de Castro R D, van Lammeren A A, Groot S P, Bino R J, Hilhorst H W
Laboratory of Experimental Plant Morphology and Cell Biology, Wageningen University, Arboretumlaan 4, 6703 BD Wageningen, The Netherlands.
Plant Physiol. 2000 Feb;122(2):327-36. doi: 10.1104/pp.122.2.327.
We studied cell cycle events in embryos of tomato (Lycopersicon esculentum Mill. cv Moneymaker) seeds during imbibition in water and during osmoconditioning ("priming") using both quantitative and cytological analysis of DNA synthesis and beta-tubulin accumulation. Most embryonic nuclei of dry, untreated control seeds were arrested in the G(1) phase of the cell cycle. This indicated the absence of DNA synthesis (the S-phase), as confirmed by the absence of bromodeoxyuridine incorporation. In addition, beta-tubulin was not detected on western blots and microtubules were not present. During imbibition in water, DNA synthesis was activated in the radicle tip and then spread toward the cotyledons, resulting in an increase in the number of nuclei in G(2). Concomitantly, beta-tubulin accumulated and was assembled into microtubular cytoskeleton networks. Both of these cell cycle events preceded cell expansion and division and subsequent growth of the radicle through the seed coat. The activation of DNA synthesis and the formation of microtubular cytoskeleton networks were also observed throughout the embryo when seeds were osmoconditioned. However, this pre-activation of the cell cycle appeared to become arrested in the G(2) phase since no mitosis was observed. The pre-activation of cell cycle events in osmoconditioned seeds appeared to be correlated with enhanced germination performance during re-imbibition in water.
我们利用DNA合成和β-微管蛋白积累的定量及细胞学分析,研究了番茄(Lycopersicon esculentum Mill. cv Moneymaker)种子在吸水和渗透调节(“引发”)过程中的细胞周期事件。干燥、未经处理的对照种子的大多数胚细胞核停滞在细胞周期的G(1)期。这表明不存在DNA合成(S期),溴脱氧尿苷掺入的缺失证实了这一点。此外,在蛋白质免疫印迹上未检测到β-微管蛋白,也不存在微管。在吸水过程中,胚根尖端的DNA合成被激活,然后向子叶扩散,导致G(2)期细胞核数量增加。同时,β-微管蛋白积累并组装成微管细胞骨架网络。这两个细胞周期事件都先于细胞扩张和分裂以及随后胚根穿过种皮的生长。当种子进行渗透调节时,在整个胚中也观察到了DNA合成的激活和微管细胞骨架网络的形成。然而,由于未观察到有丝分裂,这种细胞周期的预激活似乎停滞在G(2)期。渗透调节种子中细胞周期事件的预激活似乎与再次吸水时发芽性能的提高相关。