小麦抗叶锈基因Lr45的SCAR标记.pdfVIP

中国农业科学 2009,42(1):124-129 Scientia Agricultura Sinica doi: 10.3864/j.issn.0578-1752.2009.01.015 小麦抗叶锈基因 Lr45 的SCAR标记 闫红飞，刘春燕，高士刚，杨文香，刘大群 （河北农业大学植物病理系，河北省农作物病虫害生物防治工程技术研究中心，河北保定 071001 ） 摘要： 【目的】建立小麦抗叶锈基因Lr45 SCAR（sequenced characterized amplified region序列特征扩 增区域）标记。【方法】以小麦抗叶锈病基因材料 TcLr45 和感病材料 Thatcher 为亲本，利用黑麦基因组特异的 RAPD标记引物 OPH20进行PCR 扩增，对获得的与预期大小相同的1.5kb 的特异片段进行克隆测序，并根据该序列 设计一对特异PCR引物LRYR和LRYF，对TcLr45×Thatcher F2 代单株构建的分离群体进行扩增，验证该标记与 Lr45 的连锁关系，Mapmaker 3.0软件绘制遗传连锁图。【结果】该标记在TcLr45中扩增出单一条带，片段大小为1 272 bp（命名为 Ypsc20H ），而在感病亲本中则无扩增条带。F代分离群体进行连锁分析，遗传距离为 8.2 cM，该 1272 2 标记为与 Lr45 连锁的SCAR标记。【结论】将 Lr45 的RAPD标记转化成SCAR标记。 关键词：黑麦；抗小麦叶锈基因；Lr45; SCAR 标记 A SCAR Marker for Leaf Rust Resistance Gene Lr45 YAN Hong-fei, LIU Chun-yan, GAO Shi-gang, YANG Wen-xiang, LIU Da-qun (Department of Plant Pathology, Agricultural University of Hebei, Biological Control Center of Plant Diseases and Plant Pests of Hebei Province, Baoding 071001, Hebei) Abstract: 【Objective 】 The objective was to develop a SCAR(sequenced characterized amplified region) marker for wheat leaf rust resistance gene Lr45. 【Method 】 Wheat leaf rust resistance near isogenic lines of TcLr45 and its susceptible control Thatcher were employed. PCR was carried out with the OPH20, which is a specific RAPD primer of rye. The amplified 1.5kb fragment from TcLr45 was cloned and sequenced. Subsequently a pair of specific primers (LRYR, LRYF) were designed for the amplifying F2 population derived from TcLr45×Thatcher, and then linkage was analyzed between the marker and Lr45. 【Result 】 A unique band was obtained only in TcLr