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Genetically modified entomopathogenic bacteria, recent developments, benefits and impacts: A review.The Science of the Total Environment Sep 2020Entomopathogenic bacteria (EPBs), insect pathogens that produce pest-specific toxins, are environmentally-friendly alternatives to chemical insecticides. However, the... (Review)
Review
Entomopathogenic bacteria (EPBs), insect pathogens that produce pest-specific toxins, are environmentally-friendly alternatives to chemical insecticides. However, the most important problem with EPBs application is their limited field stability. Moreover, environmental factors such as solar radiation, leaf temperature, and vapor pressure can affect the pathogenicity of these pathogens and their toxins. Scientists have conducted intensive research to overcome such problems. Genetic engineering has great potential for the development of new engineered entomopathogens with more resistance to adverse environmental factors. Genetically modified entomopathogenic bacteria (GM-EPBs) have many advantages over wild EPBs, such as higher pathogenicity, lower spraying requirements and longer-term persistence. Genetic manipulations have been mostly applied to members of the bacterial genera Bacillus, Lysinibacillus, Pseudomonas, Serratia, Photorhabdus and Xenorhabdus. Although many researchers have found that GM-EPBs can be used safely as plant protection bioproducts, limited attention has been paid to their potential ecological impacts. The main concerns about GM-EPBs and their products are their potential unintended effects on beneficial insects (predators, parasitoids, pollinators, etc.) and rhizospheric bacteria. This review address recent update on the significant role of GM-EPBs in biological control, examining them through different perspectives in an attempt to generate critical discussion and aid in the understanding of their potential ecological impacts.
Topics: Animals; Genetic Engineering; Insecta; Insecticides; Pest Control, Biological; Photorhabdus; Xenorhabdus
PubMed: 32460068
DOI: 10.1016/j.scitotenv.2020.139169 -
Frontiers in Microbiology 2023Contractile injection systems (CISs) are phage tail-related structures that are encoded in many bacterial genomes. These devices encompass the cell-based type VI... (Review)
Review
Contractile injection systems (CISs) are phage tail-related structures that are encoded in many bacterial genomes. These devices encompass the cell-based type VI secretion systems (T6SSs) as well as extracellular CISs (eCISs). The eCISs comprise the R-tailocins produced by various bacterial species as well as related phage tail-like structures such as the antifeeding prophages (Afps) of , the virulence cassettes (PVCs), and the metamorphosis-associated contractile structures (MACs) of . These contractile structures are released into the extracellular environment upon suicidal lysis of the producer cell and play important roles in bacterial ecology and evolution. In this review, we specifically portray the eCISs with a focus on the R-tailocins, sketch the history of their discovery and provide insights into their evolution within the bacterial host, their structures and how they are assembled and released. We then highlight ecological and evolutionary roles of eCISs and conceptualize how they can influence and shape bacterial communities. Finally, we point to their potential for biotechnological applications in medicine and agriculture.
PubMed: 37886057
DOI: 10.3389/fmicb.2023.1264877 -
The Journal of Parasitology Jan 2023The entomopathogenic nematode (EPN) Heterorhabditis bacteriophora infects a wide range of insect hosts with the aid of its mutualistic bacteria Photorhabdus luminescens....
The entomopathogenic nematode (EPN) Heterorhabditis bacteriophora infects a wide range of insect hosts with the aid of its mutualistic bacteria Photorhabdus luminescens. While the mutualistic relationship between H. bacteriophora and P. luminescens and the infectivity of the nematode-bacteria complex have been characterized, how nematode fitness is affected by entomopathogenic bacteria existing in association with other EPN species remains poorly understood. In this study, the survival of H. bacteriophora infective juveniles containing or lacking P. luminescens was tested against the entomopathogenic bacteria Xenorhabdus nematophila and Photorhabdus asymbiotica as well as the non-pathogenic Escherichia coli. While X. nematophila and E. coli did not significantly affect the survival of H. bacteriophora, P. asymbiotica exerted a significant effect on nematode survival, particularly on those lacking P. luminescens. These results imply that P. asymbiotica encodes factors that are pathogenic to EPNs. Future efforts will focus on the identification of the bacterial molecular components that induce these effects. This study makes an important contribution to a growing body of research aimed at exploiting the full potential of nematode-bacterial complexes for eliminating noxious insect pests and treating infectious diseases caused by parasitic nematodes.
