Tsai C K, Ch'ien Y C, Chou Y L, Wu S H
Sci Sin. 1977 Jul-Aug;20(4):458-68.
Protoplasts were isolated from the cell suspension culture derived from leaf and stem calli of tobacco (Nicotiana tabacum cv. Ko Hsin No. 1) haploid pollen-plants. After 12 hr in the liquid culture medium, the protoplasts became oval-shaped, and produced a new cell wall. The first division of the newly formed cells was completed after 24 hr in culture. After 4 weeks in culture, the yellowish calli reached 1 mm in size were then transferred to an auxophyton. 18 days later, the calli became 3-4 mm in size. After the calli were transferred to a differentiation culture medium, shoots and roots soon turned up. Regeneration of whole plants was obtained thereafter. The division and differentiation of regenerated cells were affected not only by the calli originated from different organs and their, age, but also by the basic components of the differentiation culture medium and the type of cytokinin used.
原生质体从烟草（Nicotiana tabacum cv. Ko Hsin No. 1）单倍体花粉植株的叶片和茎愈伤组织来源的细胞悬浮培养物中分离得到。在液体培养基中培养12小时后，原生质体变成椭圆形，并形成新的细胞壁。新形成的细胞在培养24小时后完成第一次分裂。培养4周后，将大小达到1毫米的淡黄色愈伤组织转移到辅助培养基上。18天后，愈伤组织大小变为3 - 4毫米。将愈伤组织转移到分化培养基后，很快就出现了芽和根。此后获得了完整植株的再生。再生细胞的分裂和分化不仅受到源自不同器官的愈伤组织及其年龄的影响，还受到分化培养基的基本成分和所用细胞分裂素类型的影响。
