Quorum-sensing Bacteria Research Articles

Quorum sensing bacteria improve microbial networks stability and complexity in wastewater treatment plants

Quorum-sensing bacteria (QSB) are crucial factors for microbial communication, yet their ecological role in wastewater treatment plants (WWTPs) remains unclear. Here, we developed a method to identify QSB by comparing 16S rRNA gene sequences. QSB in 388 activated sludge samples collected from 130 WWTPs across China primarily were identified as rare taxa and conditionally rare taxa. A co-occurrence network shared by all sludge communities revealed that QSB exhibited higher average clustering coefficient (0.46) than non-QSB (0.15). Individual sludge networks demonstrated that quorum sensing microbiomes were positively correlated with network robustness and network complexity, including average clustering coefficient and link density. We confirmed that QSB keystones and QSB nodes have a positive impact on network complexity by influencing network modularity through a structural equation model. Meanwhile, QSB communities directly contributed to maintaining network robustness (r = 0.29, P < 0.05). Hence, QSB play an important role in promoting network complexity and stability. Furthermore, QSB communities were positively associated with the functional composition of activated sludge communities (r = 0.33, P < 0.01), especially the denitrification capacity (r = 0.45, P < 0.001). Overall, we elucidated the ecological significance of QSB and provided support for QS-based regulation of activated sludge microbial communities.

Insights into the membrane fouling aggravation under polyethylene terephthalate microplastics contamination: From a biochemical point of view

This study evaluated the potential effects of polyethylene terephthalate microplastics (PET-MPs) on a membrane bioreactor. The results showed that 10–30 mg/L PET-MPs exposure showed a minor effect on reactor performances, while 50 mg/L PET-MPs significantly decreased the chemical oxygen demand and NH4+-N removal efficiencies. Long-term PET-MPs exposure resulted in sludge foaming, and significantly increased extracellular polymeric substances (EPS) contents from 463.08 ± 62.42 to 952.54 ± 23.67 mg/m2, which was the main factor in triggering severe membrane fouling. Excitation emission matrix and Fourier transform infrared spectroscopic analysis revealed significant changes in fluorescence properties and functional groups of EPS. Particularly, protein secondary structure analysis indicated that high abundant β-sheet facilitated the exposure of inner hydrophobic groups of bacterial cells and EPS, and consequently enhanced the interfacial interactions of foulants-foulants and membrane-foulants. Long-term PET-MPs exposure significantly shifted the microbial community structures. In particular, the abundances of foam-forming filamentous bacteria and quorum-sensing (QS) bacteria, such as Thiothrix, Kouleothrix and Aeromonas, were constantly increasing with the increasing PET-MPs concentration. Furthermore, PET-MPs exposure significantly up-regulated the abundances of genes encoding QS signaling molecules and exopolysaccharides, such as lasI, lsrA and algZ. Collectively, these findings suggested that PET-MPs exposure stimulated the excessive growth of fouling-causing bacteria, which contained more biofilm formation related genes, that consequently accelerated membrane fouling.

Quorum sensing bacteria in microplastics epiphytic biofilms and their biological characteristics which potentially impact marine ecosystem

Microplastics (MPs) have been shown to be a new type of pollutant in the oceans, with complex biofilms attached to their surfaces. Bacteria with quorum sensing (QS) systems are important participants in biofilms. Such bacteria can secrete and detect signal molecules. When a signal molecule reaches its threshold level, bacteria with QS systems can perform several biological functions, such as biofilm formation and antibiotic metabolite production. However, the ecological effects of QS bacteria in biofilm as MPs distribute globally with ocean currents are not to be elucidate yet. In this study, polypropylene and polyvinyl chloride were selected for on-site enrichment to acquire microplastics with biofilms. Eight culturable QS bacteria in the resulting biofilm were isolated by using biosensor assays, and their biodiversity was analyzed. The profiles of the N-acyl-homoserine lactones (AHLs) produced by these bacteria were analyzed by using thin-layer chromatography (TLC)–bioautography and gas chromatography and mass spectrometry (GC–MS). Biofilm-forming properties and several biological characteristics, such as bacteriostasis, algal inhibition, and dimethylsulfoniopropionate (DMSP) degradation, were explored along with QS quenching. Results showed that QS bacteria were mainly affiliated with class Alphaproteobacteria, particularly Rhodobacteraceae, followed by class Gammaproteobacteria. TLC–bioautography and GC–MS analyses revealed that seven AHLs, namely, C6-HSL, C8-HSL, 3-oxo-C6-HSL, 3-oxo-C8-HSL, 3-oxo-C10-HSL, and two unidentified AHLs were produced. The QS system equipped bacteria with strong biofilm-forming capacity and may contribute to the keystone roles of Rhodobacteraceae. In addition, QS bacteria may exacerbate the adverse environmental effects of MPs, such as inducing the misfeeding of planktons on MPs. This study elucidated the diversity of QS bacteria in MP-associated biofilms and provided a new perspective of the effect of key membrane-forming bacteria on the marine ecological environment.

