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The dimeric cytokine ligand Spätzle (Spz) is responsible for Toll pathway activation and antimicrobial peptide (AMP) production upon pathogen challenge in  Tenebrio molitor . Here, we indicated that  Tm Spz5 has a functional role in response to bacterial infections. We showed that the highest expression of  TmSpz5  is induced by  Candida albicans . However,  TmSpz5  knockdown reduced larval survival against  Escherichia coli  and  Staphylococcus aureus . To evaluate the molecular mechanism underlying the observed survival differences, the role of  TmSpz5  in AMP production was examined by RNA interference and microbial injection.  T. molitor  AMPs that are active against Gram-negative and -positive bacteria, including  Tm tenecins,  Tm attacins,  Tm coleoptericins,  Tm taumatin-like-proteins, and  Tm cecropin-2, were significantly downregulated by  TmSpz-5  RNAi in the Malpighian tubules (MTs) following a challenge with  E. coli  and  S. aureus . However, upon infection with  C. albicans  the mRNA levels of most AMPs in the ds TmSpz5 -injected group were similar to those in the control groups. Likewise, the expression of the transcription factors NF-κB,  TmDorX2 , and  TmRelish  were noticeably suppressed in the MTs of  TmSpz5 -silenced larvae. Moreover,  E. coli -infected  TmSpz5  knockdown larvae showed decreased antimicrobial activity in the MTs and hindgut compared with the control group. These results demonstrate that  Tm Spz5 has a defined role in  T. molitor  innate immunity by regulating AMP expression in MTs in response to  E. coli .

Critical Roles of Spätzle5 in Antimicrobial Peptide Production Against Escherichia coli in Tenebrio molitor Malpighian Tubules