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Throughout leaf development, cell expansion is dynamic and driven by the balance between local cell wall mechanical properties and the intracellular turgor pressure that overcomes the stiffness of the cell wall leading to plastic deformation. The epidermal pavement cells in most leaves begin development as small, polygonally shaped cells, but in mature leaves epidermal pavement cells are often shaped as highly lobed puzzle pieces. However, the developmental and biomechanical trajectories between these two end points have not before been fully characterized. Here we characterized how epidermal pavement cell size and shape, cell wall thickness, and hydraulic traits change during leaf expansion in the tropical understory fern Microsorum grossum (Polypodiaceae). As fronds expanded by approximately two orders of magnitude in size, epidermal pavement cells became increasingly lobed as cell walls thickened. Furthermore, the timing of these developmental changes varied across the lamina, start first near the frond base and midrib, followed by more apical and lateral regions. During expansion, fronds also underwent substantial physiological changes: as cells expanded and cell walls thickened, intracellular turgor pressure and the bulk cell wall modulus of elasticity both increased while the water potential at turgor loss and the minimum epidermal conductance to water vapor both decreased. These results highlight the dynamic coordination between anatomical and physiological traits throughout leaf development, provide valuable data for biophysical modeling of leaf development, and highlight the vulnerability of developing leaves to drought conditions.more » « lessFree, publicly-accessible full text available September 3, 2026
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