细菌细胞间通讯的群感效应*

自然界中的细菌大多数以生物膜的形式存在,这种存在方式增强了细菌对环境的适应性和病原菌的致病性。近年来研究表明,细菌群感效应(quorumsensing)是调控生物膜形成和其它生物学功能的机制。细菌能够分泌特定的信号分子并感应它的浓度,当信号分子浓度达到阈值时,细菌就能够引发包括致病基因在内的相关基因的表达以适应环境的变化。由于生物膜的形成是病原菌致病性和其它要求一定细胞密度才能产生功能的基础,所以细菌群感效应的发现为防止病原菌的毒害作用提供了新的思路。     

细菌群感效应的猝灭

细菌群感效应（quorum sensing,QS）是一种细菌种间和种内信息交流调控机制.研究证明,细菌群感效应与细菌生物膜的形成和某些人体、植物病原菌的发病机制有关,目前它已成为医学、生物工程、农业和环境工程等领域的研究热点,有望在此基础上开发出控制生物膜形成和病原菌致病性的新方法.该文介绍目前已有的细菌群感效应的抑制方法（即细菌群感猝灭,quorum quenching）,主要包括降低R蛋白活性、抑制信号分子合成,以及降解信号分子等.     

细菌群体感应信号分子与抑制剂研究进展

具有群体感应系统的细菌通过相互交换一种自动诱导（autoinducer）信号分子来实现彼此问的信息交流。当信号分子积累到一定浓度时会改变细菌特定基因的表达,如生物膜的形成、生物发光行为、毒性基因的表达、孢子的形成等。近年来,人们发现了多种天然或者人工合成的群体感应抑制剂,可以干扰群感系统的信息回路。本文系统地阐述了细菌群体感应信息系统的划分、自体诱导分子及其抑制剂的研究进展。     

吲哚作为细菌细胞间信号分子的研究进展

吲哚广泛存在于自然界,目前已知超过145种革兰氏阳性和阴性细菌能产生吲哚,其中包括许多病原菌。随着细菌密度感应系统及其信号分子作用机制研究的深入,吲哚已被证实是肠道病原菌如致病性大肠杆菌、迟缓爱德华氏菌、霍乱弧菌等一类细胞间重要的信号分子,并参与细菌的多种生理活动,如毒力、抗药性、生物膜形成、运动性、质粒稳定性、抗酸性、孢子产生等。更为重要的是,吲哚及其衍生物还参与协调菌群竞争,有益于人体肠道菌群平衡和免疫系统。本文在吲哚作为细胞间信号分子参与迟缓爱德华氏菌的毒力、抗药性、生物膜形成和运动性的研究基础上,对近年来吲哚作为细菌细胞间信号分子的研究进展进行了综述。随着吲哚作用机制的进一步揭示,将有助于新型抗病原菌感染策略的研发和生物工程方面的应用。     

群感效应与链霉菌次生代谢调控

群感效应是细菌协调群体行为的一种外界信号传递机制,在细菌中普遍存在,参与细胞的多种生理过程。链霉菌中也存在群感效应,在抗生素等次生代谢产物的生物合成中起重要的调控作用;从自诱导信号分子的结构到信号传递机制都存在一定多样性,其中以A-因子为代表的γ-丁酸内酯类信号分子的作用机制研究最为深入。近几年在链霉菌中发现的PI-因子、M-因子以及一些特定的代谢产物则代表几类结构较新颖的信号分子,通过群感效应机制调控次生代谢过程;链霉菌中还发现胆固醇氧化酶、甘油等分子具有信号分子特征,不排除是通过群感效应来参与抗生素生物合成调控。本文主要就参与链霉菌次生代谢调控的几类群感效应系统的研究状况进行综述,重点阐述各类群感信号分子的结构和信号传递机制的不同,并对链霉菌群感效应的研究趋势以及在抗生素高产菌遗传育种中的应用前景进行了展望。     

群体感应与病原菌致病性研究进展

群体感应是细菌根据细胞密度变化进行基因表达调控的一种生理行为。当细菌密度达到临界阈值时能释放一些特定的自诱导信号分子,从而调节本种群或同环境中其他种群的群体行为。细菌群体感应参与包括人类、动植物、病原菌在内的多种生物的生物学功能调节,如生物膜的形成、毒力因子的产生、病原菌的耐药性等。深入研究病原菌群体感应系统的调控机制,将提高对病原菌发病机制的认识,有利于以群体感应作为防治疾病策略的研究。系统阐述了群体感应系统的组成类型、群体感应与病原菌致病性的关系,及其在抑制病原菌致病方面的应用。     