Biofouling mitigation and microbial community dynamics in the membrane bioreactor by the indigenous quorum quenching bacterium Delftia sp. JL5

The quorum quenching (QQ) strategy has attracted increasing attention in membrane bioreactor (MBR) fouling control. However, the applicable QQ strain remains limited. This study investigated the antibiofouling performance of a new indigenous QQ bacterium, Delftia sp. JL5 (JL5) in MBR. JL5 produces intracellular acylase that irreversibly degrades N-acylhomoserine lactones (AHL), inhibited biofilm formation of quorum-sensing bacteria from activated sludge. During 120 days of operation, immobilized JL5 substantially delayed MBR biofouling by 2.1 and 2.9 times, at a flux rate of 30 L/(m2·h) and 20 L/(m2·h), respectively. A slower flux rate was favorable for effective mitigation of JL5 biofouling. JL5 reduced the AHL and extracellular polymeric substances of biocake without affecting the efficiency of waste removal. The presence of JL5 significantly changed the microbial structure of the membrane biocake, but not the activated sludge. Collectively, high activity, durability, and acid tolerance credited JL5 as a promising strain for QQ-MBR.

Effects of astragaloside IV on membrane biofouling in a membrane bioreactor from the perspective of extracellular polymeric substances and the bacteria community structure

At present, the control of membrane biofouling is mainly achieved by regulating quorum sensing. In this study, the quorum-sensing inhibitor astragaloside IV was used to prevent membrane biofouling in a membrane bioreactor. It was found that astragaloside IV at a concentration of 1 mg/L doubled the time taken for membrane contamination to occur, and decreased the energy required for filtration by 62% without affecting reactor’s processing performance. This is because the addition of astragaloside IV decreased the soluble microbial protein concentration and the loosely bound extracellular polysaccharide content. Biodiversity analysis indicated that astragaloside IV may have inhibited membrane contamination by interfering with the structure of the bacteria community, particularly the populations of quorum-sensing bacteria, bacteria related to extracellular polymeric substance production, and quorum-quenching bacteria. This is the first time the effects of astragaloside IV on membrane contamination in membrane reactors and the inhibition mechanisms involved were investigated. Using astragaloside IV could effectively decrease the energy consumed in operating a membrane bioreactor.

Diversity of Bacteria with Quorum Sensing and Quenching Activities from Hydrothermal Vents in the Okinawa Trough

Quorum sensing (QS) is a chemical communication system by which bacteria coordinate gene expression and social behaviors. Quorum quenching (QQ) refers to processes of inhibiting the QS pathway. Deep-sea hydrothermal vents are extreme marine environments, where abundant and diverse microbial communities live. However, the nature of chemical communication in bacteria inhabiting the hydrothermal vent is poorly understood. In this study, the QS and QQ activities with N-acyl homoserine lactones (AHLs) as the autoinducer were detected in bacteria isolated from hydrothermal vents in the Okinawa Trough. A total of 18 and 108 isolates possessed AHL-producing and AHL-degrading abilities, respectively. Bacteria mainly affiliated with Rhodobacterales, Hyphomicrobiales, Enterobacterales and Sphingomonadales showed QS activities; QQ was mainly associated with Bacillales, Rhodospirillales and Sphingomonadales. The results showed that the bacterial QS and QQ processes are prevalent in hydrothermal environments in the Okinawa Trough. Furthermore, QS significantly affected the activities of extracellular enzymes represented by β-glucosidase, aminopeptidase and phosphatase in the four isolates with higher QS activities. Our results increase the current knowledge of the diversity of QS and QQ bacteria in extreme marine environments and shed light on the interspecific relationships to better investigate their dynamics and ecological roles in biogeochemical cycling.

