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David Clarke - One of the best experts on this subject based on the ideXlab platform.
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making and breaking leupeptin protease inhibitors in pathogenic gammaproteobacteria
Angewandte Chemie, 2020Co-Authors: Sabine Kienesberger, David Clarke, Nam Y Kim, Ellen L Zechner, Jason M CrawfordAbstract:Leupeptin is a bacterial small molecule that is used worldwide as a protease inhibitor. However, its biosynthesis and genetic distribution remain unknown. We identified a family of leupeptins in gammaproteobacterial pathogens, including Photorhabdus, Xenorhabdus, and Klebsiella species, amongst others. Through genetic, metabolomic, and heterologous expression analyses, we established their construction by discretely expressed ligases and accessory enzymes. In Photorhabdus species, a hypothetical protein required for colonizing nematode hosts was established as a new class of proteases. This enzyme cleaved the tripeptide aldehyde protease inhibitors, leading to the formation of "pro-pyrazinones" featuring a hetero-tricyclic architecture. In Klebsiella oxytoca, the pathway was enriched in clinical isolates associated with respiratory tract infections. Thus, the bacterial production and proteolytic degradation of leupeptins can be associated with animal colonization phenotypes.
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Photorhabdus a tale of contrasting interactions
Microbiology, 2020Co-Authors: David ClarkeAbstract:Different model systems have, over the years, contributed to our current understanding of the molecular mechanisms underpinning the various types of interaction between bacteria and their animal hosts. The genus Photorhabdus comprises Gram-negative insect pathogenic bacteria that are normally found as symbionts that colonize the gut of the infective juvenile stage of soil-dwelling nematodes from the family Heterorhabditis. The nematodes infect susceptible insects and release the bacteria into the insect haemolymph where the bacteria grow, resulting in the death of the insect. At this stage the nematodes feed on the bacterial biomass and, following several rounds of reproduction, the nematodes develop into infective juveniles that leave the insect cadaver in search of new hosts. Therefore Photorhabdus has three distinct and obligate roles to play during this life-cycle: (1) Photorhabdus must kill the insect host; (2) Photorhabdus must be capable of supporting nematode growth and development; and (3) Photorhabdus must be able to colonize the gut of the next generation of infective juveniles before they leave the insect cadaver. In this review I will discuss how genetic analysis has identified key genes involved in mediating, and regulating, the interaction between Photorhabdus and each of its invertebrate hosts. These studies have resulted in the characterization of several new families of toxins and a novel inter-kingdom signalling molecule and have also uncovered an important role for phase variation in the regulation of these different roles.
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whole genome dna methylation methylome analysis and role of dam dna methyltransferase in the entomopathogenic bacterium Photorhabdus luminescens
MicrobiOccitanie 2019- Rencontre des Microbiologistes Région Occitanie, 2019Co-Authors: Amaury Payelleville, Sylvie Pages, Alain Givaudan, David Clarke, Ludovic Legrand, Dana Blackburn, Anne Lanoisnouri, Julien BrillardAbstract:Whole Genome DNA Methylation (Methylome) Analysis and Role of Dam DNA Methyltransferase in the Entomopathogenic Bacterium Photorhabdus luminescens. MicrobiOccitanie 2019- Rencontre des Microbiologistes Region Occitanie
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The expression of stlA in Photorhabdus luminescens is controlled by nutrient limitation.
PloS one, 2013Co-Authors: Lea Lango-scholey, Alexander O Brachmann, Helge B. Bode, David ClarkeAbstract:Photorhabdus is a genus of Gram-negative entomopathogenic bacteria that also maintain a mutualistic association with nematodes from the family Heterorhabditis. Photorhabdus has an extensive secondary metabolism that is required for the interaction between the bacteria and the nematode. A major component of this secondary metabolism is a stilbene molecule, called ST. The first step in ST biosynthesis is the non-oxidative deamination of phenylalanine resulting in the production of cinnamic acid. This reaction is catalyzed by phenylalanine-ammonium lyase, an enzyme encoded by the stlA gene. In this study we show, using a stlA-gfp transcriptional fusion, that the expression of stlA is regulated by nutrient limitation through a regulatory network that involves at least 3 regulators. We show that TyrR, a LysR-type transcriptional regulator that regulates gene expression in response to aromatic amino acids in E. coli, is absolutely required for stlA expression. We also show that stlA expression is modulated by σS and Lrp, regulators that are implicated in the regulation of the response to nutrient limitation in other bacteria. This work is the first that describes pathway-specific regulation of secondary metabolism in Photorhabdus and, therefore, our study provides an initial insight into the complex regulatory network that controls secondary metabolism, and therefore mutualism, in this model organism.
