Debilitation of Galleria mellonella hemocytes using CytCo a cytolytic-like protein derived from the entomopathogen Conidiobolus obscurus

Zhang, Lvhao; Yang, Tian; Su, Xiu; Zhang, Xinqi; Zhou, Xiang

doi:10.1016/j.pestbp.2023.105418

Cited by 2 publications

(5 citation statements)

References 41 publications

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Section: Discussionmentioning

confidence: 93%

“…In this study, the changes in host gene expression during the early infection phase are comparable to the impacts of purified CytCo on gene expression in the hemocytes of Galleria mellonella and Bursaphelenchus xylophilus [ 24 , 38 ]. For example, suppressing the katabolic metabolism activities maximizes the host resources available for the pathogen [ 24 ]. The upregulation of many neurosynaptic and ion-channel-related genes, which are involved in the neuro-motor system, is probably attributed to the intoxication induced by CytCo.…”

Section: Discussionmentioning

confidence: 93%

“…Protein coronatin-2 and metabolites harman and norharman, produced by Conidiobolus coronatus , are known to cause cell death of G. mellonella hemocytes, thus facilitating successful fungal infection and host killing [ 36 , 37 ]. Additionally, C. obscurus CytCo has been proven to deliberately eliminate the insect hemocytes via the induction of apoptosis and necroptosis [ 24 ].…”

Section: Discussionmentioning

confidence: 99%

“…The purified CytCo protein with a 6× His (histidine) tag at the N-terminal was prepared according to the method [ 24 ]. For total protein extraction, 2 g of healthy aphids were ground in 5 mL of 50-mM sodium phosphate buffer (pH 8.0), 150 mM sodium chloride, 1 mM β-mercaptoethanol, 2 mM magnesium chloride, 5% glycerol, and 1% polyvinyl pyrrolidone ( w / v ) supplemented with 1/2 protease cocktail inhibitor EDTA free (Calbiochem, Darmstadt, Germany), and 2 μL of benzonase nuclease (Sigma-Aldrich, Darmstadt, Germany).…”

Section: Methodsmentioning

confidence: 99%

“…Conidiobolus obscurus , a specialized entomophthoralean pathogen targeting aphids, induces over 50% mortality within 24 h at a dosage of 1000 conidia/mm 2 [ 22 ]. Its virulence factor, CytCo, a cytolytic-like protein, correlates directly with fungal virulence [ 23 ] and is implicated in hemocyte disruption, expediting aphid mortality [ 24 ]. The present study aims to elucidate the responses of the bean aphids Megoura crassicauda during the incubation period of C. obscurus infection based on transcriptomic analysis.…”

Section: Introductionmentioning

confidence: 99%

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Transcriptomic Profiling of Bean Aphid Megoura crassicauda upon Exposure to the Aphid-Obligate Entomopathogen Conidiobolus obscurus (Entomophthoromycotina) and Screening of CytCo-Binding Aphid Proteins through a Pull-Down Assay

Zhu,

Fu,

Zhang

et al. 2024

Insects

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Prolonged periods of host-lethal infection by entomopathogenic fungi pose challenges to the development of biological control agents. The obligate entomopathogen C. obscurus, however, rapidly kills aphid hosts, warranting investigation. This study investigated the interaction between C. obscurus and a bean aphid Megoura crassicauda during the incubation period of infection, using transcriptome analysis to map host gene expression profiles. Results indicate C. obscurus-inoculated aphid activation of the wound healing immune responses, alongside suppression of the key molecules involved in Toll signaling, melanization, and metabolism. Furthermore, neuromotor system-related genes were upregulated, paralleling the intoxication observed in a nematode pest treated with C. obscurus-derived CytCo protein. To deepen interaction insights, a His-tag pull-down assay coupled with mass spectrometry analysis was conducted using CytCo as a bait to screen for potential aphid protein interactors. The proteins were identified based on the assembled transcriptome, and eleven transmembrane proteins were predicted to bind to CytCo. Notably, a protein of putatively calcium-transporting ATPase stood out with the highest confidence. This suggests that CytCo plays a vital role in C. obscurus killing aphid hosts, implicating calcium imbalance. In conclusion, C. obscurus effectively inhibits aphid immunity and exhibits neurotoxic potential, expediting the infection process. This finding facilitates our understanding of the complex host–pathogen interactions and opens new avenues for exploring biological pest management strategies in agroforestry.

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Section: Discussionmentioning

confidence: 93%

Section: Discussionmentioning

confidence: 93%

Section: Discussionmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

See 3 more Smart Citations

Transcriptomic Profiling of Bean Aphid Megoura crassicauda upon Exposure to the Aphid-Obligate Entomopathogen Conidiobolus obscurus (Entomophthoromycotina) and Screening of CytCo-Binding Aphid Proteins through a Pull-Down Assay

Zhu,

Fu,

Zhang

et al. 2024

Insects

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Calcium leakage involved in nematotoxic effects of the Conidiobolus obscurus CytCo protein on the pine wood nematode, Bursaphelenchus xylophilus

Liu,

Chen

et al. 2024

Pest Management Science

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BACKGROUNDThe pine wood nematode Bursaphelenchus xylophilus, a severe invasive species, is responsible for causing widespread pine wilt disease. The CytCo protein, a pore‐forming toxin derived from Conidiobolus obscurus, exhibits nematotoxicity towards B. xylophilus.RESULTSOur present study reveals the expression variation of a range of gene products in B. xylophilus that respond to the effects of CytCo using the isobaric tags for relative and absolute quantification proteomics technology. Functional enrichment analysis indicates that many differentially expressed proteins are linked to calcium signaling system, proteasome, energy production and conversion, and the determination of adult lifespan. It suggests that the dysregulation of calcium homeostasis, energy metabolism, and apoptosis contribute to the CytCo nematotoxicity. Using the calcium ion (Ca2+)‐indicator calcein, we detected changes in Ca2+ levels in B. xylophilus, with a significantly increase in fluorescence in the nematode's intestine and pseudocoelom following CytCo treatments. Meanwhile, the apoptosis and reactive oxygen species (ROS) assays showed an enhancement of fluorescence in B. xylophilus cells, with increased CytCo concentrations.CONCLUSIONThe protein toxin CytCo triggers Ca2+ leakage, disrupts Ca2+ balance in B. xylophilus, and induces apoptosis and ROS outburst, thereby intensifying its nematotoxic effects. This finding facilitates our understanding of the modes of action of nematotoxic proteins, and contributes to the development of innovative nematode control strategies. © 2024 Society of Chemical Industry.

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Debilitation of Galleria mellonella hemocytes using CytCo a cytolytic-like protein derived from the entomopathogen Conidiobolus obscurus

Cited by 2 publications

References 41 publications

Transcriptomic Profiling of Bean Aphid Megoura crassicauda upon Exposure to the Aphid-Obligate Entomopathogen Conidiobolus obscurus (Entomophthoromycotina) and Screening of CytCo-Binding Aphid Proteins through a Pull-Down Assay

Transcriptomic Profiling of Bean Aphid Megoura crassicauda upon Exposure to the Aphid-Obligate Entomopathogen Conidiobolus obscurus (Entomophthoromycotina) and Screening of CytCo-Binding Aphid Proteins through a Pull-Down Assay

Calcium leakage involved in nematotoxic effects of the Conidiobolus obscurus CytCo protein on the pine wood nematode, Bursaphelenchus xylophilus

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