The effect of pH on the in vitro growth of

pollen tubes at different concentrations

of Boric acid
- Hibiscus rosa-sinensis and Vinca difformis
Introduction
In flowering plants, the pollen tube delivers sperm cells to the embryo sac.
Pollen tube growth proceeds through tip extension and can be affected by
many factors, including temperature, medium osmolarity and the
availability of calcium, zinc and boron (Sawidis and Reiss 1995, Taylor and
Hepler 1997). ulis et al. 2001).
The slightest changes from the ideal concentration of boron and ideal pH
cause great changes in the growth of the pollen tube and many
morphological irregularities once the plant starts to grow.
Ideal pH for the growth of pollen tubes is pH 6.8 to pH 7, for the flowers
taken.
Abstract (Overview)
Pollen from Hibiscus rosa-sinensis and Vinca difformis were used to study the
germination of pollen tube(s) in the presence of Boric acid mixed with
different pH valued 8% sucrose buffers.
2 sets of experiments were carried out with 1:1 and 2:1 concentrations of
boric acid : 8% sucrose buffer
Acidic pH 8% sucrose buffer solutions were used in the order of pH 7
(control), pH 6.4, pH 5.6, pH 4.8 and pH 4.
The growth% was calculated in a given period of time (30 minuets) and
results showed low tolerance to acidic pH conditions.
 Requirements
• Glassware – cavity slides, beakers, pipette, measuring cylinder, petri
dishes and dropper.
• Chemicals – NaOH, HCl/Acetic acid, Sucrose powder, Boric acid and
distilled water
• Microscope and a bright light source

• Biological requirements – flowers of Hibiscus rosa-sinensis and Vinca

difformis
Procedure
1. Prepare buffer solutions by taking 8 gms of sucrose and diluting with
100ml distilled water. Using HCl and NaOH the buffer is prepared
2. Add boric acid to the buffers in the 1:1 and 2:1 concentrations.

3. Place few drops of solution inside the cavity of a clean slide.

4. Dust some pollen off both flowers inside the solution.

5. Place slide in moisture chamber for 30mins.

6. Results were based on both flowers having identical growth.

 Results
% growth out of 20 in 30 minutes

pH of 8% sucrose buffer
1:1 Boric acid 2:1 Boric acid

1 2 3 4 5 mean 1 2 3 4 5 mean

7 95 100 90 100 95 96 90 90 95 100 90 93

6.4 90 80 85 95 90 88 80 80 90 80 90 84

5.6 70 65 75 70 70 70 60 75 70 65 60 66

4.8 0 0 10 0 15 5 0 0 0 5 0 1

4 0 0 0 0 0 0 0 0 0 0 0 0
Graphical Representation
pH v/s %growth
4 4.8 5.6 6.4
7
100 93
84
66 88 96
%GROWTH

80
70
60

40 0
1
20
0 5
0
2:1 boric acid

1:1 boric acid

4 4.8 5.6 6.4 7

1:1 boric acid 0 5 70 88 96
2:1 boric acid 0 1 66 84 93
pH
1:1 boric acid 2:1 boric acid
 120

100
96
93
88
84
80
% GROWTH

70
66
60

40

20

5
0 0 1
4 4.8 5.6 6.4 7
1:1 boric acid 0 5 70 88 96
2:1 boric acid 0 1 66 84 93
pH
1:1 boric acid 2:1 boric acid
 Observations
• The growth of pollen tubes were clearly seen using different microscopy techniques such as
dark field microscopy.
• Lysed cells showed the dispersal of cell matter around the pollen grain which too was clearly
visible.
• A large change in the germination of the pollen tube due to change in pH was seen.
• Boric acid in large amounts seemed to cause lysing of the pollen but helped in germination of
the pollen tube at a much faster and larger rate.
Different microscopic views of pollen, Pollen tube
germination and lysed pollen
Conclusion
• Pollen from both the flowers has a very low tolerance for the change in
pH.
• Pollen tube germination is highly dependent on external factors and
conditions and the amount of Boric acid present decides the rate at which
the pollen tube germinate thus proving that boron is highly essential and
that at different pH there is a visible difference in growth of pollen tube in
the presence of Boric acid.