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nematode parasites and entomopathogens
Insect Pathology (Second Edition), 2012Co-Authors: Edwin E. Lewis, David ClarkeAbstract:This chapter covers recent advances in the study of nematodes that are parasites of insects and focuses primarily on those that are entomopathogenic in the genera Heterorhabditis and Steinernema , characterized by their symbiotic association with the bacteria Photorhabdus and Xenorhabdus , respectively. These entomopathogenic nematodes are extremely virulent insect pathogens that have been successfully marketed as biocontrol agents for the protection of certain high-value crops from a range of insect pests. Entomopathogenic nematodes are useful models for studying the ecology of soil food webs. The obligate interaction between the bacterium and the nematode has also established entomopathogenic nematodes as a very useful model for the study of the molecular mechanisms underpinning bacteria–host interactions. Although they appear to have very similar lifestyles, the two nematode genera are not phylogenetically close, and recent developments have shown that the Heterorhabditis – Photorhabdus and Steinernema – Xenorhabdus associations are very different at the molecular level.
Noël Boemare - One of the best experts on this subject based on the ideXlab platform.
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phylogeny of Photorhabdus and xenorhabdus based on universally conserved protein coding sequences and implications for the taxonomy of these two genera proposal of new taxa x vietnamensis sp nov p luminescens subsp caribbeanensis subsp nov p luminesc
International Journal of Systematic and Evolutionary Microbiology, 2010Co-Authors: Patrick Tailliez, Sylvie Pages, Nadege Ginibre, Christine Laroui, Armelle Paule, Noël BoemareAbstract:We used the information from a set of concatenated sequences from four genes (recA, gyrB, dnaN and gltX) to investigate the phylogeny of the genera Photorhabdus and Xenorhabdus (entomopathogenic bacteria associated with nematodes of the genera Heterorhabditis and Steinernema, respectively). The robustness of the phylogenetic tree obtained by this multigene approach was significantly better than that of the tree obtained by a single gene approach. The comparison of the topologies of single gene phylogenetic trees highlighted discrepancies which have implications for the classification of strains and new isolates; in particular, we propose the transfer of Photorhabdus luminescens subsp. thracensis to Photorhabdus temperata subsp. thracensis comb. nov. (type strain CIP 108426T =DSM 15199T). We found that, within the genus Xenorhabdus, strains or isolates that shared less than 97 % nucleotide identity (NI), calculated on the concatenated sequences of the four gene fragments (recA, gyrB, dnaN and gltX) encompassing 3395 nucleotides, did not belong to the same species. Thus, at the 97 % NI cutoff, we confirm the current 20 species of the genus Xenorhabdus and propose the description of a novel species, Xenorhabdus vietnamensis sp. nov. (type strain VN01T = CIP 109945T =DSM 22392T). Within each of the three current species of the genus Photorhabdus, P. asymbiotica, P. luminescens and P. temperata, strains or isolates which shared less than 97 % NI did not belong to the same subspecies. Comparisons of the four gene fragments plus the rplB gene fragment analysed separately led us to propose four novel subspecies: Photorhabdus luminescens subsp. caribbeanensis subsp. nov. (type strain HG29T =CIP 109949T =DSM 22391T), P. luminescens subsp. hainanensis subsp. nov. (type strain C8404T = CIP 109946T =DSM 22397T), P. temperata subsp. khanii subsp. nov. (type strain C1T =NC19T =CIP 109947T =DSM 3369T), and P. temperata subsp. tasmaniensis subsp. nov. (type strain T327T = CIP 109948T =DSM 22387T).
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taxonomy of australian clinical isolates of the genus Photorhabdus and proposal of Photorhabdus asymbiotica subsp asymbiotica subsp nov and p asymbiotica subsp australis subsp nov
International Journal of Systematic and Evolutionary Microbiology, 2004Co-Authors: R J Akhurst, Noël Boemare, David Alfredson, Peter H Janssen, Margaret M Peel, C E BeardAbstract:The relationship of Photorhabdus isolates that were cultured from human clinical specimens in Australia to Photorhabdus asymbiotica isolates from human clinical specimens in the USA and to species of the genus Photorhabdus that are associated symbiotically with entomopathogenic nematodes was evaluated. A polyphasic approach that involved DNA–DNA hybridization, phylogenetic analyses of 16S rRNA and gyrB gene sequences and phenotypic characterization was adopted. These investigations showed that gyrB gene sequence data correlated well with DNA–DNA hybridization and phenotypic data, but that 16S rRNA gene sequence data were not suitable for defining species within the genus Photorhabdus. Australian clinical isolates proved to be related most closely to clinical isolates from the USA, but the two groups were distinct. A novel subspecies, Photorhabdus asymbiotica subsp. australis subsp. nov. (type strain, 9802892T=CIP 108025T=ACM 5210T), is proposed, with the concomitant creation of Photorhabdus asymbiotica subsp. asymbiotica subsp. nov. Analysis of gyrB sequences, coupled with previously published data on DNA–DNA hybridization and PCR-RFLP analysis of the 16S rRNA gene, indicated that there are more than the three subspecies of Photorhabdus luminescens that have been described and confirmed the validity of the previously proposed subdivision of Photorhabdus temperata. Although a non-luminescent, symbiotic isolate clustered consistently with P. asymbiotica in gyrB phylogenetic analyses, DNA–DNA hybridization indicated that this isolate does not belong to the species P. asymbiotica and that there is a clear distinction between symbiotic and clinical species of Photorhabdus.
