scholarly journals Tenebrionid secretions and a fungal benzoquinone oxidoreductase form competing components of an arms race between a host and pathogen

2015 ◽  
Vol 112 (28) ◽  
pp. E3651-E3660 ◽  
Author(s):  
Nicolás Pedrini ◽  
Almudena Ortiz-Urquiza ◽  
Carla Huarte-Bonnet ◽  
Yanhua Fan ◽  
M. Patricia Juárez ◽  
...  
Keyword(s):  
Galleria Mellonella ◽  
Gene Knockout ◽  
Reductase Activity ◽  
Pathogenic Fungus ◽  
Sitophilus Oryzae ◽  
Arms Race ◽  
Wild Type ◽  
Tenebrionid Beetle ◽  
Selection Processes ◽  
Greater Wax Moth

Entomopathogenic fungi and their insect hosts represent a model system for examining invertebrate-pathogen coevolutionary selection processes. Here we report the characterization of competing components of an arms race consisting of insect protective antimicrobial compounds and evolving fungal mechanisms of detoxification. The insect pathogenic fungusBeauveria bassianahas a remarkably wide host range; however, some insects are resistant to fungal infection. Among resistant insects is the tenebrionid beetleTribolium castaneumthat produces benzoquinone-containing defensive secretions. Reduced fungal germination and growth was seen in media containingT. castaneumdichloromethane extracts or synthetic benzoquinone. In response to benzoquinone exposure, the fungus expresses a 1,4-benzoquinone oxidoreductase,BbbqrA, induced >40-fold. Gene knockout mutants (ΔBbbqrA) showed increased growth inhibition, whereasB. bassianaoverexpressingBbbqrA(Bb::BbbqrAO) displayed increased resistance to benzoquinone compared with wild type. Increased benzoquinone reductase activity was detected in wild-type cells exposed to benzoquinone and in the overexpression strain. Heterologous expression and purification of BbBqrA inEscherichia coliconfirmed NAD(P)H-dependent benzoquinone reductase activity. TheΔBbbqrAstrain showed decreased virulence towardT. castaneum, whereas overexpression ofBbbqrAincreased mortality versusT. castaneum. No change in virulence was seen for theΔBbbqrAorBb::BbbqrAOstrains when tested against the greater wax mothGalleria mellonellaor the beetleSitophilus oryzae, neither of which produce significant amounts of cuticular quinones. The observation that artificial overexpression ofBbbqrAresults in increased virulence only toward quinone-secreting insects implies the lack of strong selection or current failure ofB. bassianato counteradapt to this particular host defense throughout evolution.

scholarly journals Losing the Arms Race: Greater Wax Moths Sense but Ignore Bee Alarm Pheromones

Insects ◽  
2019 ◽  
Vol 10 (3) ◽  
pp. 81 ◽  
Author(s):  
Yuan Li ◽  
Xingchuan Jiang ◽  
Zhengwei Wang ◽  
Junjun Zhang ◽  
Katrina Klett ◽  
...  
Keyword(s):  
Galleria Mellonella ◽  
Arms Race ◽  
Oviposition Site ◽  
Benzyl Acetate ◽  
Alarm Pheromones ◽  
Bee Colony ◽  
Octyl Acetate ◽  
Greater Wax Moth ◽  
Isopentyl Acetate ◽  
Wax Moth

The greater wax moth, Galleria mellonella L., is one of main pests of honeybees. The larvae burrow into the wax, damaging the bee comb and degenerating bee products, but also causes severe effects like driving the whole colony to abscond. In the present study, we used electroantennograms, a Y maze, and an oviposition site choice bioassay to test whether the greater wax moth can eavesdrop on bee alarm pheromones (isopentyl acetate, benzyl acetate, octyl acetate, and 2-heptanone), to target the bee colony, or if the bee alarm pheromones would affect their preference of an oviposition site. The results revealed that the greater wax moth showed a strong electroantennogram response to these four compounds of bee alarm pheromones even in a low concentration (100 ng/μL), while they showed the highest response to octyl acetate compared to the other three main bee alarm components (isopentyl acetate, benzyl acetate, and 2-heptanone). However, the greater wax moth behavioral results showed no significant preference or avoidance to these four bee alarm pheromones. These results indicate that bees are currently losing the arms race since the greater wax moth can sense bee alarm pheromones, however, these alarm pheromones are ignored by the greater wax moth.

