Functionally Diversified Members of the MIR165/6 Gene Family Regulate Ovule Morphogenesis in Arabidopsis thaliana.

The ovules of flowering plants consist of a central embryo sac and surrounding layers of the inner and outer integument. As these structural units eventually give rise to the embryo/endosperm and seed coat, respectively, a precisely organized ovule structure is essential for successful fertilization and seed production. In Arabidopsis thaliana, correct ovule patterning depends on the restricted expression of the CLASS III HOMEODOMAIN LEUCINE ZIPPER (HD-ZIP III) gene PHABULOSA (PHB) in the apical region of the incipient inner integument, which in turn is regulated via post-transcriptional suppression by miR165 and miR166 (miR165/6) derived from multiple MIR165/6 genes. While a common subset of MIR165/6 genes regulate PHB expression in the root meristem, leaf primordium and embryo, it is unknown whether the same MIR165/6 subset also regulate PHB expression during ovule development. Furthermore, it is unclear where in the ovule primordia miR165/6 are produced. Here, we show that a distinct set of MIR165/6 genes that are highly expressed in the small regions of early ovule primordia restrict the PHB expression domain to promote integument formation. MIR165/6 genes that function in ovule development are phylogenetically distinct from those acting in roots and leaf primordia. Taken together, our data suggest that members of the MIR165/6 gene family are diversified in their expression capacity to establish elaborate PHB expression patterns depending on the developmental context, thereby allowing HD-ZIP III transcription factors to regulate multiple aspects of plant development.