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two distinct hemolytic activities in xenorhabdus nematophila are active against immunocompetent insect cells
Applied and Environmental Microbiology, 2001Co-Authors: Julien Brillard, Noël Boemare, Carlos Ribeiro, Michel Brehelin, Alain GivaudanAbstract:The genus Xenorhabdus consists of the specific bacterial symbionts of the entomopathogenic nematodes of the family Steinernematidae (40) and was separated from the genus Photorhabdus (11), which contains the symbionts of the entomopathogenic nematodes of the family Heterorhabditidae. Both genera are entomopathogenic gram-negative bacteria belonging to the Enterobacteriaceae. The nematodes carry their bacterial symbionts monoxenically in a special vesicle of the infective stage (L3 juveniles) in Steinernematidae (8) and throughout the whole intestine of Heterorhabditidae (20). These bacteria are transported by their nematode hosts into the hemocoel of the insect prey, which is killed, probably via a combination of toxin action and septicemia. The bacterial symbionts also contribute to the symbiotic relationship by establishing and maintaining suitable conditions for nematode reproduction (31). Recently, isolation of some Photorhabdus strains from infected humans in Australia and the United States was reported (21, 30), and the strains from the United States were classified as Photorhabdus asymbiotica (23). The form of the bacterium that is normally isolated from symbiotic infective-stage nematodes is referred to as phase I. Like many pathogenic bacteria, Xenorhabdus and Photorhabdus strains spontaneously produce colonial variants which have been called phase II variants (10). The two variants of the bacteria have generally been shown to be equally pathogenic for the larvae of the greater wax moth, Galleria mellonella (3). However, Volgyi et al. (42) described for the first time a phase II variant that showed reduced virulence in the tobacco hornworm, Manduca sexta. Xenorhabdus nematophila and Photorhabdus luminescens are highly pathogenic to insects, and 50% insect mortality has been reported with direct infection with fewer than 20 bacteria per larva (5). The bacterial factors involved in killing of the insect or in overcoming the insect immune reactions are still under investigation. Following invasion of the insect host by the nematodes, both bacteria produce potential virulence factors, including lipase, protease, lecithinase, and lipopolysaccharides (LPSs), in the hemocoel (for a review, see reference 24). It was shown that purified LPS, Photorhabdus protease fractions, or Xenorhabdus lecithinase isomers showed no toxic effect following injection into insect hemocoel (12, 16, 39). Recently, a novel toxin complex with both oral and injectable activities against a wide range of insects was identified in a supernatant of P. luminescens (13). Purified toxin complex a (Tca) has specific effects on the midgut epithelium of the insect (9). In order to study Xenorhabdus and Photorhabdus virulence in insects, a genetic approach was also used. Avirulent mutants of X. nematophila have been isolated by transposon mutagenesis (Tn5). These mutants were pleiotropic, but all five mutants that tested as avirulent in G. mellonella were nonmotile and partially impaired in blood hemolysis (43). It was also shown that a homoserine lactone autoinducer restored virulence to one avirulent X. nematophila strain and stimulated the level of bacterial lipase activity (17). Recently we reported that flhDC, the flagellar master operon of X. nematophila, controls flagellin expression. Furthermore we revealed that lipolytic and extracellular hemolysin activity is flhD dependent. We also showed that the flhD null mutant displayed an attenuated virulence phenotype in the common cutworm, Spodoptera littoralis, compared to the wild-type strain (25). The recently published partial genome sequence of P. luminescens (22) revealed a diverse array of genes that putatively encodes potential virulence factors. These factors include exoenzymes (proteases, lipases, and chitinases), a type III secretion system (Yop homolog), and several classes of toxins (insecticidal toxin complex, Rtx-like toxins, and hemolysin and cytotoxin homologs) (22). Until now, studies examining hemolytic activity of both genera have not been reported. Cytolysins are proteins which cause lysis of red blood cells (RBC) as well as nucleated cell types by hydrolysis (lipases, phospholipases, or proteases) or by forming pores in the plasma membrane. Surfactants may also cause cytolysis by solubilization of the target cell membrane. Bacterial cytolysins are usually recognized as hemolysin on blood agar where a transparent zone appears around colonies. The production by a few strains of Xenorhabdus and Photorhabdus of hemolysin has been detected on agar supplemented with sheep blood (6, 21, 25). Apart from their phoretic location inside infective juvenile nematodes, these bacteria are only observed in insect hemolymph, where they enter their growth cycle. Here the bacteria are in contact with hemocytes which achieve defense reactions in insects. Some of these cells are immunocompetent cells able to engulf (phagocytosis) or to isolate and kill (nodule formation) bacteria. We hypothesize that hemolytic activities could target the immunocompetent cells in insect hemolymph. In this study, we report that different cytolytic activities were found in supernatants of Xenorhabdus whereas none was detected in supernatants of various strains of Photorhabdus. We have studied the kinetics of the production of cytolytic activities over the course of in vitro bacterial growth. We also provide evidence on the characteristics and on the specificity of each of these cytolytic activities against mammalian RBC and insect hemocyte types.
