低氧胁迫下中华圆田螺的肝脏转录组学分析  

Transcriptome Analysis of Liver Tissue of Cipangopaludina cathayensis Under Hypoxic Stress

作　　者：殷丽坤 黄凯[1] 于凯 宿志健 郭睿婕 杨旭红 吴耀庭 YIN Likun;HUANG Kai;YU Kai;SU Zhijian;GUO Ruijie;YANG Xuhong;WU Yaoting(College of Animal Science and Technology,Guangxi University,Nanning 530004,China;Decheng Market Supervision Administration,Dezhou 253011,China)

机构地区：[1]广西大学动物科学技术学院,广西南宁530004 [2]德州市德城区市场监督管理局,山东德州253011

出　　处：《渔业科学进展》2023年第4期167-178,共12页Progress in Fishery Sciences

基　　金：广西科技重大专项创新驱动发展基金(桂科AA20302019-6);广西重点研究开发项目(AB18294011)共同资助。

摘　　要：为了探究低氧胁迫下中华圆田螺(Cipangopaludina cathayensis)肝脏组织基因的差异表达,本研究通过高通量测序技术,分析中华园田螺低氧胁迫组(2.5 mg/L)和常氧组(6.9 mg/L)某些基因的差异表达,并对差异基因进行生物信息学分析,进一步采用实时荧光定量PCR(RT-qPCR)对关键差异表达基因进行验证。结果显示,测序共获得232379条基因(unigenes),与对照组比较,低氧胁迫组筛到176个差异基因,包含64个上调基因和112个下调基因。GO功能注释分析显示,差异基因主要富集在生物学过程中的几丁质代谢过程和含氨基葡萄糖的复合代谢过程,细胞组分中的胶原三聚体成分,分子功能中的几丁质结合功能和糖衍生物结合功能。KEGG通路富集分析显示,差异基因主要集中于环境信息处理、遗传信息处理、代谢和生物系统这4大类通路。6个关键差异基因的RT-qPCR结果显示,热休克蛋白70B2和热休克蛋白β-6基因表达量上调,几丁质酶蛋白4、α-1胶原蛋白(XIV)、α-4胶原蛋白(XIV)、5-磷酸酶蛋白基因表达量下调,证实了转录组测序结果的可靠性。本研究发现,低氧胁迫激活了中华圆田螺适应缺氧的生理活动,并获得了低氧胁迫下中华圆田螺肝脏组织中相关功能基因的表达信息,为深入研究中华圆田螺响应低氧胁迫的调控机制提供了基础数据和理论依据。Cipangopaludina cathayensis is a snail species unique to China.Domestic research on C.cathayensis has mainly focused on aquaculture technology,especially the paddy field breeding method,where water quality plays a critical role in the cultivation of C.cathayensis.Dissolved oxygen is one of the most important factors in the aquatic environment because it impacts a series of biological activities such as the growth and development,metabolism,reproduction,and breeding of aquatic animals.When the dissolution of oxygen in water is less than 2 mg/L,the water is in a low oxygen or anoxic state.Hypoxia can slow the growth and development of aquatic animals,reduce their disease resistance and reproductive ability,and,in serious cases,can lead to their death.In recent years,research on C.cathayensis has mainly focused on the anti-tumor mechanism,immune response,nutritional value evaluation,and antibiotic resistance.However,there is currently no report on the regulation of response to hypoxia in C.cathayensis either domestically or internationally.The purpose of this study was to explore the differential expression of genes in the liver of C.cathayensis under hypoxic stress.In this study,healthy C.cathayensis without mechanical damage were cultured in a hypoxia stress group(2.5 mg/L)and a normoxia(control)group(6.9 mg/L),with 90 C.cathayensis in each group and 3 replicates.For the low oxygen stress treatment,dissolved oxygen was decreased from 6.9 mg/L to 2.5 mg/L within 1 h and maintained for 24 h.The liver tissue was taken as the experimental material in both the hypoxia stress group and normoxic group.The total RNA was extracted and an mRNA library was constructed.The liver tissue samples of C.cathayensis from both groups were sequenced and analyzed using an Illumina HiSeq-2500 technology platform,and unigenes were compared and annotated in GO,KOG,Nr,and KEGG databases.The differentially expressed genes were analyzed using DESeq.Bioinformatics analysis was performed on the function of GO and KEGG of differentially expres

关 键 词：中华圆田螺 低氧胁迫 转录组 差异表达基因 热休克蛋白 

分 类 号：S917[农业科学—水产科学]