cell wall function. These data open new perspectives for the development of antioomycete drugs and further studies of the molecular mechanisms involved in the recognition of pathogenic oomycetes by the host plants.

* Corresponding author. Mailing address: UMR 5546, CNRS-Université Paul-Sabatier, 24 Chemin de Borde-Rouge, BP 42617, Auzeville, F-31326 Castanet-Tolosan, France. Phone: 33 (0)5 621 935 18. Fax: 33 (0)5 621 935 02. E-mail: bottin{at}scsv.ups-tlse.fr

{triangledown} Published ahead of print on 19 September 2008.

Eukaryotic Cell, November 2008, p. 1980-1993, Vol. 7, No. 11
1535-9778/08/$08.00+0     doi:10.1128/EC.00091-08
Copyright © 2008, American Society for Microbiology. All Rights Reserved.





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