BARONE LUMAGA, M. R.*, J. E. MICKLE, AND P. DE LUCA. Orto Botanico, Universita' di Napoli Federico II, via Foria, 223, 80139 Napoli, Italy, Department of Botany, North Carolina State University, Box 7612, Raleigh, NC 27695-7612 USA. - Stomatal and epidermal morphology in developing axes of Psilotum nudum.
Apical regions of developing aerial shoots of Psilotum nudum
(L.) Beauv. were studied using both light and scanning electron
microscopy (SEM) with the aim of improving our understanding of early
stages in stomatal and epidermal ontogenesis. Samples for SEM were
fixed in 3% gluteraldehyde in phosphate buffer (0.065M, pH 7),
dehydrated in a graded ethanol series, critical point dried in
CO2 and coated with ca. 30 nm gold-palladium alloy.
Samples for light microscopy were fixed in FAA, dehydrated in a graded
ethanol series and embedded in paraffin with a melting point of 51-53
C. Sections were stained with 0.05% toluidine blue, a general protein
stain. Developing stomata in P. nudum grow from less than 20 µm to
about 70 µm long. SEM observations of specimens show 20 µm-long domed
meristemoid cells as the first detectable stage of differentiation of
the guard cell mother cells (GCMCs). First detection of a furrow
dividing the guard cells is at ca. 30 µm long. Deposition of wax that
will eventually cover the entire stomatal aperture begins In 70 µm
long stomata. Longitudinal sections of the shoot show GCMCs with an
unusual hourglass-like appearance and the presence of a cytoplasmic
protein net surrounding the nucleus and occupying all of the
developing guard cell. The cytoplasmic net appears more compact when
the developing furrow becomes deeper, and is probably involved as an
organizer of cell morphology. This staining pattern was detected only
in the cytoplasm of developing guard cells and not in surrounding
epidermal cells. This information will be used as a basis for ongoing
studies comparing Psilopsida, whose systematics are still equivocal,
to other taxa.
Key words: ontogeny, Psilotum, stomata