华山新麦草2Ns染色体SCAR标记的开发

Development of a New SCAR Marker on Chromosome 2Ns of Psathyrostachys huashanica Keng

为准确快速地建立小麦-华山新麦草杂交后代的分子标记鉴定方法,利用180条长度为10 bp的随机引物R1~R180对小麦-华山新麦草全套(1Ns~7Ns)二体附加系及其亲本华山新麦草和普通小麦7182共9个材料进行了RAPD分析。结果表明,R131在华山新麦草和小麦-华山新麦草2Ns二体附加系中可以扩增出特异条带。将该特异条带回收并测序,发现其全长为1126 bp,对其进行序列比对分析后设计SCAR引物S131,然后利用S131重新对9份材料进行了SCAR分析。结果显示,S131只在华山新麦草和小麦-华山新麦草2Ns二体附加系中扩增出特异性条带,表明RAPD标记R131已成功转化为可靠、特异的SCAR标记S131。这个新的SCAR标记可用于检测普通小麦背景下华山新麦草2Ns染色体。

In order to establishan effective way for rapid identification of wheat-Psathyrostachys huashanica Keng hybrids by molecular maker,the analysis of RAPD was conducted by using 180 random primers R1-R180(10 bp)in the initial screening of a complete set of wheat-P.huashanica disomic addition lines and its parents P.huashanica and common wheat 7182.Among the 180 primers,we confirmed that R131 can amplify a clear and legible DNA fragment,which was only present in P.huashanica and the 2Ns addition line.A SCAR marker(S131)was designed based on a sequence alignment of the R131 amplicons(1126 bp).Further screening of this SCAR marker using the nine test materials verified its specificity for P.huashanica and the P.huashanica 2Ns addition line,thereby indicating that the R131 RAPD marker was successfully converted into the reliable and specific SCAR marker S131.This novel SCAR marker for detecting the 2Ns chromosome of P.huashanica in a common wheat background will provide a scientific basis to facilitate the molecular identification of wheat-P.huashanica hybrids.