A Wox3-patterning module organizes planar growth in grass leaves and ligules

Grass leaves develop from a ring of primordial initial cells within the periphery of the shoot apical meristem, a pool of organogenic stem cells that generates all of the organs of the plant shoot. At maturity, the grass leaf is a flattened, strap-like organ comprising a proximal supportive sheath surrounding the stem and a distal photosynthetic blade. The sheath and blade are partitioned by a hinge-like auricle and the ligule, a fringe of epidermally derived tissue that grows from the adaxial (top) leaf surface. Together, the ligule and auricle comprise morphological novelties that are specific to grass leaves. Understanding how the planar outgrowth of grass leaves and their adjoining ligules is genetically controlled can yield insight into their evolutionary origins. Here we use single-cell RNA-sequencing analyses to identify a 'rim' cell type present at the margins of maize leaf primordia. Cells in the leaf rim have a distinctive identity and share transcriptional signatures with proliferating ligule cells, suggesting that a shared developmental genetic programme patterns both leaves and ligules. Moreover, we show that rim function is regulated by genetically redundant Wuschel-like homeobox3 (WOX3) transcription factors. Higher-order mutations in maize Wox3 genes greatly reduce leaf width and disrupt ligule outgrowth and patterning. Together, these findings illustrate the generalizable use of a rim domain during planar growth of maize leaves and ligules, and suggest a parsimonious model for the homology of the grass ligule as a distal extension of the leaf sheath margin. Grass ligules comprise the boundary between the leaf base and the photosynthetic blade; their homology has been debated for over 200 years. Cell-specific gene expression analyses suggest that ligules are homologous to the margins of the leaf base.

Automated summary

Grass leaves develop from a ring of initial cells and comprise a sheath and a blade separated by a hinge-like auricle and the ligule. Single-cell RNA-sequencing analyses reveal a distinct cell type at the margins of maize leaf primordia, which shares transcriptional signatures with proliferating ligule cells. Wuschel-like homeobox3 (WOX3) transcription factors regulate the function of the rim domain, affecting leaf width and ligule outgrowth.