Department of Epigenetics, Institute of Biophysics of the Czech Academy of Sciences, Královopolská, Brno, Czech Republic.
Methods Mol Biol. 2023;2672:337-349. doi: 10.1007/978-1-0716-3226-0_21.
Meiosis is a very essential cell division resulting in the formation of four haploid gametes in plants. The preparation of meiotic chromosomes is a key step in plant meiotic research. Well-spread chromosomes, low background signal, and effective cell wall elimination give the best hybridization results. Dogroses (Rosa, section Caninae) are allopolyploids and frequently pentaploids (2n = 5x = 35) with asymmetrical meiosis. Their cytoplasm is enriched with organic compounds such as vitamins, tannins, phenols, essential oils, and many more. The cytoplasm is often a huge problem, avoiding successful cytogenetic experiments using fluorescence staining techniques. Here, we present a protocol with modifications for the preparation of male meiotic chromosomes suitable for fluorescence in situ hybridization (FISH) and immunolabeling with a major focus on dogroses.
减数分裂是一种非常重要的细胞分裂，导致植物中形成四个单倍体配子。减数分裂染色体的制备是植物减数分裂研究的关键步骤。良好分散的染色体、低背景信号和有效的细胞壁去除可获得最佳的杂交结果。狗蔷薇（Rosa，狗蔷薇组）是异源多倍体，经常是五倍体（2n=5x=35），具有不对称的减数分裂。它们的细胞质富含各种有机化合物，如维生素、单宁、酚类、精油等。细胞质通常是一个巨大的问题，使用荧光染色技术避免了成功的细胞遗传学实验。在这里，我们提出了一个经过修改的雄性减数分裂染色体制备方案，该方案适用于荧光原位杂交（FISH）和免疫标记，主要针对狗蔷薇。
