First report of Phoma betae on Aloe vera in India

Leaf spot disease of A. vera was observed in nurseries of Gwalior city afterthe post-rainy season. As the disease progressed, the tip of the leaf shrank, then dried and eventually broke. The causal agent was identified as Phoma betae A.B. Frank. This is the first report of leaf spot disease on Aloe vera caused by P. betae in India.     

Some observations on assessing Phoma betae infection of sugar-beet seed

Near-ultraviolet or ‘black’ light applied continuously from the start of incubation, facilitates tests for Phoma betae on sugar-beet seed by stimulating the production of pycnidia and restricting mycelial growth of P. betae and other fungi. Pretreatment of the seed with dilute sodium hypochlorite decreases the number of seeds with P. betae by removing superficial infection, but some of this is of significance in the field. Rubbing beet seed also decreased counts of P. betae in the laboratory and increased field emergence, primarily by removing the fungus. Griseofulvin sprayed on beet-seed plants either 2 wk or 2 days before harvest significantly decreased seed infection with P. betae, but not to a level at which further seed treatment could be omitted.     

Interference by common leaf saprophytic fungi with the development of Phoma betae lesions on sugarbeet leaves

When sugarbeet leaves were inoculated simultaneously with the parasitic Phoma betae and the saprophytic Aureobasidium pullulans or Torulopsis Candida, the development of expanding lesions caused by P. betae was curtailed. Two other common saprophytes, Sporobolomyces pararoseus and Cladosporium cladosporioides, also decreased numbers and sizes of expanding lesions in the presence of pollen. The numbers of saprophyte cells applied to inoculum areas were similar to those occurring naturally on sugarbeet leaves in the summer. Lesion development was not affected by a mixture of heat-killed cells of the four saprophytes.     

A Phytotoxin,Betaenone C,and Its Related Metabolites of Phoma betae Fr

For rough quantitative analysis of genetically modified maize contents, rapid methods for measurement of the copy numbers of the cauliflower mosaic virus 35S promoter region (P35S) and MON810 construct-specific gene (MON810) using a combination of a capillary-type real-time PCR system with a plasmid DNA were established. To reduce the characteristic differences between the plasmid DNA and genomic DNA, we showed that pretreatment of the extracted genomic DNA by a combination of sonication and restriction endonuclease digestion before measurement is effective. The accuracy and reproducibility of this method for MON810 content (%) at a level of 5.0% MON810 mixed samples were within a range from 4.26 to 5.11% in the P35S copy number quantification. These methods should prove to be a useful tool to roughly quantify GM maize content.     

Effect of Microwaves on Escherichia coli and Bacillus subtilis

Suspensions of Escherichia coli and Bacillus subtilis spores were exposed to conventional thermal and microwave energy at 2,450 MHz. The degrees of inactivation by the different energy sources were compared quantitatively. During the transient heating period by microwave energy, approximately a 6 log cycle reduction in viability was encountered for E. coli. This reduction was nearly identical to what is expected for the same time-temperature exposure to conventional heating. Heating of B. subtilis spores by conventional and microwave energy was also carried out at 100 C, in ice and for transient heating. The degree of inactivation by microwave energy was again identical to that by conventional heating. In conclusion, inactivation of E. coli and B. subtilis by exposure to microwaves is solely due to the thermal energy, and there is no per se effect of microwaves.     

Effect of Bromouracil-containing Deoxyribonucleic Acid on Bacillus subtilis

Gimlin, Dixie M. (Oklahoma State University, Stillwater), Sue D. Hardman, Betty N. Kelley, Grace C. Butler, and Franklin R. Leach. Effect of bromouracil-containing deoxyribonucleic acid on Bacillus subtilis. J. Bacteriol. 92:366-374. 1966.-Replacement of one-half of the thymine with bromouracil in Bacillus subtilis transforming deoxyribonucleic acid (DNA) resulted in a slight decrease in transforming activity, but, when used at high concentrations, this DNA preparation inhibited cell growth. Acid-hydrolyzed DNA, or addition of equivalent concentrations of the free base bromouracil in a transforming mixture, was without effect on cell growth. Treatment of the DNA preparation with deoxyribonuclease completely destroyed transforming activity and killing effect, whereas treatments with ribonuclease and trypsin were without effect on either transformation or killing activity. Growth of competent B. subtilis cells in test tubes was inhibited by high concentrations of both normal and bromouracil-containing DNA, with the bromouracil-containing DNA being significantly more inhibitory. This type of inhibition was also reflected in the time of division of the cells. The inhibitory effect was not due to viscosity, or to mutagenicity. The time course of killing paralleled transformation, and competency was required. These results can be interpreted as being due to uptake of homologous but imperfect DNA (containing bromouracil instead of thymine) by means of the systems involved in transformation, followed by either integration (resulting in lethal transformation, activation of a defective, nonlytic but lethal prophage) or interference with the recombination mechanism.     

