2. Material and methods
Particular attention was paid to the variability of the vegetative
organs in different periods of ontogeny, which is of diagnostic
importance for determining the sectional and species affiliation (e.g.,
Levichev 1999a, 1999b, 2011, Peterson et al. 2008; Peruzzi et al. 2008).
Also, these features were analyzed on all herbarium sheets found in TARI
and LE. To obtain mature seeds, live samples were placed in a container
and observed in a greenhouse. Identification was carried out according
to existing literature sources (Wendelbo and Rechinger 1990; Levichev
2006; Ali and Levichev 2007; Zarrei et al. 2007, 2011). Additionally,
the anatomical features of the bulb, bulbil and leaves were studied from
the material fixed in ethyl alcohol (70%). Manually, with a razor
blade, longitudinal sections of the bulb, bulbil and cross sections of
various parts of the basal leaf and peduncle were made. Sections were
bleached in sodium hypochlorite for several minutes, then stained twice
with cellulose carmine (phloem) and dilute methyl green (xylem). The
anatomical features of the leaf were examined under a light microscope
Nikon BX53 (Japan) Nikon BX53 (in TARI) and Stemi 2000 SDA (in LE) with
a digital camera mounted on it. The images were then converted into
graphics using Adobe Illustrator CC. 17.0.