Molecular insights into septin 2 protein in rohu (Labeo rohita): revealing expression dynamics, antimicrobial activity and functional characteristics.

Septins are evolutionarily conserved GTP-binding proteins mediating innate immunity, autophagy and inflammation in higher animals; however, they are yet to be fully characterized in fish. The study encompasses cloning of complete septin 2 cDNA from the rohu carp (Labeo rohita) that consisted of an open reading frame of 1050 bp and phylogenetic amino acid similarity of 99.43 % to cyprinid Onychostoma macrolepis. Septin 2 was ubiquitously expressed in different tissues of healthy rohu, and during early developmental stages. Septin 2 transcript levels were increased in response to three infection models i.e. Aeromonas hydrophila, poly I:C, and Argulus siamensis, indicating its role in immunity. A synthetic antimicrobial peptide derived from the septin 2 gene revealed in vitro bactericidal activity. A produced recombinant protein of septin 2 (~40 kDa) when injected into rohu modulated the expression of various immune-related genes. Further, in vivo studies of this protein demonstrated protection against A. hydrophila (71 % relative percent survival) and delayed mortality against ectoparasite A. siamensis. A developed sandwich ELISA revealed enhanced septin 2 level post A. hydrophila infection. The present study provides a new understanding of the septin 2 gene's multifunctional role in rohu and its importance in fish antimicrobial defence.