Department of Biology, Kangwon National University, 200-701, Chuncheon, Republic of Korea.
Plant Cell Rep. 1996 Nov;16(1-2):18-21. doi: 10.1007/BF01275441.
Leaf mesophyll protoplasts ofDianthus superbus were cultured at a density of 5 × 10(4) protoplasts/ml and divided at about 18% plating efficiency in MS liquid medium supplemented with 0.5 mg/L BAP, 2.0 mg/L NAA and 9% mannitol after 2 weeks. Protocolonies formed after 3 to 4 weeks of culture in the dark at 27°C. These colonies were transferred to continuous illumination (21.5 μE m(-2) sec(-1)) for 2 weeks where most of the colonies divided to form microcalli, about 2 mm in diameter. Subsequently, green microcalli were transferred to MS solidified medium with 2.0 mg/L 2,4-D that induced shoot-forming calli after 4 weeks. These calli were transferred onto N6-2 medium containing 0.1 mg/L 2,4-D, 0.1 mg/L NAA, 2.0 mg/L kinetin and 2.0 g/L casein hydrolysate and were cultured under light. After 5 weeks the calli gave rise to multiple shoots (10 to 15 per callus). Upon transfer to MS medium containing 2.0 mg/L NAA, individual shoots were rooted in 4 weeks. The regenerants were successfully transplanted into potting soil.
石竹叶片原生质体以 5×10(4)个原生质体/ml 的密度培养，在补充了 0.5mg/L BAP、2.0mg/L NAA 和 9%甘露醇的 MS 液体培养基中，约 18%的原生质体在 2 周后达到约 18%的平板效率。3 到 4 周后，在黑暗中于 27°C 下培养可形成原球茎。这些原球茎在连续光照（21.5μE m(-2) sec(-1))下培养 2 周，大多数原球茎分裂形成直径约 2mm 的小愈伤组织。随后，绿色小愈伤组织转移到 MS 固体培养基上，用 2.0mg/L 2,4-D 诱导小愈伤组织形成芽，4 周后形成。这些愈伤组织转移到含有 0.1mg/L 2,4-D、0.1mg/L NAA、2.0mg/L 激动素和 2.0g/L 水解酪蛋白的 N6-2 培养基中，在光照下培养。5 周后，愈伤组织产生多个芽（每个愈伤组织 10-15 个）。转移到含有 2.0mg/L NAA 的 MS 培养基中，单个芽在 4 周内生根。再生植株成功移栽到盆栽土壤中。