Topics: Animals; Photorhabdus; Escherichia coli; Nematoda; Symbiosis
PubMed: 36805240
DOI: 10.1645/22-55 -
Journal of Invertebrate Pathology Feb 2022Recovery, yield, and dispersal are crucial developmental and behavioral indices for the infective juveniles of entomopathogenic nematodes, which are used as biocontrol...
Recovery, yield, and dispersal are crucial developmental and behavioral indices for the infective juveniles of entomopathogenic nematodes, which are used as biocontrol agents against a variety of agricultural pests. Ascarosides and isopropylstilbene (ISO) function as nematode pheromones with developmental and behavioral effects. In this study, 11 synthesized ascarosides identified from Caenorhabditis elegans, together with ISO identified from Photorhabdus luminescens, were used to determine their influence on the IJ recovery, growth on agar plates, and dispersal of S. carpocapsae All, H. bacteriophora H06 and H. indica LN2 nematodes. Compared with the controls, significant differences in IJ recovery of three nematode species were detected from the supernatants of their corresponding bacterial cultures with almost all ascarosides or isopropylstilbene (ISO) at 0.04 nM in 6 days. The highest IJ recovery percentages was obtained from ISO and ascr#3 for All strain, ascr#5 and ascr#6 for LN2 strain, and ISO and ascr#12 for H06 strain. The ISO detected from Photorhabdus bacteria also induced IJ recovery of S. carpocapsae All. IJ yields was significantly stimulated by all synthesized compounds for S. carpocapsae All, and by most compounds for H. bacteriophora H06. The higher IJ yields varied with ascarosides. Ascr#7 and DMSO was common for the improved IJ yields of both nematode species. The three nematode species showed marked differences in dispersal behavior. In response to the ascarosides or ISO, S. carpocapsae All IJs actively moved with different dispersal rates, H. indica LN2 IJs in very low dispersal rates, and H. bacteriophora H06 IJs in variable and even suppressed rates on the agar plates at least during the assay period. Based on the synthesized standards, ascr#1, ascr#9 and ascr#10 were detected from three nematode species, ascr#5 and ascr#11 also from S. carpocapsae All and H. bacteriophora H06, and ascr#12 also from H. bacteriophora H06 and H. indica LN2. Ascr#9 was most abundant in three nematode species. Compared with the sterile PBS, significantly more ascr#1, ascr#9 and ascr#10 were detected from S. carpocapsae All and H. indica LN2, but less ascr#5 and ascr#11 from S. carpocapsae All, ascr#1, ascr#5, ascr#11 and ascr#12 from H. bacteriophora H06, in the corresponding bacterial supernatant. It seems that the bacterial supernatants could regulate the ascaroside secretion by the three nematode species. These results will provide useful clues for selecting suitable ascarosides to induce the recovery, improve the yield, and enhance the dispersal of the IJs of these nematodes.
Topics: Agar; Animals; Nematoda; Pheromones; Photorhabdus
PubMed: 35031295
DOI: 10.1016/j.jip.2022.107717 -
Insects Oct 2019and are entomopathogenic bacterial symbionts that produce toxic proteins that can interfere with the immune system of insects. Herein, we show that outer membrane...
and are entomopathogenic bacterial symbionts that produce toxic proteins that can interfere with the immune system of insects. Herein, we show that outer membrane proteins (OMPs) could be involved as bacterial virulence factors. Purified totals OMPs of both bacterial species were injected into fifth instar larvae of Hübner. Larvae were surveyed for cellular defenses fluctuations in total haemocyte counts (THC) and granulocyte percentage and for the humoral defenses protease, phospholipase A2 (PLA), and phenoloxidase (PO) activities at specific time intervals. Changes in the expression of the three inducible antimicrobial peptides (AMPs), cecropin, attacin, and spodoptericin, were also measured. Larvae treated with OMPs of both bacterial species had more haemocytes than did the negative controls. OMPs of caused more haemocyte destruction than did the OMPs of . The OMPs of both bacterial species initially activated insect defensive enzymes post-injection, the degree of activation varying with enzyme type. The AMPs, attacin, cecropin, and spodoptericin were up-regulated by OMP injections compared with the normal larvae. The expression of these three AMPs was maximal at four hours post injection (hpi) with OMPs treatment. Expression of the three AMPs in treated insects was irregular and lower than in the OMPs treatment. These findings provide insights into the role of OMPs of entomopathogenic nematode bacterial symbionts in countering the physiological defenses of insects.
