Structure and Function Analyses of the Thioredoxin 2 and Thioredoxin Reductase Gene in Pacific White Shrimp (Litopenaeus vannamei)

The thioredoxin (Trx) system is one of the most significant systems in living organisms as it regulates cellular redox reactions and plays a pivotal protective role within the cell by promoting redox homeostasis. Trx and thioredoxin reductase (TrxR) are the core oxidoreductases of the Trx system. In this study, the novel full-length cDNAs of LvTrx2 and LvTrxR were cloned from Litopenaeus vannamei. The ORFs of LvTrx2 and LvTrxR were 453 bp and 1785 bp, encoding polypeptides consisting of 150 and 596 amino acids. Sequence alignment analysis revealed that the amino acid sequence of LvTrx2 shared a high degree of identity (93%) with that of Penaeus chinensis, while in LvTrxR, it exhibited a similarity level of 95% with previously submitted Penaeus chinensis and Penaeus monodon sequences. Regarding tissue-specific expression patterns, LvTrx2 showed its highest expression levels in hepatopancreas and gill. For LvTrxR, the highest expression was observed in gill followed by hepatopancreas and intestine. During exposure to ammonia-N, there was a significant upregulation in the relative mRNA levels of LvTrx2 and LvTrxR in hepatopancreas and gill, with the peak values occurring at 24 h or 48 h of exposure. After LPS injection, the LvTrx2 and LvTrxR transcripts in hepatopancreas and gill had different upregulated levels. These findings suggest that LvTrx2 and LvTrxR play pivotal roles in enhancing stress resistance and bolstering antibacterial defense mechanisms in L. vannamei. To explore the roles, LvTrx2 expression was knocked down in vivo to verify the defense mechanism against 4-NP stress. LvTrx2 silencing in 4-NP-challenged shrimp could significantly induce the gene expression of antioxidant-related genes (except for LvTrxR) and aggravate the oxidative damage of lipids. This study suggests that the Trx system is involved in regulating the antioxidant processes, and LvTrx2 and LvTrxR play a vital role in defense responses against environmental stress.