Living zygotes of tobacco (Nicotiana Tabacum L.) SR-1 were isolated and cultured in vitro by the microculture technique. Fertile plants were regenerated from the calli derived from cultured zygotes via organogenesis. Ovules were collected 120 h after pollination and used as feeder. MS combined with KM8p was selected as basic medium in the experiment. Zygotes isolated from ovules 108 h after pollination turned out to be suitable material for in vitro culture. Over 80% such zygotes could divide and around 10% of them could grow into calli and regenerate fertile plants.