盐胁迫下豌豆幼苗的超弱发光

通过钠盐对豌豆 (pisumsativum)萌发时超弱发光影响的比较研究表明 ,在等渗条件下豌豆幼苗受激发光强度顺序为Na2 CO3 >NaCl>Na2 SO4 ;Na2 CO3 盐胁迫对植物的伤害程度高于其他盐类 ;多盐比单盐存在时的毒害小 ,受激发光作用降低。利用超弱发光方法 ,可以分析盐害对植物的作用 ,进一步搞清胁迫条件下植物生理反应的变化规律。     

成熟小麦抗穗发芽能力与超弱发光关系的研究

生物体的超弱发光表现与其本身的生理活动密切相关,利用超弱发光为指标,测定和比较了小麦不同品种在成熟时抗穗发芽的能力．为其进一步应用,解决农业实际问题提供方法和依据．     

New Criteria for the Evaluation of Soil Bacterial Complexes

The initial concentration of prokaryotic microorganisms, the type of their growth, doubling time, and the growth dynamics of bacteria and actinomycetes in three types of soil (meadow, chestnut, and soddy forest) were evaluated by the luminescence microscopic analysis of soil samples incubated in a humid chamber for 1 day. Soddy forest and chestnut soils differed in most of the parameters analyzed. Meadow soil was close to soddy forest soil in some parameters and to chestnut soil in other parameters. All soil suspensions exhibited high growth rates of bacteria and actinomycetes, indicating that the fraction of viable microorganisms in the soils was high.     

New criteria for evaluation of soil bacterial complexes

The initial concentration of prokaryotic microorganisms, the type of their growth, doubling time, and the growth dynamics of bacteria and actinomycetes in three types of soil (meadow, chestnut, and soddy forest) were evaluated by the luminescence microscopic analysis of soil samples incubated in a humid chamber for 1 day. Soddy forest and chestnut soils differed in most of the parameters analyzed. Meadow soil was close to soddy forest soil in some parameters and to chestnut soil in other parameters. All soil suspensions exhibited high growth rates of bacteria and actinomycetes, indicating that the fraction of viable microorganisms in the soils was high.     

Chemiluminescent assay for detection of viable microorganisms

The redox reaction between quinone and viable microorganisms produces active oxygen species. In this study, the production rates of active oxygen species were determined by a luminol chemiluminescent assay, and the luminescence intensity was found to be proportional to the viable cell number. The high sensitivity of the luminol chemiluminescent assay was achieved with Mo-ethylenediaminetetraacetate complex and menadione or coenzyme Q1. The detectable cell densities of bacteria and yeasts were found to be approximately several thousand colony-forming units (CFU/ml) when assays were performed with a 96-well microplate luminometer. The chemiluminescent assay requires 10 min for incubation of quinone and microorganisms and 2s for photon counting. Single Escherichia coli was detected after 4h of cultivation and centrifugation (5 min x 2). This simple chemiluminescent assay is expected to be useful for the rapid detection of viable bacteria and yeast.     

Growth and luminescence of luminous bacteria promoted by agents of microbial origin

The examination of four species of luminous bacteria Photobacterium leiognathi, Photobacterium phosphoreum, Vibrio fischeri and Vibrio harveyi has enabled us to reveal some nutrient medium components effecting growth, luminescence intensity and luciferase synthesis. These agents are nucleic components (nucleotides, nucleosides and amine bases), amino acids and vitamins, which are part of hydrolysates from the biomass of various lithotrophic microorganisms, hydrogen-oxidizing, ironoxidizing and carboxydobacteria. The effect of promoting agents essentially alters the physiological state and ultrastructure of the cells of luminous bacteria and increases luciferase biosynthesis two- to three-fold compared to a control.     

