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| Cloning of Osmotic Pressure-related Genes from Cherax quadricarinatus and Their Expression Under Different Salinity |
| PENG Bo-Hao, ZHANG Yan, LIU Chang, ZHONG Xiao, FU De-Zheng, ZHANG Kai-Yi, WANG Yi-Lei* |
| Fisheries College, Jimei University, Xiamen 361021, China |
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Abstract Salinity is an important environmental factor affecting the growth and survival of crustaceans. The red claw crayfish (Cherax quadricarinatus) is a freshwater species. To increase the waters available for cultivating this species, saline waters and brackish waters may be potential waters for cultivating this species. Thus, understanding the osmotic regulation mechanism of the red claw crayfish is needed to provide a theoretical basis for farming feasibility in brackish water and improve the flavor by changing the growth environment (temporary culture in brackish water). In this study, 3 genes related to osmolality, Na+/K+-ATPase (NKA), Na+-K+-2Cl--Cotransporter (NKCC) and 14-3-3, were screened by the NCBI published transcriptome database of C. quadricarinatus, and their ORF regions were confirmed by RT-PCR. The protein structure was predicted using an online bioinformatics website, and the homologous protein sequences of different species were subjected to multiple sequence alignment and evolutionary tree construction. The expression of NKA, NKCC and 14-3-3 in different tissues and the changes of expression in gill and hepatopancreas after 24, 48 and 96 h under different salinity in the red claw crayfish were analyzed by qPCR. The results showed that NKA, NKCC and 14-3-3 proteins were highly conserved among different species. NKA, NKCC and 14-3-3 genes were expressed in the gill, muscle, heart, hepatopancreas and eyestalk tissues of red claw crayfish. Under different salinity stresses, the expression of the NKA gene in gill tissues did not change significantly at 5‰ and 10‰ salinity but increased significantly in 15‰ and 20‰ salinity groups at 48 h (P<0.05). In the hepatopancreas, the expression of NKA gene was only significantly increased in the 20‰ salinity group at 48 h (P<0.05). The expression of NKCC gene in gill tissue was significantly up regulated in 10‰, 15‰ and 20‰ salinity groups at 24, 48 and 96 h, and only significantly increased in 15‰ and 20‰ salinity groups at 48 h in hepatopancreas. The expression of the 14-3-3 gene was significantly up regulated in gill tissue in 10‰, 15‰ and 20‰ salinity groups at 48 h (P<0.05), and significantly up regulated in hepatopancreas in 5‰, 10‰ and 15‰ salinity groups at 48 h (P<0.05) and the expression significantly up regulated at 24 and 48 h in 20‰ salinity group (P<0.05). The results showed that the expression of NKA, NKCC and 14-3-3 genes was related to the change of salinity. 20‰ salinity stress after 48 h may be the key point of osmotic regulation of gill and hepatopancreas under high salinity. This study provides a reference for further exploring the role of NKA, NKCC and 14-3-3 genes in the osmoregulatory mechanism of the red claw crayfish.
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Received: 08 December 2021
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Corresponding Authors:
* ylwang@jmu.edu.cn
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