Unknown,Transcriptomics,Genomics,Proteomics

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Profiling of small RNAs in Arabidopsis ovules

ABSTRACT: We used profiling of Arabidopsis small RNA populations present in the mature ovules to define the potential genome targets of small RNAs during reproductive development. Defining the contributions and interactions of paternal and maternal genomes during embryo development is critical to understand the fundamental processes involved in hybrid vigor, hybrid sterility, and reproductive isolation. To determine the parental contributions and their regulation during Arabidopsis embryogenesis we combined deep-sequencing-based RNA profiling and genetic analyses. At the 2-4 cell stage there is a strong, genome-wide dominance of maternal transcripts, although transcripts are contributed by both parental genomes. At the globular stage the relative paternal contribution is higher, largely due to a gradual activation of the paternal genome. We identified two antagonistic maternal pathways that control these parental contributions. Paternal alleles are initially down-regulated by the chromatin siRNA pathway, linked to DNA and histone methylation, while transcriptional activation requires maternal activity of the histone chaperone complex CAF1. Our results define maternal epigenetic pathways controlling the parental contributions in plant embryos, which are distinct from those regulating genomic imprinting. Profiling of small RNAs in wild type ovules of Arabidopsis

ORGANISM(S): Arabidopsis thaliana

SUBMITTER: Daniel Grimanelli

PROVIDER: E-GEOD-28627 | biostudies-arrayexpress |

REPOSITORIES: biostudies-arrayexpress

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Publications

Maternal epigenetic pathways control parental contributions to Arabidopsis early embryogenesis.

Autran Daphné D Baroux Célia C Raissig Michael T MT Lenormand Thomas T Wittig Michael M Grob Stefan S Steimer Andrea A Barann Matthias M Klostermeier Ulrich C UC Leblanc Olivier O Vielle-Calzada Jean-Philippe JP Rosenstiel Phillip P Grimanelli Daniel D Grossniklaus Ueli U

Cell 20110501 5

Defining the contributions and interactions of paternal and maternal genomes during embryo development is critical to understand the fundamental processes involved in hybrid vigor, hybrid sterility, and reproductive isolation. To determine the parental contributions and their regulation during Arabidopsis embryogenesis, we combined deep-sequencing-based RNA profiling and genetic analyses. At the 2-4 cell stage there is a strong, genome-wide dominance of maternal transcripts, although transcripts ...[more]

PMID: 21620136

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Project description:In animals, maternal gene products deposited into eggs regulate embryonic development before activation of the zygotic genome. In plants, an analogous period of prolonged maternal control over embryogenesis is thought to occur based on gene-expression studies. However, other gene-expression studies and genetic analyses show that some transcripts must derive from the early zygotic genome, implying that the prevailing model does not fully explain the nature of zygotic genome activation in plants. To determine the maternal, paternal and zygotic contributions to the early embryonic transcriptome, we sequenced the transcripts of hybrid embryos from crosses between two polymorphic inbred lines of Arabidopsis thaliana and used single-nucleotide polymorphisms (SNPs) diagnostic of each parental line to quantify parental contributions. Although some transcripts appeared to be either inherited from primarily one parent or transcribed from imprinted embryonic loci, the vast majority of transcripts were produced in near-equal amounts from both maternal and paternal alleles, even during the initial stages of embryogenesis. Results of reporter experiments and analyses of transcripts from genes that are not expressed in sperm and egg indicate early and widespread zygotic transcription. Thus, in contrast to early animal embryogenesis, early plant embryogenesis is mostly under zygotic control. Col-0 and Cvi-0 seed stocks were obtained from the Arabidopsis Biological Resource Center. Plants were grown at 22M-BM-0 C in a Conviron growth chamber with a 16-h light/8-h dark cycle. Flowers were emasculated one day before crossing, and pools of twenty 1-cell/2-cell, 8-cell and ~32-cell embryos were hand-dissected approximately 40, 64 and 78 hours after pollination, respectively. Embryo dissections, RNA isolation, linear amplification of poly(A) RNA and strand-specific RNA-Seq was as described (Nodine and Bartel, 2010, Genes & Development), except that embryos were extensively washed prior to RNA isolation. After removing adaptor sequences, reads were mapped to both the A. thaliana genome and transcript models (Col-0 genome reference, TAIR10) with the Bowtie short-read aligner. Reads were also mapped to a M-bM-^@M-^XpseudoM-bM-^@M-^Y Cvi-0 genome and Cvi-0 transcript models, in which SNPs in the Col-0 genome and transcript models were replaced with Cvi-0 variants (ftp://ftp.arabidopsis.org/home/tair/Sequences/Ecker_Cvi_snps.txt) generated by the 1,001 Genomes Project. Reads that both perfectly and uniquely matched either the genomes or transcript models were retained, and those overlapping transcribed regions of annotated genes were evaluated for overlap with SNPs. To normalize for differences in library sizes, the numbers of reads representing each transcript were divided by the total number of reads matching the genome and transcript models. SNP-overlapping reads were assigned to one of the parental genomes and tallied for each transcript, combining tallies for multiple SNPs within the same transcript.

Dataset Information

Profiling of small RNAs in Arabidopsis ovules

Publications

Maternal epigenetic pathways control parental contributions to Arabidopsis early embryogenesis.

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