Bringing Antimicrobial Strategies to a New Level: The Quorum Sensing System as a Target to Control Streptococcus suis.

Streptococcus suis (S. suis) is an important zoonotic pathogen. It mainly uses quorum sensing (QS) to adapt to complex and changeable environments. QS is a universal cell-to-cell communication system that has been widely studied for its physiological functions, including the regulation of bacterial adhesion, virulence, and biofilm formation. Quorum sensing inhibitors (QSIs) are highly effective at interfering with the QS system and bacteria have trouble developing resistance to them. We review the current research status of the S. suis LuxS/AI-2 QS system and QSIs. Studies showed that by inhibiting the formation of AI-2, targeting the LuxS protein, inhibiting the expression of luxs gene can control the LuxS/AI-2 QS system of S. suis. Other potential QSIs targets are summarized, which may be preventing and treating S. suis infections, including AI-2 production, transmission, LuxS protein, blockage of AI-2 binding to receptors, AI-2-mediated QS. Since antibiotics are becoming increasingly ineffective due to the emergence of resistant bacteria, including S. suis, it is thus critical to find new antibacterial drugs with different mechanisms of action. QSIs provide hope for the development of such drugs.

Protocol: a simple method for biosensor visualization of bacterial quorum sensing and quorum quenching interaction on Medicago roots

BackgroundDefining interactions of bacteria in the rhizosphere (encompassing the area near and on the plant root) is important to understand how they affect plant health. Some rhizosphere bacteria, including plant growth promoting rhizobacteria (PGPR) engage in the intraspecies communication known as quorum sensing (QS). Many species of Gram-negative bacteria use extracellular autoinducer signal molecules called N-acyl homoserine lactones (AHLs) for QS. Other rhizobacteria species, including PGPRs, can interfere with or disrupt QS through quorum quenching (QQ). Current AHL biosensor assays used for screening and identifying QS and QQ bacteria interactions fail to account for the role of the plant root.MethodsMedicago spp. seedlings germinated on Lullien agar were transferred to soft-agar plates containing the broad-range AHL biosensor Agrobacterium tumefaciens KYC55 and X-gal substrate. Cultures of QS and QQ bacteria as well as pure AHLs and a QQ enzyme were applied to the plant roots and incubated for 3 days.ResultsWe show that this expanded use of an AHL biosensor successfully allowed for visualization of QS/QQ interactions localized at the plant root. KYC55 detected pure AHLs as well as AHLs from live bacteria cultures grown directly on the media. We also showed clear detection of QQ interactions occurring in the presence of the plant root.ConclusionsOur novel tri-trophic system using an AHL biosensor is useful to study QS interspecies interactions in the rhizosphere.

Exploring the Function of Quorum Sensing Regulated Biofilms in Biological Wastewater Treatment: A Review.

Quorum sensing (QS), a type of bacterial cell–cell communication, produces autoinducers which help in biofilm formation in response to cell population density. In this review, biofilm formation, the role of QS in biofilm formation and development with reference to biological wastewater treatment are discussed. Autoinducers, for example, acyl-homoserine lactones (AHLs), auto-inducing oligo-peptides (AIPs) and autoinducer 2, present in both Gram-negative and Gram-positive bacteria, with their mechanism, are also explained. Over the years, wastewater treatment (WWT) by QS-regulated biofilms and their optimization for WWT have gained much attention. This article gives a comprehensive review of QS regulation methods, QS enrichment methods and QS inhibition methods in biological waste treatment systems. Typical QS enrichment methods comprise adding QS molecules, adding QS accelerants and cultivating QS bacteria, while typical QS inhibition methods consist of additions of quorum quenching (QQ) bacteria, QS-degrading enzymes, QS-degrading oxidants, and QS inhibitors. Potential applications of QS regulated biofilms for WWT have also been summarized. At last, the knowledge gaps present in current researches are analyzed, and future study requirements are proposed.