scholarly journals Differential Roles of the Pseudomonas aeruginosaPA14 rpoN Gene in Pathogenicity in Plants, Nematodes, Insects, and Mice

2001 ◽  
Vol 183 (24) ◽  
pp. 7126-7134 ◽  
Author(s):  
Erik L. Hendrickson ◽  
Joulia Plotnikova ◽  
Shalina Mahajan-Miklos ◽  
Laurence G. Rahme ◽  
Frederick M. Ausubel
Keyword(s):  
Galleria Mellonella ◽  
Mesophyll Cell ◽  
Wild Type ◽  
Gene Encoding ◽  
Disease Symptoms ◽  
Infectivity Assay ◽  
Greater Wax Moth ◽  
Injury Model ◽  
Similar Disease ◽  
Wax Moth

ABSTRACT We cloned the rpoN (ntrA, glnF) gene encoding the alternate sigma factor ς54 from the opportunistic multihost pathogen Pseudomonas aeruginosastrain PA14. A marker exchange protocol was used to construct the PA14rpoN insertional mutationrpoN::Genr. PA14rpoN::Genr synthesized reduced levels of pyocyanin and displayed a variety of phenotypes typical of rpoN mutants, including a lack of motility and the failure to grow on nitrate, glutamate, or histidine as the sole nitrogen source. Compared to wild-type PA14,rpoN::Genr was ca. 100-fold less virulent in a mouse thermal injury model and was significantly impaired in its ability to kill the nematode Caenorhabditis elegans. In an Arabidopsis thaliana leaf infectivity assay, althoughrpoN::Genr exhibited significantly reduced attachment to trichomes, stomata, and the epidermal cell surface, did not attach perpendicularly to or perforate mesophyll cell walls, and proliferated less rapidly in Arabidopsisleaves, it nevertheless elicited similar disease symptoms to wild-typeP. aeruginosa PA14 at later stages of infection.rpoN::Genr was not impaired in virulence in a Galleria mellonella (greater wax moth) pathogenicity model. These data indicate that rpoN does not regulate the expression of any genes that encode virulence factors universally required for P. aeruginosa pathogenicity in diverse hosts.

scholarly journals Establishing an invertebrate Galleria mellonella greater wax moth larval model of Neisseria gonorrhoeae infection

Virulence ◽  
2021 ◽  
Vol 12 (1) ◽  
pp. 1900-1920
Author(s):  
Aiste Dijokaite ◽  
Maria Victoria Humbert ◽  
Emma Borkowski ◽  
Roberto M La Ragione ◽  
Myron Christodoulides
Keyword(s):  
Neisseria Gonorrhoeae ◽  
Galleria Mellonella ◽  
Greater Wax Moth ◽  
Wax Moth

scholarly journals The Alopecia Areata Phenotype Is Induced by the Water Avoidance Stress Test In cchcr1-Deficient Mice

Biomedicines ◽  
2021 ◽  
Vol 9 (7) ◽  
pp. 840
Author(s):  
Qiaofeng Zhao ◽  
Satoshi Koyama ◽  
Nagisa Yoshihara ◽  
Atsushi Takagi ◽  
Etsuko Komiyama ◽  
...  
Keyword(s):  
Alopecia Areata ◽  
Gene Knockout ◽  
Stress Test ◽  
Coiled Coil ◽  
Risk Haplotype ◽  
Gene Expression Microarray ◽  
Wild Type ◽  
Gene Knockout Mice ◽  
Deficient Mice ◽  
Gene Expression Microarray Analysis

We recently discovered a nonsynonymous variant in the coiled-coil alpha-helical rod protein 1 (CCHCR1) gene within the alopecia areata (AA) risk haplotype. We also reported that the engineered mice with this risk allele exhibited. To investigate more about the involvement of the CCHCR1 gene in AA pathogenesis, we developed an AA model using C57BL/6N cchcr1 gene knockout mice. In this study, mice (6–8 weeks) were divided into two groups: cchcr1−/− mice and wild-type (WT) littermates. Both groups were subjected to a water avoidance stress (WAS) test. Eight weeks after the WAS test, 25% of cchcr1−/− mice exhibited non-inflammatory foci of alopecia on the dorsal skin. On the other hand, none of wild-type littermates cause hair loss. The foci resembled human AA in terms of gross morphology, trichoscopic findings and histological findings. Additionally, gene expression microarray analysis of cchcr1−/− mice revealed abnormalities of hair related genes compared to the control. Our results strongly suggest that CCHCR1 is associated with AA pathogenesis and that cchcr1−/− mice are a good model for investigating AA.

