小麦T、K、V型胞质不育系和杂种mtDNA的RAPD分析及育性相关片段的克隆

RAPD Analysing of CMS-T, K, V mtDNAs and Cloning of mtDNA Fragments Associated with CMS K in Wheat

用 RAPD技术对细胞质分别来源于粘果山羊草 (Ae.kotschyi)、偏凸山羊草 (Ae.ventricosa)、提莫菲维小麦 (T.timopheevii)的三种普通小麦雄性不育系 - - K型、 V型、 T型及相应的保持系、恢复系以及杂种 F1代的线粒体 DNA(mt DNA)进行了比较分析。结果如下 :(1)发现在 mt DNA组织结构上 K型、V型、T型不育系之间以及与保持系之间均呈现差异 ,这从 DNA水平上提供了三类不育系胞质来源不同的证据 ;(2 )利用 2 3个引物的扩增结果进行了聚类分析 ,绘制出了 T型、K型、V型三种不育系、保持系和 K、V型共同的恢复系及杂种 F1代树式遗传关系图 ;(3)对两个特异的多态性DNA片段进行了回收 ,克隆、并制备探针 ,Southern杂交结果显示不育系与保持系间存在多态性 ,有两个片段可能与育性有一定的联系。

Mitochondrial DNAs(mtDNAs) from three types of cytoplasmic male sterility lines (CMS K, V and T) in wheat, of which the cytoplasm derived from Ae. kotschyi , Ae. ventricosa and T. timopheevii, respectively, were compared among CMSs and their maintainer, restorer and hybrids of CMS K and V. The results indicated that: (1) the mtDNAs from CMS K, V and T were different from each other and from their maintainer, this provided some evidence at molecular level for three kinds of CMS lines with different cytoplasms; (2) the genetic dendrogram was made and the relative genetic distance among 7 different wheat materials was calculated by cluster analysis with amplification products from 23 primers; (3) two specific fragments of mtDNA were then cloned into plasmid vector and converted into RFLP probes.Southern hybridization patterns revealed polymorphism between sterile line and maintainer line .