Acceleration of start-up of moving bed biofilm reactor at low temperature by adding specialized quorum sensing bacteria

This study aims to accelerate biofilm formation and operational performance of moving bed biofilm reactor (MBBR) at 5 ℃ by adding specialized Quorum Sensing bacteria (sphingomonas rubra BH3T). Results showed that bio augmented MBBR (RS) achieved a higher chemical oxygen demand and NH4+-N removal rate (93% and 75%), which in accordance with its increased biofilm thickness, higher biofilm activity, and nitrifying bacteria abundance (Nitrospira). The increased biofilm thickness (60.23 %) during the whole operating time, accompanied by more potent adhesion force (61.59 %), was related to increased polysaccharides and proteins in the biofilm. Pyrosequencing analysis indicated that BH3T contributed to higher species richness and triggered the rapid growth of precursor microorganisms (Nakamurella, Micropruina, and Zoogloea) and the enrichment of multifunctional microorganisms (Pseudomonas, Aeromonas, Arcobacter, Dechloromonas, and Flavobacterium) at low temperatures. This study provides an economical and practical new insight into accelerating start-up of MBBR system at low temperature.

Diversity and functions of quorum sensing bacteria in the root environment of the Suaeda glauca and Phragmites australis coastal wetlands.

The quorum sensing (QS) system plays a significant role in the bacteria-bacteria or plant-bacteria relationships through signal molecules. However, little is known about the distribution and functional diversity of QS bacteria in the root environment of Suaeda glauca and Phragmites australis in coastal wetlands. We explored the bacterial community by amplicon sequencing and isolated 1050 strains from the rhizosphere soil and root tissues of S. glauca and P. australis in northern China to investigate the bacterial community and AHL producers. AHL activity was found in 76 isolates, and 22 distinct strains were confirmed by 16S rRNA gene sequencing. A substantial number of AHL producers clustered in rhizobiales and sphingomonadale, which derived from the root tissues. AHL producers in the rhizosphere soil mostly belonged to rhodobacterales. The different taxa of AHL producers in the rhizosphere soil and root tissues resulted in a variation of AHL profiles that C6-HSL dominated the AHL profiles in root bacteria compared to the C8-HSL in rhizobacteria, implying different ecological roles for AHL producers in the rhizosphere soil and root tissues. Many AHL producers may form biofilms, and some can degrade DMSP and oil, demonstrating that QS bacteria in the root environment have a wide ecological roles. In our study, for one of the first times here, we explore the distribution and functional variety of AHL producers in the root environment of S. glauca-P. australis. This study expands current knowledge of the relationship between QS bacteria and coastal plants (S. glauca and P. australis), and vital roles of QS bacterial in maintaining the health of coastal wetlands.

Acceleration of Start-Up of Moving Bed Biofilm Reactor at Low Temperature by Adding Specialized Quorum Sensing Bacteria

Acceleration of Start-Up of Moving Bed Biofilm Reactor at Low Temperature by Adding Specialized Quorum Sensing Bacteria

Quorum Sensing Bacteria in the Phycosphere of HAB Microalgae and Their Ecological Functions Related to Cross-Kingdom Interactions.

It has been proven that the relationship between microalgae and bacteria affects the dynamic process of harmful algal blooms (HABs). Microalgae-associated microorganisms widely exist in the phycosphere and play an essential role in algae-bacteria cross-kingdom interactions. Among these processes, quorum sensing (QS), as a communication system of bacteria, is thought to participate in algae-bacteria interactions. However, the species of QS bacteria in the phycosphere and their ecological function are still unknown. In this study, microalgae-associated microorganisms with a QS system were screened by the biosensor method and identified based on 16S rRNA gene analysis. The types and number of acyl-L-homoserine lactone (AHL) signalling molecules produced by QS bacteria were analysed by thin layer chromatography (TLC) bioautography and gas chromatography-mass spectrometer (GC-MS). The film formation, β-dimethylmercaptopropionic (DMSP) degradation and algae growth effects of QS bacteria were investigated. The results showed that 113 QS bacteria were isolated from 842 microalgae-associated bacteria. Detection of AHL molecules in 10 different species of QS bacteria showed that most of them were N-(3-Oxodecanoyl)-L-homoserine lactone (OC10-HSL), N-Octanoyl-L-homoserine lactone (C8-HSL) and N-(3-Oxooctanoyl)-L-homoserine lactone (OC8-HSL). All 10 QS bacteria had film-forming ability, and they could degrade DMSP (except strain E26). The crude metabolic extracts of the 10 QS bacteria can inhibit or promote microalgae growth to different degrees. Our study is helpful to understand the role of microalgae-associated microorganisms with the QS system in algae-bacteria interactions and community succession of HAB microalgae.

