Abstract

Strongylocentrotus intermedius is an economically valuable sea urchin species in China. However, its growth and survival are severely constrained by ocean warming and the hypoxia that often accompanies high water temperatures. MicroRNAs (miRNAs) are important regulators of gene expression in response to environmental change. In this study, high-throughput RNA sequencing was used to investigate changes in miRNA expression in S. intermedius under heat (25°C), hypoxia (2 mg/L O2), and combined heat and hypoxia stresses. Twelve small RNAs libraries were constructed and 17, 14, and 23 differentially expressed miRNAs (DEMs) were identified in the heat, hypoxia, and combined stress groups (P<0.05), respectively. Gene ontology and Kyoto Encyclopedia of Genes and Genomes pathway functional analyses of putative target genes of the DEMs suggested that these miRNAs were important in basal metabolism, apoptosis, oxidative stress, and immune-related pathways. By co-analysis with published transcriptome data, key DEMs (miR-193, miR-184, miR-133, miR-125, miR-2008) and their key target genes (EGF3, ABCB4, CYCL, PAN2, CALN) were identified. Quantitative real-time PCR analysis of the expression of 10 DEMs and their key target genes confirmed the RNA sequencing results. These results provide information on gene expression regulation of the molecular mechanisms underlying the response of S. intermedius to multi-cause environmental stresses.

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