人工合成甘蓝型油菜及其亲本的甲基化变异模式分析  

Analysis of DNA Methylation Patterns in Resynthesized Brassica napus and Diploid Parents

作　　者：谢涛[1] 戎浩[1] 蒋金金[1] 孔月琴[1] 冉丽萍[1] 吴健[1] 王幼平[1] 

机构地区：[1]扬州大学生物科学与技术学院,江苏扬州225009

出　　处：《作物学报》2016年第4期513-524,共12页Acta Agronomica Sinica

基　　金：国家重点基础研究计划(973计划)项目(2015CB150201);中国和江苏省博士后基金(2014M561719;2015T80591;1401078B)资助~~

摘　　要：甘蓝型油菜作为多倍体起源和发生的历史较短,遗传背景较为狭窄,人工合成甘蓝型油菜可作为植物多倍化研究的优选模型,本文以人工合成的甘蓝型油菜为材料,通过HPLC分析发现白菜型油菜和甘蓝的甲基化率分别为8.33%和15.88%,2个杂种株系的全基因组甲基化水平介于双亲之间,分别为10.29%和12.83%。MSAP分析发现杂种F_1代及其亲本的甲基化水平存在明显差异(白菜型油菜<杂种F_1<甘蓝),杂种F_1代的甲基化变异(23.71%)中来自A、C基因组的变异分别占6.60%和10.16%。MSAP差异性条带的序列分析发现多倍化过程中与甲基化变化相关的基因参与了多种生物学过程,且差异甲基化基因在人工合成甘蓝型油菜及其亲本间的表达差异与甲基化修饰模式是一致的。本研究为了解甘蓝型油菜多倍化过程中发生的表观变异奠定了基础。The genetic background of Brassica napus, one of the most important oil crops in China, is relatively narrow due to the short history of its polyploid origin. Resynthesized B. napus provides an optimal model for researches on plant polyploidization. In the present study, we compared the DNA methylation levels in synthesized B. napus（F1 generation） and diploid parents using high-performance liquid chromatography（HPLC） and DNA methylation-sensitive amplification polymorphism（MSAP） analysis. HPLC analysis indicated methylation rates of 8.33% and 15.88% in B. rapa and B. oleracea, respectively. While the methylation rate of two hybrids was 10.29% and 12.83%, which were in-between of the parents＇ values. MSAP analysis proved the different methylation levels in F1 generation and diploids, with the lowest and highest methylation levels identified in B. rapa and B. oleracea, respectively. Variance of the DNA methylation in F1 was 23.71%, among which 6.60% and 10.16% were inherited from A and C genome, respectively. Sequence analysis of MSAP polymorphic fragments indicated genes involved in multiple molecular functions were changed during polyploidization. Expression analysis of these genes agreed to the corresponding methylation changes. This study provides preliminary basis for understanding epigenetic variations during B. napus polyploidization.

关 键 词：甘蓝型油菜 人工合成种 多倍化 甲基化 

分 类 号：S565.4[农业科学—作物学]