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polyphasic classification of the genus Photorhabdus and proposal of new taxa p luminescens subsp luminescens subsp nov p luminescens subsp akhurstii subsp nov p luminescens subsp laumondii subsp nov p temperata sp nov p temperata subsp temperata subs
International Journal of Systematic and Evolutionary Microbiology, 1999Co-Authors: Marion Fischerle Saux, Brigitte Brunel, Veronique Viallard, Phillippe Normand, Noël BoemareAbstract:The taxonomic position of Photorhabdus strains was examined through the results of DNA relatedness (S1 nuclease method) studies associated with the determination of ΔT m, 16S rRNA phylogenetic inferences and phenotypic characterization, including morphological, auxanographic, biochemical and physiological properties. Three genomic species were delineated on a consensus assessment. One of these species corresponded to Photorhabdus luminescens, since strains were at least 50% related to the type strain of this species with ΔT m less than 7 °C. The two other species were novel genomic species II and III, which were less than 40% related to each other with ΔT m higher than 9 °C. A comparison of the complete 16S rDNA sequences of several representatives of genomic species II and genomic species III revealed that each of them formed a stable lineage independent of the cluster generated by P. luminescens strains. The genomic species differed in their maximum temperatures for growth. A correlation with the ecological origin of the bacterial samples was noticed. The heat-tolerant group I (maximum growth temperature 35–39 °C) corresponded to the symbionts of Heterorhabditis bacteriophora groups Brecon and HP88 and Heterorhabditis indica, nematodes living in warm and tropical countries, respectively. Group II (maximum growth temperature 33–35 °C) encompassed symbionts from Heterorhabditis megidis, Heterorhabditis zealandica and group NC1 of H. bacteriophora, nematodes isolated in temperate climates. Group III were bacteria isolated from human specimens. Two new species, Photorhabdus temperata sp. nov. (type strain CIP 105563T and Photorhabdus asymbiotica sp. nov. (type strain ATCC 43950T), are proposed for genomic species II and III, respectively. Species I and II can be separated into sub-groups on the basis of high DNA-DNA relatedness (more than 80% DNA binding with ΔTum < 1·5 °C), 16S rDNA branching and phenotypic characters. Therefore, we propose that the two species P. luminescens and P. temperata should be subdivided into subspecies as follows: P. luminescens subsp. luminescens subsp. nov. (type strain ATCC 29999T), P. luminescens subsp. akhurstii subsp. nov. (type strain CIP 105564T), P. luminescens subsp. laumondii subsp. nov. (type strain CIP 105565T) and P. temperata subsp. temperata subsp. nov.
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fast and accurate identification of xenorhabdus and Photorhabdus species by restriction analysis of pcr amplified 16s rrna genes
Applied and Environmental Microbiology, 1997Co-Authors: Brigitte Brunel, Anne Lanois, Alain Givaudan, Noël BoemareAbstract:Thirteen bacterial strains of Xenorhabdus and 14 strains of Photorhabdus originating from a wide range of geographical and nematode host sources were typed by analyzing 16S rRNA gene (rDNA) restriction patterns obtained after digestion of PCR-amplified 16S rDNAs. Eight tetrameric restriction endonucleases were examined. A total of 17 genotypes were identified, forming two heterogeneous main clusters after analysis by the unweighted pair-group method using arithmetic averages: group I included all Xenorhabdus species and strains, symbionts of Steinernema, whereas group II encompassed the Photorhabdus strains, symbionts of Heterorhabditis. To identify the four valid species of Xenorhabdus and unclassified strains and all the genotypes of Photorhabdus luminescens, three restriction enzymes are required: CfoI, AluI, and HaeIII. Our results, in substantial agreement with DNA-DNA pairing and 16S rDNA sequence data, indicate that amplified 16S rDNA restriction analysis is a simple and accurate tool for identifying entomopathogenic nematode bacterial symbionts.
Richard H Ffrenchconstant - One of the best experts on this subject based on the ideXlab platform.
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dissecting the immune response to the entomopathogen Photorhabdus
Trends in Microbiology, 2010Co-Authors: Ioannis Eleftherianos, David Clarke, Richard H Ffrenchconstant, Andrea J Dowling, Stuart E ReynoldsAbstract:Bacterial pathogens either hide from or modulate the host's immune response to ensure their survival. Photorhabdus is a potent insect pathogenic bacterium that uses entomopathogenic nematodes as vectors in a system that represents a useful tool for probing the molecular basis of immunity. During the course of infection, Photorhabdus multiplies rapidly within the insect, producing a range of toxins that inhibit phagocytosis of the invading bacteria and eventually kill the insect host. Photorhabdus bacteria have recently been established as a tool for investigating immune recognition and defense mechanisms in model hosts such as Manduca and Drosophila. Such studies pave the way for investigations of gene interactions between pathogen virulence factors and host immune genes, which ultimately could lead to an understanding of how some Photorhabdus species have made the leap to becoming human pathogens.
