Microtubule reorganization in tobacco BY-2 cells stably expressing GFP-MBD

被引:91
作者
Granger, CL [1 ]
Cyr, RJ [1 ]
机构
[1] Penn State Univ, Dept Biol, University Pk, PA 16802 USA
基金
美国国家航空航天局;
关键词
copper-inducible promoter; cytoskeleton; green fluorescent protein (GFP); microtubule-associated protein (MAP4); microtubule; Nicotiana;
D O I
10.1007/s004250050037
中图分类号
Q94 [植物学];
学科分类号
071001 ;
摘要
Microtubule organization plays an important role in plant morphogenesis; however, little is known about how microtubule arrays transit from one organized state to another. The use of a genetically incorporated fluorescent marker would allow long-term observation of microtubule behavior in living cells. Here, we have characterized a Nicotiana tabacum L. cv. Bright Yellow 2 (BY-2) cell line that had been stably transformed with a gfp-mbd construct previously demonstrated to label microtubules (J. Mare et al., 1998, Plant Cell 10: 1927-1939). Fluorescence levels were low. but interphase and mitotic microtubule arrays, as well as the transitions between these arrays, could be observed in individual,gfp-mbd-transformed cells. By comparing several attributes of transformed and untransformed cells it was concluded that the transgenic cells are not adversely affected by low-level expression of the transgene and that these cells will serve as a useful and accurate model system for observing microtubule reorganization in vivo. Indeed, some initial observations were made that are consistent with the involvement of motor proteins in the transition between the spindle and phragmoplast arrays. Our observations also support the role of the perinuclear region in nucleating microtubules at the end of cell division with a progressive shift of these microtubules and/or nucleating activity to the cortex to form the interphase cortical array.
引用
收藏
页码:502 / 509
页数:8
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