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细胞质雄性不育是小麦杂种优势利用的重要途径,为了鉴定3例小麦雄性不育系的细胞质类型,对其线粒体DNA(mtDNA)进行扩增片段长度多态性(Amplified fragment length polymorphism,AFLP)分析。文中利用差速离心法和不连续蔗糖密度梯度超速离心法提取纯化小麦线粒体。结果表明:通过该提取方法获得的mtDNA,其质量和纯度能够满足PCR反应和遗传学分析。在64对选扩引物中,筛选到了4对特异性引物,其中引物E1/M7在ms(Kots)-90-110不育系扩增出3条特异条带;引物E4/M2在ms(Ven)-90-110不育系扩增出2条特异条带;引物E7/M6在ms(S)-90-110不育系中扩增出2条特异条带;引物E6/M4在ms(Kots)-90-110不育系中扩增出2条特异条带。这些特异引物可以用来作为鉴定具有粘果山羊草Aegilops kotschyi、偏凸山羊草Ae.ventricosa、斯卑尔脱小麦Triticum spelta 3类不育细胞质型小麦雄性不育系的细胞质分子标记,为研究小麦细胞质雄性不育机理奠定了分子基础。
Cytoplasmic male sterility (CMS) is an important pathway for heterosis utilization of wheat. In order to identify the cytoplasmic types of three male sterile lines of wheat, the mtDNA was analyzed for the amplified fragment length polymorphism (AFLP) . Purification of wheat mitochondria by differential centrifugation and discontinuous sucrose density gradient ultracentrifugation. The results showed that the quality and purity of mtDNA obtained by this method can meet the needs of PCR reaction and genetics analysis. Among 64 pairs of primers, 4 pairs of specific primers were screened, among which, three specific bands were amplified from the elite lines of E1 / M7 in ms (Kots) -90-110. The primer E4 / ) -90-110. Two specific bands were amplified from the male sterile lines of MS (S) -90-110 by primer E7 / M6. Kots) -90-110 male sterile lines amplified two specific bands. These specific primers can be used as a marker for identification of cytoplasmic molecular markers in CMS lines Aegilops kotschyi, Ae.ventricosa, and Triticum spelta. Cytoplasmic male sterility mechanism laid the molecular basis.