In flowering induction [1,2], little is known about the dynamics of epigenetic regulation during phase transitions in plants. The plant life cycle is characterized by major transitions in response to integrated internal and environmental cues that require extensive reprogramming of the transcriptome. For seed plants, the transition from a dormant to a non-dormant state of the seed is of major importance for the plant’s success in establishing a new generation. Seed dormancy is defined as the inability of a viable seed to complete germination under favorable conditions [3,4]; the transition to germination requires a specific environmental trigger. Transcriptomic analyses reveal strict spatial and temporal regulation of gene expression during the dormancy-togermination transition [5,6]. However, it is largely unknown how the major dormancy regulators are themselves regulated in response to environmental cues. DNA in the eukaryotic nucleus is organized to form chromatin. The minimal unit, the nucleosome, is composed of the DNA wrapped around complexes of eight histone cores. Histones can be modified at their N-terminus at a number of residues and with POR8 custom synthesis different modifications [7]. Methylation of histones is one mechanism that contributes to the spatial and temporal regulation of gene expression on which growth and development depends in all eukaryotes [8,9]. Depending on the amino acid residue that is modified and the number of methyl groups added to the residue, histone methylation changes (`marks’) can contribute to transcriptional activation or repression. For example, trimethylation of histone H3 lysine 4 (H3K4me3) is an `activating mark’, while trimethylation of lysine 27 of histone 3 (H3K27me3) leads to repression of transcription [7,8]. Histone modifications are important in regulating various life cycle transitions in plants, including the transition from vegetative to reproductive growth in flowering plants [10,11], and from gametophyte to sporophyte development in the moss Physcomitrella patens [12]. To date chromatin modifications have not been explored in mature seeds. Mutants in the polycomb repressive complex 2 (PRC2) display a significant delay in germination and express markers of seed development and dormancy in seedlings [13], leading to the hypothesis that histone methylation pathways play a role in the transition from dormancy to germination and seedling growth. PRC2 acts as a histone methyltransferase (HMTase) on lysine 27 of histone 3 (H3K27me3) leading to gene silencing. Moreover, the Arabidopsis pickle (pkl) mutant, which is affected in Ergocalciferol site H3K27me3 deposition [14,15], exhibits germination-associated phenotypes and expresses markers associated with seed development and dormancy during seedling growth [15]. In this work, we elucidated an important mechanistic aspect underlying the global transcriptional network re-programming during seed dormancy breakage and the transition to germination. This was achieved by following the chromatin dynamics of key regulatory genes with a focus on the two antagonistic marks, H3K4me3 and H3K27me3.Histone Methylation Dynamics in SeedsFigure 1. Schematic representation of Arabidopsis Cvi seed treatments and physiological status of the seeds. (A) The inception of primary seed dormancy occurs during seed maturation. Mature dry seeds are dormant (1) and are maintained in this state when imbibed at 22uC, even for 14 d (2). A sufficiently long period of moist chilling (4uC) will break dormancy (.In flowering induction [1,2], little is known about the dynamics of epigenetic regulation during phase transitions in plants. The plant life cycle is characterized by major transitions in response to integrated internal and environmental cues that require extensive reprogramming of the transcriptome. For seed plants, the transition from a dormant to a non-dormant state of the seed is of major importance for the plant’s success in establishing a new generation. Seed dormancy is defined as the inability of a viable seed to complete germination under favorable conditions [3,4]; the transition to germination requires a specific environmental trigger. Transcriptomic analyses reveal strict spatial and temporal regulation of gene expression during the dormancy-togermination transition [5,6]. However, it is largely unknown how the major dormancy regulators are themselves regulated in response to environmental cues. DNA in the eukaryotic nucleus is organized to form chromatin. The minimal unit, the nucleosome, is composed of the DNA wrapped around complexes of eight histone cores. Histones can be modified at their N-terminus at a number of residues and with different modifications [7]. Methylation of histones is one mechanism that contributes to the spatial and temporal regulation of gene expression on which growth and development depends in all eukaryotes [8,9]. Depending on the amino acid residue that is modified and the number of methyl groups added to the residue, histone methylation changes (`marks’) can contribute to transcriptional activation or repression. For example, trimethylation of histone H3 lysine 4 (H3K4me3) is an `activating mark’, while trimethylation of lysine 27 of histone 3 (H3K27me3) leads to repression of transcription [7,8]. Histone modifications are important in regulating various life cycle transitions in plants, including the transition from vegetative to reproductive growth in flowering plants [10,11], and from gametophyte to sporophyte development in the moss Physcomitrella patens [12]. To date chromatin modifications have not been explored in mature seeds. Mutants in the polycomb repressive complex 2 (PRC2) display a significant delay in germination and express markers of seed development and dormancy in seedlings [13], leading to the hypothesis that histone methylation pathways play a role in the transition from dormancy to germination and seedling growth. PRC2 acts as a histone methyltransferase (HMTase) on lysine 27 of histone 3 (H3K27me3) leading to gene silencing. Moreover, the Arabidopsis pickle (pkl) mutant, which is affected in H3K27me3 deposition [14,15], exhibits germination-associated phenotypes and expresses markers associated with seed development and dormancy during seedling growth [15]. In this work, we elucidated an important mechanistic aspect underlying the global transcriptional network re-programming during seed dormancy breakage and the transition to germination. This was achieved by following the chromatin dynamics of key regulatory genes with a focus on the two antagonistic marks, H3K4me3 and H3K27me3.Histone Methylation Dynamics in SeedsFigure 1. Schematic representation of Arabidopsis Cvi seed treatments and physiological status of the seeds. (A) The inception of primary seed dormancy occurs during seed maturation. Mature dry seeds are dormant (1) and are maintained in this state when imbibed at 22uC, even for 14 d (2). A sufficiently long period of moist chilling (4uC) will break dormancy (.