The gynogenic plants, growing in field, were used for ploidy
determination. Young actively growing tendrils (1–3 cm long)
from growing plantlets were used to determine chromosome
number of plants derived from ovary culture according to the
methods of Chen and Qian (2002). Tendrils were pretreated in 8-
hydroxyguinoline for 2 h at 18 8C. Tendrils were then fixed in
Carnoy’s solution (3 vol. glacial acetic acid:2 vol. chloroform:5 -
vol. ethanol) for 24 h, and stored at 4 8C in 70% (v/v) alcohol until
examination. Tendrils were hydrolyzed in 1N HCl for 10 min
at 60 8C and then stained with acetocarmine. Stained tendrils
were squashed in 45% acetic acid. At least 30 meristematic cells
with well-spread chromosomes were observed in each tendril
examined.
2.6. SSR