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      Dynamic DNA methylation reconfiguration during seed development and germination

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          Abstract

          Background

          Unlike animals, plants can pause their life cycle as dormant seeds. In both plants and animals, DNA methylation is involved in the regulation of gene expression and genome integrity. In animals, reprogramming erases and re-establishes DNA methylation during development. However, knowledge of reprogramming or reconfiguration in plants has been limited to pollen and the central cell. To better understand epigenetic reconfiguration in the embryo, which forms the plant body, we compared time-series methylomes of dry and germinating seeds to publicly available seed development methylomes.

          Results

          Time-series whole genome bisulfite sequencing reveals extensive gain of CHH methylation during seed development and drastic loss of CHH methylation during germination. These dynamic changes in methylation mainly occur within transposable elements. Active DNA methylation during seed development depends on both RNA-directed DNA methylation and heterochromatin formation pathways, whereas global demethylation during germination occurs in a passive manner. However, an active DNA demethylation pathway is initiated during late seed development.

          Conclusions

          This study provides new insights into dynamic DNA methylation reprogramming events during seed development and germination and suggests possible mechanisms of regulation. The observed sequential methylation/demethylation cycle suggests an important role of DNA methylation in seed dormancy.

          Electronic supplementary material

          The online version of this article (doi:10.1186/s13059-017-1251-x) contains supplementary material, which is available to authorized users.

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          Most cited references49

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          TopHat2: accurate alignment of transcriptomes in the presence of insertions, deletions and gene fusions

          TopHat is a popular spliced aligner for RNA-sequence (RNA-seq) experiments. In this paper, we describe TopHat2, which incorporates many significant enhancements to TopHat. TopHat2 can align reads of various lengths produced by the latest sequencing technologies, while allowing for variable-length indels with respect to the reference genome. In addition to de novo spliced alignment, TopHat2 can align reads across fusion breaks, which can occur after genomic translocations. TopHat2 combines the ability to identify novel splice sites with direct mapping to known transcripts, producing sensitive and accurate alignments, even for highly repetitive genomes or in the presence of pseudogenes. TopHat2 is available at http://ccb.jhu.edu/software/tophat.
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            Differential gene and transcript expression analysis of RNA-seq experiments with TopHat and Cufflinks.

            Recent advances in high-throughput cDNA sequencing (RNA-seq) can reveal new genes and splice variants and quantify expression genome-wide in a single assay. The volume and complexity of data from RNA-seq experiments necessitate scalable, fast and mathematically principled analysis software. TopHat and Cufflinks are free, open-source software tools for gene discovery and comprehensive expression analysis of high-throughput mRNA sequencing (RNA-seq) data. Together, they allow biologists to identify new genes and new splice variants of known ones, as well as compare gene and transcript expression under two or more conditions. This protocol describes in detail how to use TopHat and Cufflinks to perform such analyses. It also covers several accessory tools and utilities that aid in managing data, including CummeRbund, a tool for visualizing RNA-seq analysis results. Although the procedure assumes basic informatics skills, these tools assume little to no background with RNA-seq analysis and are meant for novices and experts alike. The protocol begins with raw sequencing reads and produces a transcriptome assembly, lists of differentially expressed and regulated genes and transcripts, and publication-quality visualizations of analysis results. The protocol's execution time depends on the volume of transcriptome sequencing data and available computing resources but takes less than 1 d of computer time for typical experiments and ∼1 h of hands-on time.
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              Establishing, maintaining and modifying DNA methylation patterns in plants and animals.

              Cytosine DNA methylation is a stable epigenetic mark that is crucial for diverse biological processes, including gene and transposon silencing, imprinting and X chromosome inactivation. Recent findings in plants and animals have greatly increased our understanding of the pathways used to accurately target, maintain and modify patterns of DNA methylation and have revealed unanticipated mechanistic similarities between these organisms. Key roles have emerged for small RNAs, proteins with domains that bind methylated DNA and DNA glycosylases in these processes. Drawing on insights from both plants and animals should deepen our understanding of the regulation and biological significance of DNA methylation.
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                Author and article information

                Contributors
                ecker@salk.edu
                Journal
                Genome Biol
                Genome Biol
                Genome Biology
                BioMed Central (London )
                1474-7596
                1474-760X
                15 September 2017
                15 September 2017
                2017
                : 18
                : 171
                Affiliations
                [1 ]ISNI 0000 0001 0662 7144, GRID grid.250671.7, Plant Biology Laboratory, , The Salk Institute for Biological Studies, ; La Jolla, CA 92037 USA
                [2 ]ISNI 0000 0001 0662 7144, GRID grid.250671.7, Genomic Analysis Laboratory, , The Salk Institute for Biological Studies, ; La Jolla, CA 92037 USA
                [3 ]ISNI 0000 0001 2222 0432, GRID grid.416835.d, Institute of Agrobiological Sciences, , National Agriculture and Food Research Organization, ; Tsukuba, Ibaraki 305-8602 Japan
                [4 ]ISNI 0000 0001 0662 7144, GRID grid.250671.7, Howard Hughes Medical Institute, The Salk Institute for Biological Studies, ; La Jolla, CA 92037 USA
                Article
                1251
                10.1186/s13059-017-1251-x
                5599895
                28911331
                545b1587-0d8a-45bb-ad2d-15d9196c1087
                © The Author(s). 2017

                Open AccessThis article is distributed under the terms of the Creative Commons Attribution 4.0 International License ( http://creativecommons.org/licenses/by/4.0/), which permits unrestricted use, distribution, and reproduction in any medium, provided you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The Creative Commons Public Domain Dedication waiver ( http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated.

                History
                : 10 February 2017
                : 7 June 2017
                Funding
                Funded by: FundRef http://dx.doi.org/10.13039/501100001691, Japan Society for the Promotion of Science;
                Award ID: 17H05851
                Award Recipient :
                Funded by: FundRef http://dx.doi.org/10.13039/100000936, Gordon and Betty Moore Foundation;
                Award ID: GBMF 3034
                Award Recipient :
                Categories
                Research
                Custom metadata
                © The Author(s) 2017

                Genetics
                arabidopsis thaliana,dna methylation,embryogenesis,germination,dry seed
                Genetics
                arabidopsis thaliana, dna methylation, embryogenesis, germination, dry seed

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