Light dazzles from the black box: whole-cell biosensors are ready to inform on fundamental soil biological processes

Renella, Giancarlo

doi:10.1186/s40538-016-0059-3

Cited by 16 publications

(7 citation statements)

References 147 publications

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“…In recent years, the synthetic biology toolbox has expanded rapidly and the use of genetically engineered molecular circuits to sense molecules and conditions of interest has gained increased attention. These developments have given rise to whole-cell biosensors that utilize natural regulatory systems engineered to detect metabolites and small molecules (14,15).…”

Section: Importancementioning

confidence: 99%

A Whole-Cell Biosensor for Detection of 2,4-Diacetylphloroglucinol (DAPG)-Producing Bacteria from Grassland Soil

Hansen

Wibowo

et al. 2021

Appl Environ Microbiol

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Fluorescent Pseudomonas spp. producing the antibiotic 2,4-diacetylphloroglucinol (DAPG) are ecologically important in the rhizosphere as they can control phytopathogens and contribute to disease suppressiveness. DAPG can also trigger a systemic resistance response in plants and stimulate root exudation and branching as well as induce plant-beneficial activities in other rhizobacteria. While studies of DAPG-producing Pseudomonas have predominantly focused on rhizosphere niches, the ecological role of DAPG as well as the distribution and dynamics of DAPG-producing bacteria remains less well understood for other environments such as bulk soil and grassland, where the level of DAPG producers are predicted to be low. Here, we construct a whole cell biosensor for detection of DAPG and DAPG-producing bacteria from environmental samples. The constructed biosensor contains a phlF response module and either lacZ or lux genes as output modules assembled on a pSEVA plasmid backbone for easy transfer to different host species and to enable easy future genetic modifications. We show that the sensor is highly specific toward DAPG, with a sensitivity in the low nanomolar range (>20 nM). This sensitivity is comparable to the DAPG levels identified in rhizosphere samples by chemical analysis. The biosensor enables guided isolation of DAPG-producing Pseudomonas. Using the biosensor, we probed the same grassland soil sampling site to isolate genetically related DAPG-producing Pseudomonas kilonensis strains over a period of 12 months. Next, we used the biosensor to determine the frequency of DAPG-producing Pseudomonads within three different grassland soil sites and show that DAPG producers can constitute part of the Pseudomonas population in the range of 0.35-17% at these sites. Finally, we show that the biosensor enables detection of DAPG produced by non-Pseudomonas species. Our studies show that a whole-cell biosensor for DAPG detection can facilitate isolation of bacteria that produce this important secondary metabolite and provide insight into the population dynamics of DAPG producers in natural grassland soil. IMPORTANCE The interest has grown for bacterial biocontrol agents as biosustainable alternatives to pesticides to increase crop yields. To date, we have a broad knowledge of antimicrobial compounds, such as DAPG, produced by bacteria growing in the rhizosphere surrounding plant roots. However, compared to the rhizosphere niches, the ecological role of DAPG as well as the distribution and dynamics of DAPG-producing bacteria remains less well understood for other environments such as bulk and grassland soil. Currently, we are restricted to chemical methods with detection limits and time-consuming PCR-based and probe-hybridization approaches to detect DAPG and its respective producer. In this study, we have developed a whole-cell biosensor, which can circumvent the labor-intensive screening process, as well as increase the sensitivity at which DAPG can be detected. This enables quantification of relative amounts of DAPG-producers, which in turn increases our understanding of the dynamics and ecology of these producers in natural soil environments.

