《植物生理学报》 2010, 46(11): 1114-1120

毛白杨悬浮细胞系的建立及再生植株的获得

姚娜^1,2, 安新民¹, 杨凯³, 张志毅^1,*

1 北京林业大学林木育种国家工程实验室, 林木花卉遗传育种教育部重点实验室, 国家林业局树木花卉育种与生物工程重 点开放实验室, 北京 100083; 2 中国林业科学研究院林业研究所国家林业局林木培育重点实验室, 北京 100091; 3 北京农学院 农业应用新技术北京市重点实验室, 北京 102206

通信作者：张志毅；E-mail: zhangzy@bjfu.edu.cn；Tel: 010-62338502

摘　要：

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+1.5 mg•L-1 2,4-D+30 g•L-1蔗糖的液体培 养基中振荡培养, 12 d 可建立悬浮细胞系; 悬浮细胞系继代培养基为MS+0.8 mg•L-1 2,4-D+30 g•L-1 蔗糖, 继代周期为7 d, 悬 浮细胞在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+0.6 mg•L-1 IBA+20 g•L-1蔗糖+5.5 g•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+1.0 mg•L-1 6-BA+0.1 mg•L-1 NAA+1.0 mg•L-1 ZT+30 g•L-1 蔗糖+5 g•L-1 琼脂的固体培养基上, 不定芽分化率可达到70.00%。

关键词：毛白杨; 悬浮细胞; 再生; 组织培养

收稿：2010-05-05   修定：2010-09-26

资助：国家林业局“948” 项目(2006-4-72)、国家“ 十一五” 科技支撑计划课题(2006BAD24B04)和国家“863” 计划项目(2009AA10Z107)。

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

YAO Na^1,2, AN Xin-Min¹, YANG Kai³, ZHANG Zhi-Yi^1,*

¹National Engineering Laboratory for Tree Breeding, Key Laboratory for Genetics and Breeding in Forest Trees and Ornamental Plants, Ministry of Education, The Tree and Ornamental Plant Breeding and Biotechnology Laboratory of State Forestry Administration, Beijing Forestry University, Beijing 100083, China; ²Key Laboratory of Tree Breeding and Cultivation, State Forestry Administration, Research Institute of Forestry, Chinese Academy of Forestry, Beijing 100091, China; ³Key Laboratory of Agricultural New Technology and Application in Beijing, Beijing Agricultural College, Beijing 102206, China

Corresponding author: ZHANG Zhi-Yi; E-mail: zhangzy@bjfu.edu.cn; Tel: 010-62338502

Abstract:

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 medium 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 adventitious buds could be acquired when the cells were cultured in the liquid medium of MS+1.0 mg·L^-1 6-BA+0.1mg·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 wouldregenerate 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^-1sucrose+5 g·L^-1 agar. The regeneration rate was 70.00%.

Key words: Populus tomentosa; suspension cell; regeneration; tissue culture