Characterization of thioredoxin-like PROTEIN-5 (TRXLP-5) and its differential response to grafting challenge in the black coloured selected line and control stocks of Pinctada fucata martensii.

ABSTRACT

Thioredoxin-like protein 5 (Trxlp-5) is a thioredoxin isoform associated with cellular redox homeostasis through the activity of thiol-disulfide reductase. In our study, Trxlp-5 was identified and characterized in Pinctada fucata martensii. The expression of PmTrxlp-5 was detected in response to polyinosinic polycytidylic acid (poly IC) and lipopolysaccharides (LPS) stimulation. The differences in PmTrxlp-5 expression were evaluated between the black coloured selected line and the control stock after grafting operation. The open reading frame (ORF) consisted of 1167bp encoding a 388 amino acid, 5'-UTR of 41bp and a 3'-UTR of 846bp. PmTrxlp-5 exhibited a conserved WCXXC functional motif similar to thioredoxins from other species. Tissue analysis showcased the highest relative mRNA expressions of PmTrxlp-5 in the haemocytes. Interestingly, after the grafting operation, mRNA expression of PmTrxlp-5 in the haemocytes was differentially expressed post grafting with a peak 6 h after grafting suggesting the high involvement of the gene in immune response in the early stage after grafting. The black coloured selected line group (BS) had significantly higher expression than the control group (CG) at 24 h, 6 d and 30 d after grafting operation. PmTrxlp-5 also showed a wave-like pattern in mRNA expression after bacterial endotoxin LPS and viral mimic poly IC. These results suggested that PmTrxlp-5 plays a vital function in cellular redox homeostasis and immune response against grafting operation and pathogenic infections and can be used as a gene marker for selective breeding programs.