Zolan Miriam E, Pukkila Patricia J
Department of Biology, Indiana University, Bloomington, IN, USA.
Methods Mol Biol. 2009;558:115-27. doi: 10.1007/978-1-60761-103-5_8.
The basidiomycete fungus Coprinus cinereus has naturally synchronous meiosis and is amenable to analysis using an array of well-developed genetic and molecular tools. In this chapter, we explain in detail the two methods most commonly employed for C. cinereus, staining of intact gill segments and chromosome spreads, with an example of the application of each. We describe iron-hematoxylin staining of intact gill segments for the brightfield examination of meiotic progression, and the use of surface spreads and fluorescence in situ hybridization (FISH) to investigate meiotic chromosome pairing. Gill segments can alternatively be stained with DAPI for the determination of meiotic stage, or propidium iodide for the quantitation of nuclear DNA content, and the chromosome fixation and spreading techniques used for FISH are also suitable for immunolocalization studies of chromosomal proteins.
灰盖鬼伞担子菌具有自然同步减数分裂，并且适合使用一系列成熟的遗传和分子工具进行分析。在本章中，我们将详细解释两种最常用于灰盖鬼伞的方法，完整鳃片染色和染色体铺展，并各举一个应用实例。我们描述了用于减数分裂进程明场检查的完整鳃片铁苏木精染色，以及用于研究减数分裂染色体配对的表面铺展和荧光原位杂交（FISH）。鳃片也可以用DAPI染色以确定减数分裂阶段，或用碘化丙啶定量核DNA含量，并且用于FISH的染色体固定和铺展技术也适用于染色体蛋白的免疫定位研究。
