Abstract
Blunt snout bream (Megalobrama amblycephala) is sensitive to hypoxia environment. Hypoxia-inducible factor (HIF) is the most critical factor in the HIF pathway, which strictly regulates the hypoxia stress process of fish. In this study, we found six hifα genes in blunt snout bream that demonstrated different expressions under hypoxia conditions. In HEK293T cells, all six hifαs were detected to activate the HRE region by luciferase reporter assay. More importantly, we identified two linkage-disequilibrium SNP sites at exon 203 and 752 of the hif2αb gene in blunt snout bream. Haplotype II (A203A752) and its homozygous diplotype II (A203A203A752A752) appeared frequently in a selected strain of blunt snout bream with hypoxia tolerance. Diplotype II has a lower oxygen tension threshold for loss of equilibrium (LOEcrit) over a similar range of temperatures. Moreover, its erythrocyte number increased significantly (p < 0.05) than those in diplotype I and diplotype III strains at 48 h of hypoxia. The enzymes related with hypoxia tolerant traits, i.e., reduced glutathione, superoxide dismutase, and catalase, were also significantly (p < 0.05) induced in diplotype II than in diplotype I or III. In addition, the expression of epo in the liver of diplotype II was significantly (p < 0.01) higher than that in the diplotype I or III strains at 48 h of hypoxia. Taken together, our results found that the hypoxia-tolerant-related diplotype II of hif2αb has the potential to be used as a molecular marker in future genetic breeding of hypoxia-tolerant strain.
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This work was supported by the National Key Research and Development Plan Program (No. 2022YFD2400602; Grant Recipient: Shu-Ming Zou), the National Natural Science Foundation of China (No. 32273100; Grant Recipient: Shu-Ming Zou), the Key Projects of Revitalizing Agriculture through Science and Technology from Shanghai Agricultural and Rural Commission (No. 2022–02-08–00-12-F01179; Grant Recipient: Shu-Ming Zou), and the Research on the Frontier of Aquatic Peak-Innovation Strategy Source (Award Number: A1-3201–21-0004–1; Grant Recipient: Guo-Dong Zheng).
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Shan-Shan Zhao, Xiao-Lei Su, Guo-Dong Zheng, and Shu-Ming Zou have participated in conception and design; Shan-Shan Zhao and Hui-Qi Yang completed the experiments; Shan-Shan Zhao and Xiao-Lei Su analyzed and interpreted the data; Shan-Shan Zhao and Xiao-Lei Su drafted the article or revised it critically for important intellectual content; and Guo-Dong Zheng and Shu-Ming Zou approved the final version.
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Zhao, SS., Su, XL., Yang, HQ. et al. Functional exploration of SNP mutations in HIF2αb gene correlated with hypoxia tolerance in blunt snout bream (Megalobrama amblycephala). Fish Physiol Biochem 49, 239–251 (2023). https://doi.org/10.1007/s10695-023-01173-w
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DOI: https://doi.org/10.1007/s10695-023-01173-w