Characterization of the glyoxalase I gene from the vascular wilt fungus Verticillium dahliae.
A glyoxalase I gene homologue (VdGLO1) was identified in the vascular wilt fungus Verticillium dahliae by sequence tag analysis of genes expressed during resting structure development. The results of the current study show that the gene encodes a putative 345 amino acid protein with high similarity to glyoxalase I, which produces S-D-lactoylglutathione from the toxic metabolic by-product methylglyoxal (MG). Disruption of the V. dahliae gene by Agrobacterium tumefaciens-mediated transformation resulted in enhanced sensitivity to MG. Mycelial growth of disruption mutants was severely reduced in the presence of 5 mmol/L MG. In contrast, spore production in liquid medium was abolished at 1 mmol/L MG, although not at physiologically relevant concentrations of <or=100 micromol/L. In this first report on the characterization of a glyoxalase I gene in a vascular wilt pathogen, we found that disruption of VdGLO1 had no discernable effect on the pathogenicity of V. dahliae. These data suggest that while the glyoxalase system is necessary for effectively dealing with catastrophic levels of MG, under normal conditions of growth and infection, other MG detoxification pathways in V. dahliae are able to compensate for the absence of the glyoxalase system.
Klimes
A
A
Agriculture and AGri-Food Canada, Southern Crop Protection and Food Research Centre, London, ON, Canada.
Neumann
M J
MJ
Grant
S J
SJ
Dobinson
K F
KF
D016960:Agrobacterium tumefaciens / Q000235:genetics; D015139:Blotting, Southern; D003001:Cloning, Molecular; D004271:DNA, Fungal / Q000737:chemistry / Q000235:genetics; D015971:Gene Expression Regulation, Enzymologic; D015966:Gene Expression Regulation, Fungal; D005978:Glutathione / Q000031:analogs & derivatives / Q000378:metabolism; D007791:Lactoylglutathione Lyase / Q000235:genetics* / Q000378:metabolism; D008826:Microbial Sensitivity Tests / Q000379:methods; D008969:Molecular Sequence Data; D009154:Mutation / Q000235:genetics; D011765:Pyruvaldehyde / Q000378:metabolism / Q000494:pharmacology; D020133:Reverse Transcriptase Polymerase Chain Reaction; D017422:Sequence Analysis, DNA; D014170:Transformation, Genetic / Q000235:genetics; D020172:Verticillium / Q000187:drug effects / Q000201:enzymology / Q000235:genetics*
D004271:DNA, Fungal; D011765:Pyruvaldehyde; D007791:Lactoylglutathione Lyase; D005978:Glutathione; C013585:S-lactoylglutathione