Populus alba × P. glandulosa (은수원사시나무)에 대한 速生長의 이질 3배체 육성연구 Ⅱ. 엽육조직의 원형질체 분리와 microcolony 형성에 미치는 요인

Breeding for allotriploids with quick grower of hybrid aspen (populus alba × P. glandulosa) Ⅱ. Factors affecting of protoplasts from mesophyll cells and microcolony formation

The isolation and culture of protoplasts in mesophyll tissues were conducted to induce further somatic cell hybrids in Populus aiba×P. glandulosa F₁. The results obtained were as follows: Optimum enzymes and its concentrations for mesophyll protoplast isolation and viability were 2.0% cellulase, 1.0% macerozyme and 1.2% hemicellulase, and four-hour incubation showed 5.3×10⁶ protoplasts/g mesophyll which were the best. The 6M of mannitol was the most effective cocentration to protoplast isolation and viability. The 3.4×10⁶ protoplats/g mesophyll were isolated with its viability of 96.5%. Media containing 10mg/ℓ of gibberellin were more effective to preplasmolysis than those containing 30 mM of cystain, and showed 175% in protoplast isolation. the incubation at 4℃ for 24 hours was the most adequate condition. Modified MS medium was the most effective to budding, first cell division and microcolony formation among the media. Among the treatments with various hormones added to liquid media, the treatment of 2.0mg/ℓ NAA, 1.0mg/ℓ IAA, 0.2mg/ℓ 2,4-D and 0.5mg/ℓ BAP showed the lowest cell division and microcolony formation. The medium added with 0.2mg/ℓ 2,4-D only was effective to microcolony formation.