Sliwinska Elwira
University of Technology and Agriculture, Bydgoszcz, Poland.
Curr Protoc Cytom. 2006 Feb;Chapter 7:Unit 7.29. doi: 10.1002/0471142956.cy0729s35.
Procedures describing the utilization of seeds or their parts for flow cytometric determination of plant ploidy and endopolyploidy, genome size, and cell cycle activity are presented. The methods have been developed for a single-fluorescence-parameter flow cytometer, equipped with light sources for 488-nm and UV-light illumination. The procedures presented in this unit utilize the two most widely used fluorochromes for plant DNA content analysis, propidium iodide (PI) and 4',6-diamidino-2-phenylindole (DAPI). These methods provide an alternative to estimation of DNA content based on the fluorescence of DNA in cell nuclei isolated from plant leaves. In some instances seeds are more suitable for analysis than leaves, e.g., when plant material must be transported for a long distances or stored for prolonged periods before flow cytometric analysis, or when leaves contain fluorochrome-staining inhibitors. In addition, flow cytometric determination of nuclear replication stages in seeds gives information about their physiological status (e.g., maturity, advancement of germination), which is valuable to seed producers and technologists.
本文介绍了利用种子或其部分进行流式细胞术测定植物倍性和内多倍性、基因组大小及细胞周期活性的方法。这些方法是针对配备488纳米和紫外光照明光源的单荧光参数流式细胞仪开发的。本单元介绍的方法使用了植物DNA含量分析中最常用的两种荧光染料，碘化丙啶（PI）和4',6-二脒基-2-苯基吲哚（DAPI）。这些方法为基于从植物叶片分离的细胞核中DNA荧光来估计DNA含量提供了一种替代方法。在某些情况下，种子比叶片更适合分析，例如，当植物材料必须长途运输或在流式细胞术分析前长时间储存时，或者当叶片含有荧光染料染色抑制剂时。此外，通过流式细胞术测定种子中的核复制阶段可提供有关其生理状态（如成熟度、发芽进程）的信息，这对种子生产者和技术人员很有价值。