PubMed: 31627300
DOI: 10.3390/insects10100352 -
Natural Product Research Dec 2022Anthraquinones (AQ), unveiling large structural diversity, among polyketides demonstrate a wide range of applications. The hydroxy anthraquinones (HAQ), a group of... (Review)
Review
Anthraquinones (AQ), unveiling large structural diversity, among polyketides demonstrate a wide range of applications. The hydroxy anthraquinones (HAQ), a group of anthraquinone derivatives, are secondary metabolites produced by bacteria and eukaryotes. Plant-based HAQ are well-studied unlike bacterial HAQ and applied as herbal medicine for centuries. Bacteria are known to synthesize a wide variety of structurally diversified HAQ through polyketide pathways using polyketide synthases (I, II & III) principally through polyketide synthase-II. The actinobacteria especially the genus and represent a rich source of HAQ, however novel HAQ are reported from the rare actinobacteria genera (, and . Though several reviews are available on AQ produced by plants and fungi, however none on bacterial AQ. The current review focused on sources of bacterial HAQ and their structural diversity and biological activities along with toxicity and side effects.
Topics: Plants; Polyketide Synthases; Polyketides; Streptomyces
PubMed: 35175877
DOI: 10.1080/14786419.2022.2039920 -
Microorganisms Mar 2023In bacteria, group-coordinated behavior such as biofilm formation or virulence are often mediated via cell-cell communication, a process referred to as quorum sensing...
In bacteria, group-coordinated behavior such as biofilm formation or virulence are often mediated via cell-cell communication, a process referred to as quorum sensing (QS). The canonical QS system of Gram-negative bacteria uses -acyl homoserine lactones (AHLs) as communication molecules, which are produced by LuxI-type synthases and sensed by cognate LuxR-type receptors. These receptors act as transcriptional regulators controlling the expression of specific genes. Some bacteria harbor LuxR-type receptors lacking a cognate LuxI-type synthases, designated as LuxR solos. Among many other LuxR solos, the entomopathogenic enteric bacterium harbors a SdiA-like LuxR solo containing an AHL signal-binding domain, for which a respective signal molecule and target genes have not been identified yet. Here we performed SPR analysis to demonstrate that SdiA acts as a bidirectional regulator of transcription, tightly controlling its own expression and the adjacent () gene in a gene supposed to be involved in the colonization of eukaryotes. Via qPCR we could further determine that in deletion mutant strains, is upregulated, indicating that SdiA negatively affects expression of . Furthermore, the Δ deletion mutant exhibited differences in biofilm formation and motility compared with the wild-type. Finally, using nanoDSF analysis we could identify putative binding ability of SdiA towards diverse AHLs, but also to plant-derived signals, modulating the DNA-binding capacity of SdiA, suggesting that this LuxR solo acts as an important player in interkingdom signaling between and plants.
PubMed: 37110313
DOI: 10.3390/microorganisms11040890 -
Microbial Cell Factories Apr 2024Bacteria of the genus Photorhabdus and Xenorhabdus are motile, Gram-negative bacteria that live in symbiosis with entomopathogenic nematodes. Due to their complex life...
BACKGROUND
Bacteria of the genus Photorhabdus and Xenorhabdus are motile, Gram-negative bacteria that live in symbiosis with entomopathogenic nematodes. Due to their complex life cycle, they produce a large number of specialized metabolites (natural products) encoded in biosynthetic gene clusters (BGC). Genetic tools for Photorhabdus and Xenorhabdus have been rare and applicable to only a few strains. In the past, several tools have been developed for the activation of BGCs and the deletion of individual genes. However, these often have limited efficiency or are time consuming. Among the limitations, it is essential to have versatile expression systems and genome editing tools that could facilitate the practical work.
RESULTS
In the present study, we developed several expression vectors and a CRISPR-Cpf1 genome editing vector for genetic manipulations in Photorhabdus and Xenorhabdus using SEVA plasmids. The SEVA collection is based on modular vectors that allow exchangeability of different elements (e.g. origin of replication and antibiotic selection markers with the ability to insert desired sequences for different end applications). Initially, we tested different SEVA vectors containing the broad host range origins and three different resistance genes for kanamycin, gentamycin and chloramphenicol, respectively. We demonstrated that these vectors are replicative not only in well-known representatives, e.g. Photorhabdus laumondii TTO1, but also in other rarely described strains like Xenorhabdus sp. TS4. For our CRISPR/Cpf1-based system, we used the pSEVA231 backbone to delete not only small genes but also large parts of BGCs. Furthermore, we were able to activate and refactor BGCs to obtain high production titers of high value compounds such as safracin B, a semisynthetic precursor for the anti-cancer drug ET-743.
