Gandhi R, Khurana P
Department of Plant Molecular Biology, University of Delhi, India.
Indian J Exp Biol. 2001 Jul;39(7):705-9.
Protoplasts (2 x 10(7)/g fresh wt) were isolated from leaves of A. thaliana ecotype estland, with a viability of more than 90%. Protoplasts cultured in calcium alginate beads or layers showed division while culture in liquid or agarose beads failed to elicit any division. Effect of culture density showed highest frequency of division occurring at 5 x 10(5) while no division was seen when cultured at a density of 5 x 10(4). Culture in MS medium resulted in higher division frequency and better sustenance of microcolonies as compared to B5 medium. Under optimized conditions, macrocolonies were formed at a frequency of 1.8%. Shoot regeneration was seen in 50% of microcalli transferred to shoot induction medium for regeneration. Shoots were rooted and plantlets transferred to pots. The plants produced flowers and were fertile.
从拟南芥爱沙尼亚生态型的叶片中分离出原生质体（2×10⁷/克鲜重），其活力超过90%。在海藻酸钙珠或层中培养的原生质体出现了分裂，而在液体或琼脂糖珠中培养则未引发任何分裂。培养密度的影响表明，分裂频率最高出现在5×10⁵时，而在5×10⁴的密度下培养时未观察到分裂。与B5培养基相比，在MS培养基中培养导致更高的分裂频率和微菌落更好的维持。在优化条件下，大菌落形成的频率为1.8%。将50%的微愈伤组织转移到芽诱导培养基上进行再生时可见芽再生。芽生根后，将小植株转移到花盆中。这些植株开花且可育。
