Producing Haploid Rose (Rosa hybrida L.) Plants by Anther Culture in Single and Double Layer Media

In this study, the effects of cultivar, plant growth regulators, various culture media (single and double layer), temperature stresses and carbohydrate sources (sucrose, glucose, maltose and their combinations) on callus/embryo formation from microspors in four rose cultivars i.e. „Elegant‟, „Velvet‟, „Candy‟ and „Exotic‟ were investigated. According to the results two rose cultivars „Exotic‟ and „Velvet‟ and combination of BAP (0.5 and 1 mg l-1) and IAA (4 and 8 mg l-1) significantly induced callus higher than other treatments. Frequency of callus induction in double layer media especially in MS and Nitsch and Nitsch as lower media with upper AT3 (with sucrose and lactalbomin hydrolysate) liquid medium (MS/AT3-SL and NN/AT3-SL) was higher than single layer solid medium. Examination of the effects of temperature shocks on callus induction did not improve callus formation campared to control (25°C). Results also showed that callus formation in sucrose-containing Nitsch and Nitsch media was significantly higher than other carbohydrates. In the present study, for the first time, seven embryos were formed in „Exotic‟ cultivar by shed microspore culture in double layer media which were regenerated successfully and ploidy level analysis confirmed that they were haploids.