Production of Virus-free Clones from Unfertilized Ovules of Satsuma Mandarin Cultivars

In order to produce Tristeza virus-free plants from Satsuma mandarin (Citrus unshiu) cultivars (Ishikawa, Sugiyama and Myogawa), unfertilized ovules of plants suspected of CTV (Citrus Tristeza virus), from Mazandaran Mahdasht orchard were cultured on 4 media. Embryogenesis of nucellus tissue of ovules from Ishikawa and Sugiyama cultivars on MS medium were 31% and 25% and on MT medium from Ishikawa was 40.5%. Unfertilized ovules of Myogawa did not germinate on all media. Nucellar embryos of Ishikawa and Sugiyama developed on MT+GA3+ME were 83% and 95%, respectively. Embryo differentiation, leaf and plantlet formation were observed on MT+GA3+ADE+ME medium. In the stage of two leaves, the plantlets’ growth under 1500 lux illumination was twice its growth under 800 lux illumination. Budding of in vitro plantlets from both cultivars on susceptible to Tristeza and virus-free seedlings of Key Lime did not show any infection by Tristeza.