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Nanopore-Sequencing Characterization of the Gut Microbiota of Melolontha melolontha Larvae: Contribution to Protection against Entomopathogenic Nematodes?

ABSTRACT: This study focused on the potential relationships between midgut microbiota of the common cockchafer Melolontha melolontha larvae and their resistance to entomopathogenic nematodes (EPN) infection. We investigated the bacterial community associated with control and unsusceptible EPN-exposed insects through nanopore sequencing of the 16S rRNA gene. Firmicutes, Proteobacteria, Actinobacteria, and Bacteroidetes were the most abundant bacterial phyla within the complex and variable midgut microbiota of the wild M. melolontha larvae. The core microbiota was found to include 82 genera, which accounted for 3.4% of the total number of identified genera. The EPN-resistant larvae differed significantly from the control ones in the abundance of many genera belonging to the Actinomycetales, Rhizobiales, and Clostridiales orders. Additionally, the analysis of the microbiome networks revealed different sets of keystone midgut bacterial genera between these two groups of insects, indicating differences in the mutual interactions between bacteria. Finally, we detected Xenorhabdus and Photorhabdus as gut residents and various bacterial species exhibiting antagonistic activity against these entomopathogens. This study paves the way to further research aimed at unravelling the role of the host gut microbiota on the output of EPN infection, which may contribute to enhancement of the efficiency of nematodes used in eco-friendly pest management.

SUBMITTER: Sajnaga E

PROVIDER: S-EPMC8067285 | biostudies-literature |

REPOSITORIES: biostudies-literature

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Project description:IntroductionBelow ground orientation in insects relies mainly on olfaction and taste. The economic impact of plant root feeding scarab beetle larvae gave rise to numerous phylogenetic and ecological studies. Detailed knowledge of the sensory capacities of these larvae is nevertheless lacking. Here, we present an atlas of the sensory organs on larval head appendages of Melolontha melolontha. Our ultrastructural and electrophysiological investigations allow annotation of functions to various sensory structures.ResultsThree out of 17 ascertained sensillum types have olfactory, and 7 gustatory function. These sensillum types are unevenly distributed between antennae and palps. The most prominent chemosensory organs are antennal pore plates that in total are innervated by approximately one thousand olfactory sensory neurons grouped into functional units of three-to-four. In contrast, only two olfactory sensory neurons innervate one sensillum basiconicum on each of the palps. Gustatory sensilla chaetica dominate the apices of all head appendages, while only the palps bear thermo-/hygroreceptors. Electrophysiological responses to CO(2), an attractant for many root feeders, are exclusively observed in the antennae. Out of 54 relevant volatile compounds, various alcohols, acids, amines, esters, aldehydes, ketones and monoterpenes elicit responses in antennae and palps. All head appendages are characterized by distinct olfactory response profiles that are even enantiomer specific for some compounds.ConclusionsChemosensory capacities in M. melolontha larvae are as highly developed as in many adult insects. We interpret the functional sensory units underneath the antennal pore plates as cryptic sensilla placodea and suggest that these perceive a broad range of secondary plant metabolites together with CO(2). Responses to olfactory stimulation of the labial and maxillary palps indicate that typical contact chemo-sensilla have a dual gustatory and olfactory function.

Project description:Tomato (Solanum esculentum) is one of the vegetable crops grown by both smallholder and commercial farmers in the Kingdom of Eswatini. Tomato leaf miner, Tuta absoluta M. is a major insect pest of tomatoes resulting in reduced tomato yields throughout the country. The study investigated the virulence of two sub-tropical EPN species on T. absoluta larvae. Steinernema yirgalemense and S. jeffreyense at different concentrations (0, 20, 40, 60 IJs/insect) were screened for efficacy (i.e. mortality rate) against larvae of T. absoluta in laboratory bioassays. Results obtained showed that S. yirgalemense and S. jeffreyense were able to kill T. absoluta larvae without significant differences between the two EPN species in 24-well bioassay trays. Significantly higher (p?<?0.05) mortality was observed when 60 IJs/insect was used. The combination of S. yirgalemense at 60 IJs/insect (100%) and S. jeffreyense at 60 IJs/insect (98.3%) resulted in significantly higher (p?<?0.05) mortality, compared with the other four combinations of EPN and concentration tested. In the leaf bioassays, S. yirgalemense (58.8%) resulted in significantly higher (p?<?0.05) mean mortality compared to S. jeffreyense (46.4%). Steinernema yirgalemense at 60 IJs/insect resulted in significantly higher mean mortality compared to the other EPN and concentration combinations in the leaf bioassay. The results indicated that both EPNs tested were effective against T. absoluta larvae. Steinernema yirgalemense at 60 IJs/insect can effectively find T. absoluta larvae inside leaf mines, but large-scale field trials are recommended to demonstrate the potential use of the biocontrol agent within integrated pest management programs.Tomato (Solanum esculentum) is one of the vegetable crops grown by both smallholder and commercial farmers in the Kingdom of Eswatini. Tomato leaf miner, Tuta absoluta M. is a major insect pest of tomatoes resulting in reduced tomato yields throughout the country. The study investigated the virulence of two sub-tropical EPN species on T. absoluta larvae. Steinernema yirgalemense and S. jeffreyense at different concentrations (0, 20, 40, 60 IJs/insect) were screened for efficacy (i.e. mortality rate) against larvae of T. absoluta in laboratory bioassays. Results obtained showed that S. yirgalemense and S. jeffreyense were able to kill T. absoluta larvae without significant differences between the two EPN species in 24-well bioassay trays. Significantly higher (p?<?0.05) mortality was observed when 60 IJs/insect was used. The combination of S. yirgalemense at 60 IJs/insect (100%) and S. jeffreyense at 60 IJs/insect (98.3%) resulted in significantly higher (p?<?0.05) mortality, compared with the other four combinations of EPN and concentration tested. In the leaf bioassays, S. yirgalemense (58.8%) resulted in significantly higher (p?<?0.05) mean mortality compared to S. jeffreyense (46.4%). Steinernema yirgalemense at 60 IJs/insect resulted in significantly higher mean mortality compared to the other EPN and concentration combinations in the leaf bioassay. The results indicated that both EPNs tested were effective against T. absoluta larvae. Steinernema yirgalemense at 60 IJs/insect can effectively find T. absoluta larvae inside leaf mines, but large-scale field trials are recommended to demonstrate the potential use of the biocontrol agent within integrated pest management programs.

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Nanopore-Sequencing Characterization of the Gut Microbiota of Melolontha melolontha Larvae: Contribution to Protection against Entomopathogenic Nematodes?

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