A hypomethylated population of Brassica rapa for forward and reverse epi-genetics.
A hypomethylated population of Brassica rapa for forward and reverse epi-genetics.
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DOI：
10.1186/1471-2229-12-193
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发表时间：
2012-10-20
影响因子：
5.3
通讯作者：
King GJ
中科院分区：
文献类型：
--
作者：
Amoah S;Kurup S;Rodriguez Lopez CM;Welham SJ;Powers SJ;Hopkins CJ;Wilkinson MJ;King GJ
叠加在真核生物染色体 DNA 序列上的表观遗传标记在调节基因表达、本体论和对环境的响应方面提供了灵活性和可塑性。调节胞嘧啶的甲基化状态可以产生表观等位基因，这些表观等位基因已在多个植物系统的特定位点上被检测到和表征，并且有可能产生新颖且相对稳定的表型。还没有系统地尝试探索和利用表观等位基因变异，从而扩大可用于作物改良选择的表型范围。我们开发了一种通过扰动 DNA 甲基化状态来生成新型表观等位基因的方法。 5-氮杂胞苷 (5-AzaC) 提供 5mCG 的选择性靶向，5mCG 在植物中与外显子 DNA 相关。使用 5-AzaC 进行靶向化学干预比甲基转移酶的转基因或突变体调节具有优势，可以在整个基因组中随机生成表观等位基因。我们证明了随机化学诱导的低甲基化为作物改良产生新颖且有价值的变异的潜力。对白菜中 5-AzaC 剂量反应的系统分析指导了自交随机低甲基化群体的产生，用于前向筛选几个农艺性状。几个性状的剂量反应呈 S 形，类似于化学诱变剂（如 EMS）观察到的情况。我们证明了一些表型的跨代遗传。 BraRoAZ 是一个独特的低甲基化群体，包含 1000 个 E2 同胞系。与未处理的对照相比，5-Aza C 处理的品系表现出粗线期染色体上 5mC 免疫染色的减少，以及甲基化敏感扩增多态性 (MSAP) 谱的差异和更多变化。这些品系之间的一系列种子产量和组成性状存在一致的表型变异，包括种子蛋白质含量增加和油含量减少，以及芥酸减少和亚油酸和/或棕榈酸相应增加。每个 5-AzaC 处理的品系代表低甲基化表观等位基因的独特组合。所开发的方法和群体可用于表等位基因变异的正向和反向筛选以及随后的功能和遗传研究。随机低甲基化群体的产生可用于多种作物的表观等位基因发现，并且作为作物改良的干预策略具有相当大的潜力。
Epigenetic marks superimposed on the DNA sequence of eukaryote chromosomes provide agility and plasticity in terms of modulating gene expression, ontology, and response to the environment. Modulating the methylation status of cytosine can generate epialleles, which have been detected and characterised at specific loci in several plant systems, and have the potential to generate novel and relatively stable phenotypes. There have been no systematic attempts to explore and utilise epiallelic variation, and so extend the range of phenotypes available for selection in crop improvement. We developed an approach for generating novel epialleles by perturbation of the DNA methylation status. 5- Azacytidine (5-AzaC) provides selective targeting of 5mCG, which in plants is associated with exonic DNA. Targeted chemical intervention using 5-AzaC has advantages over transgenic or mutant modulation of methyltransferases, allowing stochastic generation of epialleles across the genome. We demonstrate the potential of stochastic chemically-induced hypomethylation to generate novel and valuable variation for crop improvement. Systematic analysis of dose–response to 5-AzaC in B. rapa guided generation of a selfed stochastically hypomethylated population, used for forward screening of several agronomic traits. Dose–response was sigmoidal for several traits, similar to that observed for chemical mutagens such as EMS. We demonstrated transgenerational inheritance of some phenotypes. BraRoAZ is a unique hypomethylated population of 1000 E2 sib lines. When compared to untreated controls, 5-Aza C-treated lines exhibited reduced immuno-staining of 5mC on pachytene chromosomes, and Methylation Sensitive Amplified Polymorphism (MSAP) profiles that were both divergent and more variable. There was coincident phenotypic variation among these lines for a range of seed yield and composition traits, including increased seed protein content and decreased oil content, as well as decreased erucic acid and corresponding increases in linoleic and/or palmitic acid. Each 5-AzaC-treated line represents a unique combination of hypomethylated epialleles. The approach and populations developed are available for forward and reverse screening of epiallelic variation and subsequent functional and inheritance studies. The generation of stochastically hypomethylated populations has utility in epiallele discovery for a wide range of crop plants, and has considerable potential as an intervention strategy for crop improvement.
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影响因子：
5.1
作者：
Fujimoto, Ryo;Sasaki, Taku;Nishio, Takeshi
通讯作者：
Nishio, Takeshi
影响因子：
7.4
作者：
Beaudoin, Frederic;Wu, Xianzhong;Kunst, Ljerka
通讯作者：
Kunst, Ljerka
DOI：
10.1073/pnas.0908755106
发表时间：
2009-11-24
影响因子：
11.1
作者：
Hauben, Miriam;Haesendonckx, Boris;De Block, Marc
通讯作者：
De Block, Marc
影响因子：
7.4
作者：
Barker, Guy C.;Larson, Tony R.;King, Graham J.
通讯作者：
King, Graham J.
影响因子：
4.8
作者：
Brzeski, J;Jerzmanowski, A
通讯作者：
Jerzmanowski, A