Abstract

DNA methylation has the potential to influence plant growth and development through its influence on gene expression. To date, however, the evidence from plant systems is mixed as to whether patterns of DNA methylation vary significantly among tissues and, if so, whether these differences affect tissue-specific gene expression. To address these questions, we analyzed both bisulfite sequence (BSseq) and transcriptomic sequence data from three biological replicates of two tissues (leaf and floral bud) from the model grass species Brachypodium distachyon. Our first goal was to determine whether tissues were more differentiated in DNA methylation than explained by variation among biological replicates. Tissues were more differentiated than biological replicates, but the analysis of replicated data revealed high (>50%) false positive rates for the inference of differentially methylated sites (DMSs) and differentially methylated regions (DMRs). Comparing methylation to gene expression, we found that differential CG methylation consistently covaried negatively with gene expression, regardless as to whether methylation was within genes, within their promoters or even within their closest transposable element. The relationship between gene expression and either CHG or CHH methylation was less consistent. In total, CG methylation in promoters explained 9% of the variation in tissue-specific expression across genes, suggesting that CG methylation is a minor but appreciable factor in tissue differentiation.

Highlights

  • The term ‘epigenetics’ refers to processes beyond genetics and, more concretely, to heritable chromosomal modifications that have the potential to vary during development and stress [1,2]

  • There is a widespread belief that methylation affects gene expression during development [1], relatively few studies have contrasted methylation and gene expression between tissues on a genomic scale

  • We were able to detect a significant signal of differentiation between tissue samples based on two methodological approaches and two measures of variation (DMSs and differentially methylated regions (DMRs))

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Summary

Introduction

The term ‘epigenetics’ refers to processes beyond (epi-) genetics and, more concretely, to heritable chromosomal modifications that have the potential to vary during development and stress [1,2]. Epigenetic modifications include histone variants and DNA methylation. The methylation of cytosines occurs in three sequence contexts: CG, CHG and CHH, where H = A, C or T. All three contexts are usually methylated in repetitive sequences, which serves to limit the transcription and proliferation of transposable elements (TEs) [3]. Genes are often methylated, but typically only in the CG context [4,5,6]. The function of this genebody methylation (gbM) is not yet clear, but potential functions include the exclusion of PLOS ONE | DOI:10.1371/journal.pone.0150002. The function of this genebody methylation (gbM) is not yet clear, but potential functions include the exclusion of PLOS ONE | DOI:10.1371/journal.pone.0150002 March 7, 2016

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