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probing the tri trophic interaction between insects nematodes and Photorhabdus
Parasitology, 2010Co-Authors: Ioannis Eleftherianos, David Clarke, Susan A Joyce, Richard H Ffrenchconstant, Stuart E ReynoldsAbstract:Photorhabdus sp. are entomopathogenic bacteria which, upon experimental infection, interact with the insect immune system, but little is known about the roles of their symbiotic nematode partners Heterorhabditis sp. in natural infections. Here, we investigated the respective contributions of nematodes and bacteria by examining humoral and cellular immune reactions of the model lepidopteran insect Manduca sexta against Heterorhabditis carrying Photorhabdus, nematodes free of bacteria (axenic nematodes) and bacteria alone. Insect mortality was slower following infection with axenic nematodes than when insects were infected with nematodes containing Photorhabdus, or the bacteria alone. Nematodes elicited host immune responses to a lesser extent than bacteria. Transcription of certain recognition and antibacterial genes was lower when insects were naturally infected with nematodes carrying no bacteria compared to insects that received bacteria, either with or without nematodes. Axenic nematodes also did not elicit such high levels of phenoloxidase activity and haemocyte aggregates as did treatments involving Photorhabdus. By contrast, the phagocytic capability of host haemocytes was decreased by both axenic and bacteria-associated nematodes, but not by Photorhabdus alone. These results imply that both bacteria and nematodes contribute separately to the pathogenic modulation of host immune responses during natural infections by the mutualistic Heterorhabdus-Photorhabdus complex.
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pirab toxin from Photorhabdus asymbiotica as a larvicide against dengue vectors
Applied and Environmental Microbiology, 2009Co-Authors: Arunee Ahantarig, Nicholas R. Waterfield, Richard H Ffrenchconstant, Nantarat Chantawat, Pattamaporn KittayapongAbstract:We have evaluated Photorhabdus insect-related protein (Pir) from Photorhabdus asymbiotica against dengue vectors. PirAB shows larvicidal activity against both Aedes aegypti and Aedes albopictus larvae but did not affect the Mesocyclops thermocyclopoides predator. PirAB expressed the strongest toxicity compared to PirA, PirB, or the mixture of PirA plus PirB. Whether the presence of an enterobacterial repetitive intergenic consensus sequence in PirAB, but not in PirA, PirB, or the mixture of PirA plus PirB, has any impact on biological control efficacy needs further investigation.
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a single locus from the entomopathogenic bacterium Photorhabdus luminescens inhibits activated manduca sexta phenoloxidase
Fems Microbiology Letters, 2009Co-Authors: Ioannis Eleftherianos, Nicholas R. Waterfield, Richard H Ffrenchconstant, Peter Bone, Sam Boundy, Stuart E ReynoldsAbstract:Insect blood (hemolymph) contains prophenoloxidase, a proenzyme that is activated to protective phenoloxidase when the insect is damaged or challenged with microorganisms. The Gram-negative bacterium Photorhabdus luminescens kills the lepidopteron insect Manduca sexta by using a variety of toxins. We screened P. luminescens and Photorhabdus asymbiotica cosmid libraries in an Escherichia coli host against previously activated M. sexta hemolymph phenoloxidase and identified three overlapping cosmid clones from P. luminescens and five from P. asymbiotica that suppressed the activity of the enzyme both in vitro and in vivo. Genome alignments of cosmid end sequences from both species confirmed that they contained orthologous loci. We examined one of the cosmids from P. luminescens in detail: it induced the formation of significantly fewer melanotic nodules, proliferated faster within the insect host and was significantly more virulent towards fifth-stage larvae than E. coli control bacteria. Insertional mutagenesis of this cosmid yielded 11 transposon mutants that were no longer inhibitory. All of these were insertions into a single 5.5-kb locus, which contained three ORFs and was homologous to the maltodextrin phosphorylase locus of E. coli. The implications of this novel inhibitory factor of insect phenoloxidase for Photorhabdus virulence are discussed.
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insecticidal toxins from Photorhabdus bacteria and their potential use in agriculture
Toxicon, 2007Co-Authors: Richard H Ffrenchconstant, Andrea J Dowling, Nicholas R. WaterfieldAbstract:Abstract Most of the insecticidal toxins used in agriculture come from a single bacterium Bacillus thuringiensis or ‘Bt’. Here we review our work on the array of toxins produced by Photorhabdus and Xenorhabdus bacteria that are symbiotic with entomopathogenic nematodes, and discuss their potential for use in agriculture as alternatives to Bt. Despite the fact that both Photorhabdus and Xenorhabdus are introduced directly into the insect blood stream by their nematode vectors, they produce a range of toxins with both oral and injectable insecticidal activity. The toxin complexes (Tc's) are large orally active toxins that are displayed on the outer surface of the bacterium. They require three components (A–C) for full toxicity and one ‘A’ component has been successfully expressed in transgenic Arabidopsis to confer insect resistance. One such group of Tc's, the PirAB binary toxins, have oral activity against mosquitoes and some caterpillar pests. Their mode of action is not known but they show significant sequence similarity to a recently described neurotoxin β-leptinotarsin-h isolated from the blood of the Colorado potato beetle. Other toxins such as ‘makes caterpillars floppy’ (Mcf) and proteins encoded by the ‘Photorhabdus virulence cassettes’ (PVCs) only show injectable activity. Mcf1 promotes apoptosis in a wide range of cells and appears to mimic mammalian BH3 domain-only proteins in the mitochondrion whereas the mode of action of the PVCs remains undetermined. The likely biological reasons for the massive functional redundancy in Photorhabdus insecticidal toxins are discussed.
