A replication stress-induced synchronization method for Arabidopsis thaliana root meristems
A replication stress-induced synchronization method for Arabidopsis thaliana root meristems
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DOI：
10.1111/j.1365-313x.2010.04361.x
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发表时间：
2010-11-01
期刊：
影响因子：
7.2
通讯作者：
De Veylder, Lieven
中科院分区：
文献类型：
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作者：
Cools, Toon;Iantcheva, Anelia;De Veylder, Lieven
P>同步细胞培养物是识别和了解细胞周期关键调节因子不可或缺的工具。然而，细胞培养物的使用也有其缺点，因为它代表了一种人工系统，不能完全模拟有组织的分生组织中发生的内源条件。在这里，我们提出了一种通过羟基脲处理拟南芥根尖同步化的新且简单的方法。该方法的一个主要优点是可以研究可用的拟南芥细胞周期突变体，而无需生成细胞培养物。作为概念证明，我们测试了 B 型细胞周期蛋白依赖性激酶 CDKB1;1 基因的显性失活等位基因的过度表达对细胞周期进展的影响。先前观察到的延长的 G(2) 相得到证实，但发现可以通过减少的 G(1) 相来补偿。此外，S 相动力学的改变表明 CDKB1;1 在复制过程中发挥功能作用。
P>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 G(2) phase was confirmed, but was found to be compensated for by a reduced G(1) phase. Furthermore, altered S-phase kinetics indicated a functional role for CDKB1;1 during the replication process.