Expression profiles of two thaumatin‐like protein (<i>TmTLP</i>) genes in responses to various micro‐organisms from<i>Tenebrio molitor</i>

Kim, Dong Hyun; Noh, Mi Young; Park, Ki Beom; Jo, Yong Hun

doi:10.1111/1748-5967.12197

Cited by 8 publications

(7 citation statements)

References 32 publications

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“…In order to determine whether TmRelish knockdown affected AMP regulation, the expression profile of fourteen AMPs including members of the Tenecin family (TmTenecin-1 (TmTene1), TmTene2, TmTene3, and TmTene4) [29][30][31][32] ; Attacin family (TmAttacin-1a (TmAtta1a), TmAtta1b, and TmAtta2) 33 ; TmDefensin-1 (TmDef1); TmDef2; TmColeptericin-1 (TmCole1); TmCole2; TmCecropin-2 (TmCec2); and thaumatin-like protein (TmTLP1 and TmTLP2) 34,35 was examined in TmRelish-silenced T. molitor larvae following microbial challenge. The immune tissues of the insect (fat body, hemocytes, gut, and MTs) were dissected 24 h post microbial challenge.…”

Section: Effect Of Tmrelish Knockdown On Amp Expression Post-microbiamentioning

confidence: 99%

TmRelish is required for regulating the antimicrobial responses to Escherichia coli and Staphylococcus aureus in Tenebrio molitor

Keshavarz

Patnaik

et al. 2020

Sci Rep

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Relish, a transcription factor, is a critical downstream component of the immune deficiency (Imd) pathway and regulates host defense against bacterial infection by mediating antimicrobial peptide (AMP) synthesis. Understanding the immunological function of the mealworm beetle, Tenebrio molitor Relish (TmRelish) will be instructive in understanding insect immunity. In the present study, full-length oRf of TmRelish was retrieved from T. molitor-expressed sequence tags and RNA-seq database. The predicted TmRelish amino acid sequence contained an N-terminal Rel-homology domain; an Ig-like, plexin, and transcription factor domain; ankyrin repeat motifs; a nuclear localization signal; and a C-terminal death domain and shared the highly conserved structure of the Relish proteins of other insect species. TmRelish mRNA was detected in all developmental stages of the insect; however, the highest levels were detected in the larval gut tissue and adult hemocytes. TmRelish mRNA level was upregulated in the fat body, hemocyte, and gut tissue 9 h after infection of T. molitor larvae by the gram-negative bacteria, Escherichia coli. Furthermore, TmRelish knockdown led to significantly higher mortality of the E. coli-infected larvae, and significantly lower mortality of larvae infected with Staphylococcus aureus or Candida albicans. To elucidate the possible cause of mortality, we measured AMP transcription in the fat body, hemocytes, gut, and Malpighian tubules (MTs) of T. molitor larvae. TmRelish knockdown suppressed the expression of nine AMP genes in the larval fat body and gut tissue during E. coli infection, suggesting that TmRelish positively regulates AMP expression in both immunerelated tissues, in response to E. coli challenge. Furthermore, negative regulation of some AMPs by TmRelish in the MTs, gut and hemocytes in response to C. albicans infection suggests a crosstalk between the Toll and Imd pathways.

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Section: Effect Of Tmrelish Knockdown On Amp Expression Post-microbiamentioning

confidence: 99%

TmRelish is required for regulating the antimicrobial responses to Escherichia coli and Staphylococcus aureus in Tenebrio molitor

Keshavarz

Patnaik

et al. 2020

Sci Rep

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“…Expression patterns of 14 AMP genes including TmTenecin1, 2, 3 , and 4 ( TmTene1 , − 2 , − 3 , and − 4 ) ( Kim et al, 1998 ; Chae et al, 2012 ; Yang et al, 2017 ), TmDefensin and TmDefensin-like ( TmDef and TmDef-like ) ( Jang et al, 2020b ), TmColeoptericin-A and -B ( TmColeA and − B ) ( Zhu et al, 2014 ; Jang et al, 2020a ), TmAttacin-1a, −1b and -2 ( TmAtt1a , − 1b and − 2 ) ( Jo et al, 2018 ), TmCecropin-2 ( TmCec2 ) ( Ali Mohammadie Kojour et al, 2021 ), and TmThaumatin - like protein-1 and − 2 ( TmTLP1 and − 2 ) ( Noh and Jo, 2016 ; Kim et al, 2017 ), were examined by qRT-PCR with the AMP gene-specific primers ( Table 1 ). A relative quantitative PCR was performed as detailed above in an AMP-specific primer.…”

Section: Methodsmentioning

confidence: 99%

Tenebrio molitor Spätzle 1b Is Required to Confer Antibacterial Defense Against Gram-Negative Bacteria by Regulation of Antimicrobial Peptides

Bae

Patnaik

et al. 2021

Front. Physiol.

