Abstract

Genomic imprinting is an epigenetic phenomenon established in the gametes prior to fertilization that causes differential expression of parental alleles, mainly in the endosperm of flowering plants. The overlap between previously identified panels of imprinted genes is limited. To investigate imprinting, we used high-resolution sequencing data acquired with sequence-capture technology. We present a bioinformatics pipeline to assay parent-of-origin allele-specific expression and report more than 300 loci with parental expression bias in Arabidopsis (Arabidopsis thaliana). In most cases, the level of expression from maternal and paternal alleles was not binary, instead supporting a differential dosage hypothesis for the evolution of imprinting in plants. To address imprinting regulation, we systematically employed mutations in regulative epigenetic pathways suggested to be major players in the process. We established the mechanistic mode of imprinting for more than 50 loci regulated by DNA methylation and Polycomb-dependent histone methylation. However, the imprinting patterns of most genes were not affected by these mechanisms. To this end, we also demonstrated that the RNA-directed DNA methylation pathway alone does not substantially influence imprinting patterns, suggesting that more complex epigenetic pathways regulate most of the identified imprinted genes.

Highlights

  • Genomic imprinting is an epigenetic phenomenon established in the gametes prior to fertilization that causes differential expression of parental alleles, mainly in the endosperm of flowering plants

  • We report that even though the majority of paternally expressed genes are imprinted by Polycomb Repressive Complex2 (PRC2), only one-third of the maternally biased genes can be explained by MET1, suggesting the involvement of further mechanisms

  • Total RNA from seeds was harvested from dissected siliques at 4 d after pollination (DAP) and reverse transcribed to complementary DNA

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Summary

Introduction

Genomic imprinting is an epigenetic phenomenon established in the gametes prior to fertilization that causes differential expression of parental alleles, mainly in the endosperm of flowering plants. The importance of a balanced parental gene expression for proper development of progeny has been demonstrated in both mammals and plants (Barton et al, 1984; Birchler, 1993) In this respect, and in analogy to the mammalian placenta, the endosperm is a site of genomic imprinting, an epigenetic phenomenon that leads to parent-of-origin-dependent expression of genes due to non-DNA sequence-based mechanisms established in the male and female germline (Feil and Berger, 2007; Nowack et al, 2010). Due to the limited overlap between large-scale screens for imprinted genes, the number of imprinted genes and the fraction of maternally and paternally biased genes are still controversial (Schon and Nodine, 2017)

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