Negaard K, Hoffmann F
Department of Developmental and Cell Biology, University of California, Irvine 92717.
Biotechniques. 1989 Sep;7(8):808-12.
The isolation and regeneration of tobacco mesophyll protoplasts from fully developed leaves, an important methodological step in plant genetic engineering as well as in plant cell biology and physiology, has been proven unreliable to the extent that it has become a significant setback to basic research. This unfortunate situation is primarily due to the suboptimal physiological state of greenhouse-grown protoplast donor plants. A technically simple and inexpensive method, based on the utilization of commercial Phototron units, is described for the production of suitable tobacco donor plants. Furthermore, a modified version of such a culture unit can be used to regenerate plants from protoplast-derived calli.
从完全成熟的叶片中分离和再生烟草叶肉原生质体,这在植物基因工程以及植物细胞生物学和生理学中是一个重要的方法步骤,但已被证明不可靠,以至于成为基础研究的重大挫折。这种不幸的情况主要是由于温室种植的原生质体供体植物的生理状态欠佳。本文描述了一种基于使用商用光生物反应器装置的技术简单且成本低廉的方法,用于培育合适的烟草供体植物。此外,这种培养装置的改良版本可用于从原生质体来源的愈伤组织再生植株。