Identification of plant-inducible genes in Erwinia chrysanthemi 3937.
We present a method for identifying plant-inducible genes of Erwinia chrysanthemi 3937. Mutagenesis was done with the Mu dIIPR3 transposon, which carries a promoterless neomycin phosphotransferase gene (nptI), so upon insertion, the truncated gene can fuse to E. chrysanthemi promoters. Mutants containing insertions in plant-inducible genes were selected for their sensitivity to kanamycin on minimal plates and for their acquired resistance to this antibiotic when an S. ionantha plant extract was added to kanamycin minimal plates. The selection allowed the identification of E. chrysanthemi promoters inducible by host factors present in the S. ionantha plant extract. Using this method, we isolated 30 mutants and characterized 10 of them. Two mutants were defective in cation uptake, one was defective in the galacturonate degradation pathway, and another was altered in the production of the acidic pectate lyase. The functions of the other mutated genes are still unknown, but we show that most of them are involved in pathogenicity.
Beaulieu
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C
Département de Biologie Moléculaire, Université Libre de Bruxelles, Rhode-St-Genèse, Belgium.
Van Gijsegem
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F
D003090:Coliphages / Q000235:genetics; D004251:DNA Transposable Elements; D004279:DNA, Viral / Q000235:genetics / Q000302:isolation & purification; D004885:Erwinia / Q000187:drug effects / Q000235:genetics*; D004926:Escherichia coli / Q000235:genetics; D015964:Gene Expression Regulation, Bacterial; D005798:Genes, Bacterial; D019868:Kanamycin Kinase; D010770:Phosphotransferases / Q000235:genetics*; D010936:Plant Extracts / Q000494:pharmacology; D018521:Plant Physiological Phenomena; D010957:Plasmids; D015183:Restriction Mapping
D004251:DNA Transposable Elements; D004279:DNA, Viral; D010936:Plant Extracts; D010770:Phosphotransferases; D019868:Kanamycin Kinase