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小麦叶锈病抗性基因Lr1的高分辨率定位及Lr1位点BAC克隆的特征分析

High-resolution mapping of the leaf rust disease resistance gene Lr1 in wheat and characterization of BAC clones from the Lr1 locus.

作者信息

Ling H-Q, Zhu Y, Keller B

机构信息

Institute of Plant Biology, University of Zürich, Zollikerstrasse 107, Switzerland.

出版信息

Theor Appl Genet. 2003 Mar;106(5):875-82. doi: 10.1007/s00122-002-1139-2. Epub 2002 Oct 30.

Abstract

Leaf rust is the most common disease in wheat production. There are more than 45 specific resistance genes described and used in wheat breeding to control epidemics of leaf rust, but none of them has been cloned. The leaf rust disease resistance gene 1 ( Lr1) is a good model gene for isolation by map-based cloning because it is a single, dominant gene which is located in the distal region of chromosome 5DL of wheat. As the first step towards the isolation of this gene we constructed a high-resolution genetic map in the region of the Lr1 locus by saturation mapping of two large segregating F(2) populations (Thatcher Lr1 x Thatcher, Thatcher Lr1 x Frisal). The resistance gene Lr1 was delimited in a 0.16-cM region between the RFLP markers ABC718 and PSR567 (0.12 cM from ABC718 and 0.04 cM from PSR567). A genomic BAC library of Aegilops tauschii (D genome) was screened using the RFLP markers ABC718 and PSR567. Five positive BAC clones were identified by ABC718 and four clones by PSR567. Two NBS-LRR type of resistance gene analogs, which encode proteins highly homologous to the bacterial blight disease resistance protein Xa1 of rice, were identified on BAC clones isolated with PSR567. Polymorphic BAC end probes were isolated from both ends of a 105-kb large BAC clone identified by ABC718. The end probes were mapped at the same locus as ABC718, and no recombination event was found within 105 kb around ABC718 in our analysis of more than 4,000 gametes.

摘要

叶锈病是小麦生产中最常见的病害。在小麦育种中,已有超过45个特定的抗病基因被描述并用于控制叶锈病的流行,但它们均未被克隆。叶锈病抗病基因1(Lr1)是通过图位克隆进行分离的一个很好的模式基因,因为它是一个单显性基因,位于小麦5DL染色体的远端区域。作为分离该基因的第一步,我们通过对两个大型分离F2群体(撒切尔Lr1×撒切尔、撒切尔Lr1×弗里萨尔)进行饱和作图,构建了Lr1基因座区域的高分辨率遗传图谱。抗病基因Lr1被定位在RFLP标记ABC718和PSR567之间0.16厘摩的区域内(距ABC718 0.12厘摩,距PSR567 0.04厘摩)。使用RFLP标记ABC718和PSR567筛选了节节麦(D基因组)的基因组BAC文库。通过ABC718鉴定出5个阳性BAC克隆,通过PSR567鉴定出4个克隆。在用PSR567分离的BAC克隆上鉴定出两个NBS-LRR类型的抗病基因类似物,它们编码的蛋白质与水稻白叶枯病抗病蛋白Xa1高度同源。从通过ABC718鉴定出的一个105 kb大的BAC克隆的两端分离出多态性BAC末端探针。末端探针被定位在与ABC718相同的位点,并且在我们对4000多个配子的分析中,在ABC718周围105 kb范围内未发现重组事件。

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