Functional characterisation of glyoxalase I from the fungal wheat pathogen Stagonospora nodorum

Abstract

During an expressed sequence tag sequencing project, a gene encoding a methylglyoxal lyase (glyoxalase I) was identified, cloned and characterised from the necrotrophic wheat pathogen Stagonospora nodorum. Sequence analysis identified the gene, named Gox1, as having reasonable identity to GLO1 from yeast and hypothetical proteins identified in fungal genome sequencing projects. Expression analysis in vitro revealed Gox1 to be up-regulated in the presence of methylglyoxal and salt but not affected by starvation conditions. Analysis of Gox1 transcription in planta showed its highest expression was in ungerminated spores and during sporulation, suggesting a role for the glycolytic bypass pathway in sporulation. The gene was inactivated by homologous recombination, resulting in a S. nodorum strain with no detectable glyoxalase I activity. The gox1 mutants exhibited no discernable phenotype, with the exception of being more sensitive to the presence of methylglyoxal. Infection assays demonstrated the mutants retained full pathogenicity and sporulation was unaffected. This is the first report describing the characterisation of a glyoxalase I from a pathogen of any description. The gene has been sequenced, functionally characterised and shown not to be required for the infection of wheat by S. nodorum.