| 运用MSAP技术测定谷子基因组DNA胞嘧啶甲基化水平 |
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| 引用本文： | 张雨欣,阮亚男,赵宸,薛敏敏,李博,王晶晶,刘洋,王凯玺,王红艳.运用MSAP技术测定谷子基因组DNA胞嘧啶甲基化水平[J].生物工程学报,2019,35(2):263-269. |
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| 作者姓名： | 张雨欣 阮亚男 赵宸 薛敏敏 李博 王晶晶 刘洋 王凯玺 王红艳 |
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| 作者单位： | 1 辽宁大学 生命科学院 植物表观遗传与进化实验室，辽宁 沈阳 110036,1 辽宁大学 生命科学院 植物表观遗传与进化实验室，辽宁 沈阳 110036,1 辽宁大学 生命科学院 植物表观遗传与进化实验室，辽宁 沈阳 110036,1 辽宁大学 生命科学院 植物表观遗传与进化实验室，辽宁 沈阳 110036,1 辽宁大学 生命科学院 植物表观遗传与进化实验室，辽宁 沈阳 110036,1 辽宁大学 生命科学院 植物表观遗传与进化实验室，辽宁 沈阳 110036,1 辽宁大学 生命科学院 植物表观遗传与进化实验室，辽宁 沈阳 110036,2 辽宁省水土保持研究所，辽宁 朝阳 122000,1 辽宁大学 生命科学院 植物表观遗传与进化实验室，辽宁 沈阳 110036 |
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| 基金项目： | 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。 |
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| 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。
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| 关 键 词： | DNA甲基化，谷子，甲基化敏感扩增多态性 (MSAP) |
| 收稿时间： | 2018/5/25 0:00:00 |
Analysis of genomic DNA methylation level in foxtail millet by Methylation Sensitive Amplified Polymorphism |
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| Yuxin Zhang,Yanan Ruan,Chen Zhao,Minmin Xue,Bo Li,Jingjing Wang,Yang Liu,Kaixi Wang and Hongyan Wang.Analysis of genomic DNA methylation level in foxtail millet by Methylation Sensitive Amplified Polymorphism[J].Chinese Journal of Biotechnology,2019,35(2):263-269. |
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| Authors: | Yuxin Zhang Yanan Ruan Chen Zhao Minmin Xue Bo Li Jingjing Wang Yang Liu Kaixi Wang and Hongyan Wang |
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| Institution: | 1 Laboratory of Plant Epigenetics and Evolution, School of Life Sciences, Liaoning University, Shenyang 110036, Liaoning, China,1 Laboratory of Plant Epigenetics and Evolution, School of Life Sciences, Liaoning University, Shenyang 110036, Liaoning, China,1 Laboratory of Plant Epigenetics and Evolution, School of Life Sciences, Liaoning University, Shenyang 110036, Liaoning, China,1 Laboratory of Plant Epigenetics and Evolution, School of Life Sciences, Liaoning University, Shenyang 110036, Liaoning, China,1 Laboratory of Plant Epigenetics and Evolution, School of Life Sciences, Liaoning University, Shenyang 110036, Liaoning, China,1 Laboratory of Plant Epigenetics and Evolution, School of Life Sciences, Liaoning University, Shenyang 110036, Liaoning, China,1 Laboratory of Plant Epigenetics and Evolution, School of Life Sciences, Liaoning University, Shenyang 110036, Liaoning, China,2 Liaoning Institute of Soil and Water Conservation, Chaoyang 122000, Liaoning, China and 1 Laboratory of Plant Epigenetics and Evolution, School of Life Sciences, Liaoning University, Shenyang 110036, Liaoning, China |
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| 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 EcoR 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. |
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| Keywords: | DNA methylation foxtail millet (Setaria italica) methylation-sensitive amplification polymorphism (MSAP) |
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