群体感应抑制剂研究进展

细菌群体感应(quorum sensing,QS)是一种依赖于菌体密度而普遍存在于微生物之间的沟通协调机制,控制着细菌的生长、增殖、致病性、生物膜形成及相关群体活动行为。通过阻断细菌群体感应信号,可以抑制病原菌的致病性。本文从群体感应角度总结了抑制病原菌的方法和研究进展,为临床新型抗菌药物的设计提供理论依据和新策略。     

细菌群体感应及其干扰策略的研究进展

群体感应（QS）广泛存在于细菌中,是细菌根据细胞密度变化调控基因表达的一种机制。许多植物病原菌依赖QS调控致病基因和毒性因子的表达,导致植物发病,因此通过抑制QS效应就为控制细菌病害提供了一种有效的方法。目前发现许多途径可以干扰细菌的QS,如：产生可使信号分子降解的酶,产生病原菌信号分子的类似物与信号分子受体蛋白竞争结合来阻断病原菌的群体感应,利用QS中信号分子来诱发寄主抗性。系统阐述了细菌QS及其干扰策略。     

细菌群体感应及其在病原菌防治中的应用

细菌分泌一种或多种化学信号分子,这些化学信号分子作为诱导因子感知和判断菌群密度和周围环境的变化。当菌群达到一定阈值时会启动一系列相关基因的表达以调控菌体的群体行为,细菌的这种生理行为称为群体感应。大量的研究表明,不同类型的细菌具有不同的群体感应系统。群体感应机制广泛存在于病原菌中,并与其侵染过程、毒力基因表达及致病性密切相关。利用这种群体感应机制作为靶点进行病原菌的防治是医学领域广泛关注的问题。在此就细菌群体感应及其在病原菌防治中的应用进行阐述。     

微生物信号分子降解酶研究进展

微生物细胞之间存在的信息交流称为群体感应。群体感应在实现微生物的生物学功能方面具有重要作用,包括调节致病性、参与生物膜的形成等。微生物能够分泌特定的信号分子,通过对信号分子的检测及应答,调控目的基因的表达。抑制信号分子的积累,能够干扰群体感应系统,使微生物丧失生物学功能。研究较为全面的一类信号分子是酰基高丝氨酸内酯(acylhomoserine lactone,AHL),此类信号分子可以通过酶法降解。目前已鉴定出的AHL降解酶主要分为AHL内酯酶和AHL酰化酶两类。综述了信号分子降解酶的来源、筛选方法、纯化技术、酶学性质、作用机制及在病害防治方面的应用。对信号分子降解酶的研究有助于完善群体感应系统的调控机制,并为微生物疾病的防治提供新策略。     

Characterization of regulatory pathways in Xylella fastidiosa: genes and phenotypes controlled by algU

Many virulence genes in plant bacterial pathogens are coordinately regulated by "global" regulatory genes. Conducting DNA microarray analysis of bacterial mutants of such genes, compared with the wild type, can help to refine the list of genes that may contribute to virulence in bacterial pathogens. The regulatory gene algU, with roles in stress response and regulation of the biosynthesis of the exopolysaccharide alginate in Pseudomonas aeruginosa and many other bacteria, has been extensively studied. The role of algU in Xylella fastidiosa, the cause of Pierce's disease of grapevines, was analyzed by mutation and whole-genome microarray analysis to define its involvement in aggregation, biofilm formation, and virulence. In this study, an algU::nptII mutant had reduced cell-cell aggregation, attachment, and biofilm formation and lower virulence in grapevines. Microarray analysis showed that 42 genes had significantly lower expression in the algU::nptII mutant than in the wild type. Among these are several genes that could contribute to cell aggregation and biofilm formation, as well as other physiological processes such as virulence, competition, and survival.     

