高粱甲基化连锁群A、B的构建及甲基化位点、甲基化模式的分析  

Construction of Methylation Linkage Groups A and B in Sorghum with MSAP and SSR Markers and Analysis of Methylation Sites and Patterns

作　　者：段永红[1,2] 王铭[2] 孙毅[2,3] 杨武德[1] 

机构地区：[1]山西农业大学农学院,山西太谷030801 [2]山西省农业科学院生物技术研究中心,太原030031 [3]农业部黄土高原作物基因资源与种质创制重点实验室,太原030031

出　　处：《中国农业科学》2012年第18期3699-3708,共10页Scientia Agricultura Sinica

基　　金：国家转基因生物新品种培育重大专项(2009ZX08003-017B);山西农业大学博士后基金(112118);山西农业大学博士科研启动项目(xb2011011)

摘　　要：【目的】构建高粱甲基化连锁群A和B,并分析其上的甲基化位点分布及甲基化模式变化。【方法】从高粱品种强优势组合B2V4×1383-2杂交组合获得的F2分离群体(共150个体)为材料,以SSR标记为锚定标记,采用SSR和MSAP标记技术,并用Mapmaker/Exp(Version 3.0)和Map/Draw 2.1软件进行分析。【结果】构建出高粱甲基化连锁群A-a和A-b及甲基化连锁群B-a和B-b,其中,甲基化连锁群A覆盖高粱基因组93.7 cM,包含10个SSR标记、20个MSAP标记,甲基化连锁群B覆盖高粱基因组90.4 cM,包含4个SSR标记、39个MSAP标记;连锁群A-a上甲基化位点仅来源于EcoRⅠ/MspⅠ酶切组合,而其它连锁群上甲基化位点来源于EcoRⅠ/MspⅠ和EcoRⅠ/HpaⅡ2种酶切组合;甲基化连锁群A-b和B-b上各存在一个稍密集的甲基化位点区域,而连锁群B-a上存在一个较密集的甲基化位点区域;基于同一酶切的高粱亲本间有多态性差异,F2群体存在分离,高粱亲本与杂交种F1的甲基化模式有2种类型的变化。【结论】MSAP标记可以检测到大量的甲基化差异片段,结合锚定的SSR标记,能够有效地构建植物基因组的甲基化遗传连锁群或遗传连锁图谱;在连锁群上检测到3个密集的甲基化位点区域,分别位于SSR标记Xtxp 302、Xtxp 96及Xtxp 304附近;在获得的连锁群中,发生去甲基化反应的甲基化位点数高于甲基化水平提高的位点数。[Objective] The genetic linkage groups of A and B with methylation sensitive markers of Sorghum bicolor L. were constructed, and the methylation sites and methylation patterns were analyzed. [Method] F2 segregating population with 150 individuals derived from the sorghum cross BEV4 X 1383-2 was analyzed based on MSAP and SSR markers, and linkage groups were constructed using Map maker/EXP （version 3.0） and Map/Draw 2.1. [ Result ] Methylation linkage groups were constructed. Group A was composed of 30 loci covering 93.7 cM, of which 20 loci were MSAP markers, including 13 loci from EcoR I/Msp I enzyme digestion, 7 loci from EcoR I/Hpa II enzyme digestion, and 10 loci were SSR markers. Group B was composed of 43 loci covering 90.4 cM, of which 39 loci were MSAP markers, including 19 loci from EcoR I/Msp I enzyme digestion, 20 loci from EcoR I /Hpa II enzyme digestion and 4 loci were SSR markers. There were only methylation marker products existed from EcoR I/Msp I enzyme digestion on linkage group A-a, but on other linkage groups, the methylation markers from both EcoR I/Hpa II and EcoR I/Msp I enzyme digestion were found. Additionally, Group A-b and Group B-b each had a slightly dense methylation resign, nearby Xtxp302 and Xtxp304, while, a highly dense methylation regions, nearby Xtxp 296 in linkage group B-a, with clusters of the EcoR I/Msp I and EcoR I/HpalIenzyme digestion MSAP markers were revealed. Based on polymorphism of methylation fragments between parents and their segregation among the F2 population, cytosine methylation patterns between hybrid and their parents were divided into two major groups. [Conclusion] MSAP markers can detect a large number of different methylated fragment, with anchor SSR markers, it is an efficient technique in constructing methylation genetic linkage groups or methylation genetic linkage map. High-density methylation regions were revealed near SSR markers Xtxp 302 on Group A-b, Xtxp 96 on Group B-a and Xtxp 304 on Group B-b. Among these loci the number of demethylated l

关 键 词：高粱 SSR MSAP DNA甲基化 

分 类 号：S514[农业科学—作物学]