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Innate immunity is the ultimate line of defense against invading pathogens in insects. Unlike in the mammalian model, in the insect model, invading pathogens are recognized by extracellular receptors, which activate the Toll signaling pathway through an extracellular serine protease cascade. In the Toll-NF-κB pathway, the extracellular spätzle protein acts as a downstream ligand for Toll receptors in insects. In this study, we identified a novel Spätzle isoform (TmSpz1b) from RNA sequencing database of Tenebrio molitor. TmSpz1b was bioinformatically analyzed, and functionally characterized for the antimicrobial function by RNA interference (RNAi). The 702 bp open reading frame of TmSpz1b encoded a putative protein of 233 amino acid residues. A conserved cystine-knot domain with seven cysteine residues in TmSpz1b was involved in three disulfide bridges and the formation of a spätzle dimer. TmSpz1b was mostly expressed in the hemocytes of T. molitor late instar larvae. The mRNA expression of TmSpz1b was highly induced in the hemocytes after Escherichia coli, Staphylococcus aureus, and Candida albicans stimulation of T. molitor larvae. TmSpz1b silenced larvae were significantly more susceptible to E. coli infection. In addition, RNAi-based functional assay characterized TmSpz1b to be involved in the positive regulation of antimicrobial peptide genes in hemocytes and fat bodies. Further, the TmDorX2 transcripts were downregulated in TmSpz1b silenced individuals upon E. coli challenge suggesting the relationship to Toll signaling pathway. These results indicate that TmSpz1b is involved in the T. molitor innate immunity, causes the sequestration of Gram-negative bacteria by the regulatory action of antimicrobial peptides, and enhances the survival of T. molitor larvae.

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“…To assess the participation of TmRelish in humoral immunity and to determine whether high susceptibility of TmRelish-silenced larvae is related to AMP gene regulation, the mRNA expression levels of 14 identified AMPs, comprising TmTenecin family (TmTenecin-1, TmTenecin-2, TmTenecin-3, TmTenecin-4) [25][26][27][28], TmAttacin family (TmAttacin-1a, TmAttacin-1b, TmAttacin-2) [29], TmDefensin-1, TmDefensin-2, TmColeptericin-1, TmColeptericin-2, TmCecropin-2, TmThaumatin like protein family (TmThaumatin like protein-1, and TmThaumatin like protein-2) [30,31] were measured by qRT-PCR in fat body, hemocytes, and gut of dsTmRelish-injected larvae compared with dsEGFP-injected larvae after microbial infection. At 24 h post-L. monocytogenes infection, immune defense tissues were dissected in cold PBS solution, total RNAs were extracted and cDNA was synthesized as described above.…”

Section: Effect Of Tmrelish Gene Silencing On Amp Expression After Bamentioning

confidence: 99%

Two Roles for the Tenebrio molitor Relish in the Regulation of Antimicrobial Peptides and Autophagy-Related Genes in Response to Listeria monocytogenes

Keshavarz

Edosa

2020

Insects

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Relish is a key NF-κB transcription factor of the immune-deficiency (Imd) pathway that combats infection by regulating antimicrobial peptides (AMPs). Understanding of the fundamental role of Tenebrio molitor Relish (TmRelish) in controlling of Listeria monocytogenes virulence through the regulation of both AMPs and autophagy-related (ATG) genes is unclear. Here, we show that TmRelish transcripts were highly abundant in the larval fat body and hemocytes compared to the gut upon L. monocytogenes infection. Furthermore, significant mortality was observed in TmRelish-silenced larvae after intracellular insult. To investigate the cause of this lethality, we measured the induction of AMPs and ATG genes in the TmRelish dsRNA-treated T. molitor larvae. The expression of TmTenecin-1, TmTenecin-4, TmColeptericin-1, TmAttacin-2, and TmCecropin-2 were suppressed in the fat body and hemocytes of dsTmRelish-injected larvae during L. monocytogenes infection. In addition, TmRelish knockdown led to a noticeable downregulation of TmATG1 (a serine-threonine protein kinase) in the fat body and hemocytes of young larvae 6 h post-infection (pi). The notable increase of autophagy genes in the early stage of infection (6 h pi), suggesting autophagy response is crucial for Listeria clearance. Taken together, these results suggest that TmRelish plays pivotal roles in not only regulation of AMP genes but also induction of autophagy genes in response to L. monocytogenes challenge in fat body and hemocytes of T. molitor larvae. Furthermore, negative regulation of several AMPs by TmRelish in the fat body, hemocytes, and gut leaves open the possibility of a crosstalk between Toll and Imd pathway.

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Expression profiles of two thaumatin‐like protein (TmTLP) genes in responses to various micro‐organisms fromTenebrio molitor

Cited by 8 publications

References 32 publications

TmRelish is required for regulating the antimicrobial responses to Escherichia coli and Staphylococcus aureus in Tenebrio molitor

TmRelish is required for regulating the antimicrobial responses to Escherichia coli and Staphylococcus aureus in Tenebrio molitor

Tenebrio molitor Spätzle 1b Is Required to Confer Antibacterial Defense Against Gram-Negative Bacteria by Regulation of Antimicrobial Peptides

Two Roles for the Tenebrio molitor Relish in the Regulation of Antimicrobial Peptides and Autophagy-Related Genes in Response to Listeria monocytogenes

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