家蚕不同化性品系间DNA甲基转移酶基因转录特性及胚胎早期甲基化水平差异分析  

Transcriptional Characteristics of DNA Methyltransferase Genes and DNA Methylation Levels in Early Embryonic Stages of Different Voltinism Strains of Bombyx mori

作　　者：胡超超 朱娟[1,2] 陈艳花 王梅仙 鲁月 蒋涛 沈兴家[1,2] Hu Chaochao;Zhu Juan;Chen Yanhua;Wang Meixian;Lu Yue;Jiang Tao;Shen Xingjia(Jiangsu Key Laboratory of Sericulture Biology and Biotechnology,Jiangsu University of Science and Technology,Zhenjang Jiangsu 212100,China;Key Laboratory of Silkworm and Mulberry Genetic Improvement,Ministry of Agriculture and Rural Affairs,Sericultural Research Institute,Chinese Academy of Agricultural Sciences,Zhenjiang Jiangsu 212100,China;College of Marine and Bioengineering,Yancheng Institute of Technology,Yancheng Jiangsu 224000,China)

机构地区：[1]江苏科技大学江苏省蚕桑生物学与生物技术重点实验室,江苏镇江212100 [2]中国农业科学院蚕业研究所农业农村部蚕桑遗传改良重点实验室,江苏镇江212100 [3]盐城工学院海洋与生物工程学院,江苏盐城224000

出　　处：《蚕业科学》2023年第1期30-36,共7页ACTA SERICOLOGICA SINICA

基　　金：国家自然科学基金项目(32072791,32102609);江苏省研究生科研创新项目(KYCX21-3436)。

摘　　要：DNA甲基化修饰参与昆虫胚胎发育。家蚕是典型的胚胎滞育昆虫,为探明家蚕DNA甲基化在胚胎发育及滞育调控中的作用,以不同化性家蚕品系为材料,包括一化性品系安康4号(V^(1)AK4)和鲁一(V^(1)Lu1)、二化性品系秋丰(V^(2)QF)和P50(V^(2)P50)、有滞育多化性品系四海(V^(3)SH)、无滞育多化性品系Nistari(V^(3)Nistari),利用qRT-PCR技术分析家蚕甲基化酶基因BmDnmt在家蚕不同化性品系的表达水平,并利用免疫荧光法测定胚胎发育早期DNA甲基化水平。结果显示,在胚胎发育早期BmDnmt1和BmDnmt2在V^(1)AK4、V^(2)P50和V^(3)Nistari品系中表达水平较高,在V^(3)Nistari品系中BmDnmt1的表达水平显著高于V^(3)SH品系;在幼虫期,BmDnmt1和BmDnmt2在多化性家蚕卵巢或精巢组织中表达水平显著高于二化性家蚕,且它们在V^(3)Nistari中的表达水平显著高于V^(3)SH。采用免疫荧光法测定子代蚕卵的DNA甲基化水平,发现不同化性品系中DNA甲基化水平整体呈上升趋势,在产卵后96 h,多化性品系的DNA甲基化水平显著高于二化性品系。以上结果表明,BmDnmt在家蚕不同化性品系胚胎发育早期的表达水平和DNA甲基化水平均存在差异,推测家蚕幼虫期卵巢组织及胚胎发育早期的DNA甲基化水平与家蚕滞育性及化性有关联。研究结果为进一步解析DNA甲基化在家蚕胚胎发育和滞育中的调控机制提供了新的见解。DNA methylation involves in the embryonic development of insect.Bombyx mori is a typical insect with diapause occurring in the early embryonic development stage.To explore the relationship between DNA methylation level and embryonic development and diapause in B.mori,by using different voltinism silkworm strains as materials,including univoltine strains Ankang4(V^(1) AK4)and Luyi(V^(1)Lu1),bivoltine strains Qiufeng(V^(2)QF)and P50(V^(2)P50),polyoltine diapause strain Sihai(V^(3)SH),as well as diapause-free polyvoltine strain Nistari(V^(3) Nis-tari),we analyzed the expression levels of DNA methyltransferase genes(BmDnmts)in different voltinism strains by qRT-PCR technique,and determined the DNA methylation level in early embryonic development by immunofluorescence method.The results showed that in the early embryonic stage BmDnmt1 and BmDnmt2 were expressed at higher levels in V^(1)AK4,V^(2)P50 and V^(3)Nistari strains,and the BmDnmt1 expression level in Vs Nistari was significantly higher than that in V^(3)SH.In the larval stage,the expression levels of BmDnmt1 and BmDnmt2 in the ovary or testis tsse of polyoltine stralns were signlflcantly hlgher than those of bivsltine surains,and their expression levels In V^(3)Nistari were signlilcantly higher than those in V^(3) SH.The DNA methylation level in silkworm egg was measured by immunofluorescence method.The rooulto ohowod that tho DNA mothylation lovolo in difforont voltiniom otraino oxhibitod an ovorall upward trond.At 96 h after oviposition,the DNA methylation levels in polyvoltine strains were significantly higher than that in bivoltine strains.These results indicated that the expression levels of BmDnmts and DNA methylation levels in early embryonic develop-ment differed among different voltinism strains,implying there might be a certain relationship of the DNA methylation levels in the ovary tissues of silkworm larvae and in the early embryonic development with the diapause and voltinism of the silkworm.Our study provides new insights into the further underst

关 键 词：家蚕 DNA甲基化 DNA甲基转移酶 化性 滞育 胚胎发育 

分 类 号：S881.2[农业科学—特种经济动物饲养]