Department of Plant Systems Biology, VIB, Technologiepark 927, 9052 Gent, Belgium.
Plant J. 2010 Nov;64(4):705-14. doi: 10.1111/j.1365-313X.2010.04361.x. Epub 2010 Oct 26.
Synchronized cell cultures are an indispensable tool for the identification and understanding of key regulators of the cell cycle. Nevertheless, the use of cell cultures has its disadvantages, because it represents an artificial system that does not completely mimic the endogenous conditions that occur in organized meristems. Here, we present a new and easy method for Arabidopsis thaliana root tip synchronization by hydroxyurea treatment. A major advantage of the method is the possibility of investigating available Arabidopsis cell-cycle mutants without the need to generate cell cultures. As a proof of concept, the effects of over-expression of a dominant negative allele of the B-type cyclin-dependent kinase CDKB1;1 gene on cell-cycle progression were tested. The previously observed prolonged G₂ phase was confirmed, but was found to be compensated for by a reduced G₁ phase. Furthermore, altered S-phase kinetics indicated a functional role for CDKB1;1 during the replication process.
同步细胞培养是鉴定和理解细胞周期关键调节因子的不可或缺的工具。然而，细胞培养有其缺点，因为它代表了一种人工系统，不能完全模拟发生在有组织的分生组织中的内源性条件。在这里，我们提出了一种通过羟基脲处理进行拟南芥根尖同步化的新方法。该方法的一个主要优点是可以在不需要生成细胞培养物的情况下研究现有的拟南芥细胞周期突变体。作为概念验证，测试了过表达 B 型细胞周期蛋白依赖性激酶 CDKB1;1 基因的显性负等位基因对细胞周期进程的影响。先前观察到的 G₂ 期延长得到了证实，但发现 G₁ 期缩短了。此外，改变的 S 期动力学表明 CDKB1;1 在复制过程中具有功能作用。