Role of nano-Fe3O4 particle on improving membrane bioreactor (MBR) performance: Alleviating membrane fouling and microbial mechanism

This study would investigate the effect of nano-Fe3O4 particles on the performance of membrane bioreactor (MBR), including membrane fouling, membrane rejection and microbial community. It can effectively alleviate membrane fouling and improve the effluent quality in MBR by bio-effect rather than nanoparticle adsorption. The lowest membrane fouling resistance was achieved at R4-MBR (sludge and membrane surface with nano-Fe3O4), which decreased by 46.08%. Meanwhile, R3-MBR (sludge with nano-Fe3O4) had the lowest concentration of COD in effluent which was below 20 mg/L in the stable phase of MBR operation. After applying nano-Fe3O4, the content of extracellular polymeric substances (EPS) and soluble microbial products (SMP) were both reduced with a lower molecular weight. From the microbial community analysis, the abundance of Proteobacteria increased from 25.06 to 45.11% at the phylum level in R3-MBR. It contributed to removing organic substances in MBRs. Moreover, the nano-Fe3O4 restricted Bacteroidetes growth, especially in R4-MBR, leading to a more excellent performance of membrane flux. Besides, the applied nano-Fe3O4 promoted the abundance of Quorum Quenching (QQ) microorganism, and declined the percentage of Quorum Sensing (QS) bacteria. Then, a lower content of N-Acyl-l-Homoserine Lactones (AHLs) in containing nano-Fe3O4 sludge. That was also prone to control membrane fouling. Overall, this study indicates the nano-Fe3O4 particle is appropriate for elevating MBR performance, such as membrane fouling and effluent quality, by bio-effect.

Bacterial assembly and succession in the start-up phase of an IFAS metacommunity: The role of AHL-driven quorum sensing.

Quorum sensing (QS) plays an important role in biofilm formation and the start-up of biofilm-based reactors, while its involvement in bacterial assembly throughout biofilm development remains underexplored. We investigated the assembly and succession of the bacterial community in a full-scale integrated fixed-film activated sludge (IFAS) process, with emphasis on N-acylhomoserine lactone (AHL)-driven QS. Biofilm development could be divided into two major periods, (i) young biofilm formation phase and (ii) biofilm maturity and update phase. Mature biofilms exhibited lower levels of AHLs compared with young biofilms (p>0.05). A structural equation model, constructed to assess the linkages between AHL level and bacterial community composition as well as environmental factors, indicated that pH significantly influenced both bacterial community composition and AHL content. Along with biofilm development, there was a negative correlation between AHL concentration and community composition variation (coefficients of -0.367 and -0.329). Regarding the lower AHL level in mature biofilms, these results were consistent with the phylogenetic molecular ecological networks (pMENs) analysis, indicating that quorum-quenching (QQ) bacteria occur in keystone taxa in mature biofilms. In addition, based on the pMENs results, the proportion of positive interactions increased from 77.64 to 82.39% in mature biofilms compared to young biofilms, indicating that bacterial cooperation was strengthened in mature biofilms. The QS bacteria were predominant in the keystone taxa of pMENs, with proportions being increased to 62% in mature biofilms, which is conducive for biofilm development. Overall, this study improves our understanding of the involvement of AHL-mediated QS in biofilm development.

AHLs-mediated quorum sensing threshold and its response towards initial adhesion of wastewater biofilms

