당근의 약배양으로부터 직접 및 간접 배형성을 통한 식물체 생산 체계 확립

Establishment of Plant Production System through Direct and Indirect Embryogenesis from Anther Culture of Carrot

We tried to develop the protocol for embryogenesis and plant regeneration from anther culture of carrot (Daucus carota L.) genotype ‘S&P2342’. Anthers were cultured on MS medium with B5 vitamins containing different combinations of 2,4-D and NAA for 18 weeks in the dark. The best induction of callus and embryo was obtained in the medium containing 0.1 mg/L 2,4-D and 0.1 mg/L NAA, on which 22.0% callus and 2.0% embryo were induced. When primary embryos induced directly from anther culture were transferred to the regeneration medium, secondary embryos were initiated from primary embryos after 4 of culture and 62.5% converted into plantlets after 8 weeks of culture. The plantlets with true leaves were obtained after 12 weeks of culture. When the calli derived from anther culture were transferred to the regeneration medium, 38.8% of the calli produced primary embryos and plantlets after 8 weeks of culture. The plantlets with 2 or more leaves cultured on the regeneration under the different light intensity for the growth of in vitro plantlets. The plantlets cultured at 100 μmol･m-2･s-1 showed the highest growth rate. For the acclimatization, the in vitro plantlets with 4 or more leaves cultivated under the different light intensity and temperature, respectively. The survival rate and growth of plantlets was best at 15℃ and 100 μmol･m-2･s-1, respectively. The plants were successfully acclimatized and had a normal phenotype. The anther culture system could be used to prepare the doubled haploid lines as an appropriate breeding material for F1 hybrid breeding program.