Effect of microwave radiation on Bacillus subtilis spores

AIMS: To compare the killing efficacy and the effects exerted by microwaves and conventional heating on structural and molecular components of Bacillus subtilis spores. METHODS AND RESULTS: A microwave waveguide applicator was developed to generate a uniform and measurable distribution of the microwave electric-field amplitude. The applicator enabled the killing efficacy exerted by microwaves on B. subtilis spores to be evaluated in comparison with conventional heating at the same temperature value. The two treatments produced a similar kinetics of spore survival, while remarkably different effects on spore structures were seen. The cortex layer of the spores subjected to conductive heating was 10 times wider than that of the untreated spores; in contrast, the cortex of irradiated spores did not change. In addition, the heated spores were found to release appreciable amounts of dipicolinic acid (DPA) upon treatment, while extracellular DPA was completely undetectable in supernatants of the irradiated spores. These observations suggest that microwave radiation may promote the formation of stable complexes between DPA and other spore components (i.e. calcium ions); thus, making any release of DPA from irradiated spores undetectable. Indeed, while a decrease in measurable DPA concentrations was not produced by microwave radiation on pure DPA solutions, a significant lowering in DPA concentration was detected when this molecule was exposed to microwaves in the presence of either calcium ions or spore suspensions. CONCLUSIONS: Microwaves are as effective as conductive heating in killing B. subtilis spores, but the microwave E-field induces changes in the structural and/or molecular components of spores that differ from those attributable only to heat. SIGNIFICANCE AND IMPACT OF THE STUDY: This study provides information on the effect of microwaves on B. subtilis spore components.     

Effect of Sulfhydryl Reagents on the Ribosomes of Bacillus subtilis

The effect of various sulfhydryl reagents on the ribosomes of Bacillus subtilis was studied. The 70S ribosomes were completely dissociated into 30S and 50S subunits by appropriate concentrations of p-chloromercuribenzoic acid (PCMB) and 5,5'-dithio-bis-(2-nitro-benzoic acid). The N-ethylmaleimide and iodoacetamide failed to dissociate the ribosomes even at relatively high concentrations. The rate of dissociation of ribosomes by PCMB varied with the concentration of ribosomes. A progressive decrease in the rate of dissociation was observed as the concentration of ribosomes in the reaction mixture was increased. The PCMB-induced ribosomal subunits were unable to reassociate into 70S monomers unless they were dialyzed against buffer containing beta-mercaptoethanol. On the average, four molecules of PCMB per 70S ribosome and two molecules of PCMB per each 30S and 50S subunit were bound. The number of PCMB molecules bound per ribosome did not change with increasing concentrations of PCMB, even though higher concentrations of PCMB resulted in dissociation of ribosomes into subunits.     

Effect of caffeine on the recombination process of Bacillus subtilis

Summary The effect of caffeine on the recombination process was studied by using transformation and transfection of Bacillus subtilis as genetic systems. Data were obtained showing that caffeine reduces strongly both transformation and transfection. The inhibitory effect seems ascribable to an interference of caffeine with the process of recombination.     

Effect of Temperature on the Cannibalistic Behavior of Bacillus subtilis

Bacillus subtilis resorts to cannibalism to delay sporulation under severe nutritional limitation. We report the effect of temperature on the dynamics of cannibalism demonstrated by B. subtilis. A model consisting of a delay differential equation may explain the effect of temperature on the dynamics of cannibalism.     

枯草芽孢杆菌胞外氨肽酶对菌体生长的作用研究

【目的】利用基因敲除技术构建突变菌株BS-AP-K来研究枯草芽孢杆菌的分泌型氨肽酶对菌体生长的作用。【方法】基于Xer/dif重组系统敲除Bacillus subtilis 168基因组中ywaD基因,研究比较野生型与BS-AP-K菌株在不同培养基中的生长情况。【结果】通过比较两菌株的生长情况,发现敲除分泌型氨肽酶会对菌体生长带来不利影响,而这种影响可以通过在培养基中添加多种游离氨基酸来弥补。【结论】研究结果表明胞外氨肽酶通过酶切外源蛋白质以及多肽来为细胞生长提供营养所需。     

Sporulation in Bacillus subtilis. Effect of medium on the form of chromosome replication and on initiation to sporulation in Bacillus subtilis

Thymine-requiring mutants of Bacillus subtilis and mutants that are temperature-sensitive for initiation of chromosome replication have been used to study the relationship between sporulation and chromosome formation. The DNA synthesis that normally occurs when cells are transferred to sporulation medium is essential for spore induction. This is shown by the fact that thymine-starved cells are unable to form spores and are unable to perform even the earlier steps of sporulation, such as septum formation or synthesis of alkaline phosphatase. The nature of the medium in which the cells are growing while the DNA is being completed is also important because it determines both the shape and the position of the daughter chromosomes. If the cells are in a rich medium, the newly synthesized chromosomes are discrete and compact bodies: the cells are primed for growth, and sporulation cannot be induced by transferring them at this stage to a spore-inducing medium. If DNA synthesis was completed with the cells in a poor medium the daughter chromosomes, by the time DNA synthesis has ceased, are spread in a single filamentous band and the cells are morphologically already in stage I of sporulation.     

Effect of Monosodium n-Alkylsalicylates on Spores of Bacillus subtilis

The effects of several monosodium n-alkylsalicylates on washed spores of Bacillus subtilis have been examined. None of the compounds was sporicidal, but in general they prevented dormancy induced by incubating the spores in water. This effect was related to the position in the ring and size of the alkyl groups substituted on the salicylic acid nucleus.     

Bactericidal Effect of ADP and Acetic Acid on Bacillus subtilis

Bacillus subtilis is a ubiquitous soil bacterium used for measuring the β-lysin activity and in other bioassays. We observed a complete bactericidal effect of ADP on B. subtilis at concentrations of 50–100 μM at pH values <5.5, which disappeared at pH values above 6. The effect was also found for acetic acid at concentrations >17.4 μM and similar pH values. ATP, adenosine, and HCl were not bactericidal. We used BCECF-AM, a pH-sensitive probe, and found that the killing of B. subtilis was due to a change in the intracellular pH caused by the passage across the cell membrane of these weak organic acids when incubated with B. subtilis at pH values near the pK. More experiments are needed to determine the biological meaning of these in vitro findings. Received: 14 June 1996 / Accepted: 19 July 1996