The life cycle of higher plants alternates between a haploid gametophytic generation and a diploid sporophytic generation. The female and male gametes formed during the gametophytic phase fuse during fertilization to generate the sporophytic phase of the life cycle. Most sexually reproducing diploid plants undergo double fertilization, during which the egg cell and the homodiploid central cell are each fertilized by a sperm cell and give rise to the embryo and the triploid endosperm, respectively (Grossniklaus and Schneitz 1998).In analogy to the Greek myth in which the priestess Medea killed her children Pheres and Meidos (Euripides 431 BC; Wolf 1996), a maternal effect mutant identified in a screen for gametophytic mutants in Arabidopsis thaliana was named medea (mea; . Additional alleles of the MEA gene, termed FER-TILIZATION INDEPENDENT SEED DEVELOPMENT (FIS1), as well as mutations at two other loci, FIS2 and FERTILISATION INDEPENDENT ENDOSPERM (FIE or FIS3), were identified in genetic screens for mutants displaying seed development in the absence of fertilization (Ohad et al. 1996;Chaudhury et al. 1997). The three fisclass mutants show a gametophytic maternal effect: all seeds derived from a mutant female gametophyte (50% in a heterozygote) abort irrespective of the paternal allele. Early development of fis embryos is morphologically indistinguishable from that of wild-type siblings. However, fis embryogenesis is delayed after the globular stage and eventually arrests with oversized heart-shaped embryos surrounded by an abnormally proliferated en-

C.Ko Èhler and L.Hennig contributed equally to this workSeed development in angiosperms initiates after double fertilization, leading to the formation of a diploid embryo and a triploid endosperm. The active repression of precocious initiation of certain aspects of seed development in the absence of fertilization requires the Polycomb group proteins MEDEA (MEA), FERTILIZATION-INDEPENDENT ENDOSPERM (FIE) and FERTILIZATION-INDEPENDENT SEED2.Here we show that the Arabidopsis WD-40 domain protein MSI1 is present together with MEA and FIE in a 600 kDa complex and interacts directly with FIE. Mutant plants heterozygous for msi1 show a seed abortion ratio of 50% with seeds aborting when the mutant allele is maternally inherited, irrespective of a paternal wild-type or mutant MSI1 allele. Furthermore, msi1 mutant gametophytes initiate endosperm development in the absence of fertilization at a high penetrance. After pollination, only the egg cell becomes fertilized, the central cell starts dividing prior to fertilization, resulting in the formation of seeds containing embryos surrounded by diploid endosperm. Our results establish that MSI1 has an essential function in the correct initiation and progression of seed development.

Genomic imprinting is an epigenetic phenomenon leading to parent-of-origin specific differential expression of maternally and paternally inherited alleles. In plants, genomic imprinting has mainly been observed in the endosperm, an ephemeral triploid tissue derived after fertilization of the diploid central cell with a haploid sperm cell. In an effort to identify novel imprinted genes in Arabidopsis thaliana, we generated deep sequencing RNA profiles of F1 hybrid seeds derived after reciprocal crosses of Arabidopsis Col-0 and Bur-0 accessions. Using polymorphic sites to quantify allele-specific expression levels, we could identify more than 60 genes with potential parent-of-origin specific expression. By analyzing the distribution of DNA methylation and epigenetic marks established by Polycomb group (PcG) proteins using publicly available datasets, we suggest that for maternally expressed genes (MEGs) repression of the paternally inherited alleles largely depends on DNA methylation or PcG-mediated repression, whereas repression of the maternal alleles of paternally expressed genes (PEGs) predominantly depends on PcG proteins. While maternal alleles of MEGs are also targeted by PcG proteins, such targeting does not cause complete repression. Candidate MEGs and PEGs are enriched for cis-proximal transposons, suggesting that transposons might be a driving force for the evolution of imprinted genes in Arabidopsis. In addition, we find that MEGs and PEGs are significantly faster evolving when compared to other genes in the genome. In contrast to the predominant location of mammalian imprinted genes in clusters, cluster formation was only detected for few MEGs and PEGs, suggesting that clustering is not a major requirement for imprinted gene regulation in Arabidopsis.

The maternally expressed Arabidopsis thaliana Polycomb group protein MEDEA (MEA) controls expression of the MADS-box gene PHERES1 (PHE1). Here, we show that PHE1 is mainly paternally expressed but maternally repressed and that this maternal repression of PHE1 breaks down in seeds lacking maternal MEA activity. Because Polycomb group proteins control parental imprinting in mammals as well, the independent recruitment of similar protein machineries for the imprinting of genes is a notable example of convergent evolution.