摘要
为探究罗氏沼虾(Macrobrachium rosenbergii)急性低温应激响应的基因表达模式,研究以罗氏沼虾成虾为研究对象,分别设置实验组(16℃)和对照组(24℃),实验组从24℃急性降温(2℃/1h)至16℃后的0、1h、3h、6h、12h、24h和回温至24℃后共7个时间点采集肝胰腺组织进行转录组测序分析。差异表达基因(DEGs)筛选结果显示,胁迫3h与回温后的DEGs数量(5062个)几乎是1h与回温后的1.5倍(3516个),随着胁迫时间的延长,各时间点与回温后的DEGs数量逐渐降低。KEGG富集发现,DEGs显著富集在溶酶体、淀粉和蔗糖代谢、抗原加工与呈递等途径中。此外,罗氏沼虾在急性低温应激下,黏着斑、ECM-受体互作、细胞色素P450对异生物质的代谢、谷胱甘肽代谢、氧化磷酸化、p53信号通路等相关基因也发生了显著变化。WGCNA分析发现各组间的共有DEGs聚类在与细胞功能及免疫相关模块和与能量物质代谢相关模块上。另外,花生四烯酸代谢信号通路中的Cytochrome P4502L1-like基因、谷胱甘肽代谢信号通路中的NADP-specific isocitrate dehydrogenase基因及淀粉和蔗糖代谢信号通路中的Hexokinase基因等的表达量随低温应激时间增加呈先增后减的趋势。这些通路及基因可能在罗氏沼虾急性低温应激下的能量代谢及免疫调节等活动中起重要作用。研究为揭示罗氏沼虾急性低温应激响应的分子调控机制提供了基础数据。
The giant freshwater prawn(GFP)Macrobrachium rosenbergii is one of the important economically freshwater shrimp species,and its annual production in China is dominant in the world.However,its tolerance to low temperatures is extremely poor,and acute low temperatures can lead to large-scale deaths and cause huge economic losses.In order to explore genes related to the GFP response to acute low temperature,comparative transcriptomic analyses were performed for the hepatopancreas of adult GFPs exposed to low temperature.The low temperature stress group(16℃)and the control group(24℃)were set up.The water temperature of the stress group was decreased from 24℃to 16℃at a rate of around 2℃/h through adding ice cubes.The hepatopancreas were collected for transcriptomic analyses from exposed shrimps respectively at 1h,3h,6h,12h,24h after acute cooling at 16℃,rewarming to 24℃and control group.The results of differentially expressed genes(DEGs)showed that 1702 DEGs were identified between samples of cooling for 1h and the control group(M1 vs.C0);the number of DEGs between the stress group and the control group began to increase gradually with stress time,and reached the maximum at 6h(M3 vs.C0,a total of 2899),then gradually decreased,maintaining the homeostasis under low temperature stress.After rewarming to 24℃,the number of DEGs(M6 vs.C0,1969)returned to the level of 1h stress.Additionally,the DEGs number(5062)between samples of stress for 3h and rewarming to 24℃(M2 vs.M6)was almost 1.5 times of that(3516)between samples of stress for 1h and rewarming(M1 vs.M6).With the extension of time,the number of DEGs decreased gradually,suggesting that the homeostasis of the GFP had drastic changes in the first 3h of acute low temperature stress,but the adaptability to low temperature gradually increased with stress time,and a steady state under low temperature was established probably after being stressed from 3h to 6h,and after rewarming,the homeostasis returned to the equilibrium of short-term cold stress.KEGG enrichment analysis showed that DEGs was enriched in lysosome,starch and sucrose metabolism,antigen processing and presentation.Adhesion spots,ECM-receptor interaction,metabolism of cytochrome P450 to isobiotic substances,glutathione metabolism,oxidative phosphorylation,and p53 signaling pathway were also involved in the regulation of acute cold stress in the GFP.The common DEGs among all groups were clustered as the cell function and immunity modules and energy metabolism modules.In addition,the up-regulated cytochrome P4502L1-like gene in the arachidonic acid metabolic signaling pathway was screened,and its expression increased firstly and then decreased with the increase of cold stress time.NADP-specific isocitrate dehydrogenase,an upregulated gene in glutathione metabolic signaling pathway,was firstly reduced and then increased with time.Expression of hexokinase,a down-regulated gene in starch and sucrose metabolic signaling pathways,increased and then decreased over time.The above mentioned enriched pathways indicated that the metabolic function of the GFP might have been seriously affected under acute low temperature stress,with a large amount of reactive oxygen species being produced,the balance of energy circulation being disrupted,and the immune system being also damaged,which was corroborated with the phenomena of fasting,slow movement,susceptibility to diseases and even death of the GFP under acute low temperature.Consequently,these screened pathways and genes may play important roles in energy metabolism and immune regulation during acute cold stress in the GFP.The present study provides basic data for revealing the molecular regulatory mechanism of the response of the GFP to acute low temperature stress,also provides a theoretical basis for the selective breeding of new cold-tolerant GFP varieties.
作者
杨旻珉
屠海慧
邢钱钱
唐琼英
易少奎
夏正龙
蔡缪荧
陈国柱
蓝璇
钟镇霄
黄笑
高权新
杨国梁
YANG Min-Min;TU Hai-Hui;XING Qian-Qian;TANG Qiong-Ying;YI Shao-Kui;XIA Zheng-Long;CAI Miao-Ying;CHEN Guo-Zhu;LAN Xuan;ZHONG Zhen-Xiao;HUANG Xiao;GAO Quan-Xin;YANG Guo-Liang(Zhejiang Provincial Key Laboratory of Aquatic Resources Conservation and Development,Key Laboratory of Aquatic Animal Genetic Breeding and Nutrition,Chinese Academy of Fishery Sciences,College of Life Sciences,Huzhou University,Huzhou 313000,China;Jiangsu Shufeng Prawn Breeding Co.LTD,Gaoyou 225654,China)
出处
《水生生物学报》
CAS
CSCD
北大核心
2023年第4期581-593,共13页
Acta Hydrobiologica Sinica
基金
江苏省科技厅重点研发专项资金(现代农业)项目(BE2019352)
国家重点研发计划“蓝色粮仓科技创新”重点专项(2018YFD0901300)
国家现代农业产业技术体系资助项目(CARS-48)
浙江省农业(水产)新品种选育重大科技专项(2021C02069-4-3)资助。
关键词
急性低温应激
肝胰腺
转录组
差异表达基因
能量代谢
免疫调节
罗氏沼虾
Acute cold stress
Hepatopancreas
RNA-seq
Differentially expressed genes
Energy metabolism
Immune regulation
Macrobrachium rosenbergii
