Cell cycle synchronization in plant root meristems
Cell cycle synchronization in plant root meristems
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DOI：
10.1023/a:1009876621187
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发表时间：
1999-01-01
期刊：
影响因子：
--
通讯作者：
Lucretti, S.
中科院分区：
文献类型：
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作者：
Dolezel, J.;Cihalikova, J.;Lucretti, S.
在细胞周期的特定阶段对细胞的结构和代谢进行分析往往很困难，因为体细胞组织中的细胞周期进程是不同步的，只有一部分细胞处于细胞周期中。获得富含细胞周期单一阶段的细胞群体的一个巧妙解决方案是人工诱导同步化。已经使用了不同的系统来获得植物细胞的同步化群体，包括悬浮培养细胞、叶肉原生质体和根尖分生组织。根尖在从染色体分析到细胞周期及其调控的各种研究中经常被使用。在大多数植物物种中都可以获得具有活跃生长根系的幼苗，它们易于操作，实验系统明确、可重复，并且可以很容易地针对不同物种进行调整。本文描述了一种基于使用DNA合成抑制剂羟基脲（18）使蚕豆根尖细胞周期同步化的实验方案。还给出了针对豌豆、紫花苜蓿、大麦、黑麦、小麦和玉米对该方案的修改。流式细胞术数据表明，大约90%的根尖细胞实现了同步化。通过这种方法，平均可获得超过50%的有丝分裂指数。使用有丝分裂纺锤体抑制剂可以使同步化细胞在中期积累，从而使中期指数超过50%。
The analysis of structure and metabolism of a cell at a defined phase of cell cycle is often difficult because cell cycle progression in somatic tissues is asynchronous and only a fraction of cells are cycling. An elegant solution to obtain populations of cells enriched for single stage of the cell cycle is to impose the synchrony artificially. Different systems have been used to obtain synchronized populations of plant cells, including suspension-cultured cells, leaf mesophyll protoplasts and root tip meristems. Root tips have been frequently used in a variety of studies ranging from chromosome analysis to cell cycle and its regulation. Seedlings with actively growing roots may be obtained in most plant species, they are easy to handle, the experimental system is well defined, reproducible and can be easily modified for different species. This paper describes a protocol for cell cycle synchronization in root tips of Vicia faba, which is based on the use of DNA synthesis inhibitor hydroxyurea (18). Modifications of the protocol for Pisum sativum, Medicago sativa, Hordeum vulgare, Secale cereale, Triticum aestivum, and Zea mays are also given. Flow cytometric data indicate that about 90% of root tip cells are synchronized. On average, mitotic indices exceeding 50% are obtained with the method. Synchronized cells may be accumulated at metaphase using a mitotic spindle inhibitor to achieve metaphase indices exceeding 50%.