Study on the differential expression of temperature tolerance related genes in Apostichopus japonicus between south-cultivated in Fujian Province and north-cultivated groups

doi:10.14012/j.cnki.fjsc.2024.01.001

Abstract

Abstract:

In this study,juvenile A.japonicus (experimental group) were purchased from northern China and reared in southern China under natural water temperature for one year.The same age seedlings of juvenile A.japonicus (control group) were directly collected from northern China to compare their thermal resistance with those of the experimental group.The study of thermal limits showed that the sublethal temperature of A.japonicus in the experimental group was 32 °C,significantly higher than the 30 °C in the control group.The ULTs₅₀ of A.japonicus in the experimental and control groups were 33.1 °C and 31.9 °C,respectively.These results demonstrated that the thermal history influenced the upper thermal limit of A.japonicus,and A.japonicus undergoing continuous thermal shock would possess stronger thermal resistance.Additionally,the expression patterns of HSP90a,gp96,HSP70,HSP26,and proeintl(2)efl genes after heat shock for 1,2,and 3 hours between the experimental and control groups were analyzed using qRT-PCR.The expression patterns of HSP90a,gp96,HSP70,HSP26,and proeintl(2)efl genes were different between the experimental and control groups.At 30 °C heat shock,the gene expressions of HSP90a,gp96,HSP70,and HSP26 in the control group did not noticeably increase after stimulation for 1 and 2 hours but were significantly up-regulated at 3 hours,with expression values reaching (9.801±1.303),(2.508±0.910),(8.649±1.936),and (34.787±4.978),respectively.However,all of the HSP90a,gp96,HSP70,and HSP26 gene expressions in the experimental group were dramatically up-regulated at 1 hour at 30 °C,with expression values reaching (42.000±8.798),(20.019±6.224),(218.750±78.701),and (93.710±5.674),respectively.These four temperature-tolerant related genes detected in the present study in the experimental group were up-regulated earlier than those in the control group,indicating a higher ability of A.japonicus in the experimental group to respond adaptively to heat shock than that in the control group.Therefore,the research showed that A.japonicus undergoing high temperature acclimation could respond earlier to external environmental heat conditions.

Key words: Apostichopus japonicus, high temperature acclimation, heat shock protein, high temperature tolerance

摘要：

本研究将仿刺参幼苗从北方南移至福建进行养殖,在自然水温下饲养1年,并作为南移福建养殖的实验组,同时以北方相同苗龄的仿刺参作为对照组,比较两组间的抗热性能;采用荧光定量法分析经过热诱导1、2、3 h后的实验组和对照组仿刺参的HSP90a、gp96、HSP70、HSP26和proeintl(2)efl温度耐受性相关基因的转录表达变化情况。研究发现:1)实验组仿刺参的亚致死温度为32 ℃,明显高于对照组的30 ℃,实验组和对照组仿刺参的半致死温度分别为33.1、31.9 ℃,表明高温驯化能够提高仿刺参的耐热能力。2)实验组与对照组仿刺参的5种温度耐受性相关基因的表达存在差异。在30 ℃高温刺激下,对照组仿刺参的HSP90a、gp96、HSP70、HSP26基因表达在温度刺激1、2 h后没有明显增加,而刺激3 h后才出现上调,表达量分别达到(9.801±1.303)、(2.508±0.910)、(8.649±1.936)、(34.787±4.978);实验组仿刺参的HSP90a、gp96、HSP70、HSP26表达在温度刺激1 h后即出现显著上调,表达量分别达到(42.000±8.798)、(20.019±6.224)、(218.750±78.701)、(93.710±5.674)。实验组仿刺参的4种温度耐受性相关基因的上调时间早于对照组,表明实验组仿刺参对外界高温的适应能力强于对照组。因此,高温驯化后的仿刺参能够更早地对外界高温变化做出反应。

关键词: 仿刺参, 高温驯化, 热冲击蛋白, 高温耐受性

CLC Number:

S968.9

ZHU Zhihuang, LIN Qi, WU Jianshao, YANG Qiuhua. Study on the differential expression of temperature tolerance related genes in Apostichopus japonicus between south-cultivated in Fujian Province and north-cultivated groups[J]. Journal of Fisheries Research, 2024, 46(1): 1-10.

朱志煌, 林琪, 吴建绍, 杨求华. 仿刺参南移养殖温度耐受性相关基因的差异表达分析[J]. 渔业研究, 2024, 46(1): 1-10.

Figures/Tables 7

Tab.1 The primer used in the study

基因 Genes	引物 Primers	序列 Sequences (5'-3')
HSP90a	HSP90a-F	TTGTTGAAAGGGAGGAGG
HSP90a	HSP90a-R	GGCATCAGAGGCGTTAGA
HSP26	HSP26-F	TCCATCATTTGGCCTTCAGCGTA
HSP26	HSP26-R	CATCAAACTCGTCATCGGCAAGC
HSP70	HSP70-F	GACACCACTTCAAAGACACC
HSP70	HSP70-R	ATCATCAACGAACCTACCGC
gp96	gp96-F	GTTGAAAGGGAGGAGGAAG
gp96	gp96-R	GGCATCAGAGGCGTTAGA
proeintl- (2)efl	proeintl- (2)efl-F	TGGCATTAGCCGAACAGT
proeintl- (2)efl	proeintl- (2)efl-R	TTCGTCGTCGGATTCATAAC
β-actin	β-actin-F	CGGGAAATCGTTCGTGACA
β-actin	β-actin-R	AGGACAAAGTTGGCGTACA

Fig.1 Survival rate of different populations of A. japonicus seedlings after high temperature and heat stimulation

Fig.2 Changes of HSP90a gene expression in different populations of A. japonicus seedlings after heat stimulation Notes:Different lowercase letters indicated significant difference (P<0.05),while the same lowercase letter meant no significant difference (P>0.05).The same as below.

Fig.3 Changes of gp96 gene expression in different populations of A. japonicus seedlings after heat stimulation

Fig.4 Changes of HSP70 gene expression in different populations of A. japonicus seedlings after heat stimulation

Fig.5 Changes of HSP26 gene expression in different populations of A. japonicus seedlings after heat stimulation

Fig.6 Changes of proeintl(2)efl gene expression in different populations of A. japonicus seedlings after heat stimulation

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