虾夷扇贝选育群体与野生群体基因组DNA甲基化的MSAP分析  

MSAP ANALYSIS ON GENOME-WIDE DNA METHYLATION IN SELECTED AND WILD JAPANESE SCALLOP PATINOPECTEN YESSOENSIS

作　　者：吴彪[1] 杨爱国[1] 董迎辉[2] 郑利兵[1,3] 刘志鸿[1] 周丽青[1] 孙秀俊[1] 郑言鑫 

机构地区：[1]农业部海洋渔业可持续发展重点实验室中国水产科学研究院黄海水产研究所,青岛266071 [2]浙江省水产种质资源高效利用技术研究重点实验室浙江万里学院,宁波315100 [3]上海海洋大学水产与生命学院,上海201306

出　　处：《海洋与湖沼》2015年第4期976-982,共7页Oceanologia Et Limnologia Sinica

基　　金：黄海水产研究所基本科研科业务费;20603022013012号;浙江省水产种质资源高效利用技术研究重点实验室开放基金;KL2013-3号;山东省自主创新专项;2013CXC80202号

摘　　要：基因组DNA甲基化作为表观遗传学研究的重要手段之一,在水产科学领域得到越来越广泛的应用。为从表观遗传学角度探讨虾夷扇贝选育群体-"玉贝"和普通虾夷扇贝群体的DNA甲基化差异,本文运用甲基化敏感扩增多态性(Methylation-sensitive amplification polymorphism,MSAP)技术分析了两群体的DNA序列中CCGG位点的甲基化情况。结果表明,筛选得到11对引物组合能够在两个群体中得到稳定扩增、清晰且多态性丰富的片段;11对引物在"玉贝"和对照群体中分别共检测到1432和1442个位点,其中发生甲基化的位点分别为306和341个,总甲基化率分别为21.5%和23.65%,"玉贝"总甲基化率下降2.15%;全甲基化位点分别为152和236个,全甲基化率为10.68%和16.37%,"玉贝"全甲基化率下降5.69%;半甲基化位点分别为154和105个,半甲基化率为10.82%和7.28%,"玉贝"半甲基化率上升3.54%;"玉贝"群体的甲基化多态性条带所占比例高于对照群体,分别为31%和29.167%,尤其是B类型条带链,"玉贝"群体明显高于对照群体。这说明,通过对虾夷扇贝生长和耐热性状的累代选育导致了DNA甲基化水平和模式的改变,本研究为进行虾夷扇贝抗逆基因的筛查提供了参考数据,丰富了表观遗传学在扇贝中的研究资料。DNA methylation is one of key epigenetic modifications in eukaryotes genomes and has been studied widely in fishery science in recent years. However, characterization of DNA methylation in invertebrate remains unclear. We therefore employed methylation-sensitive amplification polymorphism（MSAP） technique to investigate the genome-wide DNA methylation profiles in selected Patinopecten yessoensis ＂Yubei＂ and control populations. The P. yessoensis ＂Yubei＂, a new strain with fast-growing and heat-resistant traits, was selected via successive breeding of multi-generation. The results showed that the DNA methylation level and pattern between the two populations were significantly different. Eleven pairs of primer combinations were selected to amplify the enzyme products, and total of 1432 and 1442 fragments were detected in the ＂Yubei＂ and control populations, respectively. Comparing to the control group, the total methylation rate（21.5%） and full-methylation rate（10.68%） of ＂Yubei＂ decreased by 2.15% and 5.69%, respectively, while hemi-methylation rate（10.82） increased by 3.54%. Furthermore, the rate of methylation polymorphic fragments in ＂Yubei＂ was higher than that in the control group, and class B was the dominant type. Therefore, in the selected breeding of the scallop, DNA methylation level and pattern modified correspondingly, which implies that the DNA methylation plays an important role in regulating functional gene expression associated with the growth and resistance traits. This study may provide new insights into understanding of epigenetic regulatory mechanism in bivalve.

关 键 词：虾夷扇贝 “玉贝” DNA甲基化 甲基化敏感扩增多态性(MSAP) 

分 类 号：Q75[生物学—分子生物学]