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作物学报 ›› 2011, Vol. 37 ›› Issue (12): 2285-2292.doi: 10.3724/SP.J.1006.2011.02285

• 研究简报 • 上一篇    下一篇

四种细胞质来源的烟草不育系线粒体SSR位点差异

李凤霞,杨爱国,崔萌萌,龚达平,王卫锋,孙玉合*   

  1. LI Feng-Xia,YANG Ai-Guo,CUI Meng-Meng,GONG Da-Ping,WANG Wei-Feng,SUN Yu-He*
  • 收稿日期:2011-05-23 修回日期:2011-09-18 出版日期:2011-12-12 网络出版日期:2011-09-29
  • 通讯作者: 孙玉合, E-mail: yhsun@163.com
  • 基金资助:

    本研究由国家烟草专卖局重点科技项目(110200601001)和中国农业科学院作物科学研究所基本科研业务费资助。

Mitochondrial Microsatellite Variability of Tobacco CMS Lines with Four Different Cytoplasms

中国农业科学院烟草研究所 / 中国农业科学院烟草遗传改良与生物技术重点开放实验室，山东青岛 266101   

  1. Key Laboratory of Tobacco Genetic Improvement and Biotechnology, Tobacoo Research Institute, Chinese Academy of Agricultural Sciences, Qingdao 266101, China
  • Received:2011-05-23 Revised:2011-09-18 Published:2011-12-12 Published online:2011-09-29
  • Contact: 孙玉合, E-mail: yhsun@163.com

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摘要： 采用线粒体SSR(mtSSR)方法对普通烟草4种不育类型的不育系、不育胞质来源的烟草野生种和保持系进行线粒体位点差异分析，探讨mtSSR位点差异与胞质不育的关系。在烟草线粒体基因组上检索到24个能重复稳定扩增出片段的mtSSR，对烟草属植物的这些mtSSR分析结果显示，普通烟草种内mtSSR较为保守，多态性比率仅10.5%；4种不育类型的各自不育系与保持系、不育胞质上存在多个差异位点，在Nta(sua.)S类型中差异位点在线粒体编码基因orf138a、orf138c以及orf102~orf210的基因间区；在Nat(gla.)S类型中，差异位点在线粒体orf138a、orf138c两个编码基因以及nad2~sdh3、rps12~orf125b、orf116~orf101b、orf102~orf210的4个基因间区；在Nta(rep.)S类型中，差异位点在线粒体编码基因orf190以及orf104c~orf202的基因间区；在Nta(rus.)S类型中，差异位点在线粒体编码基因orf137以及rps12~orf125b的基因间区。这些位点差异可能与各自的雄性不育有关。另外，mtSSR引物TMS20在Nat(sua.)S、Nat(gla.)S、Nat(rep.)S和Nat(rus.)S类型的雄性不育系中分别扩增出了184、172、212和225 bp的片段，表明可用该引物区分4种烟草CMS类型。

关键词: 烟草, 不育胞质类型, 线粒体DNA, SSR

Abstract: Sequence variation of the mitochondrial DNA has a direct relation with cytoplasmic male sterility (CMS). In this study, we analyzed mtSSR variability offour sterile cytoplasm types, including four CMS lines, their maintainer lines and sterial cytoplasm sources, and explored the relationship between mtSSR variability and CMS. Based on the entire mitochondrial DNA sequence of Nicotiana tabacum, we identified 39 mtSSR loci having more than ten mononucleotide repeats and 24 mtSSR markers that can be used to analyze mitochondrial DNA variability of four sterile cytoplasm types. The results showed that the mtSSR polymorphism was very low in N. tabacum, which was 10.5%, and there were some different loci between the CMS lines and their maintainer lines, the wild tobaccoprovided the sterile cytoplasm. Such as, in the Nat(sua.)S, the different loci were located in coding regions of orf138a, orf138c and intergenic region orf102–orf210; in the Nat(gla.)S, the different loci were located in two coding regions of orf138a, orf138c and four intergenic regionsnad2–sdh3, rps12–orf125b, orf116–orf101b, orf102–orf210; in the Nat(rep.)S, the different loci werelocated in coding regionorf190 and intergenic region orf104c–orf202; in the Nat(rus.)S, the different loci werelocated in coding regions oforf137 and rps12–orf125b. These different loci likely related to their CMS. The results also showed that the fragments of 184, 172, 212 and 225 bp were amplified by mtSSR primer TMS20 in four types of CMS of ms G-28, 86-6, 98-43, 116, respectively, and using this primer we could distinguish the four types of CMS of Nat(sua.)S, Nat(gla.)S, Nat(rep.)S, Nat(rus.)S between each other.

Key words: Tobacco, Sterile cytoplasm type, Mitochondrial DNA, SSR

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