Characterization and mapping of NBS-LRR resistance gene analogs in apricot (Prunus armeniaca L.)
Characterization and mapping of NBS-LRR resistance gene analogs in apricot (Prunus armeniaca L.)
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DOI：
10.1007/s00122-005-1920-0
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发表时间：
2005-02
影响因子：
5.4
通讯作者：
Jose Miguel Soriano;Santiago Vilanova;C. Romero;G. Llácer;M. Badenes
中科院分区：
文献类型：
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作者：
Jose Miguel Soriano;Santiago Vilanova;C. Romero;G. Llácer;M. Badenes
从杏（Prunus armeniacaL.）基因组DNA中分离并克隆了与NBS-LRR（nucleotide binding site-leucine-rich repeat）抗病基因同源的序列。使用从NBS结构域的保守区域设计的简并引物的PCR方法。限制性酶切和序列分析的扩增片段导致43个独特的氨基酸序列的鉴定，分为6个家庭的抗性基因类似物（RGA）。所鉴定的RGA均属于植物抗病基因（R-基因）的Toll-白细胞介素受体（TIR）组。基于NBS结构域的非保守区域的RGA特异性引物从每个RGA家族的共有序列开发。这些引物被用来开发扩增片段长度多态性（AFLP）-RGA标记的AFLP-修改的程序，其中一个标准引物被取代的RGA特异性引物的装置。使用这种方法，27个多态性标记，其中6个共享的同源性与TIR类的NBS-LRR-基因，从17个不同的引物组合获得。在这27个标记中，有16个标记在先前构建的杏遗传图谱中，该遗传图谱是由品种利托的自花授粉构建的。AFLP-RGA标记的建立为杏抗病基因的标记辅助选择和图位克隆奠定了基础。
Genomic DNA sequences sharing homology with the NBS-LRR (nucleotide binding site-leucine-rich repeat) resistance genes were isolated and cloned from apricot (Prunus armeniacaL.) using a PCR approach with degenerate primers designed from conserved regions of the NBS domain. Restriction digestion and sequence analyses of the amplified fragments led to the identification of 43 unique amino acid sequences grouped into six families of resistance gene analogs (RGAs). All of the RGAs identified belong to the Toll-Interleukin receptor (TIR) group of the plant disease resistance genes (R-genes). RGA-specific primers based on non-conserved regions of the NBS domain were developed from the consensus sequences of each RGA family. These primers were used to develop amplified fragment length polymorphism (AFLP)-RGA markers by means of an AFLP-modified procedure where one standard primer is substituted by an RGA-specific primer. Using this method, 27 polymorphic markers, six of which shared homology with the TIR class of the NBS-LRR R-genes, were obtained from 17 different primer combinations. Of these 27 markers, 16 mapped in an apricot genetic map previously constructed from the self-pollination of the cultivar Lito. The development of AFLP-RGA markers may prove to be useful for marker-assisted selection and map-based cloning of R-genes in apricot.