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Molecular Characterization and Functional Analysis of MgNLP, the Sole NPP1 Domain--Containing Protein, from the Fungal Wheat Leaf Pathogen Mycosphaerella graminicola

July 2009 , Volume 22 , Number  7
Pages  790 - 799

Juliet Motteram,1 Isabell Küfner,2 Siân Deller,1 Frédéric Brunner,2 Kim E. Hammond-Kosack,1 Thorsten Nürnberger,2 and Jason J. Rudd1

1Centre for Sustainable Pest and Disease Management, Department of Plant Pathology and Microbiology, Rothamsted Research, Harpenden, Herts AL5 2JQ, U.K.; 2University of Tübingen ZMBP--Plant Biochemistry, Auf der Morgenstelle 5, D-72076 Tübingen, Germany

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Accepted 3 March 2009.

Analysis of the fully sequenced genome of the wheat leaf-specific fungal pathogen Mycosphaerella graminicola identified only a single gene encoding a member of the necrosis- and ethylene-inducing peptide 1 (Nep1)-like protein family (NLP). NLP proteins have frequently been shown to trigger cell death and the activation of defense signaling reactions in dicotyledonous plants. However, complete loss-of-function reverse genetics analyses for their importance in the virulence of eukaryotic plant pathogens are generally lacking. Real-time quantitative polymerase chain reaction on MgNLP demonstrated the gene to be specifically expressed in planta. Peak expression was observed during the immediate presymptomatic phase of colonization of a susceptible host genotype. This was followed by a dramatic decrease during disease lesion formation which, in this system, exhibits characteristics of host programmed cell death (PCD). No comparable peak in transcript levels was seen during an incompatible interaction with a host genotype exhibiting gene-for-gene--based disease resistance. Heterologously expressed MgNLP protein induced necrotic cell death and the activation of defense-related genes when infiltrated into Arabidopsis leaves but not in leaves of a susceptible wheat genotype. MgNLP infiltration also failed to stimulate wheat mitogen-activated protein kinase activities. Finally, targeted deletion of M. graminicola MgNLP caused no detectable reduction in plant pathogenicity or virulence, suggesting that this protein is not a major virulence determinant during fungal infection of its host plant. To our knowledge, this represents the first complete loss-of-function analysis of NLP in a eukaryotic plant pathogen and we discuss our findings in the context of possible functions for NLP in pathogens which only infect monocotyledonous plants.

© 2009 The American Phytopathological Society