Abstract
BackgroundTilapia (Oreochromis niloticus) cultures are frequently infected by Vibrio vulnificus, causing major economic losses to production units. Previously, tilapia expressing recombinant delta-5 desaturase and delta-6 desaturase (D56) were found to be resistant to V. vulnificus infection. In this report, we profile the D56-mediated molecular changes underlying this resistance in tilapia. A comparative transcriptome analysis was performed on V. vulnificus-infected wild-type and D56-transgenic tilapia using Illumina’s sequencing-by-synthesis approach. Gene enrichment analysis on differentially expressed unigenes was performed, and the expression patterns were validated by real-time PCR.ResultsComparative transcriptome analysis was performed on RNA-sequence profiles obtained from wild-type and D56-transgenic tilapia at 0, 6 and 24 h post-infection with V. vulnificaus. GO and KEGG gene enrichment analyses showed that D56 regulates several pathways and genes, including fatty acid (FA) metabolism associated, and inflammatory and immune response. Expression of selected FA metabolism-associated, inflammatory and immune responsive genes was validated by qPCR. The inflammatory and immune responsive genes that are modulated by FA-associated D56 likely contribute to the enhanced resistance against V. vulnificus infection in Tilapia.ConclusionsTranscriptome profiling and filtering for two-fold change variation showed that 3795 genes were upregulated and 1839 genes were downregulated in D56-transgenic tilapia. These genes were grouped into pathways, such as FA metabolism, FA elongation, FA biosynthesis, biosynthesis of unsaturated FA, FA degradation, inflammation, immune response, and chemokines. FA-associated genes and immune-related genes were modulated by D56 at 6 h and 24 h post infection with V. vulnificus. The expression patterns of FA-related genes, inflammatory genes, antimicrobial peptide genes and immune responsive genes at 0, 3, 6, 12, 24 and 48 h post-infection suggests these genes are involved in the enhanced resistance of D56 transgenic tilapia to V. vulnificus.
Highlights
Tilapia (Oreochromis niloticus) cultures are frequently infected by Vibrio vulnificus, causing major economic losses to production units
Expression of recombinant delta-6 desaturase and delta-5 desaturase alters the transcriptome in tilapia Wild-type and dual expression of SsFadsd5 and SsFadsd6 (D56)-transgenic tilapia were infected with V. vulnificus, and RNA was extracted from liver at 0, 6 and 24 h post-infection
We found that Toll-like receptor (TLR)-2, Toll- like receptor 5 (TLR-5) and Atypical chemokine receptor 4 (ACKR4) were induced in the D56-transgenic line
Summary
Tilapia (Oreochromis niloticus) cultures are frequently infected by Vibrio vulnificus, causing major economic losses to production units. Tilapia (Oreochrombis niloticus) is an important commercial aquaculture species throughout the world, and its production is severely affected by the pathogenic bacteria Vibrio vulnificus, which causes septicemia in fish and humans [1,2,3,4]. N-3 PUFAs show positive ionotropic effects and minimize tachyarrhythmia in animal models [7]. Many of these effects may be mediated by alterations in the proinflammatory cytokines, TNF-α, IL-1β, IL-6, prostaglandin (PG) E2, and PGF1α, which modulate the immune response in model organisms [8,9,10]. Transgenic expression of n-3 PUFA biosynthesis genes from Atlantic salmon, i.e., Fatty acyl desaturase synthase delta (Fadsd) and Fadsd, in zebrafish limits infection with Vibrio alginolyticus and V. vulnificus [5, 14]
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