Feuillet C, Messmer M, Schachermayr G, Keller B
Department of Plant Breeding, Swiss Federal Research Station for Agronomy, Zürich, Switzerland.
Mol Gen Genet. 1995 Sep 20;248(5):553-62. doi: 10.1007/BF02423451.
The objective of this study was to characterize the leaf rust resistance locus Lr1 in wheat. Restriction fragment length polymorphism (RFLP) analysis was performed on the resistant line Lr1/6* Thatcher and the susceptible varieties Thatcher and Frisal, as well as on the segregating F2 populations. Seventeen out of 37 RFLP probes mapping to group 5 chromosomes showed polymorphism between Lr1/6* Thatcher and Frisal, whereas 11 probes were polymorphic between the near-isogenic lines (NILs) Lr1/6* Thatcher and Thatcher. Three of these probes were linked to the resistance gene in the segregating F2 populations. One probe (pTAG621) showed very tight linkage to Lr1 and mapped to a single-copy region on chromosome 5D. The map location of pTAG621 at the end of the long arm of chromosome 5D was confirmed by the absence of the band in the nulli-tetrasomic line N5DT5B of Chinese Spring and a set of deletion lines of Chinese Spring lacking the distal part of 5DL. Twenty-seven breeding lines containing the Lr1 resistance gene in different genetic backgrounds showed the same band as Lr1/6* Thatcher when hybridized with pTAG621. The RFLP marker was converted to a sequence-tagged-site marker using polymerase chain reaction (PCR) amplification. Sequencing of the specific fragment amplified from both NILs revealed point mutations as well as small insertion/deletion events. These were used to design primers that allowed amplification of a specific product only from the resistant line Lr1/6* Thatcher. This STS, specific for the Lr1 resistance gene, will allow efficient selection for the disease resistance gene in wheat breeding programmes. In addition, the identification of a D-genome-specific probe tightly linked to Lr1 should ultimately provide the basis for positional cloning of the gene.
本研究的目的是对小麦叶锈病抗性基因座Lr1进行特征分析。对抗病品系Lr1/6Thatcher、感病品种Thatcher和Frisal以及分离的F2群体进行了限制性片段长度多态性(RFLP)分析。在定位到第5组染色体的37个RFLP探针中,有17个在Lr1/6Thatcher和Frisal之间表现出多态性,而11个探针在近等基因系(NILs)Lr1/6Thatcher和Thatcher之间表现出多态性。其中3个探针在分离的F2群体中与抗性基因连锁。一个探针(pTAG621)与Lr1表现出非常紧密的连锁,并定位到5D染色体上的一个单拷贝区域。通过中国春的缺体-四体N5DT5B和一组缺失5DL远端部分的中国春缺失系中该条带的缺失,证实了pTAG621在5D染色体长臂末端的图谱位置。27个在不同遗传背景下含有Lr1抗性基因的育种系与pTAG621杂交时,显示出与Lr1/6Thatcher相同的条带。利用聚合酶链反应(PCR)扩增将RFLP标记转化为序列标签位点标记。从两个近等基因系扩增的特定片段测序揭示了点突变以及小的插入/缺失事件。这些被用于设计引物,使得仅能从抗病品系Lr1/6*Thatcher中扩增出特定产物。这个对Lr1抗性基因特异的STS,将有助于在小麦育种计划中对抗病基因进行高效选择。此外,鉴定出与Lr1紧密连锁的D基因组特异探针最终应为该基因的图位克隆提供基础。