Production of Microspore-Derived Plants by Anther Culture of Cyclamen Coum

Abstract

Cyclamen is one of the most important ornamental crops sold worldwide as a potted flower for winter production. Cyclamen species take up a wide swathe of habitats across Türkiye. Ten wild Cyclamen species grow naturally in Türkiye and some of them are endemic. This study aimed to produce haploid plants of C. coum using anther culture. The microspore developmental stage was evaluated by staining anther with acetocarmine (%2), and then the stage was correlated with bud size. It was determined that the buds between 7.64 and 8.23 mm had the appropriate bud size for the late uninuclear stage. Anthers were cultured in B5 medium containing different levels of 1-Naphthaleneacetic acid (NAA) (0.1, 1, 2 mgL-1), 2,4-Dichlorophenoxyacetic acid (2,4-D) (0.1, 1, 2 mgL-1), and kinetin (0, 1 mgL-1), 90 gL-1 sucrose and 3 gL-1 gelrite for haploid embryo production. Anthers were kept at 4°C for 4 days after culture. The explants were incubated at 24°C in a completely dark condition until the embryo was formed, then embryos were transferred to hormone-free media in 16:8 hours (light (75 µmolm-2s-1): dark) photoperiod. The experiment was carried out for two years. In the first year, 12 different media were examined in view of regeneration and the experiments were continued with selected 7 media in the second year. The highest callus regeneration rates were %5.71 and 14.5% and the highest embryo induction rates varied between 8.57% and 4.0% in the first and second year respectively. Embryo/callus formation was observed in 7 of a total of 12 different media tested for haploid plant production, and the best media were kinetin (1 mgL-1) + NAA (1, 2 mgL-1) and kinetin (1 mgL-1) +2,4-D (2 mgL-1). Our findings indicated that cold pre-treated anther explants collected at appropriate flower bud size resulted in embryo production. Additionally, B5 medium supplemented with NAA and kinetin ensured successful embryo regeneration from anther explants in wild C. coum.