Bacterial species specificity in proU osmoinducibility and nptII and lacZ expression

Reporter gene-based transcriptional fusions are increasingly being used to address questions in microbial ecology, with constitutively expressed fusions enabling microbe tracking and inducible fusions reporting the presence of environmental signals. To more readily apply this technology to a variety of bacterial species, we examined species specificity in the expression of three promoters of interest. A comparison of two potentially constitutive promoters, each fused to the reporter gene gfp, showed that the nptII promoter (P(nptII)) was expressed in a broader range of species (100% of 11 tested) than the lacZ promoter (P(lacZ)) (75% of 11 tested), and thus has broader applicability for marking bacteria than P(lacZ). For the species that expressed P(lacZ), however,P(lacZ) was expressed 3-fold more than P(nptII), on average. The Escherichia coli proU promoter, which is induced by low water potential in E. coli, Salmonella typhimurium, Pantoea agglomerans, and Pseudomonas syringae, was shown to be similarly responsive to water potential in strains of Clavibacter michiganensis, Enterobacter aerogenes, Pseudomonas fluorescens, Pseudomonas putida, and Sinorhizobium meliloti, as well as mildly osmoresponsive in Agrobacterium tumefaciens, supporting its broad use as a reporter of water potential. Surprisingly, this promoter was not regulated by water potential in strains of Staphylococcus aureus and Erwinia amylovora, illustrating heretofore unrecognized species specificity in proU inducibility, as well as potential limitations in the species that can serve as bioreporters of water potential.