Differential expression analysis of sexual and apomictic Boechera uncovers FAS4 as crucial for gametogenesis

During sexual reproduction of higher plants, seed formation is initiated by double fertilization of egg and central cell. In contrast, pseudogamous apomicts form embryos asexually by parthenogenesis of the egg, but initiation of endosperm development still depends on central cell fertilization. It can be envisioned that these differences are determined during gametogenesis and specification of gametophytic cells. To deepen the understanding of the transcriptional basis underlying sexual and apomictic reproduction, we applied tissue type-specific RNA-Seq. We compared expression in reproductive tissues of different Boechera accessions at distinct developmental stages. This confirmed previous evidence for an enrichment of RNA helicases at onset of reproductive development. We further identified a small number of members of this gene family as differentially expressed in female reproductive ovule tissues harbouring mature gametophytes from apomictic and sexual accessions. This included homologues of A. thaliana FASCIATED STEM 4 ( FAS4 ) and of ENHANCED SILENCING PHENOTYPE 3 ( ESP3 ), which have previously been identified as potential candidates for gametogenesis and apomixis, respectively. Unlike in A. thaliana , for either of them additional homologues or copies of related genes are present in Boechera , indicating complex evolutionary histories. As the expression patterns implied potential roles of FAS4 during gametogenesis, we first studied A. thaliana lines carrying mutant alleles. Indeed, we observed defects during male and female gametogenesis and severely reduced transmission efficiencies through both parents. In conclusion, our study identifies FAS4 as crucial for plant reproduction and suggests the potential for sub-functionalization of additional homologous genes in Boechera to shape reproductive development. ### Competing Interest Statement The authors have declared no competing interest.