簇毛麦基因组特异DNA序列标记的建立和应用

Development and Application of A Dasypyrum Genome-specific DNA Marker

为建立簇毛麦基因组特异DNA序列的分子标记,以普通小麦中国春、绵阳11、川农17和R111以及多年生簇毛麦、硬粒小麦-多年生簇毛麦双二倍体TDB-3为材料,用120条随机引物进行RAPD分析,筛选出一个多年生簇毛麦的特异性RAPD片段,克隆测序表明该片段全长为937 bp(记为OPM2937)。以OPM2937的DNA序列为基础设计了一对特异引物,并用该特异引物对多年生簇毛麦、二倍体簇毛麦以及中国春-二倍体簇毛麦附加系CSDA1V^CSDA7V与小麦-多年生簇毛麦衍生后代进行了扩增,结果表明,凡具有簇毛麦染色体的材料均可扩增出目标DNA片段,而不具簇毛麦染色体的材料均未能得到扩增。将OPM2937在NCBI中进行序列比对,发现它是一个新的序列,说明OPM2937为簇毛麦基因组的一个特异序列,该DNA序列标记可用于快速跟踪检测小麦背景中的簇毛麦染色体。

Random amplified polymorphic DNA （RAPD） analysis was performed on common wheat lines Chinese Spring, Mianyang11, Chuannong 17, and Rl11, as well as Dasypyrurn breviaristaturn, Triticurn durum- D. breviaristaturn amphiploid with 120 random primers, and a D. breviaristaturn specific RAPD fragment was obtained. The fragment of 937 bp was revealed by cloning and sequencing, which was thus designated as OPM2937. Based on the sequence of OPM2937, A pair of primer was developed, and used to amplify the D. breviaristaturn, D. villosurn, T. aestivurn Chinese Spring-D. villosurn addition lines CSDA1V to CSDA7V, and wheat-D. breviaristaturn derivatives. The results indicated that the target product of 900bp was produced only from the lines with the Dasypyrurn chromosomes, while it was not observed in the lines belong to the common wheat. The OPM29~7 was a new sequence when Blast it in NCBI gene bank, which further confirmed that it was a Dasypyrurn genome-specific sequence. It could be used a novel molecular marker for detecting and tracing the Dasypyrurn chromatin introduced in wheat background.