Enhancing ATP-based bacteria and biofilm detection by enzymatic pyrophosphate regeneration

The manufacturing processes of many electronic and medical products demand the use of high-quality water. Hence the water supply systems for these processes are required to be examined regularly for the presence of microorganisms and microbial biofilms. Among commonly used bacteria detection approaches, the ATP luminescence assay is a rapid, sensitive, and easy to perform method. The aim of this study is to investigate whether ATP regeneration from inorganic pyrophosphate, a product of the ATP luminescence assay, can stabilize the bioluminescence signals in ATP detection. ADPglc pyrophosphorylase (AGPPase), which catalyzes the synthesis of ATP from PP_i in the presence of ADPglc, was selected because the system yields much lower luminescence background than the commercially available ATP sulfurylase/adenosine 5′-phosphosulfate (APS) system which was broadly used in pyrosequencing technology. The AGPPase-based assay could be used to measure both PP_i and ATP quantitatively and shows 1.5- to 4.0-fold slight increases in a 10-min assay. The method could also be used to stabilize the luminescence signals in detection of Escherichia coli, Pseudomonas aeruginosa, and Bacillus cereus in either broth or biofilm. These findings suggest that the AGPPase-based ATP regeneration system will find many practical applications such as detection of bacterial biofilm in water pipelines.     

Continuous culture of photobacterium

The design and performance characteristics of a small volume (20 ml) continuous culture device for the cultivation of luminous bacteria are described. This simply constructed device can be used to supply luminescent bacteria with constant properties for either laboratory use or the assay of environmental pollutants. Furthermore, bacteria can be deployed to make sensitive (<1 nM) oxygen measurements. The culture device may be configured, alone or in combination, as a chemostat, turbidostat or a "bioluminostat" where bacterial bioluminescence becomes the controlling variable. Over extended periods (>1 week) it was possible to maintain steady-state luminescence within 5% of a pre-set value, although occasionally a non-bioluminescent "mutant" became dominant; in this case light emission was irreversibly lost. A secondary chamber provided additional flexibility and easy manipulation of the cultivated bacteria for testing. The continuous culture system is also suitable for the growth of recombinant microorganisms that either constitutively express luciferase, or do so in response to stress promoter activity. The non-standard control methodologies reported may have important research and industrial applications, for example in providing immediate process control or as an inferential method to optimize biomass: product yield ratios.     

羧酶体的组成、结构、功能和检测及其在脱氮细菌中的意义

羧酶体(Carboxysome)是一种具有CO2浓缩功能的"类细胞器",它存在于自养型脱氮细菌中,可增强细菌的自养生长能力。硝化细菌、厌氧氨氧化细菌和部分反硝化细菌都是重要的自养型脱氮细菌,探明其羧酶体的组成、结构和功能,将有助于揭示自养型脱氮菌的生长规律,进而强化生物脱氮过程。基于文献阅读和相关研究,本文对自养型细菌中羧酶体在组成、结构、功能和检测等方面的研究进展进行综述,以期为自养生物脱氮过程的深入理解和优化改进提供参考。     

The uncultured luminous symbiont of Anomalops katoptron (Beryciformes: Anomalopidae) represents a new bacterial genus

Flashlight fishes (Beryciformes: Anomalopidae) harbor luminous symbiotic bacteria in subocular light organs and use the bacterial light for predator avoidance, feeding, and communication. Despite many attempts anomalopid symbionts have not been brought into laboratory culture, which has restricted progress in understanding their phylogenetic relationships with other luminous bacteria, identification of the genes of their luminescence system, as well as the nature of their symbiotic interactions with their fish hosts. To begin addressing these issues, we used culture-independent analysis of the bacteria symbiotic with the anomalopid fish, Anomalops katoptron, to characterize the phylogeny of the bacteria and to identify the genes of their luminescence system including those involved in the regulation of luminescence. Analysis of the 16S rRNA, atpA, gapA, gyrB, pyrH, recA, rpoA, and topA genes resolved the A. katoptron symbionts as a clade nested within and deeply divergent from other members of Vibrionaceae. The bacterial luminescence (lux) genes were identified as a contiguous set (luxCDABEG), as found for the lux operons of other luminous bacteria. Phylogenetic analysis based on the lux genes confirmed the housekeeping gene phylogenetic placement. Furthermore, genes flanking the lux operon in the A. katoptron symbionts differed from those flanking lux operons of other genera of luminous bacteria. We therefore propose the candidate name Candidatus Photodesmus (Greek: photo = light, desmus = servant) katoptron for the species of bacteria symbiotic with A. katoptron. Results of a preliminary genomic analysis for genes regulating luminescence in other bacteria identified only a Vibrio harveyi-type luxR gene. These results suggest that expression of the luminescence system might be continuous in P. katoptron.     

