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运用MSAP技术测定谷子基因组DNA胞嘧啶甲基化水平

Analysis of genomic DNA methylation level in foxtail millet by Methylation Sensitive Amplified Polymorphism

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【作者】 张雨欣阮亚男赵宸薛敏敏李博王晶晶刘洋王凯玺王红艳

【Author】 Yuxin Zhang;Yanan Ruan;Chen Zhao;Minmin Xue;Bo Li;Jingjing Wang;Yang Liu;Kaixi Wang;Hongyan Wang;Laboratory of Plant Epigenetics and Evolution, School of Life Sciences, Liaoning University;Liaoning Institute of Soil and Water Conservation;

【通讯作者】 王红艳;

【机构】 辽宁大学生命科学院植物表观遗传与进化实验室辽宁省水土保持研究所

【摘要】 DNA甲基化是真核生物一种重要的表观修饰形式。为了探讨谷子基因组DNA胞嘧啶甲基化的水平和模式,以谷子Setaria italica的两个品种朝谷58号和豫谷1号为实验材料,利用Eco RⅠ和HpaⅡ/MspⅠ双酶切建立适合于谷子基因组的甲基化敏感扩增多态性(MSAP)分析体系。结果表明,从100对MSAP选扩引物中,筛选出32对MSAP引物组合,在朝谷58号和豫谷1号中分别扩增产生1 615、1 482条清晰可辨且可重复的DNA条带,其中包括3种类型的甲基化条带,朝谷58号和豫谷1号的基因组中CCGG序列胞嘧啶甲基化水平分别为6.93%和8.77%。这种谷子不同品种间甲基化水平和分布位点的差异为从表观遗传学的角度培育新品种提供了初步的理论依据和参考。

【Abstract】 DNA methylation is an important type of epigenetic modification in eukaryotes. In order to research genome-wide methylation levels and patterns in foxtail millet(Setaria italica), the Methylation Sensitive Amplified Polymorphism(MSAP) analysis(employing double digestion with Eco R I and Hpa II/Msp I) was established and applied in two foxtail millet cultivars(Chaogu 58 and Yugu 1). The results showed that 32 pairs of MSAP primers were selected from 100 MSAP primers, and 1 615 and 1 482 clearly distinguishable and reproducible bands were amplified from Chaogu 58 and Yugu 1 respectively, including 3 types of methylation patterns. Cytosine methylation levels of CCGG context in Chaogu 58 and Yugu 1 were characterized as 6.93% and 8.77% respectively. Such different genomic DNA methylation levels between two foxtail millet varieties may provide a preliminary reference for the cultivation of this crop from a novel epigenetic viewpoint.

【基金】 国家自然科学基金(No.31100172);辽宁省高等学校优秀人才支持计划(No.LJQ2013003);辽宁省科技厅自然基金指导计划(No.20180550686);沈阳市科技局项目(No.F16-205-1-49)资助~~
  • 【文献出处】 生物工程学报 ,Chinese Journal of Biotechnology , 编辑部邮箱 ,2019年02期
  • 【分类号】S515
  • 【被引频次】3
  • 【下载频次】273
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