苎麻不同来源原生质体的分离和培养的比较研究

COMPARATIVE STUDIES ON ISOLATION AND CULTURE OF DIFFERENT PROTOPLASTS OF RAMIE

比较了苎麻子叶与子叶悬浮细胞两种材料的原生质体在分离和培养时的差异。取真叶初露期的绿色子叶，用３％纤维素酶、０．５％离析酶的０．６Ｍ甘露醇混合溶液处理５小时，原生质体产量可达５×１０￣６个／ｇｆｒ·ｗｔ，但其原生质体以海藻酸钠包埋方式培养在附加２，４—Ｄ０．５ｍｇ／Ｌ、ＫＴ０．５ｍｇ／ＬＫＭ＿８Ｐ培养基上，原生质体仅发生二次分裂；子叶悬浮细胞只有在纤维素酶４．５％，离析酶０．８％、半纤维素酶０．８％的０．５５Ｍ甘露醇混合液中处理１１小理，原生质体才达最大程度释放，每克悬浮细胞可得原生质体２×１０￣６个，但其原生质体易于诱导分裂，在与前者相同的培养条件下，５０天左右可形成肉眼可见的小愈伤组织，该愈伤组织经过扩增，在不同的培养基上可诱导芽、根的分化，从而形成完整植株。

The differences of isolation and culture between cotyledon protoplast and protoplast from cotyledon cutured cells of ramie were studied. The protoplasts (5×10￣6/g fr. wt)were isolated from green cotyledon swhen the ture leaves begin to emerge with the treatment of 3% Cellulase R-10,0. 5% Macerozyme R-10 in 0. 6 mol/L minnitol solution for 5hr. But they only divide 2 times when cultured on KM_8 Pmedium containing 2,4-D 0. 5 mg/L,KT 0. 6 mg/L in alginate embedding method. Protoplasts derived from cotyledon cultured cells can reach highest (2×10￣6/g cells)only treated with 4. 5% Cellulase, 0. 8% Macerozyme .0.8% Hemicellulase in 0. 55 mol/L mannitol solution for 11 hr. But they are easy to induce to divide and can form microcalli within fifty days when cultured on the former mediu. After subcultured,the calli can induce to from shoot and root and therefore complete plants were obtained.