Sylvie Pages - One of the best experts on this subject based on the ideXlab platform.
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whole genome dna methylation methylome analysis and role of dam dna methyltransferase in the entomopathogenic bacterium Photorhabdus luminescens
MicrobiOccitanie 2019- Rencontre des Microbiologistes Région Occitanie, 2019Co-Authors: Amaury Payelleville, Sylvie Pages, Alain Givaudan, David Clarke, Ludovic Legrand, Dana Blackburn, Anne Lanoisnouri, Julien BrillardAbstract:Whole Genome DNA Methylation (Methylome) Analysis and Role of Dam DNA Methyltransferase in the Entomopathogenic Bacterium Photorhabdus luminescens. MicrobiOccitanie 2019- Rencontre des Microbiologistes Region Occitanie
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Photorhabdus heterorhabditis sp nov., a symbiont of the entomopathogenic nematode Heterorhabditis zealandica
International Journal of Systematic and Evolutionary Microbiology, 2014Co-Authors: Tiarin Ferreira, Sylvie Pages, Patrick Tailliez, Carol A. Van Reenen, Akihito Endo, Cathrin Sproeer, Leon M. T. DicksAbstract:The bacterial symbionts SF41(T) and SF783 were isolated from populations of the insect pathogenic nematode Heterorhabditis zealandica collected in South Africa. Both strains were closely related to strain Q614 isolated from a population of Heterorhabditis sp. collected from soil in Australia in the 1980s. Sequence analysis based on a multigene approach, DNA-DNA hybridization data and phenotypic traits showed that strains SF41(T), SF783 and 0614 belong to the same species of the genus Photorhabdus with Photorhabdus temperata subsp. cinerea as the most closely related taxon (DNA DNA hybridization value of 68%). Moreover, the phylogenetic position of Photorhabdus temperata subsp. cinerea DSM 19724(T) initially determined using the gyrB sequences, was reconsidered in the light of the data obtained by our multigene approach and DNA-DNA hybridization experiments. Strains SF41(T)., SF783 and 0614 represent a novel species of the genus Photorhabdus, for which the name Photorhabdus heterorhabditis sp. nov. is proposed (type strain SF41(T)=ATCC BAA-2479(T)=DSM 25263(T)).
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Photorhabdus luminescens subsp noenieputensis subsp nov., a symbiotic bacterium associated with a novel Heterorhabditis species related to Heterorhabditis indica
International Journal of Systematic and Evolutionary Microbiology, 2013Co-Authors: Tiarin Ferreira, Sylvie Pages, Patrick Tailliez, Carol Van Reenen, Antoinette P. Malan, Leon M. T. DicksAbstract:The bacterial symbiont AM7(T), isolated from a novel entomopathogenic nematode species of the genus Heterorhabditis, displays the main phenotypic traits of the genus Photorhabdus and is highly pathogenic to Galleria mellonella. Phylogenetic analysis based on a multigene approach (16S rRNA, recA, gyrB, dnaN, gltX and infB) confirmed the classification of isolate AM7(T) within the species Photorhabdus luminescens and revealed its close relatedness to Photorhabdus luminescens subsp. caribbeanensis, P. luminescens subsp. akhurstii and P. luminescens subsp. hainanensis. The five concatenated protein-encoding sequences (4197 nt) of strain AM7(T) revealed 95.8, 95.4 and 94.9% nucleotide identity to sequences of P. luminescens subsp. caribbeanensis HG29(T), P. luminescens subsp. akhurstii FRG04(T) and P. luminescens subsp. hainanensis C8404(T), respectively. These identity values are less than the threshold of 97% proposed for classification within one of the existing subspecies of P. luminescens. Unlike other strains described for P. luminescens, strain AM7(T) produces acid from adonitol, sorbitol and xylitol, assimilates xylitol and has no lipase activity on medium containing Tween 20 or 60. Strain AM7(T) is differentiated from P. luminescens subsp. caribbeanensis by the assimilation of N-acetylglucosamine and the absence of haemolytic activity. Unlike P. luminescens subsp. akhurstii, strain AM7(T) does not assimilate mannitol, and it is distinguished from P. luminescens subsp. hainanensis by the assimilation of trehalose and citrate, the inability to produce indole from tryptophan and the presence of acetoin production and urease activity. Strain AM7(T) (=ATCC BAA-2407(T) =DSM 25462(T)) belongs to a novel subspecies, and is proposed as the type strain of Photorhabdus luminescens subsp. noenieputensis sp. nov.