Characterization of Regulatory Pathways in Xylella fastidiosa: Genes and Phenotypes Controlled by algU

Many virulence genes in plant bacterial pathogens are coordinately regulated by “global” regulatory genes. Conducting DNA microarray analysis of bacterial mutants of such genes, compared with the wild type, can help to refine the list of genes that may contribute to virulence in bacterial pathogens. The regulatory gene algU, with roles in stress response and regulation of the biosynthesis of the exopolysaccharide alginate in Pseudomonas aeruginosa and many other bacteria, has been extensively studied. The role of algU in Xylella fastidiosa, the cause of Pierce's disease of grapevines, was analyzed by mutation and whole-genome microarray analysis to define its involvement in aggregation, biofilm formation, and virulence. In this study, an algU::nptII mutant had reduced cell-cell aggregation, attachment, and biofilm formation and lower virulence in grapevines. Microarray analysis showed that 42 genes had significantly lower expression in the algU::nptII mutant than in the wild type. Among these are several genes that could contribute to cell aggregation and biofilm formation, as well as other physiological processes such as virulence, competition, and survival.     

Future perspective on host-pathogen interactions during bacterial biofilm formation within the nasopharynx

Nasopharyngeal colonization provides bacteria with a place of residence, a platform for person-to-person transmission and for many opportunistic pathogens it is a prerequisite event towards the development of invasive disease. Therefore, how host factors within the nasopharynx contribute to, inhibit or otherwise shape biofilm formation, the primary mode of existence for colonizing bacteria, and how biofilm bacteria subvert the acute inflammatory response that facilitates clearance, are important topics for future microbiological research. This review proposes the examination of host components as bridging molecules for bacterial interactions during biofilm formation, altered virulence determinant production and cell wall modification as a mechanism for immunoquiescence, and the role of host factors as signals and co-opted mechanisms for bacterial dissemination, together providing an opportunity for disease.     

Food color ‘Azorubine’ interferes with quorum sensing regulated functions and obliterates biofilm formed by food associated bacteria: An in vitro and in silico approach

Quorum sensing (QS) plays a crucial role in different stages of biofilm development, virulence production, and subsequently to the growth of bacteria in food environments. Biofilm mediated spoilage of food is one of the ongoing challenge faced by the food industry worldwide as it incurs substantial economic losses and leads to various health issues. In the present investigation, we studied the interference of quorum sensing, its regulated virulence functions, and biofilm in food-associated bacteria by colorant azorubine. In vitro bioassays demonstrated significant inhibition of QS and its coordinated virulence functions in Chromobacterium violaceum 12472 (violacein) and Pseudomonas aeruginosa PAO1 (elastase, protease, pyocyanin, and alginate). Further, the decrease in the production EPS (49–63%) and swarming motility (61–83%) of the pathogens was also recorded at sub-MICs. Azorubine demonstrated broad-spectrum biofilm inhibitory potency (50–65%) against Chromobacterium violaceum, Pseudomonas aeruginosa, E. coli O157:H7, Serratia marcescens, and Listeria monocytogenes. ROS generation due to the interaction between bacteria and azorubine could be responsible for the biofilm inhibitory action of the food colorant. Findings of the in vitro studies were well supported by molecular docking and simulation analysis of azorubine and QS virulence proteins. Azorubine showed strong binding to PqsA as compared to other virulent proteins (LasR, Vfr, and QscR). Thus, it is concluded that azorubine is a promising candidate to ensure food safety by curbing the menace of bacterial QS and biofilm-based spoilage of food and reduce economic losses.     

The Rise of Pathogens: Predation as a Factor Driving the Evolution of Human Pathogens in the Environment

Bacteria in the environment must survive predation from bacteriophage, heterotrophic protists, and predatory bacteria. This selective pressure has resulted in the evolution of a variety of defense mechanisms, which can also function as virulence factors. Here we discuss the potential dual function of some of the mechanisms, which protect against heterotrophic protists, and how predation pressure leads to the evolution of pathogenicity. This is in accordance with the coincidental evolution hypothesis, which suggests that virulence factors arose as a response to other selective pressures, for example, predation rather than for virulence per se. In this review we discuss some of those environmental factors that may be associated with the rise of pathogens in the marine environment. In particular, we will discuss the role of heterotrophic protists in the evolution of virulence factors in marine bacteria. Finally, we will discuss the implications for expansion of current pathogens and emergence of new pathogens.     

