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Ogier, J.-C.

Publications and source records attributed to Ogier, J.-C..

4 recordsLinked to original sources

Priority effects drive fungal and nematode emergence from insect larvae

Priority effects, in which species arrival history influences community assembly, are increasingly recognized to affect host-parasite systems. However, priority effects across disparate groups of parasitic organisms are poorly understood despite the wide range of taxonomic groups involved. In California oak woodland, we investigated how priority effects between two insect-parasitic fungi (Metarhizium and Beauveria) influenced emergence of nematodes from insect larvae. Field and laboratory results indicated that both fungi were common, but priority effects prevented them from co-emerging from the same larva. Metarhizium- and Beauveria-infected insects did not differ in the species composition of emerging nematodes, but larvae without fungal emergence had distinct nematode communities, with Oscheius almost always emerging without fungi. Experiments indicated that none of the commonly found nematodes (Acrobeloides, Mesorhabditis, Oscheius, and Rhabditis) were entomopathogenic, but that Oscheius could exclude Beauveria if arrived early. This time-dependent exclusion was likely caused by an Ochrobactrum bacterium that Oscheius nematodes carried. Together, these findings suggest that fungi enter insects as primary arrivers, while nematodes come as secondary arrivers to exploit fungus-killed insects, with priority effects influencing both groups. We suggest that this system is a promising natural microcosm for understanding priority effects across disparate groups in host-parasite systems.

ecology↗

Partner-specific induction of Spodoptera frugiperda immune genes in response to the entomopathogenic nematobacterial complex Steinernema carpocapsae-Xenorhabdus nematophila

The Steinernema carpocapsae-Xenorhabdus nematophila association is a nematobacterial complex (NBC) used in biological control of insect crop pests. The ability of this dual pathogen to infest and kill an insect strongly depends on the dialogue between the hosts immune system and each partner of the complex. Even though this dialogue has been extensively studied from the two partners points of view in several insect models, still little is known about the structure and the molecular aspects of the insects immune response to the dual infection. Here, we used the lepidopteran pest Spodoptera frugiperda as a model to analyze the respective impact of each NBC partner in the spatiotemporal immune responses that are induced after infestation. To this aim, we first analyzed the expression variations of the insects immune genes in the fat bodies and hemocytes of infested larvae by using previously obtained RNAseq data. We then selected representative immune genes for RT-qPCR investigations of the temporal variations of their expressions after infestation and of their induction levels after independent injections of each partner. We found that the fat body and the hemocytes both produce potent and stable immune responses to the infestation by the NBC, which correspond to combinations of bacterium- and nematode-induced ones. Consistent with the nature of each pathogen, we showed that X. nematophila mainly induces genes classically involved in antibacterial responses, whereas S. carpocapsae is responsible for the induction of lectins and of genes expected to be involved in melanization and cellular encapsulation. In addition, we found that two clusters of unknown genes dramatically induced by the NBC also present partner-specific induction profiles, which paves the way for their functional characterization. Finally, we discuss putative relationships between the variations of the expression of some immune genes and the NBCs immunosuppressive strategies.\n\nAuthor summaryEntomopathogenic nematodes (EPNs) are living in the soil and prey upon insect larvae. They enter the insect by the natural orifices, and reach the hemocoel through the intestinal epithelium. There, they release their symbiotic bacteria that will develop within the insect and eventually kill it. Nematodes can then feed and reproduce on the insect cadaver. By using transcriptomic approaches, we previously showed that Lepidoptera larvae (caterpillars of the fall armyworm Spodoptera frugiperda) produce a strong immune response in reaction to infestation by EPNs. However, we do not know if this immune reaction is triggered by the nematode itself -Steinernema carpacapsae - or its symbiotic bacteria - Xenorhabdus nematophila. To answer this question, we present in this work a careful annotation of immunity genes in S. frugiperda and surveyed their activation by quantitative PCR in reaction to an injection of the bacteria alone, the axenic nematode or the associated complex. We found that the immune genes are selectively activated by either the bacteria or the nematode and we discuss the implication of which pathway are involved in the defense against various pathogens. We also show that a cluster of newly discovered genes, present only in Lepidoptera, is activated by the nematode only and could represent nematicide genes.

immunology↗

rpoB, a promising marker for analyzing the diversity of bacterial communities by amplicon sequencing

BackgroundMicrobiome composition is frequently studied by the amplification and high-throughput sequencing of specific molecular markers (metabarcoding). Various hypervariable regions of the 16S rRNA gene are classically used to estimate bacterial diversity, but other universal bacterial markers with a finer taxonomic resolution could be employed. We compared specificity and sensitivity between a portion of the rpoB gene and the V3V4 hypervariable region of the 16S rRNA gene.\n\nResultsWe first designed universal primers for rpoB suitable for use with Illumina sequencing-based technology and constructed a reference rpoB database of 45,000 sequences. The rpoB and V3V4 markers were amplified and sequenced from (i) a mock community of 19 bacterial strains from both Gram-negative and Gram-positive lineages; (ii) bacterial assemblages associated with entomopathogenic nematodes. In metabarcoding analyses of mock communities with two analytical pipelines (FROGS and DADA2), the estimated diversity captured with the rpoB marker resembled the expected composition of these mock communities more closely than that captured with V3V4. The rpoB marker had a higher level of taxonomic affiliation, a higher sensitivity (detection of all the species present in the mock communities), and a higher specificity (low rates of spurious OTU detection) than V3V4. We applied both primers to infective juveniles of the nematode Steinernema glaseri. Both markers showed the bacterial community associated with this nematode to be of low diversity (< 50 OTUs), but only rpoB reliably detected the symbiotic bacterium Xenorhabdus poinarii.\n\nConclusionsOur results confirm that different microbiota composition data may be obtained with different markers. We found that rpoB was a highly appropriate marker for assessing the taxonomic structure of mock communities and the nematode microbiota. Further studies on other ecosystems should be considered to evaluate the universal usefulness of the rpoB marker. Our data highlight two crucial elements that should be taken into account to ensure more reliable and accurate descriptions of microbial diversity in high-throughput amplicon sequencing analyses: i) the need to include mock communities as controls; ii) the advantages of using a multigenic approach including at least one housekeeping gene (rpoB is a good candidate) and one variable region of the 16S rRNA gene.

microbiology↗

Changes in rearing conditions rapidly modify gut microbiota structure in Tenebrio molitor larvae

The gut microbiota of multicellular organisms has been shown to play a key role in their host biology. In mammals, it has an invariant component, responsible for establishing a mutualistic relationship with the host. It also contains a dynamic fraction which facilitates adaptation in response to changes in the environment. These features have been well described in mammals, but little is known about microbiota stability or plasticity in insects. We assessed changes in microbiota composition and structure in a reared insect after a change in rearing conditions. We reared Tenebrio molitor (Coleoptera, Tenebrioninae) larvae for five days in soil samples from two river banks and analyzed their gut microbial communities by a metabarcoding technique, using the V3-V4 region of the 16S rRNA gene and the housekeeping gene gyrB. We found that soil-reared insects had a significantly more diverse microbiota than the control insects and that insects reared in soil from different sites had significantly different microbiota. We confirmed this trend by absolute quantification of the two mains fluctuating taxonomic groups: the Enterobacteriaceae family and the Pseudomonas genus, dominant in the soil-reared insects and in the control insects, respectively. Our results suggest the existence of a resident microbiota in T. molitor gut, but indicate that rearing changes can induce rapid and profound changes in the relative abundance of some of the members of this resident microbiota.

microbiology↗