Plant-RRBS: DNA Methylome Profiling Adjusted to Plant Genomes, Utilizing Efficient Endonuclease Combinations, for Multi-Sample Studies
Martin Schmidt , Magdalena Wołoszyńska , Michiel Van Bel , Frederik Coppens , Mieke Van Lijsebettens
AbstractIn plants, methylation at cytosines often leads to changes in gene expression and inactivation of transposable elements. Changes in cytosine methylation (epimutations) might produce epialleles with distinct phenotypes. We present a genome-wide cytosine methylation profiling method based on bisulfite conversion and next-generation sequencing, which is applicable for plant species with available reference genomes. This so-called plant-RRBS profiling method reproducibly covers specific genomic regions and enriches for coverage of cytosine positions that are suitable for comparative analyses in multi-sample studies in basic biology and breeding studies. The plant-RRBS workflow consists of genomic DNA digestion with coverage-efficient restriction endonuclease combinations followed by a performant library generation and next-generation sequencing and a straightforward, publically available methylation data processing pipeline. Plant-RRBS has a twofold higher ratio of cytosine coverage per covered genome as compared to whole-genome bisulfite sequencing, covering tens of millions of cytosine positions, and allows detection of differential cytosine methylation, which was evaluated using rice epilines.
|Publication size in sheets||0.75|
|Book||Spillane Charles, McKeown Peter (eds.): Plant Epigenetics and Epigenomics. Methods in Molecular Biology, Methods in molecular biology (Clifton, N.J.), vol. 2093, 2020, Humana Press, ISBN 978-1-0716-0178-5, [978-1-0716-0179-2], 272 p., DOI:10.1007/978-1-0716-0179-2|
|Keywords in English||Cytosine methylation, DpnII, MspI and ApeKI endonucleases, Reduced representation bisulfite sequencing (RRBS), Epiline, Breeding, Oryza sativa|
|Score||= 70.0, 15-09-2020, MonographChapterJournalSeries|
|Publication indicators||= 0; : 2018 = 0.404|
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