十字花科 植物의 原形質體 培養과 融合에 관한 硏究 Ⅰ. 油菜의 原形質體 培養과 植物體의 再生

Protoplast culture and fusion in cruciferae Ⅰ. Plant regeneration from mesophyll protoplast of rape(brassica napus L.)

The experiments were conducted to identify several factors influencing mesophyll protoplast isolation, culture and plant regeneration of rape(Brassica napus cv. ‘Hanrayuchae’) The results obtained were summarized as follows ; It was required to preplasmolyse the mesophyll tissue before enzyme treatment to increase yield and stability of protoplasts. The most healthy protoplasts could be obtained when the tissue was treated with enzymes containing macerozyme 1.0% and cellulase 2.0% for 4 hours. Sustained cell division was obtained after culturing the protoplasts in liquid 8P-KM medium supplemented with 2.4-D 0.2mg/ℓ, NAA 1.0mg/ℓ, BAP 0.5mg/ℓ, glucose 0.4M, and mannitol 0.1M. First cell division was observed 5 days after plating and was sustained to eventually produce cell colonies after another 16 days. When micro-calli 4-5 weeks after plating were transferred to B₅ basal medium with NAA 1.0mg/ℓ, BAP 0.5mg/ℓ, sucrose 30g/ℓ and agarose 6g/ℓ, green calli were proliferated to be at least 0.5-1.0cm in diameter 8 weeks after initial plating. Multiple shoots were induced on MS medium supplemented with BAP 1.0mg/ℓ and NAA 0.01mg/ℓ. After transferring differentiated shoots onto MS medium without hormones, intact plants were eventually produced.