【作者】 安晓珂；

【导师】 赵来喜；

【作者基本信息】 中国农业科学院 ， 草业科学， 2008， 硕士

【摘要】 三叶草属(TrifoliumL.)植物中有很多是世界上非常著名的优良栽培牧草,同时也是利用很广泛的草坪草,主要分布在世界温带地区。由于三叶草属植物喜温暖湿润气候,在干旱和寒冷的内蒙古地区引种红三叶和白三叶等优质三叶草属牧草至今未获成功。但是在内蒙古、新疆的北部地区,由于严冬时有积雪覆盖,野生红三叶、白三叶及同属的野火球均可以安全越冬,对这些抗寒性强的野生材料进行研究将为红三叶和白三叶在更大范围内推广奠定基础。本研究利用随机多态性DNA(RAPD)标记技术对采集于内蒙古、新疆的野生野火球、白三叶和红三叶等三种三叶草材料进行种内和种间遗传多样性分析,探讨种内不同材料间的亲缘关系,通过对材料的特异片段的筛选及鉴定,以获得材料的特异标记,以期对野生材料的多样性有初步的了解,为三叶草种质资源和育种研究提供依据。本试验对100条随机引物进行了筛选,最后确定了7条引物,通过优化试验因子确定了最优组合的RAPD反应体系,对20份材料进行了扩增。利用7个引物扩增的总谱数为121条,其中多态性条带为117条,多态性比率为96.7%,不同种扩增的谱带数分别为野火球58、白三叶88、红三叶70,其多态性比率分别为77.6%,75%,81.4%。用非加权组平均法(UPGMA)构建系统发生树,探讨了各材料之间的遗传关系。种内分析显示,三个种的聚类结果与其地理分布及生境有很大的相关性;作为对照的栽培品种与野生材料间有较大的差异;在白三叶材料中,采集于牙克石的白三叶与其它白三叶材料差异很大,可以作为特殊材料进行研究。种间聚类显示白三叶和红三叶的亲缘关系较为接近。通过分析DNA图谱,得到3个重复性好的特异标记片断,OPB08-257,OPC11-745可以将三种三叶草区分开来,OPB08-300可以鉴别海发白三叶与野生白三叶。更多还原

【Abstract】 Clovers (Trifolium) are an important group of species of one genus in the world as famous cultivated forage grass as well as lawn grass. In general, clovers inhabit temperature regions of the world. Because of this, clovers cannot grow in Inner Mongolia Autonomous Region with drought and cold climate. But in some area in the north of Inner Mongolia Autonomous Region and Xinjiang Uygur Autonomous Region, for heavy snows in the winter, three species of clovers, white clover, red clover and T. lupinaster can survive in severe winter. In order to expand the clovers, three speices of clovers were used to detect the genetic diversity.20 materials belong to three species of clovers, which were collected from Inner Mongolia Autonomous Region and Xinjiang Uygur Autonomous Region, were analysed by Random Amplified Polymorphism DNA (RAPD). Based on analysis data, discuss the genetic diversity in and among species, and plants’blood relationship, through to special fragment screening appraisal, so that can obtain the special mark to lay the foundation for germplasm resource and breeding improved of clovers.20 clover materials were analyzed by RAPD markers to determine their genetic relationship. Of 100 random nucleotide primers screened, 7 showed steady banding patterns and produced 121 bands with 117 polymorphisms. Using un-weighted pair-group method with arithmetic means (Abbreviation UPGMA), constructs the phylogeny tree and acts according the phylogeny tree, according to UPGMA gathers cluster. The analysis result in species indicated that, the result of gathers cluster were more relative with the geography and the climate; it showed more different between cultivated variaties and wild materials; among the white clover, the material collected from Yakeshi had further distance from others. Another gather among species indicated that white clover and red clover were more similar compared to T. lupinaster. Obtain 3 special mark bands with good duplication and strong signal, OPB08-257 and OPC11-745 can take special mark bands to separate three clovers, and OPB08-300 may identify Haifa clover and other wild white clover.更多还原