Embryological Observation of Flower Development

For embryological processes observation, 23 flowers (KAM, KON, PIL, RUR) were cleared using methyl salicylate method, and divided into two groups. I. Flowers at preanthesis and anthesis stages were cleared in a mixture of 100% ethanol and methyl salicylate (Sigma-Aldrich) in proportions 3:1; 1:1; 1:3, 0:3 two hours each change. II. At postanthesis stages prior to clearing, pre-softening of the tissues using Schiff reagent was applied. Cleared pistils and/or anthers were transferred onto Ray chambers in a drop of methyl salicylate and observed under Nikon Eclipse Ni light microscope with Nomarski differential interference contrast (DIC) optics, equipped with camera Nikon DS-Filc and NIS-Elements BR Viewer imaging software ver. 4.10 (^{57 (link)}, slightly modified). For the study, 10 randomly selected ovules from each flower were analyzed.