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The volume of solution differed for each species (dilution depended on the
pollen quantity) and was established empirically. We used Burker counting
camera for the pollen counts after standard methods for leucocytes blood cell
counts. However instead of 100 squares we counted the pollen grains in all 144
squares (in 0,576
Ml
volume) which reflects in the calculation formula. To
reduce bios for each flower we counted 5 doubles (in each of the two counting
nets of the camera). The total number of pollen grains per flower of each
species is calculated considering the dilution following the standard formula:
P = PJ
0,576. lOOO.d,
where
P
is the total number of pollen grains per flower of each species;
P{
-
the mean number of pollen grains for all the counts
(n
= 10) per flower of each
species;
d
is dilution.
RESULTS AND DISCUSSION
The total number of pollen grains vary significantly within the investigated
Gentianaceae
taxa. Most numerous are the pollen grains of
Gentiana lutea
ssp.
symphiandra. Gentianella bulgarica
produce lowest number of pollen grains.
The difference between the two is about 81 times. The number of ovules is
more permanent. Most numerous are the ovules of
Gentiana verna,
and fewest
are the ovules of
Gentianella bulgarica.
The difference between the two is
about 11 times (Table 1, Fig. 1).
P :0 values ratio vary significantly (Table 1). Our results are compared to
those published by Cruden (1973, 1976) and Dafni (1992). Their P :0 ratio
data is correlated with the breeding systems (Table 2). On this base are
discussed the breeding systems of the taxa studied by us (Table 3, Figs. 2-10).
The interpretation of the breeding systems data obtained by the P:0 ratio
indirect method (Table 3, Figs. 2-10) is compared to the breeding systems
data obtained by direct methos such as exclusion techniques, and hand-
pollination in the field published by Kozuharova and co-authors presented in
tables Tables 3 and 4 ( К о ж у х а р о в а , 1994; K o z u h a r o v a , 1994;
K o z u h a r o v a , 1998; K o z u h a r o v a , 2004; K o z u h a r o v a et al., 1994;
K o z u h a r o v a , An c h e v , 2002)
The P :0 ratio values of the investigated flowers of
Gentiana lutea
ssp.
symphyandra
correspond with no doubt to obligate xenogamy (Tables land 3,
Fig. 2). According to the results from direct methos there is no spontaneous
pollination in this subspecies but the flowers are self-compatible (Tables 3, 4).
For the typical subspecies G.
lutea ssp. lutea
L. is reported self-incompatiblelity
(Kery, et al. 2000). The self-compatibility of G.
lutea ssp. symphyandra,
the
inflorescence specifics and pollinator behaviour indicate probability for
geitonogamy ( К о ж у х а р о в а , 1994; K o z u h a r o v a , 1994; K o z u h a r o v a
et al., 1994; K o z u h a r o v a , 1998, 1999).
The P :0 ratio values of the investigated flowers of
Gentiana punctata
coincide with the maximal values of facultative xenogamy and minimal values
30
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