烟草不育系与保持系线粒体DNA的重测序研究

Re-sequencing Study on the Mitochondrial DNA of Tobacco Sterile Line and Maintainer Line

基于重测序技术,对两对不育系和对应的保持系材料的线粒体DNA进行重测序,统计并分析了不育系与保持系线粒体DNA编码区序列的突变位点,探讨了这些突变位点与烟草胞质不育的关系。对重测序结果比对分析可知:不育系和保持系在orf106b上具有一致的1个In Del位点,在orf138a、orf121b、orf129b和orf110b上具有一致的SNP位点,这些突变位点没有造成不育系与保持系编码区序列的差异,推测这些突变位点可能与CMS无关;不育系MSK326和MSK394在orf112a和orf103c上有相同的In Del位点;MSK326和MSK394在65个基因片段或开放阅读框上有一致SNP位点,且这些突变没有出现在保持系线粒体编码区。这些基因片段或开放阅读框上的In Del和SNP突变都导致了基因或开放阅读框编码序列的改变,并且造成了氨基酸的突变,推测可能与烟草CMS密切的相关。其中atp1、atp4、atp6、coxl、cox3、atp9、orf25、nad6、nad7、orf108、orf239、orf129、sdh4等基因或开放阅读框上的突变位点与报道研究一致,均被报道与烟草CMS存在密切的联系。本研究所挖掘的这些大量基因片段上的In Del和SNP位点,不仅有助于烟草CMS机理的进一步研究,也为烟草不育系育种研究提供有力的遗传工具。

Based on the technology of re-sequencing in this study, resequencd the mitochondrial DNA of two pairs of tobacco material of sterile line and maintainer line, statisticed analyzed the differences in mitochondrial DNA sequence of sterile line and maintainer line, discussed the relationship between the mutation site and CMS. For the comparison analysis of resequencing date in this samples,can know that： the sterile line and maintainer line had one same In Del site in orf106 b.The same SNP site were located in orf138 a,orf121b, orf129 b and orf110 b. These mutations had not make differences on coding sequence of sterile line and maintainer line, these mutations may had nothing to do with the CMS. The sterile lines MSK326 and MSK394 have the same In Del site in orf112 a and orf103 c. And they had the same SNP site in 65 genes or open reading frame. But the maintainer line had no mutations in these genes or open reading frame. These In Del and SNP had led to the changes of coding sequence in genes or the open reading frame. These mutant sites likely related to their CMS.Some of these mutant sites, including atp1, atp4, atp6, coxl, cox3, atp9, orf25, nad6, nad7,orf108, orf239, orf129 and sdh4 have been reported. This report have confirmed that these mutant sites likely related to their CMS.We had detected the large number of In Del and SNP sites in our research, this found not only helps to further study, also would offer powerful tools for tobacco germplasm identification and marker-assisted selectionin tobacco breeding.