大葱雄性不育系及保持系atp6基因的克隆及表达分析  

Cloning and Expression Analysis of atp6 Genes between Cytoplasmic Male Sterile Line and Maintainer Line in Welsh Onion

作　　者：王清华 张荣亭 贾述娟 高莉敏 Wang Qinghua;Zhang Rongting;Jia Shujuan;Gao Limin(Vegetable Science Observation and Experimental Station in Huang-Huai District of Ministry of Agriculture,Shandong Branch of National Vegetable Improvement Center,Shandong Key Laboratory of Greenhouse Vegetable Biology,Vegetable and Flower Research Institute of Shandong Academy of Agricultural Sciences,Jinan,250100;Jinan Institute of Agricultural Sciences,Jinan,250316;Jiyuan Agricultural Comprehensive Development Office,Jiyuan,459000)

机构地区：[1]山东省农业科学院蔬菜花卉研究所,山东省设施蔬菜生物学重点实验室,国家蔬菜改良中心山东分中心,农业部黄淮地区蔬菜科学观测实验站,济南250100 [2]济南市农业科学研究院,济南250316 [3]济源市农业综合开发办公室,济源459000

出　　处：《分子植物育种》2021年第7期2163-2168,共6页Molecular Plant Breeding

基　　金：山东省农业良种工程项目(2019LZGC01703);济南农业科学研究院科研项目(yy202001)共同资助。

摘　　要：为进一步研究atp6基因与大葱细胞质雄性不育的关系,以章丘大葱不育系和保持系作为本研究的实验材料,利用同源克隆的方法获得大葱atp6基因序列,并对其在蛋白质二级结构和生长发育各时期atp6基因表达量进行了比较。结果表明:不育系与保持系atp6基因开放阅读框第171碱基处存在一个A/T多态性位点,这一多态性位点使蛋白质二级结构的比例发生了轻微变化,但并未改变ATP6蛋白跨膜结构。利用实时荧光定量聚合酶链反应检测atp6基因苗期、抽薹期、开花期以及开花后期在根系和叶/花蕾中的表达水平,结果表明:在植株整个发育过程中,根系atp6基因表达量表现出先升高后降低的趋势,在开花期达到最大值,保持系是不育系的1.14倍;叶/花atp6基因表达量在整个生育期逐渐降低,苗期最高,保持系是不育系的1.96倍。In order to further study the relationship between atp6 gene and Cytoplasm male sterility(CMS)of bunching onion,the sequence of atp6 gene was obtained from male sterile line and maintainer line of Zhangqiu onion using homologous cloning.The protein secondary structure and the expression of atp6 gene in different growth stages were compared.The results showed that there was one polymorphic locus(A/T)at the 171 st base of the open reading frame of atp6 gene in CMS line and maintainer line,which changed the proportion of protein secondary structure slightly,but did not change the transmembrane structure of ATP6 protein.The expression of atp6 gene in roots and leaves/flowers at seedling,bolting,flowering and late flowering stage was detected by real-time fluorescence quantitative polymerase chain reaction.The results showed that the expression of atp6 gene in root showed a trend of increasing at first and then decreasing during the whole development of the plant,reaching the maximum at the flowering stage,and the expression of the maintainer line was 1.14 times higher than that of the CMS;the expression of atp6 gene in leaves/flowers decreased gradually during the whole growth period,and reached the highest level at the seedling stage,and the maintainer line was 1.96 times higher than that of the CMS.

关 键 词：大葱 atp6基因 克隆 表达分析 

分 类 号：S633.1[农业科学—蔬菜学]