RNA-Seq analysis of the pyloric caecum, liver, and muscle reveals molecular mechanisms regulating fillet color in rainbow trout

Abstract

Background: The characteristic pink-reddish color in the salmonids fillet is an important, appealing quality trait for consumers and producers. The color results from diet supplementation with carotenoids, which accounts for up to 20-30% of the feed cost. Pigment retention in the muscle is a highly variable phenotype. In this study, we aimed to understand the molecular basis for the variation in fillet color when rainbow trout (Oncorhynchus mykiss) fish families were fed an Astaxanthin-supplemented diet. We used RNA-Seq to study the transcriptome profile in the pyloric caecum, liver, and muscle from fish families with pink-reddish fillet coloration (red) versus those with lighter pale coloration (white).

Results: More DEGs were identified in the muscle (5,148) and liver (3,180) than in the pyloric caecum (272). Genes involved in lipid/carotenoid metabolism and transport, ribosomal activities, mitochondrial functions, and stress homeostasis were uniquely enriched in the muscle and liver. For instance, the two beta carotene genes (BCO1 and BCO2) were significantly under-represented in the muscle of the red fillet group favoring more carotenoid retention. Enriched genes in the pyloric caecum were involved in intestinal absorption and transport of carotenoids and lipids. In addition, the analysis revealed the modulation of several genes with immune functions in the pyloric caecum, liver, and muscle.

Conclusion: The results from this study deepen our understanding of carotenoid dynamics in rainbow trout and can guide us on strategies to improve Astaxanthin retention in the rainbow trout fillet.

Keywords: Antioxidant; Aquaculture; Beta-carotene; Micellization; Pathways; Pigmentation.

Fig. 1

a: Principal component analysis (PCA) performed on the gene expression data from pyloric caeca samples showing no apparent clustering. b: PCA performed on the expression data of muscle samples showing the separation of the white (Low) and red (High) fillet groups. c: PCA performed on the gene expression data of liver samples showing the separation of the white (Low) and red (High) fillet groups. d: PCA performed on the expression data of pyloric caecum, liver, and muscle samples of rainbow trout. The PCA shows a clear separation of the three tissues

Fig. 2

Enriched GO biological function terms for the upregulated genes in the liver comparing the white versus red fillet groups

Fig. 3

The topmost significant canonical pathways for the muscle DEGs

Fig. 4

Modulated diseases and functions from the DEGs in the muscle (a) Genes related to lipid metabolism (b) Genes related to immunity

Fig. 5

Enriched KEGG pathways for upregulated genes in the muscle comparing the white versus red fillet groups [40]

Fig. 6

Enriched GO Biological process terms for the upregulated genes in the muscle comparing the white versus red fillet groups

Fig. 7

Enriched KEGG pathways for the downregulated genes in the muscle comparing the white versus red fillet group [40]

Fig. 8

Enriched GO biological process for the downregulated genes in the muscle comparing white versus red fillet group