微生物腐蚀及腐蚀机理研究进展

在不同的环境中,不同种类的微生物能在材料上附着繁殖,其生命活动会引起或加剧材料的腐蚀。根据种类及功能的不同,腐蚀微生物可以分为硫酸盐还原菌、硫氧化菌、产酸菌、铁氧化细菌、铁还原细菌、硝酸盐还原菌以及产粘液细菌等。微生物腐蚀几乎能使所有现用的材料受到严重影响,破坏材料的结构与性能,在建筑、运输管道、工业环境(石油化工等)以及海洋环境中造成巨大的安全隐患和财产损失。本文概述了目前发现的腐蚀相关微生物的类群和特性,以及相对应的微生物腐蚀机理,为防护和控制材料的微生物腐蚀提供理论指导。     

Superweak effects of chemical compounds and physical factors on biological systems

The results of extensive investigations of the responses of living systems to superweak influences are summarized. The results of the effect of ultralow concentrations of various biologically active substances and ultraweak physical fields (primarily electromagnetic) on biological systems of different level of organization, from molecular to population, were considered. Several single-type regularities peculiar to all systems were revealed: polymodal dose dependences, the efficiency of influences at doses below the background level and its dependence on the state of the system, the modification of the sensitivity of a system to subsequent influences, etc. Several hypotheses of the mechanisms of superweak influences and the role of water as a universal intermediate in these processes are presented. Possible consequences of the phenomenon and its applications in practice are discussed.     

Novel sensitive high-throughput screening strategy for nitrilase-producing strains

Nitrilases have found wide use in the pharmaceutical industry for the production of fine chemicals, and it is important to have a method by which to screen libraries of isolated or engineered nitrilase variants (including bacteria and fungi). The conventional methods, such as high-performance liquid chromatography, liquid chromatography-mass spectrometry, capillary electrophoresis, or gas chromatography, are tedious and time-consuming. Therefore, a direct and sensitive readout of the nitrilase's activity has to be considered. In this paper, we report a novel time-resolved luminescent probe: o-hydroxybenzonitrile derivatives could be applied to detect the activity of the nitrilases. By the action of nitrilases, o-hydroxybenzonitrile derivatives can be transformed to the corresponding salicylic acid derivatives, which, upon binding Tb(3+), serve as a photon antenna and sensitize Tb(3+) luminescence. Because of the time-resolved property of the luminescence, the background from the other proteins (especially in the fermentation system) in the assay could be reduced and, therefore, the sensitivity was increased. Moreover, because the detection was performed on a 96- or 384-well plate, the activity of the nitrilases from microorganisms could be determined quickly. Based on this strategy, the best fermentation conditions for nitrilase-producing strains were obtained.     

Quenching of the luminescence of phosphorescent bacteria as a test for assessing the toxicity of the phenol components of sewage

The object of this work was to estimate whether the luminescence of luminescent bacteria could be used as a biological test for assessment of the toxicity of phenol compounds in sewage. The toxicity of phenol compounds for luminescent bacteria was compared in terms of three indices: the quenching of luminescence, the inhibition of dehydrogenase activity and the ability to grow. Among the three indices, the quenching of luminescence was characterized by the highest sensitivity and the most rapid response.     

液质发酵食品中微生物群落及与乳酸菌间相互作用研究进展

液质发酵食品发酵过程中微生物组成复杂,复杂的微生物发酵体系会影响微生物的生长和代谢,继而改变微生物的群落结构与功能,最终影响液质发酵食品的品质.乳酸菌在食品发酵中对形成风味物质、提高营养价值、抑制腐败菌生长具有重要的作用.本文主要对近年来食醋、酱油和饮料酒等液质发酵食品中微生物群落及与乳酸菌间相互作用关系进行综述,了解...     

