饥饿胁迫对拟穴青蟹大眼幼体的比较转录组分析  

Comparative transcriptome study of starvation stress on the megalopae of Scylla paramamosain

作　　者：胡伟胜 王伟[1] 马凌波[1] 张凤英[1] 刘志强 赵明[1] 马春艳[1] HU Weisheng;WANG Wei;MA Lingbo;ZHANG Fengying;LIU Zhiqiang;ZHAO Ming;MA Chunyan(Laboratory of Genetics,Breeding and Biotechnology,East China Sea Fisheries Research Institute,Chinese Academy of Fishery Sciences,Shanghai200090,China;National Demonstration Center for Experimental Fisheries Science Education,College of Fishery and Life Science,Shanghai Ocean University,Shanghai201306,China)

机构地区：[1]中国水产科学研究院东海水产研究所遗传育种与生物技术实验室,上海200090 [2]上海海洋大学水产与生命学院,水产科学国家级实验教学示范中心,上海201306

出　　处：《海洋渔业》2023年第4期460-471,共12页Marine Fisheries

基　　金：上海市农委“科技兴农”项目(2018-02-08-00-07-F01550);宁波市科技创新2025重大专项(2019B10010);中国水产科学研究院基本科研业务费专项(No.2020TD28);国家虾蟹产业技术体系建设-虾蟹(CARS-48)。

摘　　要：为了探究甲壳类幼体应对饥饿胁迫的分子机理,以拟穴青蟹(Scylla paramamosain)为研究对象,设置正常投喂组和饥饿组,处理3 d后对不同组别的拟穴青蟹大眼幼体进行转录组测序并开展生物信息学分析。De novo组装后共获得79581个unigene,平均长度1065.44 bp。Nr数据库共注释20061个unigene。DESeq分析筛选出5621个差异表达基因,饥饿组相比对照组2641个上调、2980个下调。基因本体论(GO)富集分析表明,共254条GO Term显著富集(矫正后的P值小于0.05)。京都基因与基因组百科全书(KEGG)富集分析表明,共60条KEGG pathway显著富集(矫正后的P值小于0.05)。基于KEGG富集分析结果筛选了一系列与饥饿胁迫相关的代谢通路,如DNA复制、细胞凋亡、缬氨酸,亮氨酸和异亮氨酸代谢、亚油酸代谢、淀粉和蔗糖代谢。研究表明,饥饿胁迫干扰了拟穴青蟹大眼幼体的细胞分裂和凋亡,影响了体内的氨基酸代谢、脂质代谢和碳水化合物代谢。研究结果可为拟穴青蟹苗种的健康培育提供理论参考,并丰富拟穴青蟹的分子生物学基础数据。In nature,food scarcity is one of the environmental stresses that crustaceans often face.Crustaceans often adjust metabolism,behavior and utilize internal storage to maintain basic physiological functions and achieve limited growth and reproduction in short-term or long-term nutritional limitations.Scylla paramamosain is an important marine breeding crab in the southeast coastal area of China.In this study,comparative transcriptome analysis was conducted on normally fed and starved megalopae of Scylla paramamosain to explore the effect of starvation stress on the metabolic process of megalopae,and to provide some theoretical references for the healthy cultivation of megalopae,and to enrich the molecular biological research data of Scylla paramamosain.The experiment was divided into two groups:the normal feeding group for 3 days(F3S0)and the starvation group for 3 days(S3F0).Each group had 3 replicates,and each replicate had 8 megalopaes.Transcriptome sequencing was carried out on the collected samples of megalopae.RNA was extracted separately from each sample.RNA degradation and contamination were assessed on agarose gels.RNA concentration,OD260/280,and OD260/230 ratios were measured.The RNA integrity number(RIN)was determined.Oligo(dT)magnetic beads were used to enrich mRNA with polyA tails,and the RNA was cut into fragments of about 300 bp in length by ion interruption.The first strand of cDNA was synthesized using RNA as template with 6-base random primers and reverse transcriptase.The second strand of cDNA was then synthesized using the first strand of cDNA as template.Library preparation and sequencing were then performed.Gene function was annotated based on NCBI non-redundant protein sequences(Nr),Gene Ontology(GO),Kyoto Encyclopedia of Genes and Genomes(KEGG),evolutionary genealogy of genes:Non-supervised Orthologous Groups(eggNOG),Swiss-Prot,and Pfam databases.Differential expression was assessed using transcript abundances as inputs to the DESeq.The charting of volcano map and cluster analysis about diff

关 键 词：拟穴青蟹 大眼幼体 饥饿 转录组 

分 类 号：S917.4[农业科学—水产科学]