Different species of bacteria were tested for production of extracellular autoinducer-like activities that could stimulate the expression of the luminescence genes in Vibrio harveyi. Several species of bacteria, including the pathogens Vibrio cholerae and Vibrio parahaemolyticus, were found to produce such activities. Possible physiological roles for the two V. harveyi detection-response systems and their joint regulation are discussed.At least two species of marine bacteria, Vibrio fischeri and Vibrio harveyi, express bioluminescence in response to cell density. These two vibrios are found in different environments in the ocean. V. harveyi is found free-living in the sea as well as in the gut tracts of marine animals, where it exists at high population densities in association with other species of bacteria. V. fischeri is found in these habitats and also lives in pure culture as a light organ symbiont of various fish and squid (8). V. fischeri and V. harveyi accomplish density-dependent lux regulation by the synthesis, excretion, and detection of small signal molecules called autoinducers, which accumulate in the environment (9). The autoinducer that controls light production in V. fischeri is N-(3-oxohexanoyl)-L-homoserine lactone. Two autoinducers control density-dependent lux expression in V. harveyi. One of the V. harveyi autoinducers is N-(3-hydroxybutanoyl)-L-homoserine lactone (AI-1), and the second autoinducer (AI-2) remains to be identified. Recently, a number of other bacteria have been shown to control cell density-dependent functions through the excretion of and response to acyl-homoserine lactone autoinducers. Cell density-dependent regulation of lux expression is an example of a phenomenon called quorum sensing (5).Genetic analysis of the density-sensing apparatus of V. harveyi has shown that two independent density-sensing systems exist, and each is composed of a sensor-autoinducer pair; system 1 is composed of sensor 1 and AI-1, and system 2 is composed of sensor 2 and AI-2 (1, 2). The two density-sensing systems are redundant, because a null mutation in either system alone results in Lux ϩ strains, whereas null mutations in both systems render the cells dark and incapable of density sensing. In 1979 Greenberg et al. (6) reported that V. harveyi was stimulated to produce light following the addition of cellfree culture fluid from several species of nonluminous bacteria. At the time of that study it was not known that two autoinducer-detection systems existed in V. harveyi. Because V. harveyi mutants capable of responding to only AI-1 or AI-2 now exist, it is possible to determine through which pathway(s) the signals from these other organisms flow. Now that we also appreciate that many bacteria communicate intercellularly and control gene expression through the use of autoinducers, it is of interest to understand how they might accomplish crossspecies communication and use it for survival in various niches. In this study we have used V. harveyi sensor mutants as reporters for specific autoinduc...
Bacteria and yeast frequently exist as populations capable of reaching extremely high cell densities. With conventional culturing techniques, however, cell proliferation and ultimate density are limited by depletion of nutrients and accumulation of metabolites in the medium. Here we describe design and operation of microfabricated elastomer chips, in which chemostatic conditions are maintained for bacterial and yeast colonies growing in an array of shallow microscopic chambers. Walls of the chambers are impassable for the cells, but allow diffusion of chemicals. Thus, the chemical contents of the chambers are maintained virtually identical to those of the nearby channels with continuous flowthrough of a dynamically defined medium. We demonstrate growth of cell cultures to densely packed ensembles that proceeds exponentially in a temperature-dependent fashion, and we use the devices to monitor colony growth from a single cell and to analyze the cell response to an exogenously added autoinducer.
Vibrio fischeri belongs to the Vibrionaceae, a large family of marine ␥-proteobacteria that includes several dozen species known to engage in a diversity of beneficial or pathogenic interactions with animal tissue. Among the small number of pathogenic Vibrio species that cause human diseases are Vibrio cholerae, Vibrio parahaemolyticus, and Vibrio vulnificus, the only members of the Vibrionaceae that have had their genome sequences reported. Nonpathogenic members of the genus Vibrio, including a number of beneficial symbionts, make up the majority of the Vibrionaceae, but none of these species has been similarly examined. Here we report the genome sequence of V. fischeri ES114, which enters into a mutualistic symbiosis in the light organ of the bobtail squid, Euprymna scolopes. Analysis of this sequence has revealed surprising parallels with V. cholerae and other pathogens.genomics ͉ pili ͉ symbiosis ͉ toxins ͉ toxin-coregulated pilus
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