Pollen fertility test by germination ability
Culture medium (for 50 samples)
5 g Sucrose (20 % sucrose) C12H22O11
0.25 g Bacto Agar
2.5 ml 10 x stock solution [0.3 g Ca(NO3)2·4H2O and 0.05 g H3BO3 (50 ppm) in 100 ml distilled water]
- Place components of culture medium in a beaker and fill up to 25 ml with distilled water.
- Warm to dissolve in a microwave oven.
- Pour the dissolved solution on a slide glass to an approximate size of a cover slip using a Pasteur pipette.
- Solidify the culture medium in a moisture box (a sealed plastic container with a sheet of wet paper towel on the bottom) at least for 2 hours.
- Strew pollen grains on the medium.
- Incubate pollen in a moisture box at room temperature for 3-16 hours.
- Cut off the edge of the culture medium to the size of 1 cm square.
- Place a drop of 1% acetocarmine and cover with a cover slip.
- Place the slide glass on a heater, and melt slowly the culture medium (don't boil up).
- Remove the slide glass from the heater just after the culture medium is melted.
- Observe pollen tube growth under a microscope.