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MATERIALS AND METHODS
The
Gentianaceae
is a family of annual and perennial herbs (and a few
shrubs) including the gentians and other plants with medicinal „bitter principles1.
The family is cosmopolitan. There are 80 genera with about 900 species. They
occur from sea level to high mountains (H e уwо о d, 1978). In the European
Flora 9 genera are listed (Tu t i n , 1972). On the territory of Bulgaria are
present 5 of them:
Blackstonia
Huds.,
Swertia
L.,
Centaurium
Hill,
Gentianella
Moench,
Gentiana
L. They reproduce by seeds or vegetatively by sprouts
( Пе т р о в а , К о ж у х а р о в , 1982).
The samples for this study (10 tol5 flowers per species) are collected from
their native populations in the mountains during the period May to September
2000 as follows:
Gentiana
L.
date
mountain
G. lutea
ssp.
symphiandra
(Murb.) Ha y e k
30.07.2000
Vitosha
1900 m 14 flowers
G. punctata
L.
12.06.2000
Rila
2000 m 11 flowers
G. asclepiadea
L.
03.09.2000
Vitosha
1800 m 10 flowers
G. pyrenaica
L.
14.07.2000
Rila
2300 m 13 flowers
G. vema
L.
21.05.2000
Vitosha
1900 m 15 flowers
G. utriculosa
L.
25.05.2000
Vitosha
1000 m 15 flowers
Gentianella
Moench
G. bulgarica
(V e 1e n) H о 1u b 30.07.2000
Vitosha
1800 m 11 flowers
Swertia
L.
S. perrennis
L.
30.07.2000
Vitosha
1800 m 12 flowers
Centaurium
H i l l
C. erythraea
Ra f n
30.07.2000
Lozenska
900 m 14 flowers
All the anthers of a flower were collected just before pollen exposure in
a tag numbered jelly capsule separately. The ovary of the same flower was
placed in a small tube (the same tag number as the respective pollen capsule)
and fixed with 50 % ethanol.
LABORATORY TREATMENT
Ovule counting (after Dafni 1992 with slight modification)
Protocol. We cut the ovary longitudinally with a blade on a small petry
dish. Then removed carefully the ovules with a needle and counted them under
a stereomicroscope.
Pollen counting (after Dafni 1992 with modification)
Protocol. We moved the pollen (of each flower) from the jelly capsule to
a graduated glass tube of 5 ml (20°C) using fuxin-ethanol-detergent solution.
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