Analyses of the differential expression of cloned Caspase-3 and Caspae-9 genes,of Onchidium reevesii,exposed to extreme hot and cold conditions  

作　　者：Fengshen Zhao Heding Shen 

机构地区：[1]Key Laboratory of Aquatic Germplasm Resources Exploration and Utilization of Ministry of Education,Shanghai Ocean University,Shanghai,201306,China [2]National Experimental Teaching Demonstration Center of Aquaculture Science,Shanghai Ocean University,Shanghai,201306,China [3]Classification and Evolution of Marine Animal System,Shanghai Ocean University Key Laboratory of Shanghai Universities,Shanghai,201306,China

出　　处：《Aquaculture and Fisheries》2021年第6期583-592,共10页渔业学报（英文）

基　　金：This work was supported by the Construction Project of the Double First-class Disciplines of Fisheries.

摘　　要：During low tide intertidal organisms face severe temperature fluctuations.This situation may be more pronounced in the context of climate change.To investigate the molecular response mechanism of Onchidium reevesii when stressed by hot and cold temperatures,we cloned its Caspase-3 and Caspase-9 genes and detected their expression characteristics using RT-qPCR.The results showed that the full-length of the cDNA of Caspase-3 consist of 1831 bp,and included an open reading frame(ORF)of 894 bp,a 50UTR of 155 bp and a 30 UTR of 300 bp,and encoding for 297 amino acids.The full-length of the cDNA of Caspase-9 consisted of 2425 bp,and included an ORF of 1373 bp,a 50 UTR of 290 bp,and a 30 UTR of 759 bp,and encoding for 457 amino acids.During the evolution of species the two genes have been greatly conserved.A homology analysis and phylogenetic tree reconstruction revealed that O.reevesii is most closely related to Aplysia californica.The RT-qPCR showed that both genes were expressed in all of the tissues and that expression was highest in the hepatopancreas.It is speculated that these genes play an essential role in the immunity of O.reevesii to certain diseases.Conditions of temperature stress(40�C and 10�C)showed that both heat and cold stresses could induce significant changes in both genes.Compared to the control group,under cold stress conditions the levels of expression of Caspase-3 and Caspase-9 were significantly upregulated(P<0.05)in each experimental group.When exposed to a constant temperature of 10�C,their expression peaked after 1 h(4.32-fold,P<0.05;and 5.58-fold,P<0.05,respectively)and was at its lowest after 12 h(1.93-fold,P<0.05;and 2.35-fold,P<0.05,respectively).Their expression was upregulated after 24 h(2.68-fold,P<0.05;and 4.21-fold,P<0.05,respectively).The expression of both genes showed similar trends.Under heat stress,the expressions of the two genes showed a wave pattern.When exposed to a constant temperature of 40�C,their expression was up-regulated after 1,6,and 24 h,and down-regulated a

关 键 词：Onchidium reevesii CASPASE-3 CASPASE-9 Temperature Stress 

分 类 号：S917[农业科学—水产科学]