Identification and analysis of imprinted genes in wild strawberry uncover a regulatory pathway in endosperm development

Abstract Fertilization is a fundamental process that triggers seed and fruit development, but the molecular mechanisms underlying fertilization-induced seed development are poorly understood. Previous research has established AGamous-Like62 (AGL62) activation and auxin biosynthesis in the endosperm as key events following fertilization in Arabidopsis (Arabidopsis thaliana) and wild strawberry (Fragaria vesca). To test the hypothesis that epigenetic mechanisms are critical in mediating the effect of fertilization on the activation of AGL62 and auxin biosynthesis in the endosperm, we first identified and analyzed imprinted genes from the endosperm of wild strawberry. We isolated endosperm tissues from F1 seeds of two wild strawberry Fragaria vesca subspecies, generated endosperm-enriched transcriptomes, and identified candidate Maternally-Expressed and Paternally-Expressed Genes (MEGs and PEGs). Through bioinformatic analyses, we identified four imprinted genes that may be involved in regulating the expression of FveAGL62 and auxin biosynthesis genes. We conducted functional analysis of a maternally expressed gene FveMYB98 through CRISPR-knockout and overexpression in transgenic strawberry as well as analysis in heterologous systems. FveMYB98 directly repressed FveAGL62 at stage 3 endosperm, which likely serves to limit auxin synthesis and endosperm proliferation. These results provide an inroad into the regulation of early stage seed development by imprinted genes in strawberry, suggest potential function of imprinted genes in parental conflict, and identify FveMYB98 as a regulator of a key transition point in endosperm development.