Effect of tririum on growth and bioluminescence of P. phosphoreum bacteria

Effect of tritium labeled amino acid valine (0.3-1.0 MBq/ml) on luminous bacteria P. Phosphoreum was studied. The amino acid was used as a nutrient medium for the bacteria. Tritium was found to suppress bacterial growth, but stimulate luminescence: luminescence intensity, quantum yield and time of light-emitting were increased. Activation of the luminescent function is explained by redistribution of electronic density at beta-decay, and affecting biochemical processes in the bacterial media. Effects of alpha- and beta-radiation on luminous bacteria are compared.     

Quantification of the geocarposphere and rhizosphere effect of peanut (Arachis hypogaea L.)

Roots and pods of field-grown peanut (groundnut) (Arachis hypogaea L.) were sampled at the R3, R5, and R7 developmental stages and examined in comparison to root- and pod-free soil for microbial population densities to assess the geocarposphere and rhizosphere effects. G/ S (no. geocarposphere microorganisms/no. soil microorganisms) and R/S (no. rhizosphere microorganisms/no. soil microorganisms) ratios were calculated for total fungi,Asperigillus flavus, spore-forming bacilli, coryneform bacteria, fluorescent pseudomonads, and total bacteria isolated on low- and high-nutrient media. A clear geocarposphere effect was evidenced by increased population densities of bacteria and fungi associated with developing pods compared to soil. G/S and R/S ratios were generally greater than 1.0 for all groups of microorganisms except bacilli. G/S ratios were greater for total bacteria than for total fungi at two of the three sample times, suggesting that bacteria were stimulated more than fungi in the zone around developing pods. In contrast, R/S ratios, were higher for total fungi than for total bacteria at two of three sample times. The preferential association of fungi and bacteria with early developmental stages of the pod indicates that some microorganisms are particularly well adapted for colonization of the peanut geocarposphere. These microorganisms are logical candidates for evaluation as biological control candiates forA. flavus.     

Effect of concentrating and exposing the bioluminescent bacteria to the non‐luminescent allo‐bacterial extracellular products on their luminescence

Bioluminescence is a biochemical process occurring in many organisms. Bacterial bioluminescence has been investigated extensively that lead to many applications of such knowledge. Quorum sensing in the bioluminescent bacteria is a chemical signal process to recognize the strength of its own population to start luminescence in harmony. There is a mechanism in these bacteria to also recognize inter‐species strength. When there is a higher number of these bacteria, the possibility and frequency of cell–cell physical contact will be high. In this study, the physical proximity was artificially enhanced between cells and the effect on luminescence in the concentrated cells in the normal culture medium and in the presence of other non‐bacterial cell‐free supernatants was investigated. The role of such physical contact in the quorum sensing in the bioluminescence is not known. Increase in the luminescence of V. fischeri when concentrated shows that the presence of physical proximity facilitates the quorum sensing for their bioluminescence. Copyright © 2009 John Wiley & Sons, Ltd.     

Visualization of tumors and metastases in live animals with bacteria and vaccinia virus encoding light-emitting proteins

We have shown that bacteria injected intravenously into live animals entered and replicated in solid tumors and metastases. The tumor-specific amplification process was visualized in real time using luciferase-catalyzed luminescence and green fluorescent protein fluorescence, which revealed the locations of the tumors and metastases. Escherichia coli and three attenuated pathogens (Vibrio cholerae, Salmonella typhimurium, and Listeria monocytogenes) all entered tumors and replicated. Similarly, the cytosolic vaccinia virus also showed tumor-specific replication, as visualized by real-time imaging. These findings indicate that neither auxotrophic mutations, nor vaccinia virus deficient for the thymidine kinase gene, nor anaerobic growth conditions were required for tumor specificity and intratumoral replication. We observed localization of tumors by light-emitting microorganisms in immunocompetent and in immunocompromised rodents with syngeneic and allogeneic tumors. Based on their 'tumor-finding' nature, bacteria and viruses may be designed to carry multiple genes for detection and treatment of cancer.