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Section: Importancementioning

confidence: 99%

A Whole-Cell Biosensor for Detection of 2,4-Diacetylphloroglucinol (DAPG)-Producing Bacteria from Grassland Soil

Hansen

Wibowo

et al. 2021

Appl Environ Microbiol

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“…The luminescence response of transformed E. coli proved to be a capable biosensor for measuring environmental contaminants. Furthermore, other than the biotechnological workhorse E. coli, different soil-borne bacteria, such as S. aureus, P. uorescens and P. putida, have been used in the engineering of whole-cell biosensors (Renella and Giagnoni 2016), providing the additional advantage of monitoring soil-based contaminants.…”

Section: Recent Advances Of Biosensors For Xenobiotic Compounds: Hydrocarbons and Pesticidesmentioning

confidence: 99%

In-situ monitoring of xenobiotics using genetically engineered whole-cell-based microbial biosensors: recent advances and outlook

Ali

Mittal

Kaur

2021

World J Microbiol Biotechnol

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Industrialisation, directly or indirectly, exposes humans to various xenobiotics. The increased magnitude of chemical pesticides and toxic heavy metals in the environment, as well as their intrusion into the food chain, seriously threatens human health. Therefore, the surveillance of xenobiotics is crucial for social safety and security. Online investigation by traditional methods is not su cient for the detection and identi cation of such compounds because of the high costs and their complexity. Advancement in the eld of genetic engineering provides a potential opportunity to use genetically modi ed microorganisms. In this regard, whole-cell-based microbial biosensors (WCBMB) represent an essential tool that couples genetically engineered organisms with an operator/promoter derived from a heavy metal-resistant operon combined with a regulatory protein in the gene circuit. The plasmid controls the expression of the reporter gene, such as gfp, luc, lux and lacZ, to an inducible gene promoter and has been widely applied to assay toxicity and bioavailability. This review summarises the recent trends in the development and application of microbial biosensors and the use of mobile genes for biomedical and environmental safety concerns.

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“…Progress in the development of biosensory diagnostics of bacterial and viral infections has been achieved mainly due to the latest improvements in the methods used for indication of specific markers [22,[64][65][66][67].…”

Section: Main Types Of Biosensors and Their Functionsmentioning

confidence: 99%

Label-Free Biosensors for Laboratory-Based Diagnostics of Infections: Current Achievements and New Trends

et al. 2020

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Infections pose a serious global public health problem and are a major cause of premature mortality worldwide. One of the most challenging objectives faced by modern medicine is timely and accurate laboratory-based diagnostics of infectious diseases. Being a key factor of timely initiation and success of treatment, it may potentially provide reduction in incidence of a disease, as well as prevent outbreak and spread of dangerous epidemics. The traditional methods of laboratory-based diagnostics of infectious diseases are quite time- and labor-consuming, require expensive equipment and qualified personnel, which restricts their use in case of limited resources. Over the past six decades, diagnostic technologies based on lateral flow immunoassay (LFIA) have been and remain true alternatives to modern laboratory analyzers and have been successfully used to quickly detect molecular ligands in biosubstrates to diagnose many infectious diseases and septic conditions. These devices are considered as simplified formats of modern biosensors. Recent advances in the development of label-free biosensor technologies have made them promising diagnostic tools that combine rapid pathogen indication, simplicity, user-friendliness, operational efficiency, accuracy, and cost effectiveness, with a trend towards creation of portable platforms. These qualities exceed the generally accepted standards of microbiological and immunological diagnostics and open up a broad range of applications of these analytical systems in clinical practice immediately at the site of medical care (point-of-care concept, POC). A great variety of modern nanoarchitectonics of biosensors are based on the use of a broad range of analytical and constructive strategies and identification of various regulatory and functional molecular markers associated with infectious bacterial pathogens. Resolution of the existing biosensing issues will provide rapid development of diagnostic biotechnologies.

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Light dazzles from the black box: whole-cell biosensors are ready to inform on fundamental soil biological processes

Cited by 16 publications

References 147 publications

A Whole-Cell Biosensor for Detection of 2,4-Diacetylphloroglucinol (DAPG)-Producing Bacteria from Grassland Soil

A Whole-Cell Biosensor for Detection of 2,4-Diacetylphloroglucinol (DAPG)-Producing Bacteria from Grassland Soil

In-situ monitoring of xenobiotics using genetically engineered whole-cell-based microbial biosensors: recent advances and outlook

Label-Free Biosensors for Laboratory-Based Diagnostics of Infections: Current Achievements and New Trends

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