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位于簇毛麦V染色体短臂上抗白粉病基因Pm21的AFLP标记研究
【作者】 赵红梅;
【导师】 王振英;
【作者基本信息】 天津师范大学 , 细胞生物学, 2006, 硕士
【摘要】 由白粉菌引起的小麦白粉病是中国和世界上最主要的小麦病害之一,近年来,在我国麦区的危害日益严重,已发展成为当前小麦生产上发病面积最广的病害。化学防治虽不无成效,但培育抗病品种是目前被认为防治小麦病害最为经济、安全和有效的途径。到目前为止,小麦基因组中已定名抗白粉病基因30多个,其中在已被定位在特定染色体或染色体臂上的抗病基因中来源于簇毛麦的Pm21(6AL/6VS)基因是目前最有效的抗白粉病基因之一。 分子标记技术是进行抗病基因鉴定、抗病基因积累的有效方法,现已被广泛用于小麦抗病辅助选择育种中。本研究就是通过运用AFLP技术,对抗白粉病材料簇毛麦、6A/6V代换系、6AL/6VS易位系和感白粉病材料京411进行大量的引物筛选,旨在寻找出与抗白粉病基因Pm21紧密连锁的分子标记,从而为克隆该基因以及为小麦抗白粉病的分子标记辅助选择育种奠定基础。 对杂交组合京411与6A/6V代换系的F2代分离群体进行了白粉病抗性鉴定,结果显示,抗、感分离比符合3∶1的比例,证实抗白粉病基因Pm21是属于显性单基因遗传。 通过对AFLP预扩增反应中模板浓度、Mg2+浓度及引物浓度等主要条件的优化,确认在20μl体系中,模板量为500ng、Mg2+终浓度为1.5mM、引物量为30ng时,预扩增的结果最好,从而建立了AFLP反应最佳体系,为后续实验的顺利进行提供了条件。 采用AFLP技术对抗病材料簇毛麦、6A/6V代换系、6AL/6VS易位系和感病材料京411进行100对引物组合的筛选,结果发现有20对引物组合可以在抗感材料中扩增出多态性条带,多态性率为20%,平均每对引物组合扩增的产物分子量范围均在80bp—1kb之间,扩增条带数平均为100条左右。 采用京411与6A/6V代换系的杂交F2代120株单株进行AFLP扩增来检测20对引物组合扩增出的特异性片段与抗白粉病基因Pm21之间的连锁性关系。结果表明,20对引物组合中只有3对引物组合可以扩增出特异性条带,说明这三个扩增片段是与抗病基因Pm21相连锁的。而其它17对引物组合的扩增结果均未表现出差异性,说明这些扩增片段与抗白粉病基因Pm21之间无连锁性。
【Abstract】 Powdery mildew, caused by Erysiphe graminis DC.f.sp.tritici, is one of the most important wheat diseases in China and in many regions of the world. In recent years, the damage of the wheat powdery mildew gets more and more serious in many wheat regions of our country and it made severe damage in wheat production. Though chemistry controlling is useful, breeding for resistance has proved to be the most economical and effective way for controlling wheat diseases. Up to now, more than 30 genes loci for resistance to powdery mildew have been identified. Furthermore, many resistance genes have been tagged and assigned to specific chromosomes or chromosome arms. Among these genes, the Pm21 (6AL/6VS), originating from Haynaldia villosa, is one of the most effective resistance genes.Molecular marker technology is an effective measure for the identification and combinations of the resistance genes and it has been widely used to marker-assisted selection and gene pyramiding in wheat resistance breeding. In this study, amplified fragments length polymorphism (AFLP) analysis was carried out in common wheat (AABBDD), Haynaldia villosa, 6A/6V substitution line and 6AL/6VS translocation line. The objectives of this study were to search new AFLP markers tightly linked to Pm21 gene. It will be also used for the cloning of Pm21 gene and marker-assisted selection (MAS) and wheat resistance breeding.F2 population from a cross between Jing411 and 6A/6V substitution line were identified for resistance to powdery mildew. The results showed the expected Mendelian ratio of 3:1 and indicated that the resistance was controlled by a single dominant gene.With the research of main condition including concentration of template、 Mg2+ and primer, the result of pre-amplified is good when the content of template is 500ng, Mg2+ final concentration is 1.5mM and the content of primer is 30ng in 20μl reaction system. Optimal AFLP reaction system is established and it provided advantaged condition for next study.
【Key words】 Wheat; Haynaldia villosa; Translocation line; Powdery mildew resistance gene Pm21; AFLP marker; Linkage analysis;
- 【网络出版投稿人】 天津师范大学 【网络出版年期】2007年 02期
- 【分类号】S512.1
- 【被引频次】1
- 【下载频次】83