A special feature found in Amaryllidaceae is that some guard cells of the neighboring stomata form a "connection strand" between their dorsal cell walls. In the present work, this strand was studied in terms of both its composition and its effect on the morphology and function of the stomata in Pancratium maritimum L. leaves. The structure of stomata and their connection strand were studied by light and transmission electron microscopy. FM 4-64 and aniline blue staining and application of tannic acid were performed to detect cell membranes, callose, and pectins, respectively. A plasmolysis experiment was also performed. The composition of the connection strand was analyzed by fluorescence microscopy after immunostaining with several cell-wall-related antibodies, while pectinase treatment was applied to confirm the presence of pectins in the connection strand. To examine the effect of this connection on stomatal function, several morphological characteristics (width, length, size, pore aperture, stomatal distance, and cell size of the intermediate pavement cell) were studied. It is suggested that the connecting strand consists of cell wall material laid through the middle of the intermediate pavement cell adjoining the two stomata. These cell wall strands are mainly comprised of pectins, and crystalline cellulose and extensins were also present. Connected stomata do not open like the single stomata do, indicating that the connection strand could also affect stomatal function. This trait is common to other Amaryllidaceae representatives.
Keywords: cell wall; guard cells; homogalacturonans; kidney-shaped; stomata clustering.