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The first record of entomopathogenic nematodes (Rhabiditiae: Steinernematidae and Heterorhabditidae) in natural ecosystems in Lebanon: A biogeographic approach in the Mediterranean region
Journal of Invertebrate Pathology, 2011Co-Authors: Elise Noujeim, Sylvie Pages, Patrick Tailliez, Carla Khater, Jean-claude Ogier, Mouin Hamze, Jacques Olivier ThalerAbstract:A survey of entomopathogenic nematodes in Lebanon was conducted for the first time during 2008–2009. Samples were collected on the coastal strip and in nine vegetation types extending from the coastal line to 3088 m above sea level. Wooded and herbaceous ecosystems were considered for sampling purposes. A total of 570 samples were taken, out of which 1% were positive for entomopathogenic nematodes. Approximately, 15.8% out of the 19 sites sampled revealed entomopathogenic nematodes presence (representing three samples). Two entomopathogenic nematodes species Heterorhabditis bacteriophora and Steinernema feltiae were recovered, and identification of their symbiotic bacteria revealed the presence of a Xenorhabdus bovienii, Photorhabdus temperata subsp. thracensis, Photorhabdus luminescens subsp. kayaii and Photorhabdus luminescens subsp. Laumondii
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phylogeny of Photorhabdus and xenorhabdus based on universally conserved protein coding sequences and implications for the taxonomy of these two genera proposal of new taxa x vietnamensis sp nov p luminescens subsp caribbeanensis subsp nov p luminesc
International Journal of Systematic and Evolutionary Microbiology, 2010Co-Authors: Patrick Tailliez, Sylvie Pages, Nadege Ginibre, Christine Laroui, Armelle Paule, Noël BoemareAbstract:We used the information from a set of concatenated sequences from four genes (recA, gyrB, dnaN and gltX) to investigate the phylogeny of the genera Photorhabdus and Xenorhabdus (entomopathogenic bacteria associated with nematodes of the genera Heterorhabditis and Steinernema, respectively). The robustness of the phylogenetic tree obtained by this multigene approach was significantly better than that of the tree obtained by a single gene approach. The comparison of the topologies of single gene phylogenetic trees highlighted discrepancies which have implications for the classification of strains and new isolates; in particular, we propose the transfer of Photorhabdus luminescens subsp. thracensis to Photorhabdus temperata subsp. thracensis comb. nov. (type strain CIP 108426T =DSM 15199T). We found that, within the genus Xenorhabdus, strains or isolates that shared less than 97 % nucleotide identity (NI), calculated on the concatenated sequences of the four gene fragments (recA, gyrB, dnaN and gltX) encompassing 3395 nucleotides, did not belong to the same species. Thus, at the 97 % NI cutoff, we confirm the current 20 species of the genus Xenorhabdus and propose the description of a novel species, Xenorhabdus vietnamensis sp. nov. (type strain VN01T = CIP 109945T =DSM 22392T). Within each of the three current species of the genus Photorhabdus, P. asymbiotica, P. luminescens and P. temperata, strains or isolates which shared less than 97 % NI did not belong to the same subspecies. Comparisons of the four gene fragments plus the rplB gene fragment analysed separately led us to propose four novel subspecies: Photorhabdus luminescens subsp. caribbeanensis subsp. nov. (type strain HG29T =CIP 109949T =DSM 22391T), P. luminescens subsp. hainanensis subsp. nov. (type strain C8404T = CIP 109946T =DSM 22397T), P. temperata subsp. khanii subsp. nov. (type strain C1T =NC19T =CIP 109947T =DSM 3369T), and P. temperata subsp. tasmaniensis subsp. nov. (type strain T327T = CIP 109948T =DSM 22387T).
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diversity of xenorhabdus and Photorhabdus spp and their symbiotic entomopathogenic nematodes from thailand
PLOS ONE, 2012Co-Authors: Aunchalee Thanwisai, Nicholas R. Waterfield, Sarunporn Tandhavanant, Natnaree Saiprom, Phan Ke Long, Helge B. Bode, Sharon J. Peacock, Narisara ChantratitaAbstract:Xenorhabdus and Photorhabdus spp. are bacterial symbionts of entomopathogenic nematodes (EPNs). In this study, we isolated and characterized Xenorhabdus and Photorhabdus spp. from across Thailand together with their associated nematode symbionts, and characterized their phylogenetic diversity. EPNs were isolated from soil samples using a Galleria-baiting technique. Bacteria from EPNs were cultured and genotyped based on recA sequence. The nematodes were identified based on sequences of 28S rDNA and internal transcribed spacer regions. A total of 795 soil samples were collected from 159 sites in 13 provinces across Thailand. A total of 126 EPNs isolated from samples taken from 10 provinces were positive for Xenorhabdus (n = 69) or Photorhabdus spp. (n = 57). Phylogenetic analysis separated the 69 Xenorhabdus isolates into 4 groups. Groups 1, 2 and 3 consisting of 52, 13 and 1 isolates related to X. stockiae, and group 4 consisting of 3 isolates related to X. miraniensis. The EPN host for isolates related to X. stockiae was S. websteri, and for X. miraniensis was S. khoisanae. The Photorhabdus species were identified as P. luminescens (n = 56) and P. asymbiotica (n = 1). Phylogenenic analysis divided P. luminescens into five groups. Groups 1 and 2 consisted of 45 and 8 isolates defined as subspecies hainanensis and akhurstii, respectively. One isolate was related to hainanensis and akhurstii, two isolates were related to laumondii, and one isolate was the pathogenic species P. asymbiotica subsp. australis. H. indica was the major EPN host for Photorhabdus. This study reveals the genetic diversity of Xenorhabdus and Photorhabdus spp. and describes new associations between EPNs and their bacterial symbionts in Thailand.
