The characterization, expression and activity analysis of three superoxide dismutases in Eriocheir hepuensis under azadirachtin stress. Issue 117 (October 2021)

• Record Type: Journal Article
• Authors: Liao, Yongyan; Liu, Ke; Ren, Tianjiao; Zhang, Zining; Ma, Zihang; Dan, Solomon Felix; Lan, Zhenyu; Lu, Min; Fang, Huaiyi; Zhang, Yan; Liu, Jinxia; Zhu, Peng
• Is Part Of: Fish & shellfish immunology. Issue 117(2021)
• Page Start: 228
• Page End: 239
• Publication Date: 2021-10
• Subjects: Eriocheir hepuensis -- CuZn-SOD -- cMn-SOD -- Azadirachtin -- Expression analysis; Fishes -- Immunology -- Periodicals; Shellfish -- Immunology -- Periodicals; Poissons -- Immunologie -- Périodiques; Crustacés -- Immunologie -- Périodiques; 571.9617
• DOI: 10.1016/j.fsi.2021.08.010
• Languages: English
• ISSNs: 1050-4648

Abstract

Abstract: Superoxide dismutase (SOD) can effectively eliminate of excess ROS, which causes oxidative damage to lipids, proteins, and DNA. In this study, we cloned the CuZn-SOD, cMn-SOD1, and cMn-SOD2 genes in Eriocheir hepuensis, and found that the coding sequence (CDS) lengths were 627 bp, 861 bp and 1062 bp, which encoded 208, 286, and 353 amino acids, respectively. Phylogenetic analysis indicated that all SOD genes were evolutionarily conserved, while cMn-SOD2 had an extra gap (67 amino acids) in the conserved domain compared with cMn-SOD1 without huge changes in the tertiary structure of the conserved domain, suggesting that cMn-SOD2 may be a duplicate of cMn-SOD1. qRT-PCR showed that the three SOD genes were widely expressed in all the tested tissues, CuZn-SOD and cMn-SOD1 were mostly expressed in the hepatopancreas, while cMn-SOD2 was mostly expressed in thoracic ganglia. Under azadirachtin stress, the oxidation index of surviving individuals, including the T-AOC, SOD activity, and MDA contents increased in the early stage and then remained steady except for a decrease in MDA contents in the later stage. qRT-PCR showed that the three SOD genes displayed the same trends as SOD activity in surviving individuals, and the highest expressions of CuZn-SOD in the hepatopancreas, heart, and gill were 14.16, 1.41, and 30.87 times that of the corresponding control group, respectively. The changes were 1.35, 5.77 and 3.33 fold for cMn-SOD1 and 1.62, 1.71 and 1.79 fold for cMn-SOD2, respectively. However, the activity and expression of SOD genes in dead individuals were lower than that observed in surviving individuals. These results reveal that SOD plays a significant role in the defence against azadirachtin-induced oxidative stress. Highlights: The three SOD genes in Eriocheir hepuensis were firstly cloned. cMn-SOD2 was a novel transcript isoform of cMn-SOD1. The oxidation index increased, but remained stable under azadirachtin stress. The three SOD genes have different expression patterns in normal tissues. The expression of three genes changed significantly under azadirachtin stress.