脊尾白虾不同发育期线粒体基因组甲基化特征分析  

Methylation Profile in Mitochondrial Genome of Ridgetail White Prawn Exopalaemon carinicauda in Response to Different Developmental Stages

作　　者：薛蓓[1,2,3] 张培 李志辉[1,2,3] 赵莲 赖晓芳[1,2,3] 阎斌伦 高焕[1,2,3] 

机构地区：[1]淮海工学院海洋生命与水产学院江苏省海洋生物技术重点实验室,江苏连云港222005 [2]江苏省海洋生物产业技术协同创新中心,江苏连云港222001 [3]江苏省农业种质资源保护与利用平台,江苏南京210014

出　　处：《水产科学》2017年第5期628-633,共6页Fisheries Science

基　　金：江苏高校优势学科建设工程项目(PAPD);江苏省高等学校自然科学研究重大项目(17KJA240001);江苏省"六大人才高峰"创新人才团队项目(2016-HYGC-CXTD-004);江苏省2015年度普通高校研究生科研创新计划项目(KYLX15_1486);连云港市产学研合作项目(CXY1517)

摘　　要：为研究不同发育阶段脊尾白虾线粒体基因组表观遗传学特征,本研究在筛选硫化测序引物的基础上,对不同发育期的脊尾白虾线粒体基因组甲基化特征进行了分析。经引物筛选验证,选取了4对引物在脊尾白虾的受精卵、幼体期、仔虾期和成虾期共4个发育期进行甲基化特征的分析,结果表明,除ND5基因3′端序列(21#引物扩增区)无甲基化现象外,COX3基因的起始区序列(13#引物扩增区)、COX3基因的3′端序列(16#引物扩增区)和ND5基因5′端序列(19#引物扩增区)均存在不同程度的甲基化现象。其中,COX3基因的起始区序列仅在受精卵孵化期和幼体期存在甲基化现象,COX3基因3′端序列在除受精卵外的其他3个时期均存在甲基化现象,且以上甲基化率随个体发育阶段的提高呈下降趋势;ND5基因5′端序列在4个不同发育期均存在甲基化现象,但甲基化率无显著差异。甲基化主要发生在COX3基因和ND5基因位点上,推测脊尾白虾在生长发育过程中通过调节线粒体基因组甲基化来调节机体能量代谢过程。本研究首次从整个发育周期角度探讨了线粒体基因组甲基化特征,以期为进一步揭示甲壳类生长发育相关遗传机制提供帮助。BSP（Bisulfite sequencing PCR）primer pairs were screened in order to analyze the methylation profile of the mitochondria genome in response to different developmental stages in the ridgetail white prawn Exopalaemon carinicauda.Within ten synthesized BSP primer pairs,four primer pairs were selected for further studies because of their repeatable PCR amplification merits,including 13th（located in the5′end initiation zone of COX3）,16th（located in the 3′sequence of COX3）,19th（located in the 5′sequence of ND5）and 21st（located in the 3′sequence of ND5）.The methylation profile of different individuals at different developmental stages（fertilized eggs,larval,post larval and adult stages）were analyzed,using the above 4 primer pairs.The results showed that the methylation in both CpG sites and non-CpG sites were found in the 5′and 3′sequence of COX3 and the 5′sequence of ND5,but not in the 3′sequence of ND5.The 5′sequence of COX3 was methylated at fertilized eggs and larval stage,while the 3′sequence of COX3 was methylated at larval,post larval and adult stages.Additionally,the methylation ratio was decreased with the increasing of development stage for above two sequences of COX3.The 5′sequence of ND5 was methylated in the all four developmental stages,and no significant differences for methylation ratio were found among different developmental stages.In view of the biology function of COX and ND genes which expression productions were responsible for maintaining the respiratory electron-transport chain in mitochondria,it is inferred that the function of methylation of mitochondrial genome in ridgetail white prawn is to regulate the energy metabolism during developmental stage.

关 键 词：脊尾白虾 线粒体基因组 不同发育期 甲基化 

分 类 号：S917[农业科学—水产科学]