2.4. Scarification procedures
Due to the lack of specific guidelines for diverse species, scarification
times were estimated for each accession based on the
subgenus and seed weight. (1) Control–no treatment. (2) concentrated
sulfuric acid (98+%H2SO4) in an ice bath for 30 min–3 h, then
rinsed in running water for 1 h. Then seeds were soaked for 5 min in
a solution of 3 g/L Ca(ClO)2 completely dissolved in water with 3 g/L
Ca(OH)2 and rinsed for 5min in running water. Seeds were rubbed
against a strainer before stratification to remove the carbonized
portions of testa. (3) Sodium hypochlorite (14% NaOCl) at room
temperature for 2–8 h and rinsed for 1 h in running water. Scarified
seeds were placed on germination blotters for stratification
and germination.