大葱细胞质雄性不育系及保持系线粒体基因组RFLP分析

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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F0 ATP合成酶复合物亚基6(ATP synthase F0 subunit 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 BamHⅠ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。

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	王婵
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	王永勤

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)

Abstract：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 BamHⅠ 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 HindⅢ/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 (HindⅢ/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.

Key words： Welsh onion Cytoplasmic male sterile Mitochondrial genome Restriction fragment length polymorphism (RFLP) NADH dehydrogenase subunit 5 (nad5)

收稿日期: 2014-12-18 出版日期: 2015-05-13

ZTFLH:

S633.1

基金资助:国家自然科学基金;北京市农林科学院科技创新能力建设专项

通讯作者: 王永勤 E-mail: wangyongqin@nercv.org

引用本文:

王婵赵泓张丽英裴雁曦王永勤. 大葱细胞质雄性不育系及保持系线粒体基因组RFLP分析[J]. , 2015, 23(7): 888-893.

链接本文:

http://journal05.magtech.org.cn/Jwk_ny/CN/ 或 http://journal05.magtech.org.cn/Jwk_ny/CN/Y2015/V23/I7/888

Akagi H，Sakamoto M，Shinjyo C，Shimada H，Fujimura T. A unique sequence located downstream from the rice mitochondrial atp6 may cause male sterility[J]. Curr Genet，1994, 25（1）：52-58.
Dewey R E，Levings C S III，Timothy D H. Novel recombinations in the maize mitochondrial genome produce a unique transcriptional unit in the Texas male-sterile cytoplasm[J]. Cell，1986, 44（3）：439-449.
Engelke T，Tatlioglu T. Mitochondrial genome diversity in connection with male sterility in Allium schoenoprasum L.[J]. Theoretical and Applied Genetics，2000, 100：942-948.
Ernst J M，Speel A. Detection and amplification systemsitive multiple-target DNA and RNA in situ hybridization: looking inside cells with a spectrum of colors[J]. Histochem Cell Biol，1999, 112：89-113.
Gai S P，Meng X D. 2010. Application of molecular markers linking to cytoplasmic male sterile loci to assist maintainer line selection and their selection efficiency in welsh onion(Allium fistulosum L.) [J]. Agricultural Sciences in China，2010, 9（11）：1571- 1576.
Havey M. A putative donor of S-cytoplasm and its distribution among open-pollinated populations of onion. Theoretical and Applied Genetics[J]，1993, 86：128-134.
Holford R，Croft J，Newbury H. Differences between, and possible origins of, the cytoplasms found in fertile and male-sterile onions (AIlium cepa L.) [J]. Theoretical and Applied Genetics，1991, 82：737-744.
Iwabuchi M，Kyozuka J，Shimamoto K. Processing followed by complete editing of an altered mitochondrial atp6 RNA restores fertility of cytoplasmic male sterile rice[J]. European Molecular Biology Organization Journal，1993, 12(4)：1437-1446.
Kadowaki K，Suzuki T, Kazama S. A chimeric gene containing the 5' portion of atp6 is associated with cytoplasmic male-sterility of rice[J]. Molecular General Genetics，1990, 224：10-16.
L. Homme Y, Stahl R J, Li X Q, Hameed A, Brown G G. Brassica napus cytoplasmic male sterility is associated with expression of a mtDNA region containing a chimeric gene similar to the pol CMSassociat ed orf224 gene[J]. Curr Genet ,1997, 31: 325 -335
Levings C S III，Pring D R. Restriction endonuclease analysis of mitochondrialDNA from normal and Texas cytoplasmic male-sterile maize. Science，1976, 193：158–169.
