Excretion and folding of plasmalemma function to accommodate alterations in guard cell volume during stomatal closure in Vicia faba L.

Stomatal movement results in large and repetitive changes in cell volume and consequently surface area. While endocytosis has been extensively studied and is thought to be a major mechanism for accommodating the volume changes as evidenced mainly by fluorescent labelling and confocal imaging, studies at the ultrastructural level in intact guard cells of stomata regulated by natural factors have never been reported. Here, it is reported that excretion and folding of the plasmalemma are critical for accommodating the volume alterations in intact guard cells in Vicia faba L. Using transmission electron microscopy in combination with laser confocal microscopy, it was observed that in fully opened stomata the plasmalemma was smooth and tightly adhered to the cell walls while a whole large vacuole appeared in the cell. In the closed stomata, besides vacuole fragmentation, endocytosis of the tonoplast rather than the plasmalemma commonly occurred. Importantly, in stomata where pore closure was induced by circadian rhythm or CO(2), numerous tiny vesicles were found outside the plasmalemma and, moreover, extensive folding of the plasmalemma could also be found in some regions of the cells. Additionally, an unknown structure was found at the interface between the plasmalemma and cell walls, especially in those areas of the cell where extensive folding occurred, suggesting that plasmalemma turnover is possibly associated with an interaction between the plasmalemma and cell walls. Collectively, the results strongly indicate that excretion and folding of the plasmalemma are critical for the accommodation of the cell volume alterations during stomatal movement.