毛白杨悬浮细胞系的建立及再生植株的获得

以毛白杨基因型TC152无菌苗为材料,研究毛白杨悬浮细胞系建立与植株再生,结果表明,通过悬浮培养和固体培养两种方法诱导毛白杨悬浮细胞分化不定芽,最终获得无菌生根苗。愈伤组织在MS＋1.5mg·L-12,4-D＋30g·L-1蔗糖的液体培养基中振荡培养,12d可建立悬浮细胞系;悬浮细胞系继代培养基为MS＋0.8mg·L-12,4-D＋30g·L-1蔗糖,继代周期为7d,悬浮细胞在MS＋1.0mg·L-16-BA＋0.1mg·L-1NAA＋0.5～1.0mg·L-1ZT＋30g·L-1蔗糖培养基中悬浮培养,可分化大量不定芽,每个培养瓶中可得到40～50个芽,个别不定芽玻璃化;不定芽在1/2MS＋0.6mg·L-1IBA＋20g·L-1蔗糖＋5.5g·L-1琼脂培养基上可分化不定根。悬浮细胞通过固体平板培养增殖为愈伤组织块后,在MS＋1.0mg·L-16-BA＋0.1mg·L-1NAA＋1.0mg·L-1ZT＋30g·L-1蔗糖＋5g·L-1琼脂的固体培养基上,不定芽分化率可达到70.00%。

Establishment of Suspension Cell Line of Populus tomentosa Carr.and Plant Regeneration from Cells

The establishment of suspension cell line and plant regeneration from cells were studied by using the in vitro explants of Populus tomentosa genotype TC152. Two methods for regeneration,suspension culture and solid culture were used to obtain regenerated plants. The suspension cell line could be established by inoculating calli in the medium of MS＋1.5 mg·L-1 2,4-D＋30 g·L-1 sucrose after 12-d shaking culture. The subculture me-dium was MS＋0.8 mg·L-1 2,4-D＋30 g·L-1 sucrose. And the subculture cycle was 7 d. Large numbers of adven-titious buds could be acquired when the cells were cultured in the liquid medium of MS＋1.0 mg·L-1 6-BA＋0.1 mg·L-1 NAA＋0.5–1.0 mg·L-1 ZT＋30 g·L-1 sucrose. There were 40–50 buds in every bottle. Some of the buds were vitrified. The adventitious buds could root on the medium of 1/2MS＋0.60 mg·L-1 IBA＋20 g·L-1 sucrose＋ 5.5 g·L-1 agar. The calli which were acquired after suspension cells being cultured in solid medium would regenerate adventitious buds on the medium of MS＋1.0 mg·L-1 6-BA＋0.1 mg·L-1 NAA＋1.0 mg·L-1 ZT＋30 g·L-1 sucrose＋5 g·L-1 agar. The regeneration rate was 70.00%.