In vivo vs in vitro activation of Heterorhabditis bacteriophora from the transcriptomics perspective

Result code in IS VaVaI

<a href="https://www.isvavai.cz/riv?ss=detail&h=RIV%2F00216224%3A14310%2F24%3A00139418" target="_blank" >RIV/00216224:14310/24:00139418 - isvavai.cz</a>

Result on the web

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DOI - Digital Object Identifier

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Result language

angličtina

Original language name

In vivo vs in vitro activation of Heterorhabditis bacteriophora from the transcriptomics perspective

Original language description

Entomopathogenic nematodes (EPNs), such as Heterorhabditis bacteriophora, have gained recognition as effective biocontrol agents against insect pests, offering an eco-friendly alternative to chemical insecticides. This preliminary study delves into the molecular dynamics of H. bacteriophora infection, shedding light on the responses to host molecules. Invasion strategy of H. bacteriophora involves the active penetration of the host by 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 non-active stage to the infective one. Within a few hours after the host colonisation, IJs release symbiotic bacteria of the genus Photorhabdus, which cause host septicaemia and death. Under experimental conditions, the infection process and activation of IJs are often simulated by challenging IJs with various host-derived materials (such as tissue homogenate or haemolymph). This study aims to bridge the gap in understanding how in vitro activation of H. bacteriophora corresponds to the in vivo infection. 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, 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 employ a single-cell RNA NGS library preparation strategy, followed by sequencing at the Illumina NovaSeq 6000 platform. Differential expression analysis will identify key transcripts by mapping reads to a reference transcriptome. Our objective will be to evaluate the extent to which H. bacteriophora gene expression during in vitro activation mirrors the in vivo infection dynamics. This pilot data will contribute to the understanding of H. bacteriophora in vivo molecular strategies and address the relevance of in vitro activation models in studying the infection process.

Czech name

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Czech description

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Type

O - Miscellaneous

CEP classification

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OECD FORD branch

10608 - Biochemistry and molecular biology

Project

<a href="/en/project/GA23-06457S" target="_blank" >GA23-06457S: Identification and functional characterization of bioactive molecules produced by entomopathogenic nematodes</a><br>

Continuities

P - Projekt vyzkumu a vyvoje financovany z verejnych zdroju (s odkazem do CEP)

Publication year

2024

Confidentiality

S - Úplné a pravdivé údaje o projektu nepodléhají ochraně podle zvláštních právních předpisů