流式细胞仪快速检测鸭茅与多花黑麦草染色体倍性的研究

Detection of chromosome ploidy level in Dactylis glomerata and Lolium multiflorum by flow cytometry

以鸭茅(Dactylis glomerata)和多花黑麦草(Lolium multiflorum)不同倍性的种质为材料,对流式细胞仪测定牧草多倍体的技术方法和结果进行评价,旨在利用流式细胞仪技术快速确定供试牧草的染色体倍性。采用根尖染色体计数法确定鸭茅‘北绿’品种和多花黑麦草‘Mammoth B’品种为四倍体,以这两个品种的相对核DNA含量和核内DNA复制的平均周期值作为参照,经流式细胞仪检测表明,鸭茅‘早生绿’品种和PI316209种质材料为四倍体,种质PI237602和PI368880为二倍体,PI316209种质的倍性与种质背景信息所提供的倍性存在差异;多花黑麦草‘Musashi’品种为四倍体,‘Tachimasari’和‘WasehopeⅢ’两个品种为二倍体。研究结果为流式细胞仪如何在同一物种范围内检测不同倍性样品提供了依据。

A practice of detecting chromosome ploidy levels in Dactylis glomerata and Lolium multiflorum by a flow cytometry method was reported;and aimed to discuss its application and reliability in forage polyploidy research.In order to investigate the chromosome ploidy levels of 4 D.glomerata germplasms（Wasemidori,PI316209,PI237602 and PI36880） and 3 L.multiflorum cultivars（Musashi,Tachimasari and Wasehope Ⅲ） by using the flow cytometry method,two known tetraploidy cultivars,including D.glomerata cv.’Kitamidori’ and L.multiflorum cv.’Mammoth B’,were used as control materials.Their chromosome number in root tissue cells were double checked by the optical method and used to set a standard value in a flow cytometry.The results indicated that both of ’Wasemidori’ and ’PI316209’ in D.glomerata were tetraploids,but ’PI237602’ and ’PI36880’ were diploids.For 3 tested L.multiflorum cultivars,’Musashi’ was tetraploid,while ’Tachimasari’ and ’Wasehope Ⅲ’ were diploids.All chromosome ploidy levels of the tested cultivars or germplasms were confirmed to their known ploidy levels except of ’PI316209’ in D.glomerata,which was a tetraploid instead of a diploid.The results provided a good reference to detect various chromosome levels in a same species by the flow cytomety method.