Transgenic expression of gallerimycin, a novel antifungal insect defensin from the greater wax moth Galleria mellonella, confers resistance to pathogenic fungi in tobacco

2006 ◽  
Vol 387 (5) ◽  
pp. 549-557 ◽  
Author(s):  
Gregor Langen ◽  
Jafargholi Imani ◽  
Boran Altincicek ◽  
Gernot Kieseritzky ◽  
Karl-Heinz Kogel ◽  
...  
Keyword(s):  
Transgenic Tobacco ◽  
Fungal Pathogens ◽  
Galleria Mellonella ◽  
Ectopic Expression ◽  
Pathogenic Fungi ◽  
Transgenic Expression ◽  
Greater Wax Moth ◽  
Insect Defensin ◽  
Wax Moth

Abstract A cDNA encoding gallerimycin, a novel antifungal peptide from the greater wax moth Galleria mellonella, was isolated from a cDNA library of genes expressed during innate immune response in the caterpillars. Upon ectopic expression of gallerimycin in tobacco, using Agrobacterium tumefaciens as a vector, gallerimycin conferred resistance to the fungal pathogens Erysiphe cichoracearum and Sclerotinia minor. Quantification of gallerimycin mRNA in transgenic tobacco by real-time PCR confirmed transgenic expression under control of the inducible mannopine synthase promoter. Leaf sap and intercellular washing fluid from transgenic tobacco inhibited in vitro germination and growth of the fungal pathogens, demonstrating that gallerimycin is secreted into intercellular spaces. The feasibility of the use of gallerimycin to counteract fungal diseases in crop plants is discussed.

scholarly journals KCNJ11 knockout morula re-engineered by stem cell diploid aggregation

2008 ◽  
Vol 364 (1514) ◽  
pp. 269-276 ◽  
Author(s):  
Timothy J Nelson ◽  
Almudena Martinez-Fernandez ◽  
Andre Terzic
Keyword(s):  
Metabolic Flux ◽  
Ex Vivo ◽  
Gene Knockout ◽  
Embryonic Stem ◽  
Adaptive Behaviour ◽  
Wild Type ◽  
New Paradigm ◽  
Cell Functions ◽  
Dependent Cell ◽  
Channel Dependent

KCNJ11 -encoded Kir6.2 assembles with ATP-binding cassette sulphonylurea receptors to generate ATP-sensitive K + (K ATP ) channel complexes. Expressed in tissues with dynamic metabolic flux, these evolutionarily conserved yet structurally and functionally unique heteromultimers serve as high-fidelity rheostats that adjust membrane potential-dependent cell functions to match energetic demand. Genetic defects in channel subunits disrupt the cellular homeostatic response to environmental stress, compromising organ tolerance in the adult. As maladaptation characterizes malignant K ATP channelopathies, establishment of platforms to examine progression of K ATP channel-dependent adaptive behaviour is warranted. Chimeras provide a powerful tool to assay the contribution of genetic variance to stress intolerance during prenatal or post-natal development. Here, KCNJ11 K ATP channel gene knockout↔wild-type chimeras were engineered through diploid aggregation. Integration of wild-type embryonic stem cells into zona pellucida-denuded morula derived from knockout embryos achieved varying degrees of incorporation of stress-tolerant tissue within the K ATP channel-deficient background. Despite the stress-vulnerable phenotype of the knockout, ex vivo derived mosaic blastocysts tolerated intrauterine transfer and implantation, followed by full-term embryonic development in pseudopregnant surrogates to produce live chimeric offspring. The development of adult chimerism from the knockout↔wild-type mosaic embryo offers thereby a new paradigm to probe the ecogenetic control of the K ATP channel-dependent stress response.

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