大葱细胞质雄性不育系及保持系线粒体基因组RFLP分析  

RFLP Analysis of Mitochondrial Genomes of Cytoplasmic Male Sterile Lines and Maintainer Lines in Welsh Onion(Allium fistulosum L.)

作　　者：王婵[1,2] 赵泓[1] 张丽英[1] 裴雁曦[2] 王永勤[1] 

机构地区：[1]北京市农林科学院蔬菜研究中心/农业部华北地区园艺作物生物学与种质创制重点实验室/蔬菜种质改良北京市重点实验室,北京100097 [2]山西大学生命科学学院,太原030005

出　　处：《农业生物技术学报》2015年第7期888-893,共6页Journal of Agricultural Biotechnology

基　　金：国家自然科学基金项目(No.31372066);北京市农林科学院科技创新能力建设专项(No.KJCX20140111)

摘　　要：为分析现有大葱(Allium fistulosum L.)不育系及保持系的基因多态性,本研究利用限制性片段长度多态性(restriction fragment length polymorphism,RFLP)技术对5个大葱不育系及4个保持系的线粒体基因组进行了对比分析,利用线粒体基因保守序列探针线粒体F0ATP合成酶复合物亚基6(ATP synthase F0subunit 6,atp6)、NADH脱氢酶亚基5(NADH dehydrogenase subunit 5,nad5)及细胞色素c氧化酶第Ⅲ亚基(cytochrome c oxidase subunit 3,cox3)和HindⅢ和Bam HⅠ两种酶切组合进行Southern杂交,结果显示,大葱线粒体基因组存在丰富的多态性,其中HindⅢ/nad5组合中5个雄性不育系均含有一条10 kb大小的特异条带,4个保持系均具有一条7 kb大小的保守特异条带。本研究为大葱不同胞质类型的鉴定提供新的技术手段,也为克隆大葱线粒体基因,阐明细胞质雄性不育的分子机制提供理论依据。In order to analyze genetic polymorphisms of Allium fistulosum CMS（cytoplasmic male sterile）lines and maintainer, 3 mitochondrial gene probes ATP synthase F0 subunit 6（atp6）, NADH dehydrogenase subunit 5（nad5） and cytochrome c oxidase subunit 3（cox3）, combined with 2 restriction enzymes HindⅢ and Bam HⅠ were used to detect the polymorphism of mitochondrial genomes. The Southern blot results indicated that mitochondrial genome had polymorphism between the CMS lines and the maintainer lines. By using the combination of Hind Ⅲ/nad5 in the RFLP（restriction fragment length polymorphism） experiment, 5 CMS lines showed 1 specific band of 10 kb,and the 4 maintainer lines showed another specific band of 7 kb. In summary, RFLP（Hind Ⅲ/nad5） could be used to identify the type of cytoplasm in Allium fistulosum. These results may help to reveal the molecular mechanisms underlying the CMS and will be useful for mitochondrial gene cloning studies of the Allium fistulosum.

关 键 词：大葱 细胞质雄性不育 线粒体基因组 限制性片段长度多态性(RFLP) NADH脱氢酶亚基5(nad5) 

分 类 号：S565.4[农业科学—作物学]