簇毛麦基因组特异性PCR标记的建立和应用

以普通小麦中国春、簇毛麦、中国春－簇毛麦二体附加系和代换系为材料进行RAPD分析，筛选出一个簇毛麦基因组特异性RAPD片段OPFO2757，该片段分布于簇毛麦所有染色体上。在对OPFO2757进行克隆、测序的基础上，设计一对PCR引物，建立了簇毛麦基因组特异性PCR标记。用这对PCR引物对不同普通小麦品种、不同硬粒小麦品种、不同居群的簇毛麦、中国春－簇毛麦二体附加系、中国春－簇毛麦二体代换系、普通小麦－簇毛麦双二倍体、硬粒小麦－簇毛麦双二倍体等材料进行扩增，凡具有簇毛麦染色体的材料都能扩增出一条长为677bp的DNA片段，而不具簇毛麦染色体的材料包括大麦、黑麦、长穗偃麦草、中间偃麦草等不能扩增出该片段。所以，该特异性PCR标记可用于快速跟踪检测小麦背景中的簇毛麦染色体。

Development and Application of a Genome Specific PCR Marker for Haynaldia villosa

Random amplified polymorphic DNA (RAPD) analysis was performed on common wheat Chinese Spring, H.villosa ,addition lines of H.villosa chromosome in CS,substitution line 3V of H.villosa chromosome in Triticum aestivum. A genome specific polymorphic DNA segment from H.villosa ,OPF02 757 ,was obtained.On the basis of cloning and sequencing of OPF02 757 ,two PCR primers were designed and a genome specific PCR marker for H.villosa was established.The PCR marker including 677 bp was localized on all the seven pairs of H.villosa chromosomes.The result of PCR amplification by the primers indicated that there was a specific band of 677 bp in the materials containing H.villosa Chromosome such as T.aestivum H.villosa addition, T.aestivum H.villosa substitution, T.aestivum H. villosa amphidiploid, T.durum H.villosa amphidiploid and H.villosum from different accessions,and there was no specific band of 677 bp if the materials did not contain H.villosa chromosome,such as T.aestivum,T.durum,Secale cereale,Hordeum vulgare,Thinopyrum elongatum,Thinopyrum intermedium. Therefore,the PCR maker of 677 bp is specific to H.villosa genome,and could be used as a molecular marker for detection of chromosomes of H.villosa in wheat.