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13. SUPPLEMENTARY MATERIALS
ANNEX 13.2. SEEDLING ROOT FLUORESCENCE TEST (THE GENTNER’S TEST)
Fluorescence tests were conducted according to Niemyski and Budzyńska (1972) but
with own modifications that enabled the standardization of the protocol.
Seed germination
About 30-40 seeds were hand sown in sterile Petri dishes of 10 cm in diameter. White
filter paper moistened with tap water was a germination substrate. To keep moisture the internal
side of each Petri dish top was covered with wet filter paper. Filter paper humidity was
checked daily. Seeds were incubated at 24 o C in darkness for six days. Seminal roots of 1-2
mm were visible at this moment. Then roots were checked for fluorescence under UV light. If
no fluorescence was visible, seeds were transferred onto glass plates.
Seedling root fluorescence test
Glass plates of 16- by 21 cm were covered with a sheet of white filter paper of 20- by
25 cm. The edges sticking out were folded under the glass plates. Seed places were marked
with a pencil at 6-7 cm from the top edge of the glass plate (Figure 13.2.1). Then the filter
paper was moistened with tap water.
In total 20 seeds were placed on a glass plate so that the embryos were oriented into
the bottom. Then seeds were protected from falling dawn with a wet filter paper stripe of 2.0-
by 25 cm (Figure 13.2.1A). Maximum eight glass plates were laid in glass boxes to give a tilt
65 o relative to the bottom. This enabled the roots to go down and adhere tight to the filter
paper. The last glass plate was covered with an empty glass plate to ensure the comparable
conditions for all seeds. Glass boxes were filled with tap water up to 1/4 of the volume (Figure
13.2.1B). The level of water was kept at the same level throughout the whole experiment.
The seeds were incubated at 27 o C for 14 days. The fluorescence was determined under UV
light on the 14 th day.