J Integr Plant Biol. ›› 2003, Vol. 45 ›› Issue (4): 472-478.

• Research Articles • Previous Articles     Next Articles

Isolation and Characterization of Mlo and NBS-LRR-like Gene Sequences in Wheat

LI Ai-Li, KONG Xiu-Ying, ZHOU Rong-Hua, MA Zhi-Ying, JIA Ji-Zeng   

  • Published:2003-04-15


Based on both cDNA sequence of barley powdery mildew resistance control element Mlo and DNA sequence of the known putative disease resistance gene from Triticum monococcum L., we designed some primers to amplify resistant homologous sequences in the near isogenic lines (NILs) of powdery mildew resistance using RT-PCR method. Two expressed cDNA fragments were isolated from wheat genome. One showed 83% homology to the Mlo gene of barley. The other contained two possible open reading frames (ORFs). NBS conservative domains 2, 3 of disease resistance gene and 13 LRR structures similar to rice Pib protein terminal were found respectively in the two ORFs. It indicated that the latter fragment belongs to NBS-LRR-like genes. The obvious difference of RT-PCR products was observed between the before challenged and the challenged for 72 h by Blumeria graminis f. sp. tritici, which implied that this sequence could be associated with disease resistance of wheat. Using nulli-tetrasomic lines of “Chinese Spring", the NBS-LRR-like gene had been located on chromosome 1D. Key words:Mlo; NBS-LRR; cDNA cloning; NIL; RGA

李爱丽  孔秀英 周荣华  马峙英 贾继增

(1. 中国农业科学院作物品种资源研究所,农业部作物种质资源与生物技术重点开放实验室,北京100081;2. 河北农业大学农学院,保定071001)

摘要: 根据GenBank 中公布的大麦白粉病抗性控制基因Mlo cDNA 序列及一个来源于栽培一粒小麦( Triticummonococcum  L. ) 的假定抗病基因序列分别设计引物,以携带小麦抗白粉病基因的近等基因系为材料进行RT- PCR 筛选。结果获得两个表达基因的cDNA 克隆。其中一个与大麦白粉病抗性控制基因Mlo 的同源性达83 %。另一个为非通读序列,含有两个可能的开放阅读框,分别包含抗病基因NBS 保守结构域2 和3 以及与水稻抗稻瘟病基因Pib蛋白末端相似的13 个LRR 区域,推测该序列属于NBS-LRR 类。白粉菌诱导前后,该片段RT- PCR 扩增产物存在差异,表明该片段可能与小麦抗病性相关。利用“中国春”缺体-四体系,将该NBS-LRR 类序列定位在小麦1D 染色体上。
关键词:  Mlo ; NBS-LRR ; cDNA 克隆; 近等基因系; RGA

通讯作者。E-mail: jizeng @ mimi. cnc.ac. cn。

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