Transcripts analysis of the entomopathogenic nematode Steinernema carpocapsae induced in vitro with insect haemolymph

Hao, Youjin; Montiel, Rafaél; Abubucker, Sahar; Mitreva, Makedonka; Simões, Nélson

doi:10.1016/j.molbiopara.2009.10.002

Cited by 39 publications

(27 citation statements)

References 70 publications

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“…Hemolymph plasma from Manduca sexta was selected as an activation media because M. sexta is a natural host, and its large size facilitates extraction of large quantities of hemolymph. Previous studies identified hemolymph soaking as a valid method for inducing synchronous activation of entomopathogenic IJs, while also allowing for the mass activation of IJs necessary to produce enough high quality RNA for RNA-seq [19, 22–24]. Additionally, our pilot studies used to determine the maximum concentration of IJs per milliliter hemolymph plasma showed IJs develop into J4s within 48 h of soaking and continue to adults by 72 h (data not shown).…”

Section: Discussionmentioning

confidence: 70%

Identification of candidate infection genes from the model entomopathogenic nematode Heterorhabditis bacteriophora

et al. 2017

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BackgroundDespite important progress in the field of innate immunity, our understanding of host immune responses to parasitic nematode infections lags behind that of responses to microbes. A limiting factor has been the obligate requirement for a vertebrate host which has hindered investigation of the parasitic nematode infective process. The nematode parasite Heterorhabditis bacteriophora offers great potential as a model to genetically dissect the process of infection. With its mutualistic Photorhabdus luminescens bacteria, H. bacteriophora invades multiple species of insects, which it kills and exploits as a food source for the development of several nematode generations. The ability to culture the life cycle of H. bacteriophora on plates growing the bacterial symbiont makes it a very exciting model of parasitic infection that can be used to unlock the molecular events occurring during infection of a host that are inaccessible using vertebrate hosts.ResultsTo profile the transcriptional response of an infective nematode during the early stage of infection, we performed next generation RNA sequencing on H. bacteriophora IJs incubated in Manduca sexta hemolymph plasma for 9 h. A subset of up-regulated and down-regulated genes were validated using qRT-PCR. Comparative analysis of the transcriptome with untreated controls found a number of differentially expressed genes (DEGs) which cover a number of different functional categories. A subset of DEGs is conserved across Clade V parasitic nematodes revealing an array of candidate parasitic genes.ConclusionsOur analysis reveals transcriptional changes in the regulation of a large number of genes, most of which have not been shown previously to play a role in the process of infection. A significant proportion of these genes are unique to parasitic nematodes, suggesting the identification of a group of parasitism factors within nematodes. Future studies using these candidates may provide functional insight into the process of nematode parasitism and also the molecular evolution of parasitism within nematodes.Electronic supplementary materialThe online version of this article (doi:10.1186/s12864-016-3468-6) contains supplementary material, which is available to authorized users.

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Section: Discussionmentioning

confidence: 70%

Identification of candidate infection genes from the model entomopathogenic nematode Heterorhabditis bacteriophora

et al. 2017

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“…For small RNA (sRNA) sequencing, nematodes were induced for 2 hour with hemolymph of Galleria mellonella and with buffer as control64. Nematodes were grinded under liquid nitrogen and RNA was extracted with Trizol according to the manufacturer’s instructions.…”

Section: Methodsmentioning

confidence: 99%

“…Nematodes were induced as described64 with slight modifications. A pool of approximately 25,000 nematodes were induced with either hemolymph or intestine of Galleria mellonella .…”

Section: Methodsmentioning

confidence: 99%

The genome, transcriptome, and proteome of the nematode Steinernema carpocapsae: evolutionary signatures of a pathogenic lifestyle

Rougon‐Cardoso

Flores-Ponce

Ramos-Aboites

et al. 2016

Sci Rep

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The entomopathogenic nematode Steinernema carpocapsae has been widely used for the biological control of insect pests. It shares a symbiotic relationship with the bacterium Xenorhabdus nematophila, and is emerging as a genetic model to study symbiosis and pathogenesis. We obtained a high-quality draft of the nematode’s genome comprising 84,613,633 bp in 347 scaffolds, with an N50 of 1.24 Mb. To improve annotation, we sequenced both short and long RNA and conducted shotgun proteomic analyses. S. carpocapsae shares orthologous genes with other parasitic nematodes that are absent in the free-living nematode C. elegans, it has ncRNA families that are enriched in parasites, and expresses proteins putatively associated with parasitism and pathogenesis, suggesting an active role for the nematode during the pathogenic process. Host and parasites might engage in a co-evolutionary arms-race dynamic with genes participating in their interaction showing signatures of positive selection. Our analyses indicate that the consequence of this arms race is better characterized by positive selection altering specific functions instead of just increasing the number of positively selected genes, adding a new perspective to these co-evolutionary theories. We identified a protein, ATAD-3, that suggests a relevant role for mitochondrial function in the evolution and mechanisms of nematode parasitism.

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“…In an attempt to understand the interaction between S. carpocapsae and insects, an expressed sequence tag (EST) library was created and analysed, resulting in the identification of a serine protease inhibitor [23]. This inhibitor was later detected in ESPs produced by parasitic stage nematodes, suggesting its involvement in the parasitic process.…”

Section: Introductionmentioning

confidence: 99%

A Serpin Released by an Entomopathogen Impairs Clot Formation in Insect Defense System

et al. 2013

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Steinernema carpocapsae is an entomopathogenic nematode widely used for the control of insect pests due to its virulence, which is mainly attributed to the ability the parasitic stage has to overcome insect defences. To identify the mechanisms underlying such a characteristic, we studied a novel serpin-like inhibitor (sc-srp-6) that was detected in a transcriptome analysis. Recombinant Sc-SRP-6 produced in Escherichia coli had a native fold of serpins belonging to the α-1-peptidase family and exhibited inhibitory activity against trypsin and α-chymotrypsin with Ki of 0.42×10−7 M and 1.22×10−7 M, respectively. Functional analysis revealed that Sc-SRP-6 inhibits insect digestive enzymes, thus preventing the hydrolysis of ingested particles. Moreover, Sc-SRP-6 impaired the formation of hard clots at the injury site, a major insect defence mechanism against invasive pathogens. Sc-SRP-6 does not prevent the formation of clot fibres and the activation of prophenoloxidases but impairs the incorporation of the melanin into the clot. Binding assays showed a complex formation between Sc-SRP-6 and three proteins in the hemolymph of lepidopteran required for clotting, apolipophorin, hexamerin and trypsin-like, although the catalytic inhibition occurred exclusively in trypsin-like. This data allowed the conclusion that Sc-SRP-6 promotes nematode virulence by inhibiting insect gut juices and by impairing immune clot reaction.

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Transcripts analysis of the entomopathogenic nematode Steinernema carpocapsae induced in vitro with insect haemolymph

Cited by 39 publications

References 70 publications

Identification of candidate infection genes from the model entomopathogenic nematode Heterorhabditis bacteriophora

Identification of candidate infection genes from the model entomopathogenic nematode Heterorhabditis bacteriophora

The genome, transcriptome, and proteome of the nematode Steinernema carpocapsae: evolutionary signatures of a pathogenic lifestyle

A Serpin Released by an Entomopathogen Impairs Clot Formation in Insect Defense System

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