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Dorsal, a member of the nuclear factor-kappa B (NF-κB) family of transcription factors, is a critical downstream component of the Toll pathway that regulates the expression of antimicrobial peptides (AMPs) against pathogen invasion. In this study, the full-length ORF of  Dorsal  was identified from the RNA-seq database of the mealworm beetle  Tenebrio molitor  ( TmDorX2 ). The ORF of  TmDorX2  was 1,482 bp in length, encoding a polypeptide of 493 amino acid residues.  Tm DorX2 contains a conserved Rel homology domain (RHD) and an immunoglobulin-like, plexins, and transcription factors (IPT) domain.  TmDorX2  mRNA was detected in all developmental stages, with the highest levels observed in 3-day-old adults.  TmDorX2  transcripts were highly expressed in the adult Malpighian tubules (MT) and the larval fat body and MT tissues. After challenging the larvae with  Staphylococcus aureus  and  Escherichia coli , the  TmDorX2  mRNA levels were upregulated 6 and 9 h post infection in the whole body, fat body, and hemocytes. Upon  Candida albicans  challenge, the  TmDorX2  mRNA expression were found highest at 9 h post-infection in the fat body. In addition,  TmDorX2 -knockdown larvae exposed to  E. coli ,  S. aureus , or  C. albicans  challenge showed a significantly increased mortality rate. Furthermore, the expression of 11 AMP genes was downregulated in the gut and fat body of ds TmDorX2 -injected larvae upon  E. coli  challenge. After  C .  albicans  and  S. aureus  challenge of ds TmDorX2 -injected larvae, the expression of 11 and 10 AMPs was downregulated in the gut and fat body, respectively. Intriguingly, the expression of antifungal transcripts  TmTenecin-3  and  TmThaumatin-like protein-1  and  -2  was greatly decreased in  TmDorX2 -silenced larvae in response to  C. albicans  challenge, suggesting that  TmDorX2  regulates antifungal AMPs in the gut in response to  C. albicans  infection. The AMP expression profiles in the fat body, hemocytes, gut, and MTs suggest that  TmDorX2  might have an important role in promoting the survival of  T. molitor  larvae against all mentioned pathogens.

TmDorX2 positively regulates antimicrobial peptides in Tenebrio molitor gut, fat body, and hemocytes in response to bacterial and fungal infection