Modifications of Xanthomonas axonopodis pv. citri lipopolysaccharide affect the basal response and the virulence process during citrus canker

Xanthomonas axonopodis pv. citri (Xac) is the phytopathogen responsible for citrus canker, one of the most devastating citrus diseases in the world. A broad range of pathogens is recognized by plants through so-called pathogen-associated molecular patterns (PAMPs), which are highly conserved fragments of pathogenic molecules. In plant pathogenic bacteria, lipopolisaccharyde (LPS) is considered a virulence factor and it is being recognized as a PAMP. The study of the participation of Xac LPS in citrus canker establishment could help to understand the molecular bases of this disease. In the present work we investigated the role of Xac LPS in bacterial virulence and in basal defense during the interaction with host and non host plants. We analyzed physiological features of Xac mutants in LPS biosynthesis genes (wzt and rfb303) and the effect of these mutations on the interaction with orange and tobacco plants. Xac mutants showed an increased sensitivity to external stresses and differences in bacterial motilities, in vivo and in vitro adhesion and biofilm formation. Changes in the expression levels of the LPS biosynthesis genes were observed in a medium that mimics the plant environment. Xacwzt exhibited reduced virulence in host plants compared to Xac wild-type and Xacrfb303. However, both mutant strains produced a lower increase in the expression levels of host plant defense-related genes respect to the parental strain. In addition, Xac LPS mutants were not able to generate HR during the incompatible interaction with tobacco plants. Our findings indicate that the structural modifications of Xac LPS impinge on other physiological attributes and lead to a reduction in bacterial virulence. On the other hand, Xac LPS has a role in the activation of basal defense in host and non host plants.     

The HD-GYP domain of RpfG mediates a direct linkage between the Rpf quorum-sensing pathway and a subset of diguanylate cyclase proteins in the phytopathogen Xanthomonas axonopodis pv citri

Bacteria use extracellular levels of small diffusible autoinducers to estimate local cell-density (quorum-sensing) and to regulate complex physiological processes. The quorum-sensing signal transduction pathway of Xanthomonas spp. phytopathogens has special features that distinguish it from that of other pathogens. This pathway consists of RpfF, necessary for the production of the unique autoinducer 'diffusible signalling factor' (DSF), and RpfC and RpfG, a two-component system necessary for the DSF-dependent production of extracellular pathogenicity factors and cellular dispersion. Yeast two-hybrid and direct in vitro assays were used to identify interactions involving the Rpf group of proteins. We show that RpfC, a protein consisting of N-terminal transmembrane, histidine kinase, response-regulator and C-terminal histidine phosphotransfer domains interacts with both RpfG, a protein consisting of an N-terminal response regulator domain and a C-terminal HD-GYP domain, and with RpfF. We also show that RpfC interacts with the only known homologue of 'conditioned medium factor', which is involved in quorum-sensing in Dictyostelium discoideum under conditions of nutritional stress. Furthermore, RpfCG is shown to interact with a second two-component system made up of NtrB and NtrC homologues. Finally we show that the recently characterized HD-GYP phosphodiesterase domain of RpfG interacts directly with diguanylate cyclase GGDEF domain-containing proteins coded by the Xanthomonas axonopodis pv. citri genome, which in other bacteria produce cyclic diGMP, an important second messenger involved in the regulation of complex bacterial processes including biofilm production, virulence and motility. These results demonstrate a direct physical linkage between quorum-sensing and cyclic diGMP signalling pathways in bacteria.     

Clinical and Environmental <Emphasis Type="Italic">Burkholderia</Emphasis> Strains: Biofilm Production and Intracellular Survival

Bacteria belonging to the Burkholderia species are important pulmonary pathogens in cystic fibrosis (CF) patients. Their ability to establish chronic and sometimes fatal infections seems linked to the quorum sensing-regulated expression of virulence factors. We examined 23 Burkholderia isolates, 19 obtained from CF patients and 4 from the environment, to evaluate their ability to form biofilm and to penetrate and replicate inside J774 macrophagic cells. Our results indicate that biofilm formation and intracellular survival are behavioral traits frequently expressed by Burkholderia strains isolated from CF patients. Successive isolates obtained from each of four chronically infected patients yielded bacteria consistently belonging to the same strain but showing increasing ability to replicate intracellularly and to produce biofilm, possibly due to in vivo bacterial microevolution driven by the selective lung environmental conditions. Protection against antimicrobials granted to burkholderiae by the expression of these two virulence factors might account for the frequent failures of antibiotic treatment in CF patients.