Use of an intergenic region in Pseudomonas syringae pv. syringae B728a for site-directed genomic marking of bacterial strains for field experiments.
To construct differentially-marked derivatives of our model wild-type strain, Pseudomonas syringae pv. syringae B728a (a causal agent of bacterial brown spot disease in snap bean plants), for field experiments, we selected a site in the gacS-cysM intergenic region for site-directed insertion of antibiotic resistance marker cassettes. In each of three field experiments, population sizes of the site-directed chromosomally marked B728a derivatives in association with snap bean plants were not significantly different from that of the wild-type strain. Inserts of up to 7 kb of DNA in the intergenic region did not measurably affect fitness of B728a in the field. The site is useful for site-directed genomic insertions of single copies of genes of interest.
Hirano
S S
SS
Departments of Plant Pathology, University of Wisconsin-Madison, Madison, Wisconsin 53706, USA. ssh@plantpath.wisc.edu
Willis
D K
DK
Clayton
M K
MK
Upper
C D
CD
D001426:Bacterial Proteins / Q000235:genetics / Q000378:metabolism; D015169:Colony Count, Microbial; D021901:DNA, Intergenic / Q000235:genetics*; D004352:Drug Resistance, Microbial / Q000235:genetics; D007887:Fabaceae / Q000382:microbiology*; D005819:Genetic Markers; D016680:Genome, Bacterial; D009154:Mutation; D010935:Plant Diseases / Q000382:microbiology*; D010946:Plants, Medicinal; D011549:Pseudomonas / Q000187:drug effects / Q000235:genetics / Q000254:growth & development* / Q000472:pathogenicity*; D011995:Recombination, Genetic; D014157:Transcription Factors / Q000235:genetics
D001426:Bacterial Proteins; D021901:DNA, Intergenic; D005819:Genetic Markers; D014157:Transcription Factors; C074165:lemA protein, bacterial