Suda Jan, Trávnícek Pavel
Charles University in Prague and Academy of Sciences of the Czech Republic, Prague, Czech Republic.
Curr Protoc Cytom. 2006 Nov;Chapter 7:Unit7.30. doi: 10.1002/0471142956.cy0730s38.
The power of flow cytometry in field plant research may be greatly enhanced by analysis of nonfresh tissues. Previous attempts to use chemical fixatives, however, received only little attention because of protocol complexity and limited time, after which successful assays were achieved. This unit describes simple and rapid procedures for relative nuclear DNA content estimation in dehydrated tissues of vascular plants by DAPI flow cytometry. Histograms with reasonable resolution can be obtained in several-year-old specimens. The approach here is particularly suitable for reliable determination of DNA ploidy level, although detection of small variation in nuclear DNA content is also possible in many cases. Retrospective ploidy inference in silica-dry or herbarium vouchers and simplified transport of plant material from remote areas are among the principal benefits for plant biosystematics, ecology, and population biology.
通过对非新鲜组织进行分析，可极大地增强流式细胞术在田间植物研究中的作用。然而，由于方案复杂且时间有限，此前使用化学固定剂的尝试并未受到太多关注，在此之后才实现了成功的检测。本单元介绍了通过DAPI流式细胞术在维管植物脱水组织中估算相对核DNA含量的简单快速程序。在数年生的标本中可获得分辨率合理的直方图。尽管在许多情况下也能够检测到核DNA含量的微小变化，但这里的方法特别适用于可靠地确定DNA倍性水平。对于植物生物系统学、生态学和种群生物学而言，在硅胶干燥或标本馆凭证中进行追溯倍性推断以及简化来自偏远地区的植物材料运输是主要的优势。
