Feucht W, Treutter D, Dithmar H, Polster J
Department für Pflanzenwissenschaften, Fachgebiet Obstbau, Wissenschaftszentrum Weihenstephan (WZW), Technische Universität München, 85350 Freising, Germany.
Plant Biol (Stuttg). 2005 Mar;7(2):168-75. doi: 10.1055/s-2005-837472.
Young anthers excised from closed tea flower buds ( Camellia sinensis L.) were stained as fresh tissues with p-dimethylaminocinnamaldehyde reagent to localize flavanols associated with nuclei and chromosomes, apart from those flavanols stored in vacuoles. This staining reagent yields a blue colour for flavanols. In the nonsporogenic somatic cells of developing anthers, flavanols were found to be attached to chromosomes at all mitotic stages. Male meiosis started at a bud size of about 3.5 mm in diameter in pollen mother cells which displayed generally more or less pronounced blue nuclei and cytoplasm. The meiotic divisions from prophase I to telophase II were characterized by blue stained nuclei and chromosomes, but within the cytoplasm there was, if any, a random and very poor reaction for flavanols. Metaphase and telophase of meiotic divisions showed maximally condensed chromosomes staining dark blue. Early in telophase II, the cytoplasm was again stained blue; this faded at late tetrad stage. Flavanols of young mitotic and older non-mitotic anthers were determined using high pressure liquid chromatography--chemical reaction detection (HPLC-CRD). Catechin, epicatechin, B2, and epigallocatechin were minor compounds, whereas epicatechin gallate and epigallocatechin gallate were found in higher amounts. The major flavanol compound of the anthers, epicatechin gallate, exhibited a significant affinity to histone sulphate, as shown by UV-VIS spectroscopic titration.
从闭合的茶花芽(茶树)上切下的幼嫩花药作为新鲜组织,用对二甲氨基肉桂醛试剂染色,以定位与细胞核和染色体相关的黄烷醇,除了那些储存在液泡中的黄烷醇。这种染色试剂能使黄烷醇产生蓝色。在发育中的花药的非孢子体细胞中,发现在所有有丝分裂阶段黄烷醇都附着在染色体上。在直径约3.5毫米的芽中,花粉母细胞开始进行雄配子减数分裂,这些细胞通常显示出或多或少明显的蓝色细胞核和细胞质。从减数分裂前期I到末期II的减数分裂过程中,细胞核和染色体都被染成蓝色,但在细胞质中,如果有黄烷醇反应的话,也是随机且非常微弱的。减数分裂中期和末期显示出高度浓缩的染色体染成深蓝色。在末期II早期,细胞质再次被染成蓝色;在四分体后期这种蓝色褪去。使用高压液相色谱 - 化学反应检测(HPLC - CRD)测定幼嫩有丝分裂期和较老的非有丝分裂期花药中的黄烷醇。儿茶素、表儿茶素、B2和表没食子儿茶素是次要成分,而表儿茶素没食子酸酯和表没食子儿茶素没食子酸酯的含量较高。花药中的主要黄烷醇化合物表儿茶素没食子酸酯对硫酸组蛋白表现出显著的亲和力,如紫外 - 可见光谱滴定所示。