三例小麦细胞质雄性不育系线粒体DNA的扩增片段长度多态性分析  

Analysis of three wheat cytoplasmic male sterile lines mitochondrial DNA by AFLP

作　　者：朱启迪[1] 张新钵[1] Ejaz M 张改生[1] 车会学[1] 王书平[1] 宋齐鲁[1] 杨书玲[1] 张龙雨[1] 

机构地区：[1]西北农林科技大学农学院,陕西杨凌712100

出　　处：《生物工程学报》2013年第5期646-656,共11页Chinese Journal of Biotechnology

基　　金：国家高技术研究发展计划重大专项(863计划)(No.2011AA10A106);国家自然科学基金(Nos.31071477;31171611);高等学校博士学科点专项科研基金(No.20090204110024);陕西省"13115"科技创新工程重大科技专项(No.2010ZDKG-68)资助~~

摘　　要：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 is an important way to utilize wheat heterosis. The purpose of thisstudy was to identify cytoplasmic type of three wheat male sterile lines. Amplified fragment length polymorphism （AFLP） marker technique was used to analyze the wheat mitochondrial DNA. We isolated mitochondria by differential centrifugation and density gradient ultracentrifugation. The results show that the extracted mitochondrial DNA was pure. It was suitable for PCR and genetic analysis. We got 4 pairs of specific primers from 64 primers combinations. Primer El/M7 amplified 3 specific fragments in ms（Kots）-90-110. Primer E4/M2 generated 2 specific fragments in ms（Ven）-90-110. Primer E7/M6 amplified 2 specific fragments in ms（S）-90-110. Primer E6/M4 produced 2 specific fragments in ms（Kots）-90-110. Four specific primers could be used to identify three cytoplasmic types ofAegilops kotschyi, Ae. ventricosa and Triticum spelta. It provided the molecular basis to further study the mechanism of wheat cytoplasmic male sterility.

关 键 词：小麦 细胞质雄性不育 线粒体DNA 扩增片段长度多态性 分子标记 

分 类 号：S512.1[农业科学—作物学] Q943.2[生物学—植物学]