Effects of Alkaline Stress on AKP Gene Expression and Physiology Response in Leuciscus waleckii
MI Bo-Han1,2, ZHAO Xue-Fei3, GAO Shan2,4, CHANG Yu-Mei2, SUN Bo2, LUO Liang2, ZHANG Li-Min2, LIANG Li-Qun2,*
1 National Demonstration Center for Experimental Fisheries Science Education, Shanghai Ocean University, Shanghai 201306, China; 2 Heilongjiang River Fisheries Research Institute, Chinese Academy of Fishery Sciences, Harbin 150070, China; 3 College of Wildlife and Protected Area, Northeast Forestry University,Harbin 150040, China; 4 College of Marine Life and Fisheries, Jiangsu Ocean University, Lianyungang 222000, China
Abstract：Alkaline phosphatase (AKP) can catalyze a variety of phosphorus-related hydrolysis reactions, and is a reference indicator for immune function and health status of organisms. Leuciscus waleckii is a kind of fish that is extremely resistant to the environment (especially high carbonate). In order to clarify various physiological reactions and health conditions in high alkali environments, two geographical groups of L. waleckii, which were Dalinuoer Lake L. waleckii (short for alkaline L. waleckii) from alkaline water and Songhua River Leuciscus waleckii (short for freshwater L. waleckii) from fresh water, were set to 30 and 50 mmol/L NaHCO3 for 20 d. Then the blood was taken out and placed in the Automatic microplate reader to determine the concentration of AKP in serum. Under the condition of 30 mmol/L carbonate alkalinity, the AKP activity of freshwater L. waleckii was significantly higher than that of alkaline L. waleckii (P<0.05). At the same time, based on the AKP gene sequence of the closely related species Cyprinus carpio, the AKP gene in L. waleckii was cloned, and AKP gene expression in the muscle, sputum, liver, brain and kidney was detected by real-time quantitative PCR. The results showed that the AKP ORF of L. waleckii was 876 bp in length (GenBank No. MN473375) and encoded 291 amino acids. The expression of AKP gene in different tissues of the two squids were different. The AKP gene expression in alkaline water L. waleckii increased in the liver with the increasing of alkalinity. The AKP gene expression in freshwater species increased in the kidney with the increasing of alkalinity. The AKP gene expression at different alkalinity levels in multiple tissues showed significant differences (P<0.05). In summary, the AKP gene might play a role in the alkali-resistant response in L. waleckii. This study could provide basic data for further investigation on the involvement of AKP in the alkali-resistance in L. waleckii.
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