Burrows Samuel, Dorussen Delfi, Crudgington Joseph, Di Santolo Giorgia, Simmonds James, Catoni Marco, Borrill Philippa
Department of Crop Genetics, John Innes Centre, Norwich Research Park, Norwich NR4 7UH, UK.
School of Biosciences, University of Birmingham, Birmingham B15 2TT, UK.
J Exp Bot. 2025 Jun 17;76(9):2500-2516. doi: 10.1093/jxb/eraf016.
DNA methylation plays important roles in gene expression, transposable element silencing, and genome stability. Altering DNA methylation could generate additional phenotypic variation for crop breeding, however the lethality of epigenetic mutants in crop species has hindered its investigation. Here, we exploit partial redundancy between homoeologues in polyploid wheat to generate viable mutants in the DNA methyltransferase 1-1 (MET1-1) gene with altered methylation profiles. In Triticum turgidum (tetraploid wheat) and Triticum aestivum (hexaploid wheat), we found under-representation of higher order mutants (5/6 and 6/6 mutant met1-1 copies in hexaploid wheat and 3/4 and 4/4 copies in tetraploid wheat) when genotyping segregating seeds and seedlings, due to reduced transmission of null mutant gametes from the paternal and maternal side. The loss of four or more functional copies of MET1-1 results in decreased CG methylation in hexaploid wheat. Changes to gene expression increase stepwise with the number of mutant alleles, suggesting a dosage-dependent effect. We identified heritable changes to flowering and awn phenotypes which segregate independently of MET1-1. Together our results demonstrate that polyploidy can be leveraged to generate quantitative changes to CG methylation without the lethal consequences observed in other crops.
DNA甲基化在基因表达、转座元件沉默和基因组稳定性中发挥着重要作用。改变DNA甲基化可为作物育种产生额外的表型变异,然而,作物物种中表观遗传突变体的致死性阻碍了对其的研究。在此,我们利用多倍体小麦同源基因之间的部分冗余,在DNA甲基转移酶1-1(MET1-1)基因中产生具有改变的甲基化谱的可行突变体。在硬粒小麦(四倍体小麦)和普通小麦(六倍体小麦)中,我们在对分离的种子和幼苗进行基因分型时发现,高阶突变体(六倍体小麦中5/6和六倍体小麦中6/6突变的met1-1拷贝,四倍体小麦中3/4和4/4拷贝)的代表性不足,这是由于来自父本和母本的无效突变配子的传递减少。MET1-1四个或更多功能拷贝的缺失导致六倍体小麦中CG甲基化减少。基因表达变化随着突变等位基因数量的增加而逐步增加,表明存在剂量依赖性效应。我们鉴定出开花和芒表型的可遗传变化,这些变化与MET1-1独立分离。我们的结果共同表明,多倍体可用于产生CG甲基化的定量变化,而不会出现其他作物中观察到的致死后果。