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

a 2024

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

ILGOVÁ, Jana, Sara ŠREIBR, Pavel DOBEŠ, Jiří VOREL, Jacek MARCINIAK et. al.

Základní údaje

Originální název

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

Autoři

ILGOVÁ, Jana (703 Slovensko, garant, domácí), Sara ŠREIBR (276 Německo, domácí), Pavel DOBEŠ (203 Česká republika, domácí), Jiří VOREL (203 Česká republika), Jacek MARCINIAK (203 Česká republika, domácí), Jana HURYCHOVÁ (203 Česká republika, domácí), Martin KAŠNÝ (203 Česká republika, domácí) a Pavel HYRŠL (203 Česká republika, domácí)

Vydání

35th Symposium of the European Society of Nematologists, 2024

Další údaje

Jazyk

angličtina

Typ výsledku

Konferenční abstrakt

Obor

10608 Biochemistry and molecular biology

Stát vydavatele

Španělsko

Utajení

není předmětem státního či obchodního tajemství

Organizační jednotka

Přírodovědecká fakulta

Klíčová slova anglicky

entomopathogenic nematodes; transcriptomics; infective juveniles; recovery

Změněno: 2. 5. 2024 23:26, doc. RNDr. Pavel Hyršl, Ph.D.

Anotace

V originále

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.

Návaznosti

GA23-06457S, projekt VaV

Název: Identifikace a funkční charakteristika bioaktivních molekul produkovaných entomopatogenními hlísticemi

Investor: Grantová agentura ČR, Identifikace a funkční charakteristika bioaktivních molekul produkovaných entomopatogenními hlísticemi

Citovat

ILGOVÁ, Jana, Sara ŠREIBR, Pavel DOBEŠ, Jiří VOREL, Jacek MARCINIAK, Jana HURYCHOVÁ, Martin KAŠNÝ a Pavel HYRŠL. In vivo vs in vitro activation of Heterorhabditis bacteriophora from the transcriptomics perspective. In 35th Symposium of the European Society of Nematologists. 2024.

@proceedings{2398599,
   author = {Ilgová, Jana and Šreibr, Sara and Dobeš, Pavel and Vorel, Jiří and Marciniak, Jacek and Hurychová, Jana and Kašný, Martin and Hyršl, Pavel},
   booktitle = {35th Symposium of the European Society of Nematologists},
   keywords = {entomopathogenic nematodes; transcriptomics; infective juveniles; recovery},
   language = {eng},
   title = {In vivo vs in vitro activation of Heterorhabditis bacteriophora from the transcriptomics perspective},
   year = {2024}
}

TY  - CONF
ID  - 2398599
AU  - Ilgová, Jana - Šreibr, Sara - Dobeš, Pavel - Vorel, Jiří - Marciniak, Jacek - Hurychová, Jana - Kašný, Martin - Hyršl, Pavel
PY  - 2024
TI  - In vivo vs in vitro activation of Heterorhabditis bacteriophora from the transcriptomics perspective
KW  - entomopathogenic nematodes
KW  - transcriptomics
KW  - infective juveniles
KW  - recovery
N2  - 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.
ER  -

ILGOVÁ, Jana, Sara ŠREIBR, Pavel DOBEŠ, Jiří VOREL, Jacek MARCINIAK, Jana HURYCHOVÁ, Martin KAŠNÝ a Pavel HYRŠL. In vivo vs in vitro activation of Heterorhabditis bacteriophora from the transcriptomics perspective. In \textit{35th Symposium of the European Society of Nematologists}. 2024.

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