Informace o publikaci
Unveiling molecular strategies of Heterorhabditis bacteriophora: a comparative analysis of in vitro activation and in vivo infection dynamics
| Autoři | |
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| Rok publikování | 2024 |
| Druh | Konferenční abstrakty |
| Fakulta / Pracoviště MU | |
| Citace | |
| Popis | The interplay between entomopathogenic nematodes (EPNs) and their hosts offers promising avenues for environmentally sustainable pest management strategies. Our research investigates the intricate molecular mechanisms underlying the infection dynamics of Heterorhabditis bacteriophora strain Az 148. Invasion tactics of this EPN species involve the active penetration of the host by the third-stage larvae, also called infective juveniles (IJs). Upon contact with the host or its molecules, IJs start the process called activation or recovery, which involves the transition from the dauer stage to the infective one. Shortly after the host colonization, IJs release symbiotic bacteria of the genus Photorhabdus, which induce fatal septicemia in the host. In laboratory settings, the recovery of IJs is often simulated by challenging them with various host-derived materials (such as tissue homogenate or hemolymph). Nevertheless, the extent to which in vitro activation corresponds to in vivo infection has not been elucidated. We are conducting a comparative RNA-seq analysis at various time points throughout the in vitro activation and in vivo infection of the greater wax moth larvae, Galleria mellonella. Our analysis focuses on five critical time points (3, 6, 9, 12, and 15 hours post-infection) to capture the dynamic changes in H. bacteriophora gene expression during IJs infection in vivo. In vitro activation involves exposure of IJs to G. mellonella-derived homogenates for three periods (3, 6 and 9 hours), simulating the interaction with host tissues. Recognizing the low recovery of IJs after in vivo infection, we employed a single-cell RNA NGS library preparation strategy, followed by sequencing at the DNBSEQ-G400 (MGI) platform using FCL PE100 kit. Differential expression analysis will identify key transcripts by mapping reads to a de novo assembled transcriptome. Our aim is to assess the correlation between H. bacteriophora gene expression patterns during in vitro activation and the dynamics observed during in vivo infection. This data will contribute to the understanding of H. bacteriophora molecular strategies employed in vivo and address the relevance of in vitro activation models in studying the infection process. |
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