Microtubule reorganization in tobacco BY-2 cells stably expressing GFP-MBD
Microtubule reorganization in tobacco BY-2 cells stably expressing GFP-MBD
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DOI：
10.1007/s004250050037
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发表时间：
2000-02-01
期刊：
影响因子：
4.3
通讯作者：
Cyr, RJ
中科院分区：
文献类型：
--
作者：
Granger, CL;Cyr, RJ
微管组织在植物形态发生中起着重要作用；然而，人们对微管阵列如何从一种有组织的状态过渡到另一种状态知之甚少。使用基因结合荧光标记可以长期观察活细胞中的微管行为。在这里，我们对一株烟草株系进行了鉴定。明亮的黄色2 （BY-2）细胞系，用gfp-mbd结构稳定转化，先前证明可以标记微管（J. Mare等，1998,Plant cell 10: 1927-1939）。荧光水平低。但在gfp-mbd转化的单个细胞中，可以观察到间期和有丝分裂微管阵列，以及这些阵列之间的转换。通过比较转化细胞和未转化细胞的一些特性，得出转基因细胞不受低水平表达转基因基因的不利影响的结论，这些细胞将为观察体内微管重组提供有用和准确的模型系统。事实上，一些初步的观察结果与运动蛋白参与纺锤体和片质体阵列之间的转换是一致的。我们的观察结果也支持核周区在细胞分裂末期微管成核中的作用，这些微管和/或成核活动逐渐转移到皮层，形成期间皮层阵列。
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.