Transcriptomic response of the Pacific abalone(Haliotis discus hannai)to different algae diets and preliminary functional analysis of related sulfatase gene ids-1
In order to analyze the decomposition and adaptation mechanism of Pacific abalone(Haliotis discus han-nai)to its ingested algae,the differentially expressed genes(DEGs)in the viscera mass of the Pacific abalone fed three types of algae(Laminaria japonica,Gracilaria lemaneiformis and Ulva pertusa)as well as the selected sulfa-tase gene ids-1,were investigated by transcriptome sequencing,bioinformatics analysis,gene cloning,quantitative real-time PCR,prokaryotic expression,protein purification,and enzymatic activity analysis.The results showed that a total of 104 up-regulated and 118 down-regulated DEGs were identified when comparing the group LXC to HD.A total of 80 up-regulated genes and 93 down-regulated genes were identified in the group SC to HD,while the number of up-regulated and down-regulated genes in the group SC to LXC were 77 and 72.The results of Gene On-tology(GO)showed that the DEGs were enriched to entries mostly related to substance metabolism,especially the synthesis,catabolism,and metabolism of sulfated substances,with several sulfatase genes that showed DEGs iden-tified at GO:0006027(glycosaminoglycan catabolic process).Further,the sulfatase ids-1 gene was cloned and its molecular characteristics were analyzed.The open reading frame of this gene was determined to be 1 767 bp in length,encoding 588 amino acids.The deduced protein contains an N-terminal signal peptide(1-23 aa),a con-served domain of sulfatase(28-376 aa),and 9 glycosylation sites.Phylogenetic analysis showed that IDS-1 from pacific abalone shared the highest homology with IDS from Haliotis refescens and Haliotis rubra.Multiple amino acid sequence alignment demonstrated that IDS-1 carries the typical core motif"CXPXR"of the sulfatase family as well as the highly conserved active sites of the IDS subfamily.However,the recombinant IDS-1 protein produced from Escherichia coli did not exhibit sulfatase activity.Furthermore,qPCR analysis showed the highest expression of ids-1 in the visceral mass of pacific abalone,followed by the gill and mantle,with comparatively lower expression in upper foot tentacles.Our study also demonstrated a developmental stage-specific expression pattern of ids-1.The ex-pression level is low from fertilized eggs to planktonic larvae,and increases in plantigrade larvae,with the highest level in adults.Additionally,we found that the expression of ids-1 varied significantly under different algae feeding conditions,with the brown alga-fed group exhibiting notably higher expression compared to the red and green alga-fed groups,while the green alga group displayed the lowest expression level.We speculate that ids-1 may be closely related to the nutritional patterns and dietary transitions during the development of abalone,and its expression level is potentially directly affected by differences in feeding preferences and food intake.This findings provides reference for in-depth analysis of the diet adaptation regulation mechanism in Pacific abalone.
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