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Analysis of Cytosine Methylation Status in Potato by Methylation-sensitive Amplified Polymorphisms under Low-temperature Stress

机译:低温胁迫下甲基化敏感性扩增多态性分析马铃薯胞嘧啶甲基化状态

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摘要

China is the leading producer of potatoes (Solanum tuberosum) in the world. However, low-temperature stress can severely restrict the yield and quality of potato crops in that country. We used methylation-sensitive amplified polymorphisms (MSAPs) to analyze how such stress might alter the status of cytosine methylation in potato cultivar 'Atlantic'. Over 2400 loci were identified and assessed for DNA methylation in chilling-treated and control plants. The level of cytosine methylation increased from 37.71% to 56.51% after 48 h of treatment but then decreased to 42.14% after 60 h of chilling. Stress-induced changes in the banding patterns of DNA methylation primarily resulted from full methylation of CG (internal cytosine of 5'-CCGG-3') on double DNA strands. We sequenced 78 polymorphic DNA fragments and blasted them against NCBI. Expression patterns of 4 genes were then assessed via semi-quantitative RT-PCR. The expression patterns of genes related to chilling stress appeared to be regulated by the methylation or demethylation status in potato.
机译:中国是世界领先的马铃薯(马铃薯)生产国。但是,低温胁迫会严重限制该国马铃薯作物的产量和质量。我们使用甲基化敏感的扩增多态性(MSAPs)分析了这种胁迫如何改变马铃薯品种“大西洋”中胞嘧啶甲基化的状态。在冷藏处理和对照植物中,鉴定并评估了超过2400个基因座的DNA甲基化。处理48小时后,胞嘧啶甲基化水平从37.71%增加到56.51%,但在冷却60小时后降至42.14%。应力诱导的DNA甲基化条带模式变化主要是由于双DNA链上的CG(5'-CCGG-3'的内部胞嘧啶)完全甲基化所致。我们对78个多态性DNA片段进行了测序,并将其针对NCBI进行了爆炸。然后通过半定量RT-PCR评估4种基因的表达模式。与低温胁迫相关的基因的表达方式似乎受马铃薯中甲基化或脱甲基状态的调节。

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