The oxidative stress response in Bacillus subtilis

Abstract Bacillus subtilis undergoes a typical bacterial stress response when exposed to low concentrations (0.1 mM) of hydrogen peroxide. Protection is thereby induced against otherwise lethal, challenge concentrations (10 mM) of this oxidant and a number of proteins are induced including the scavenging enzymes, catalase and alkyl hydroperoxide reductase, and a putative DNA binding and protecting protein. Induced protection against higher concentrations (10–30 mM) of hydrogen peroxide is eliminated in a catalase-deficient mutant. Both RecA and Spo0A influence the basal but not the induced resistance to hydrogen peroxide. A regulatory mutation has been characterized that affects the inducible phenotype and is constitutively resistant to high concentrations of hydrogen peroxide. This mutant constitutively overexpresses the proteins induced by hydrogen peroxide in the wild-type. The resistance of spores to hydrogen peroxide is partly attributable to binding of small acid soluble proteins by the spore DNA and partly to a second step which coincides with the depletion of the NADH pool, which may inhibit the generation of hydroxyl radicals from hydrogen peroxide.     

Evaluation of monoammonium phosphate and bacterial strains to increase tree growth and fruit yield in apple replant problem soil

The long-term effects of biological agents alone and in combination with monoammonium phosphate on tree growth and fruit production of apple trees planted on apple replant soil was studied for five years. Application of monoammonium phosphate (MAP) in the year of planting increased shoot growth, cross-sectional trunk area and fruit yield of McIntosh on M.26 rootstock for the first two years. The application of bacterial agents alone were not effective in increasing young tree growth except BACT-1 in 1987. None of the bacterial agents increased fruit yield when applied alone. The addition of certain bacterial agents to MAP application increased young tree growth in various years. The combination of bacterial agent B-10 and MAP reduced young tree growth and yield compared with the MAP treatment alone. These results suggest that the application of MAP alone may be sufficient to alleviate the replant problem and the addition of BACT-1, EBW-4 or B8 bacterial agents to this treatment may be beneficial to increase tree growth in some years. Contribution number 822. Contribution number 822.     

Plasmid instability in an industrial strain ofBacillus subtilis grown in chemostat culture

Summary A pUB110-derived plasmid/Bacillus subtilis host combination was segregationally unstable when grown in chemostat culture with complex or minimal medium and under starch, glucose or magnesium limitation. The kinetics of plasmid loss were described in terms of the difference in growth rates between plasmid-containing and plasmid-free cells (d) and the rate at which plasmid-free cells were generated from plasmid-containing cells (R). Loss of plasmid-containing cells from the population was d dominated. Changes in medium composition and the nature of growth limitation caused variations in both d and R. The plasmid was most stable in glucose-limited chemostat cultures with minimal medium and least stable under starch limitation with complex complex medium. R and d were smaller for cultures in complex media than those in minimal media. Limitation by starch induced expression of the plasmid-encoded HT amylase gene and was associated with increased values of R and d. Magnesium limitation in minimal medium caused a significant increase in d and a decrease in R.Abbreviations Cm chloramphenicol - Kan kanamycin - Cm^r cells resistant to chloramphenicol (5 mg L^–1) - Kan^r cells resistant to kanamycin (5 mg L^–1) - Cm^sKan^s cells sensitive to chloramphenicol and kanamycin     

不同饲毒法测定Bt8010毒力效价研究

以棉铃虫初孵幼虫为供试虫,用合成饲料和天然饲料两种饲毒法,CS3ab-1991（毒力效价Ha 15000IU/mg)为标准品对Bt 8010粉剂进行毒力测定,饲毒后的供试虫在30℃恒温下培育72小时,合成饲料饲毒法测定的CS3ab-1991 LC50=108.6±10.8μg/ml,Bt8010 LC50=197.1±15.6μg/ml,毒力效价8273.1±715.8Ha IU/mg,天然饲料饲毒法测定的CS3ab-1991 LC50=406.3±47.5μg/ml,Bt8010 LC50=705.0±86.8μg/ml,毒力效价8633.2±871.8Ha IU/mg。     

Recovery and evaluation of soybean plants transgenic for aBacillus thuringiensis var.Kurstaki insecticidal gene

Summary Lepidopteran insects are major defoliating pests of soybean in the southeastern United States. Soybean plants transgenic for a nativecryIA(b) gene fromBacillus thuringiensis var.kurstaki HD-1 were obtained. Embryogenic cultures were induced by plating cotyledons on a Murashige and Skoog-based medium supplemented with 40 mg/liter of 2,4-dichlorophenoxyacetic acid (2,4-D). The embryogenic cultures were maintained in liquid medium containing 5 mg/liter 2,4-D. These cultures were subjected to microprojectile bombardment, followed by selection on 50 mg/liter hygromycin. Resistant embryogenic cell lines were transferred to growth regulator-free medium to permit recovery of mature somatic embryos. After a desiccation period, the somatic embryos were returned to growth regulator-free medium for conversion into plants. Southern hybridization analysis verified transformation. Feeding assays of T₁ plants from one cell line deterred feeding, development, and survival of velvetbean caterpillar at a level comparable to that of GatIR81-296, a soybean breeding line with a high level of insect resistance. Reduced feeding on T₁ plants correlated with the presence of the transgene.     

Sterilization of various diameter dead-ended tubes

Effect of tube diameter on steam-in-place sterilization of dead-ended tubes was studied by examining temperature profiles and rates of kill of Bacillus stearothermophilus spores. Time required for sterilization was determined for 9.4-cm-long tubes with various inside diameters from 0.4 to 1.7 cm. Sterilization time increased with decreasing tube diameter. Experimentally measured kill kinetics in 1.7-cm tubes were in agreement with those predicted if measured temperatures represented saturated steam. A 12-log spore reduction was achieved in 1.7-cm diameter vertical and horizontal tubes in less than 63 minutes. For smaller diameter tubes, entrapped air remained after 2 hours and rates of kill were very dependent on position within the tube, tube diameter, and tube orientation with respect to the gravitational vector. Times to achieve a 1-log drop in spore population in the smaller tubes were as much as 10 times greater than those expected if measured temperatures represented saturated steam. Sterilization was not achieved throughout the 0.4-cm tubes. Recommendations are made for including steam bleeders or using prevaccum cycles for these smaller diameter tubes. (c) 1993 John Wiley & Sons, Inc.     

A simulation model for the continuous production of acetoin and butanediol using Bacillus subtilis with integrated pervaporation separation

The potential for producing acetoin and butanediol with a Bacillus subtilis strain was investigated with continuous culture using molasses as carbon substrate. The steady-state results were influenced by both oxygen and undetermined limiting compounds. Employing the known metabolic pathways, four overall stoichiometry relations were used with an energetic assumption on the energy requirements for biomass formation to establish a linear relations were used with an energetic assumption on the energy requirements for biomass formation to establish a linear relation between the overall rates, whose parameters were determined by linear regression. This provided a relationship for the product formation rate. The chemostat culture data were described with a growth kinetics model, which included limitation by molasses and oxygen as well as diauxic effects and product inhibition. The biokinetics model was combined with an experimentally verified model for the membrane Pervaporation. From this combined model were determined the influence of the membrane characteristics (enrichment factors and membrane area) and the dilution rate on the performance of the integrated process. Simulations revealed that an increase of the enrichment factor, possible by membrane improvement, would have counteracting influences, owing to decreased product inhibition but with lower biomass concentration. (c) 1993 Wiley & Sons, Inc.