RESUMEN
Ten strains of Xanthomonas axonopodis pv. glycines, the causal agent of bacterial pustule of soybean, which were isolated from various soybean growing regions of Thailand, produced an extracellular diffusible factor (DSF) related to a well-characterized quorum sensing molecule produced by other Xanthomonas spp. Genomic DNA of the 10 strains of X. axonopodis pv. glycines contained rpfF, a gene encoding for the biosynthesis of the DSF in X. campestris pv. campestris. The rpfF gene from one strain of X. axonopodis pv. glycines was fully sequenced, and the 289 aa product is closely related to RpfF of other Xanthomonas spp. (95 to 98% identical). Three independently generated rpfF mutants of X. axonopodis pv. glycines strain No12-2 were defective in the production of a DSF, as expected if rpfF encodes for DSF biosynthesis in X. axonopodis pv. glycines. The rpfF mutants of X. axonopodis pv. glycines exhibited reduced virulence on soybean and produced less than wild-type levels of extracellular polysaccharide and the extracellular enzymes carboxylmethylcellulase, protease, endo-beta-1,4-mannanase, and pectate lyase. Transcripts for three genes that encode for the extracellular enzymes protease, endoglucanase, and pectate lyase were at lower abundance in an rpfF mutant than in the parental strain of X. axonopodis pv. glycines. These results indicate that X. axonopodis pv. glycines produces a diffusible signal related to the DSF of X. campestris pv. campestris, which contributes to virulence and exoenzyme production by this phytopathogenic bacterium.
Asunto(s)
Proteínas Bacterianas/genética , Regulación Bacteriana de la Expresión Génica , Glycine max/microbiología , Xanthomonas axonopodis/genética , Proteínas Bacterianas/metabolismo , Celulasa/genética , Manosidasas/genética , Mutación , Péptido Hidrolasas/genética , Enfermedades de las Plantas/microbiología , Polisacárido Liasas/genética , Polisacáridos Bacterianos/metabolismo , Reacción en Cadena de la Polimerasa de Transcriptasa Inversa , Tailandia , Virulencia/genética , Xanthomonas axonopodis/metabolismo , Xanthomonas axonopodis/patogenicidadRESUMEN
Nitrogen-fixing microbial populations in a Douglas fir forest on the western slope of the Oregon Cascade Mountain Range were analyzed. The complexity of the nifH gene pool (nifH is the marker gene which encodes nitrogenase reductase) was assessed by performing nested PCR with bulk DNA extracted from plant litter and soil. The restriction fragment length polymorphisms (RFLPs) of PCR products obtained from litter were reproducibly different than the RFLPs of PCR products obtained from the underlying soil. The characteristic differences were found during the entire sampling period between May and September. RFLP analyses of cloned nifH PCR products also revealed characteristic patterns for each sample type. Among 42 nifH clones obtained from a forest litter library nine different RFLP patterns were found, and among 64 nifH clones obtained from forest soil libraries 13 different patterns were found. Only two of the patterns were found in both the litter and the soil, indicating that there were major differences between the nitrogen-fixing microbial populations. A sequence analysis of clones representing the 20 distinct patterns revealed that 19 of the patterns had a proteobacterial origin. All of the nifH sequences obtained from the Douglas fir forest litter localized in a distinct phylogenetic cluster characterized by the nifH sequences of members of the genera Rhizobium, Sinorhizobium, and Azospirillum. The nifH sequences obtained from soil were found in two additional clusters, one characterized by sequences of members of the genera Bradyrhizobium, Azorhizobium, Herbaspirillum, and Thiobacillus and the other, represented by a single nifH clone, located between the gram-positive bacteria and the cyanobacteria. Our results revealed the distinctness of the nitrogen-fixing microbial populations in litter and soil in a Douglas fir forest; the differences may be related to special requirements for degradation and mineralization processes in the plant litter.