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Photorhabdus and a host of hosts
Annual Review of Microbiology, 2009Co-Authors: Nicholas R. Waterfield, Todd A Ciche, David ClarkeAbstract:Photorhabdus is a member of the family Enterobacteriaceae that lives in a mutualistic association with a Heterorhabditis nematode worm. The nematode worm burrows into insect prey and regurgitates Photorhabdus, which goes on to kill the insect. The nematode feeds off the growing bacteria until the insect tissues are exhausted, whereupon they reassociate and leave the cadaver in search of new prey. This highly efficient partnership has been used for many years as a biological crop protection agent. The dual nature of Photorhabdus as a pathogen and mutualist makes it a superb model for understanding these apparently exclusive activities. Furthermore, recently identified clinical isolates of Photorhabdus are helping us to understand how human pathogens can emerge from the enormous reservoir of invertebrate pathogens in the environment. As Photorhabdus has never been found outside a host animal, its niche represents an entirely biotic landscape. In this review we discuss what molecular adaptations allow this bacterium to complete this fascinating and complex life cycle.
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pirab toxin from Photorhabdus asymbiotica as a larvicide against dengue vectors
Applied and Environmental Microbiology, 2009Co-Authors: Arunee Ahantarig, Nicholas R. Waterfield, Richard H Ffrenchconstant, Nantarat Chantawat, Pattamaporn KittayapongAbstract:We have evaluated Photorhabdus insect-related protein (Pir) from Photorhabdus asymbiotica against dengue vectors. PirAB shows larvicidal activity against both Aedes aegypti and Aedes albopictus larvae but did not affect the Mesocyclops thermocyclopoides predator. PirAB expressed the strongest toxicity compared to PirA, PirB, or the mixture of PirA plus PirB. Whether the presence of an enterobacterial repetitive intergenic consensus sequence in PirAB, but not in PirA, PirB, or the mixture of PirA plus PirB, has any impact on biological control efficacy needs further investigation.
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a single locus from the entomopathogenic bacterium Photorhabdus luminescens inhibits activated manduca sexta phenoloxidase
Fems Microbiology Letters, 2009Co-Authors: Ioannis Eleftherianos, Nicholas R. Waterfield, Richard H Ffrenchconstant, Peter Bone, Sam Boundy, Stuart E ReynoldsAbstract:Insect blood (hemolymph) contains prophenoloxidase, a proenzyme that is activated to protective phenoloxidase when the insect is damaged or challenged with microorganisms. The Gram-negative bacterium Photorhabdus luminescens kills the lepidopteron insect Manduca sexta by using a variety of toxins. We screened P. luminescens and Photorhabdus asymbiotica cosmid libraries in an Escherichia coli host against previously activated M. sexta hemolymph phenoloxidase and identified three overlapping cosmid clones from P. luminescens and five from P. asymbiotica that suppressed the activity of the enzyme both in vitro and in vivo. Genome alignments of cosmid end sequences from both species confirmed that they contained orthologous loci. We examined one of the cosmids from P. luminescens in detail: it induced the formation of significantly fewer melanotic nodules, proliferated faster within the insect host and was significantly more virulent towards fifth-stage larvae than E. coli control bacteria. Insertional mutagenesis of this cosmid yielded 11 transposon mutants that were no longer inhibitory. All of these were insertions into a single 5.5-kb locus, which contained three ORFs and was homologous to the maltodextrin phosphorylase locus of E. coli. The implications of this novel inhibitory factor of insect phenoloxidase for Photorhabdus virulence are discussed.
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insecticidal toxins from Photorhabdus bacteria and their potential use in agriculture
Toxicon, 2007Co-Authors: Richard H Ffrenchconstant, Andrea J Dowling, Nicholas R. WaterfieldAbstract:Abstract Most of the insecticidal toxins used in agriculture come from a single bacterium Bacillus thuringiensis or ‘Bt’. Here we review our work on the array of toxins produced by Photorhabdus and Xenorhabdus bacteria that are symbiotic with entomopathogenic nematodes, and discuss their potential for use in agriculture as alternatives to Bt. Despite the fact that both Photorhabdus and Xenorhabdus are introduced directly into the insect blood stream by their nematode vectors, they produce a range of toxins with both oral and injectable insecticidal activity. The toxin complexes (Tc's) are large orally active toxins that are displayed on the outer surface of the bacterium. They require three components (A–C) for full toxicity and one ‘A’ component has been successfully expressed in transgenic Arabidopsis to confer insect resistance. One such group of Tc's, the PirAB binary toxins, have oral activity against mosquitoes and some caterpillar pests. Their mode of action is not known but they show significant sequence similarity to a recently described neurotoxin β-leptinotarsin-h isolated from the blood of the Colorado potato beetle. Other toxins such as ‘makes caterpillars floppy’ (Mcf) and proteins encoded by the ‘Photorhabdus virulence cassettes’ (PVCs) only show injectable activity. Mcf1 promotes apoptosis in a wide range of cells and appears to mimic mammalian BH3 domain-only proteins in the mitochondrion whereas the mode of action of the PVCs remains undetermined. The likely biological reasons for the massive functional redundancy in Photorhabdus insecticidal toxins are discussed.