Parental conflict driven regulation of endosperm cellularization by a family of Auxin Response Factors

The endosperm is a reproductive tissue supporting embryo development. In most flowering plants, the initial divisions of endosperm nuclei are not succeeded by cellularization; this process occurs only after a specific number of mitotic cycles have taken place. The timing of cellularization significantly influences seed viability and size. Previous research implicated auxin as a key factor in initiating nuclear divisions and determining the timing of cellularization. Here we uncover the involvement of a family of clustered auxin response factors (cARFs) as dosage-sensitive regulators of endosperm cellularization. cARFs, maternally expressed and paternally silenced, are shown to induce cellularization, thereby restricting seed growth. Our findings align with the predictions of the parental conflict theory, suggesting that cARFs represent major molecular targets in this conflict. We further demonstrate a recurring amplification of cARFs in the Brassicaceae, suggesting an evolutionary response to parental conflict by reinforcing maternal control over endosperm cellularization. Our study highlights that antagonistic parental control on endosperm cellularization converges on auxin biosynthesis and signalling. Overall design: Each library was made from around 500 seeds harvested from different crosses. For pPHE1:ARF22, Col-0 x Col-0 and osd1 x Col-0, two biological replicates were used for library preparation. For the CRISPR/Cas9 induced arf13 arf20 mutations, Col-0 x Col-0 and Col-0 x osd1 three biological replicates were used for library preparation.