Macfarlane JL, Wahleithner JA, Wolstenholme DR. A gene for cytochrome c oxidase subunit III (COXIII) in broad bean mitochondrial DNA: structural features and sequence evolution[J]. Curr Genet. 1990, 17(1):33-40.
苗锦山，杨文才，李美芹，沈火林. 葱细胞质雄性不育与花蕾发育过程中活性氧代谢关系研究[J]. 华北农学报，2009, 24（4）：92-95(Miao Jin-shan，Yang Wen-cai，Li Mei-qin, Shen Huo-lin. Studies on the Relationships between Cytoplasmic Male Sterility and Reactive Oxygen Species Metablisms during Development of Welsh Onion( A. f istulosum) Buds. Acta Agriculturae Boreali-Sinica. 2009, 24（4）：92-95.).
苗锦山，杨文才，刘彩霞，李美芹，乔宁，沈火林. 葱胞质雄性不育花蕾生化物质含量和能量代谢酶活性的动态变化特征[J]. 西北植物学报. 2010, 30（6）：1142-1148(Miao Jin-shan，Yang Wen-cai，Liu Cai-xia，Li Mei-qin，Qiao Ning，Shen Huo-lin. Dynamic changes of physiological and biochemical substances and energy metablism enzyme activities in bud of welsh onion CMS Line. Acta Bot. Boreal.-Occident . Sin，2010, 30（6）：1142-1148.).
Nishimura Y，Shibano M. Male sterility in A. fistulosum L. Cytological and anatomical studies[J]. Journal of the Japanese Society for Horticultural Science，1972, (Suppl 1)：180–181. (in Japanese)
Schnable P S，Wise R P. The molecular basis of cytoplasmic male sterility and fertility restoration[J]. Trends in Plant Science，1998, 3，175–180.
Singh M，Brown G G. Suppression of cytoplasmic male-sterility by nuclear genes alters expression of a novel mitochondrial generegion[J]. Plant Cell，1991, 3：1349–1362.
Wang C，LI H Y，Zhang LY，Pei YX，Wang YQ. Identification of an AFLP marker and conversion to a SCAR marker to identify cytoplasmic male-sterile or normal cytoplasm in Welsh onion (Allium fistulosum L.) [J]. Journal of Horticultural Science & Biotechnology，2013, 88 （4）：409–414.
王晓静，沈火林，杨学妍，程杰山.大葱雄性不育系及保持系花药和花粉发育的细胞学比较研究[J]. 中国瓜菜，2007, （2）：7-10(Biotechnology，88 （4）：409–414(Wang Xiao-jing，Shen Huo-lin，Yang Xue-yan，Cheng Jie-shan. A cytological study on compar ison of anther and pollen development in male- ster ile and cor r esponding maintainer lines in Chinese onion, China melons and vegetables. 2007, （2）：7-10.).
习湘媛. 大葱雄性可育系及雄性不育系的花药花粉发育的比较研究[J]．植物学报，1991, 33（10）：770-771(Xi Xianyuan. Acomparative study of anther and pollen development in male sterile green onion(Allium fistulosum L.) Acta botanica sinica，1991, 33（10）：770-771.).
Young E G，Hanson M R. A fused mitochondrial gene associated with cytoplasmic male sterility is developmentally regulated[J]. Cell，1987, 50（1）：41-49.
Zhang Q P. Breeding male sterile sterile line on the welsh onion and preliminary study of its heterosis. Acta Horticulture，1995, 402-403.
张启沛，魏佑营，张琦.大葱自然群体的雄性不育性[J]. 山东农业大学学报，1987, 18（4）：1-10(Zhang Qi-pei，Wei You-ying，Zhang Qi. Male sterility of Welsh onion (Allium fistulosum L.) in a natural population. Journal of Shandong Agricultural University，1987, 18：1-10).
张启沛. 葱. 见：周长久主编. 蔬菜种质资源概论[M]. 北京：北京农业大学出版社, 1995(Zhang Qi-pei. 1995. Welsh onion. In：Zhou Changjiu Vegetable germplasm introduction. Beijing: Beijing Kasetsart University press.1995).
张晓，张锐，史计，孟志刚，孙国清，周焘，郭三堆.陆地棉胞质雄性不育系与保持系线粒体基因组RFLP分析[J]. 中国农业科学，2012, 45（2）：208-217(Zhang Xiao，Zhang Rui，Shi Ji，Meng Zhi-gang，Sun Guo-qing，Zhou Tao，Guo San-dui. RFLP Analysis of Mitochondrial Genomes Between CytoplasmicMale Sterile Line and Maintainer Line in Upland Cotton. Scientia Agricultura Sinica，2012, 45（2）：208-217.).