Bioactive molecules produced by Heterorhabditis bacteriophora after in vitro stimulation by insect tissue

ELIÁŠ, Sara, Jana ILGOVÁ, Jana HURYCHOVÁ, Paola ALBERT POYÁN, Pavel DOBEŠ, Martin KAŠNÝ a Pavel HYRŠL. Bioactive molecules produced by Heterorhabditis bacteriophora after in vitro stimulation by insect tissue. In 18th meeting IOBC/WPRS Working Group Microbial and Nematode Control of Invertebrate Pests. 2022.

Základní údaje

• Originální název: Bioactive molecules produced by Heterorhabditis bacteriophora after in vitro stimulation by insect tissue
• Název česky: Bioactive molecules produced by Heterorhabditis bacteriophora after in vitro stimulation by insect tissue
• Autoři: ELIÁŠ, Sara (276 Německo, domácí), Jana ILGOVÁ (703 Slovensko, domácí), Jana HURYCHOVÁ (203 Česká republika, domácí), Paola ALBERT POYÁN (724 Španělsko, domácí), Pavel DOBEŠ (203 Česká republika, domácí), Martin KAŠNÝ (203 Česká republika) a Pavel HYRŠL (203 Česká republika, garant, domácí).
• Vydání: 18th meeting IOBC/WPRS Working Group Microbial and Nematode Control of Invertebrate Pests, 2022.

Další údaje

• Originální jazyk: angličtina
• Typ výsledku: Konferenční abstrakt
• Obor: 10616 Entomology
• Stát vydavatele: Česká republika
• Utajení: není předmětem státního či obchodního tajemství
• WWW: URL
• Kód RIV: RIV/00216224:14310/22:00129440
• Organizační jednotka: Přírodovědecká fakulta
• Klíčová slova česky: Heterorhabditis bacteriophora; secreted/excreted products; entomopathogenic nematodes
• Klíčová slova anglicky: Heterorhabditis bacteriophora; secreted/excreted products; entomopathogenic nematodes
• Příznaky: Mezinárodní význam

Anotace

Entomopathogenic nematodes produce bioactive molecules referred to as excreted/secreted products (ESPs). The ESPs comprise a mixture of proteins, lipids, glycans, and nucleic acids that can interfere with the host immune system in order to increase the chance of entomopathogenic nematodes to reproduce. Infective juveniles (IJs), the only free-living stage of nematodes, release ESPs typically in the early phase of infection when contacted with host tissues. The spectrum of ESPs has not yet been fully described, however, its quantitative and qualitative changes during the process of nematode infection were observed. Of particular importance are the immune-modulating proteins, such as proteolytic enzymes and their inhibitors, which have not yet been largely investigated. There are several approaches how to in vitro stimulate IJs to produce ESPs. We investigated the possible effect of various induction materials and time-related parameters on the yield of ESPs. We induced IJs of Heterorhabditis bacteriophora using four differently prepared Galleria mellonella homogenates and tested two time points of IJs induction and subsequent ESPs production. Based on our findings, the time of the IJs induction does not significantly affect the protein concentration of collected ESPs. However, we observed that the induction material affects the ESPs protein quantity. The collected ESPs were further characterised by mass spectrometry to identify proteins present in ESPs and suggest their function. According to GO annotation of mass spectrometric data, lipid-binding proteins, peptidase inhibitors, peptidases, and proteins with chitinase activity are among the most abundant groups of ESP components. As one of the major fractions determined by mass spectrometry were peptidases, we further focussed on their proteolytic activity and observed the presence of serine proteases. Our preliminary mass spectrometry data allow us to continue with identification and functional characterization of candidate bioactive molecules.

Anotace česky

Entomopathogenic nematodes produce bioactive molecules referred to as excreted/secreted products (ESPs). The ESPs comprise a mixture of proteins, lipids, glycans, and nucleic acids that can interfere with the host immune system in order to increase the chance of entomopathogenic nematodes to reproduce. Infective juveniles (IJs), the only free-living stage of nematodes, release ESPs typically in the early phase of infection when contacted with host tissues. The spectrum of ESPs has not yet been fully described, however, its quantitative and qualitative changes during the process of nematode infection were observed. Of particular importance are the immune-modulating proteins, such as proteolytic enzymes and their inhibitors, which have not yet been largely investigated. There are several approaches how to in vitro stimulate IJs to produce ESPs. We investigated the possible effect of various induction materials and time-related parameters on the yield of ESPs. We induced IJs of Heterorhabditis bacteriophora using four differently prepared Galleria mellonella homogenates and tested two time points of IJs induction and subsequent ESPs production. Based on our findings, the time of the IJs induction does not significantly affect the protein concentration of collected ESPs. However, we observed that the induction material affects the ESPs protein quantity. The collected ESPs were further characterised by mass spectrometry to identify proteins present in ESPs and suggest their function. According to GO annotation of mass spectrometric data, lipid-binding proteins, peptidase inhibitors, peptidases, and proteins with chitinase activity are among the most abundant groups of ESP components. As one of the major fractions determined by mass spectrometry were peptidases, we further focussed on their proteolytic activity and observed the presence of serine proteases. Our preliminary mass spectrometry data allow us to continue with identification and functional characterization of candidate bioactive molecules.

Návaznosti

• QK1910286, projekt VaV: Název: Efektivní postupy a strategie pro zvládání včelích chorob a udržitelný chov včelstev

Investor: Ministerstvo zemědělství ČR, Efektivní postupy a strategie pro zvládání včelích chorob a udržitelný chov včelstev