Development of sequence tagged site and cleaved amplified polymorphic sequence markers for wheat stripe rust resistance gene Yr5
Development of sequence tagged site and cleaved amplified polymorphic sequence markers for wheat stripe rust resistance gene Yr5
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DOI：
10.2135/cropsci2003.2058
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发表时间：
2003-11-01
期刊：
影响因子：
2.3
通讯作者：
Dubcovsky, J
中科院分区：
文献类型：
--
作者：
Chen, XM;Soria, MA;Dubcovsky, J
Yr5基因对小麦条锈菌[Puccinia striiformis Westend]的所有小种都具有抗性。F.sp.特里奇·埃里克。(P.S.tritici)]到目前为止在美国被鉴定。Yr5的共分离抗性基因模拟多态(RGAP)标记是可用的，但它们的使用需要聚丙烯酰胺凝胶电泳术，并且可能在不同品种之间不是多态的。为了开发更好的分子标记用于Yr5抗性的分子标记辅助选择，根据来自春小麦(Triticum aestivum L.)‘Avocet Sensible’(AVS)的RGAP标记Xwgp-18(AY167598)和来自Yr5的近等基因系(NIL)的Xwgp-17(AY167597)序列设计了序列标记位点(STS)。Spelta(L.)瑟尔。CV。专辑(TSA)。开发了3组STS标记(2个共显性标记和1个显性标记)来扩增含有多态6-碱基对(BP)插入-缺失(INDel)的区域。用AVS和TSA杂交育成的114个Bc7：F-3系证实了STS标记与Yr5的共分离。STS标记在5个非Yr5小麦品种和17个非Yr5小麦品种中效果良好，但其余品种的片段大小与Yr5标记相似。因为共显性的STS标记是基于6个碱基的插入片段，所以不能用琼脂糖凝胶电泳法进行分离。裂解扩增多态序列(CAPS)标记是基于DpnII酶切位点开发的，该酶切位点在所有非Yr5品种中都存在，在Yr5近等基因系中不存在。用琼脂糖凝胶电泳法可以分离出Yr5、Nil和非Yr5品种的CAPS标记。共显性STS标记比原始RGAP标记更容易得分。CAPS标记不仅更容易评分，而且还可以用于Yr5供体与范围更广的小麦种质的杂交。这些标记将是加速Yr5导入到商业品种中的有价值的工具，并将Yr5与其他持久抗条锈病的基因相结合。
The Yr5 gene confers resistance to all races of the wheat stripe rust pathogen [Puccinia striiformis Westend. f. sp. tritici Eriks. (P. s. tritici)] identified so far in the USA. Cosegregating resistance gene analog polymorphism (RGAP) markers for Yr5 are available but their use requires skills in polyacrylamide gel electrophoresis and may not be polymorphic across various varieties. To develop better markers to be used in marker-assisted selection for the Yr5 resistance, sequence tagged site (STS) primers were designed on the basis of the sequences of RGAP markers Xwgp-18 (AY167598) from the spring wheat (Triticum aestivum L.)'Avocet Susceptible' (AVS) and Xwgp-17 (AY167597) from the Yr5 near isogenic line (NIL) in the AVS background carrying the Yr5 gene from T. aestivum subsp. spelta (L.) Thell. cv. Album (TSA). Three sets of STS markers (two codominant and one dominant) were developed to amplify a region including a polymorphic 6-base pairs (bp) insertion-deletion (indel). The cosegregation of the STS markers with Yr5 was confirmed with 114 BC7:F-3 lines developed from the cross between AVS and TSA. The STS markers worked well in five out or 17 non-Yr5 wheat varieties, but the remaining varieties had a similar size of fragment to the Yr5 marker. Because the codominant STS markers were based on a 6-bp indel, they could not be separated by agarose gel electrophoresis. Cleaved amplified polymorphic sequence (CAPS) markers were then developed on the basis of a DpnII restriction site that is present in all non-Yr5 varieties and absent in the Yr5 NIL. The CAPS markers for the Yr5 NIL and non-Yr5 varieties can be separated by agarose gel electrophoresis. The codominant STS markers are easier to score than the original RGAP markers. The CAPS markers are not only easier to score, but also can be used in crosses of an Yr5 donor with a much wider range of wheat germplasms. These markers should be valuable tools to accelerate the introgression of Yr5 into commercial cultivars and to combine Yr5 with other genes for durable resistance to stripe rust.