Quorum sensing (QS) plays a crucial role during initial biofilm formation, however the QS threshold and the response of biofilm formation towards N-acyl-homoserine lactones (AHLs) remains largely unknown due to the limitation of nondestructive online methods for monitoring bacterial adherence and the complexity of QS system, which limits the application of QS signal reagents in biofilm reactors. In this study, bacterial QS threshold and its response of biofilm formation to AHLs in purely cultured Sphingomonas rubra biofilm as well as in three different wastewater biofilms #1–3 were investigated via real time cell analysis (RTCA). The main perspective was to study the biomass adherence in response to 12 different forms of AHLs at different concentrations. Results showed that bacterial adhesion was significantly improved by exogenous AHLs with the maximum increase of 2.26-, 2.36-, 2.52-, and 2.80- times biomass production in the four respective biofilms. Although the preferred form of AHL differed for various biofilms, the long-chain AHLs (12–14 carbons) resulted in an overall improvement of bacterial adhesion due to their stronger hydrophobicity and hydrolysis resistance. In addition, bacterial QS threshold of AHLs was observed to have a wide range of concentration from 10 ng/L to 10 μg/L. Meanwhile, QS response time to AHLs also showed a significant difference in different biofilms. Biofilm #2 inoculated with bulking sludge had lower QS threshold of 10 ng/L and faster response to most AHLs that is less than 6 h. Thus, considering the improvement of biofilm adhesion by AHLs, 10 ng/L of C12-HSL, 10 ng/L of C12-HSL, and 10 ng/L of C6-HSL were preferentially selected for wastewater biofilms #1–3 respectively. Unexpectedly, adding high-concentration of AHLs detected in sludges did not significantly improved the bacterial adhesion. Infact the addition of these AHLs at low concentrations or even undetected concentrations substantially improved bacterial adhesion, which could be explained by bacterial communities composition. According to the Pearson correlation analysis, 62% of the top 50 most abundant genera in bacterial communities were significantly negatively related to the response time of multiple AHLs, representing their fast QS response. The QS bacteria, Dechloromonas and Nitrospira have fast QS response for C4-HSL and C8-HSL while, Comamonadaceae has fast QS response for 3OC8-HSL, 3OC10-HSL, 3OC12-HSL, and 3OC14-HSL. In contrast, the rest 38% of the top most abundant genera, such as Ferruginibacter, Hyphomicrobium, and Terrimonas quickly responded to only one AHL, showing significant negative relationship with the response time of C6-HSL. Overall, this study provides an effective and convenient means to select appropriate AHL reagents to promote bacterial adhesion in biofilm systems. Moreover, it also suggests that exogenous AHLs may be useful in improving the settling property of bulking sludge.

Screening of Surface-associated Bacteria from the Mexican Red Alga Halymenia floresii for Quorum Sensing Activity

Macroalgae host a dense bacterial epibiome forming surface biofilms, which act as a biological defense by protecting the surface from macrofoulers. During experimental cultivation of Halymenia floresii (Rhodophyta, Halymeniales) under Integrated MultiTrophic Aquaculture (IMTA), the culture tanks remained clean of any exogenous algal colonization, and the surface of the H. floresii thalli was remarkably free from any fouling organisms. The presence of Halymenia also appeared to restrict the establishment of opportunist green algae and the colonization of barnacles usually impairing the cultures. To date, nothing is known about the diversity and biological potential of H. floresii epibionts. Hence, to better understand their epiphytic bacterial community, surface-associated bacteria from distinct H. floresii samples, beach-cast, integrated multi trophic aquaculture cultivar, and cultivar cylinders, were isolated. Thirty-one axenic bacterial strains belonging to 3 phyla, 20 genera, and 25 species were identified by 16S rRNA gene sequencing. Following isolation, they were screened for the production of quorum sensing (QS) signals, since these communication signals play a vital role in biofilm formation. All, except one, isolates were identified as gram-negative; hence, acylated homoserine lactones (AHLs) were focused upon. Using the reporter strain Escherichia coli pSB406, the isolates were screened for their ability to induce luminescence. Of 31 isolates screened, 17 strains, including Pseudoalteromonas arabiensis, Pseudoalteromonas sp. (B5BC and B6.1BC), Pseudoalteromonas mariniglutinosa, Vibrio owensii, Tenacibaculum sp., Maribacter sp., Spongiimicrobium salis, Aquimarina sp., Kordiimonas sp., Alteromonas sp. (B12CC and B16CC), Roseobacter sp., Erythrobacter sp., Ruegeria lacuscaerulensis, and Epibacterium sp., exhibited the presence of extracellular QS signals. Thus, the presence of QS bacteria on the surface of H. floresii was demonstrated for the first time. Our results may open a new direction to investigate the host—QS bacteria interactions.

Metagenomic insights into quorum sensing in membrane-aerated biofilm reactors for phenolic wastewater treatment

ABSTRACT Quorum sensing (QS) is of crucial importance for the formation and performance of biofilms adhered to aerated membranes. In this study, the QS-related genes in membrane-aerated biofilm reactors (MABR) for phenolic wastewater treatment were investigated through high-throughput metagenomic sequencing. Results showed that numerous regulatory QS-related genes were associated with the production of signals including acyl-homoserine lactones (AHLs) and diguanylate monophosphate (c-di-GMP), indicating that the biofilms were potentially regulated by the AHL-mediated QS and c-di-GMP-mediated QS systems. Species and functional contribution analysis demonstrated that Pseudomonas, Achromobacter, Rhodococcus, Granulicoccus and Thauera were the key QS-related gene carriers. Redundancy analysis and Spearman correlation analysis showed that high influent phenolic loading gave rise to a high relative abundance of QS bacteria within the biofilm community. Thus, QS-related genes likely play an important role in strengthening biofilm resistance to phenolics, as well as the removal of phenolic contaminants.

