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作物学报 ›› 2011, Vol. 37 ›› Issue (10): 1794-1800.doi: 10.3724/SP.J.1006.2011.01794

• 作物遗传育种·种质资源·分子遗传学 • 上一篇    下一篇

以流式核型分析小麦根尖细胞的有丝分裂同步化过程

郭东伟1，张仁和1，李春莲1，陈耀锋1，闵东红1,*，陈明2，李连成2，徐兆师2，马有志2   

  1. 1西北农林科技大学农学院，陕西杨凌 712100；2中国农业科学院作物科学研究所，北京 100081
  • 收稿日期:2011-01-13 修回日期:2011-06-25 出版日期:2011-10-12 网络出版日期:2011-07-28
  • 通讯作者: 闵东红, E-mail: mdh2493@nwsuaf.edu.cn
  • 基金资助:

    本研究由陕西省自然科学基金项目(SJ08-ZD04)资助。

Analyzing Synchronization Process of Mitosis Cells in Wheat Root-tip with Flow Karyotype

GUO Dong-Wei1,ZHANG Ren-He1,LI Chun-Lian1,CHEN Yao-Feng1,MIN Dong-Hong1,*,CHEN Ming2,LI Lian-Cheng2,XU Zhao-Shi2,MA You-Zhi2   

  1. 1 Agronomy College, Northwest A & F University, Yangling 712100, China; 2 Institute of Crop Sciences, Chinese Academy of Agricultural Sciences, Beijing 100081, China
  • Received:2011-01-13 Revised:2011-06-25 Published:2011-10-12 Published online:2011-07-28
  • Contact: 闵东红, E-mail: mdh2493@nwsuaf.edu.cn

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摘要： 构建染色体特异文库是简化小麦基因组测序的有效途径，而通过同步化处理提高根尖细胞有丝分裂指数，使根尖细胞中富集高比例的中期染色体是实现染色体分选成功的前提。采用双阻断法用50 µmol L-1羟基脲和100 µmol L-1氟乐灵对普通小麦根尖细胞进行同步化处理，然后用流式核型分析法观测各周期细胞在不同时间点上的比例变化。结果显示，最佳方案为羟基脲处理10 h，恢复7 h，氟乐灵处理4 h。以此方案处理中国春小麦根尖可获得最高的有丝分裂指数，达70.1%。附加冰水处理(0°C)有助于减少染色体碎片和增加流式核型的分辨率，但对富集更多的中期染色体没有效果。

关键词: 小麦, 有丝分裂, 同步化, 流式核型, 分选

Abstract: Genomic sequencing of wheat can be simplified through constructing chromosome specific library. Synchronization of cells is propitious to high mitosis index (MI), which results in the accumulation of metaphase chromosomes and a successful flow sorting. In the present study, we used a double-blocked strategy in the synchronization treatment of wheat (Triticum aestivum L. cv. Chinese Spring) root-tip cells with 50 µmol L-1 hydroxyurea (HU) and 100 µmol L-1 trifluralin. The effects of treatments were evaluated by analyzing the dynamic proportions of cells at different stages of cell cycle. The optimal hours of treatments were 10 h for HU block, 7 h for recovery, and 4 h for trifluralin block. According to this synchronization scheme, the maximum MI was 70.1%. Additional ice water (0°C) depression was helpful for decreasing chromosome debris and rising resolution of the flow karyotype, but had no effect on the accumulation of metaphase chromosomes in root-tip cells.

Key words: Wheat, Mitosis, Synchronization, Flow karyotype, Flow sorting

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