CONCLUSIONS
The results of this study provide new inducible expression vectors and a CRISPR/CPf1 encoding vector all based on the SEVA (Standard European Vector Architecture) collection, which can improve genetic manipulation and genome editing processes in Photorhabdus and Xenorhabdus.
Topics: Xenorhabdus; Photorhabdus; Gene Editing; Biological Products; Clustered Regularly Interspaced Short Palindromic Repeats
PubMed: 38561780
DOI: 10.1186/s12934-024-02363-8 -
Gene Aug 2021The genome sequences of entomopathogenic nematode (EPN) bacteria and their functional analyses can lead to the genetic engineering of the bacteria for use as biocontrol...
Genome assembly and annotation of Photorhabdus heterorhabditis strain ETL reveals genetic features involved in pathogenicity with its associated entomopathogenic nematode and anti-host effectors with biocontrol potential applications.
The genome sequences of entomopathogenic nematode (EPN) bacteria and their functional analyses can lead to the genetic engineering of the bacteria for use as biocontrol agents. The bacterial symbiont Photorhabdus heterorhabditis strain ETL isolated from an insect pathogenic nematode, Heterorhabditis zealandica strain ETL, collected in the northernmost region of South Africa was studied to reveal information that can be useful in the design of improvement strategies for both effective and liquid production method of EPN-based pesticides. The strain ETL genome was found closely related to the type strain genome of P. australis DSM 17,609 (~60 to 99.9% CDSs similarity), but closely related to the not yet genome-sequenced type strain, P. heterorhabditis. It has a genome size of 4,866,148 bp and G + C content of 42.4% similar to other Photorhabdus. It contains 4,351 protein coding genes (CDSs) of which, at least 84% are shared with the de facto type strain P. luminescens subsp. laumondii TTO1, and has 318 unknown CDSs and the genome has a higher degree of plasticity allowing it to adapt to different environmental conditions, and to be virulent against various insects; observed through genes acquired through horizontal gene transfer mechanisms, clustered regularly interspaced short palindromic repeats, non-determined polyketide- and non-ribosomal peptide- synthase gene clusters, and many genes associated with uncharacterized proteins; which also justify the strain ETL's genes differences (quantity and quality) compared to P. luminescens subsp. laumondii TTO1. The protein coding sequences contained genes with both bio-engineering and EPNs mass production importance, of which numerous are uncharacterized.
Topics: Animals; Base Sequence; Biological Control Agents; Genes, Bacterial; Genome, Bacterial; Host-Pathogen Interactions; Molecular Sequence Annotation; Photorhabdus; Phylogeny; Strongyloidea; Virulence
PubMed: 34147570
DOI: 10.1016/j.gene.2021.145780 -
Life Science Alliance Oct 2019Engineering delivery systems for proteins and peptides into mammalian cells is an ongoing challenge for cell biological studies as well as for therapeutic approaches....
Engineering delivery systems for proteins and peptides into mammalian cells is an ongoing challenge for cell biological studies as well as for therapeutic approaches. toxin complex (PTC) is a heterotrimeric protein complex able to deliver diverse protein toxins into mammalian cells. We engineered the syringe-like nanomachine for delivery of protein toxins from different species. In addition, we loaded the highly active copepod luciferase M-Luc7 for accurate quantification of injected molecules. We suggest that besides the probable size limitation, the charge of the cargo also influences the efficiency of packing and transport into mammalian cells. Our data show that the PTC constitutes a powerful system to inject recombinant proteins, peptides, and potentially, other molecules into mammalian cells. In addition, in contrast to other protein transporters based on pore formation, the closed, compact structure of the PTC may protect cargo from degradation.
Topics: Animals; Bacterial Proteins; Bacterial Toxins; Cloning, Molecular; Copepoda; Cysteine Endopeptidases; Drug Delivery Systems; HeLa Cells; Humans; Injections; Luciferases; Nanoparticles; Photorhabdus; Protein Engineering
PubMed: 31540947
DOI: 10.26508/lsa.201900485