Linking Short-Chain N-Acyl Homoserine Lactone-Mediated Quorum Sensing and Replant Disease: A Case Study of Rehmannia glutinosa.

Rehmannia glutinosa, a perennial medicinal plant, suffers from severe replant disease under consecutive monoculture. The rhizosphere microbiome is vital for soil suppressiveness to diseases and for plant health. Moreover, N-acyl homoserine lactone (AHL)-mediated quorum sensing (QS) regulates diverse behavior in rhizosphere-inhabiting and plant pathogenic bacteria. The dynamics of short-chain AHL-mediated QS bacteria driven by consecutive monoculture and its relationships with R. glutinosa replant disease were explored in this study. The screening of QS bacteria showed that 65 out of 200 strains (32.5%) randomly selected from newly planted soil of R. glutinosa were detected as QS bacteria, mainly consisting of Pseudomonas spp. (55.4%). By contrast, 34 out of 200 (17%) strains from the diseased replant soil were detected as QS bacteria, mainly consisting of Enterobacteriaceae (73.5%). Functional analysis showed most of the QS bacteria belonging to the Pseudomonas genus showed strong antagonistic activities against Fusarium oxysporum or Aspergillus flavus, two main causal agents of R. glutinosa root rot disease. However, the QS strains dominant in the replant soil caused severe wilt disease in the tissue culture seedlings of R. glutinosa. Microbial growth assays demonstrated a concentration-dependent inhibitory effect on the growth of beneficial QS bacteria (i.e., Pseudomonas brassicacearum) by a phenolic acid mixture identified in the root exudates of R. glutinosa, but the opposite was true for harmful QS bacteria (i.e., Enterobacter spp.). Furthermore, it was found that the population of quorum quenching (QQ) bacteria that could disrupt the beneficial P. brassicacearum SZ50 QS system was significantly higher in the replant soil than in the newly planted soil. Most of these QQ bacteria in the replant soil were detected as Acinetobacter spp. The growth of specific QQ bacteria could be promoted by a phenolic acid mixture at a ratio similar to that found in the R. glutinosa rhizosphere. Moreover, these quorum-quenching bacteria showed strong pathogenicity toward the tissue culture seedlings of R. glutinosa. In conclusion, consecutive monoculture of R. glutinosa contributed to the imbalance between beneficial and harmful short-chain AHL-mediated QS bacteria in the rhizosphere, which was mediated not only by specific root exudates but also by the QQ bacterial community.

From Quorum Sensing to Positional Sensing

With Quorum Sensing (QS) bacteria adapt their physiology and metabolism when they reach a certain density and activate genes relevant for their nutrient metabolism, survival and virulence. Genes controlled by QS have relevance for the pathogenic behavior of bacteria including adherence, secretion, toxin production, immune subversion, biofilm formation and resistance to both antimicrobial and biological/chemical stressors. Most studies assess QS by growing bacteria to various densities in liquid culture. Here, we report that when bacteria are grown on agar plates at various densities, in addition to QS, positional sensing (PS) comes into play. Genes controlled by PS can be studied by cloning their promoter in front of an autonomous bioluminescent cassette (luxCDABE) and measuring their time‐ and position‐dependent expression in biofilms by using an in vivo imaging system (IVIS). The Pseudomonas putida XylR/XylS intergenic region controlling genes relevant for the metabolism and degradation of xylene and related organic molecules was cloned in front of the autonomous luciferase cassette from Photorhabdus luminescence, with the sigma 54 controlled Ps1 promoter regulating the a luxCDABE genes and the Pr promoter regulating the XylR gene. Interestingly, when E. coli carrying this construct were grown to various density on agar plates, bioluminescence was not only expressed in a density‐ and time‐dependent, but also position‐dependent manner. Such positional cues may be mediated by signaling molecules in response to cell density, growth stage, and nutrient availability. Overexpression of the sigma factor 54 (NtrA) from Pseudomonas putida modified the density‐ and time‐dependent response suggesting a regulatory role in QS and PS. The use of bioluminescence in QS and PS promises to reveal important regulatory circuits and communication systems relevant for bacterial normal physiology, pathogenicity and virulence.