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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  Tm Relish 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 immune-related 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.

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