以流式核型分析小麦根尖细胞的有丝分裂同步化过程  

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

作　　者：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] 

机构地区：[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

出　　处：《作物学报》2011年第10期1794-1800,共7页Acta Agronomica Sinica

基　　金：陕西省自然科学基金项目(SJ08-ZD04)资助

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50μmolL?1羟基脲和100μmolL?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10h,恢复7h,氟乐灵处理4h。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℃)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。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 time 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℃） 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.

关 键 词：小麦 有丝分裂 同步化 流式核型 分选 

分 类 号：S512.1[农业科学—作物学]