Asunto(s)
Bacterias/genética , Genes Bacterianos , Fijación del Nitrógeno , Nitrogenasa/genética , Oxidorreductasas , Microbiología del Suelo , Árboles/microbiología , Bacterias/enzimología , Bacterias/aislamiento & purificación , Cianobacterias/genética , Cianobacterias/aislamiento & purificación , Ecosistema , Datos de Secuencia Molecular , Oregon , Filogenia , Reacción en Cadena de la Polimerasa/métodos , Polimorfismo de Longitud del Fragmento de Restricción , Rhizobiaceae/genética , Análisis de Secuencia de ADNRESUMEN
Temporal airborne bacterial concentrations and meteorological conditions were measured above a grass seed field in the Willamette River Valley, near Corvallis, Oreg., in the summer of 1993. The concentration of airborne bacteria had a maximum of 1,368.5 CFU/m(sup3), with a coefficient of variation of 90.5% and a mean of 121.3 CFU/m(sup3). The lowest concentration of bacteria occurred during the predawn hours, with an average of 32.2 CFU/m(sup3), while sunrise and early evening hours had the highest averages (164.7 and 158.1 CFU/m(sup3), respectively). The concentrations of bacteria in the atmosphere varied greatly, with a maximum difference between two 2-min samples of 1,995 CFU/m(sup3). The concentrations of bacteria in the atmosphere could be divided into five time periods during the day that were thought to be related to the local diurnal sea breeze and Pacific Coast monsoon weather conditions as follows: (i) the nighttime minimum concentration, i.e., 2300 to 0600 h; (ii) the sunrise peak concentration, i.e., 0600 to 0800 h; (iii) the midday accumulating concentration, i.e., 0800 to 1515 h; (iv) the late-afternoon sea breeze trough concentration, i.e., 1515 to 1700 h; and (v) the evening decrease to the nighttime minimum concentration, i.e., 1700 to 2300 h. The sunrise peak concentration (period ii) is thought to be a relatively general phenomenon dependent on ground heating by the sun, while the afternoon trough concentration is thought to be a relatively local phenomenon dependent on the afternoon sea breeze. Meteorological conditions are thought to be an important regulating influence on airborne bacterial concentrations in the outdoor atmosphere in the Willamette River Valley.
RESUMEN
Freeze-dried cultures of wild-type and genetically engineered strains of Escherichia coli lost their colony-forming ability upon exposure to air, visible light, and certain relative humidity levels. Both strains could be maximally protected from these lethal effects with 100 mM trehalose, a concentration calculated to just saturate the interphospholipid spaces in the cell membrane, thus preserving the liquid-crystalline structure. The trehalose protection was observed for at least 96 h. Trehalose increased viability as much as 2000-4000% over nontreated populations. In all cases, exposure to environmental conditions was more damaging to the genetically engineered strain.
Asunto(s)
Crioprotectores/farmacología , Escherichia coli/efectos de los fármacos , Liofilización/métodos , Trehalosa/farmacología , Escherichia coli/genética , Escherichia coli/crecimiento & desarrollo , Estudios de Evaluación como Asunto , Ingeniería GenéticaRESUMEN
Because the death mechanisms of freeze-dried and air-dried bacteria are thought to be similar, freeze-drying was used to investigate the survival differences between potentially airborne genetically engineered microorganisms and their wild types. To this end, engineered strains of Escherichia coli and Pseudomonas syringae were freeze-dried and exposed to air, visible light, or both. The death rates of all engineered strains were significantly higher than those of their parental strains. Light and air exposure were found to increase the death rates of all strains. Application of death rate models to freeze-dried engineered bacteria to be released into the environment is discussed.
Asunto(s)
Escherichia coli/fisiología , Ingeniería Genética , Pseudomonas/fisiología , Aire , Exposición a Riesgos Ambientales/efectos adversos , Escherichia coli/clasificación , Liofilización , Luz/efectos adversos , Pseudomonas/clasificaciónRESUMEN
Catalase incorporation into enumeration media caused a significant increase (greater than 63%) in the colony-forming abilities of airborne bacteria. Incubation for 30 to 60 min of airborne bacteria in collection fluid containing catalase caused a greater than 95% increase in colony-forming ability. However, catalase did not have any effects on enumeration at high relative humidities (80 to 90%).
Asunto(s)
Microbiología del Aire , Bacterias Aerobias/efectos de los fármacos , Catalasa/farmacología , Bacterias Aerobias/crecimiento & desarrollo , Recuento de Colonia Microbiana , Medios de CultivoRESUMEN
Simulated droplet trajectories of a polydispersed microbial aerosol in a laminar air flow regimen were compared with observed dispersal patterns of aerosolized Bacillus subtilis subsp. niger spores in quasilaminar airflow. Simulated dispersal patterns could be explained in terms of initial droplet sizes and whether the droplets evaporated to residual aeroplanktonic size before settling to the ground. For droplets that evaporated prior to settling out, a vertical downwind size fractionation is predicted in which the microbial residue of the smallest droplets settles the least, and is found in the airstream at about sprayer height, and progressively larger droplet residues settle to progressively lower heights. Observations of spore particle size distributions downwind from a spray source support the simulation. Droplet and particle size distributions near the source had three size fractions: one containing large, presumably nonevaporated droplets of greater than or equal to 7 microns in diameter, and two smaller fractions, with diameters of 2 to 3 microns (probably the residue of droplets containing more than one spore) and 1 to 2 microns (probably the residue from single-spore droplets). As predicted by the simulation, the aerosol settled and progressed downwind, with the number of small droplets and particles increasing in proportion to the height and distance downwind.