洋葱S型细胞质雄性不育相关基因的挖掘及分子标记开发  

Gene Discovery Related to S-type Male Sterility in Onion and Its Usage for Development of Molecular Marker

作　　者：陈立[1,2] 杨亚会 霍雨猛 刘冰江 孙雨晴 杨建平 吴雄[2] 杨妍妍[2] CHEN Li1,2, YANG Yahui1,2, HUO Yumeng2, LIU Bingjiang2, SUN Yuqing2,3, YANG Jianping1, WU Xiong2,3, YANG Yanyan2(1College of Horticulture Science andEngineering, ShandongAgricultural University, Tai＇an, Shandong 271018, China; 2Institute of Vegetables and Flowers, Shandong Academy of Agricultural Sciences, Ji＇nan 250100, China; 3College of Horticulture, Jilin Agricultural University, Changchun 130118, Chin)

机构地区：[1]山东农业大学园艺科学与工程学院,山东泰安271018 [2]山东省农业科学院蔬菜花卉研究所,济南250100 [3]吉林农业大学园艺学院,长春130118

出　　处：《园艺学报》2018年第3期503-510,共8页Acta Horticulturae Sinica

基　　金：国家自然科学基金项目(31301786;31672165);国家现代农业产业技术体系建设专项资金项目(CARS-24-A-10);山东省重点研发计划项目(2017GNC10110);山东省农业科学院青年科研基金项目(2015YQN33)

摘　　要：以洋葱(Allium cepa L.)S型细胞质雄性不育系和保持系材料为试材,对洋葱线粒体全基因组中的58个开放阅读框(ORF)进行差异表达分析,发现了一个表达差异片段orf393。采用实时荧光定量PCR技术,利用洋葱S型细胞质雄性不育系和保持系的根,叶片,花茎以及花粉母细胞、四分体、单核花粉粒、成熟花粉粒等4个发育时期的花蕾对orf393进行时空表达分析。结果表明:orf393在洋葱S型细胞质雄性不育系的根、叶片、花茎和4个发育时期的花蕾中均有表达,尤其是在四分体时期花蕾中表达量最高,之后恢复到营养生长时期的表达水平,但在保持系中未见其表达,这表明orf393是组成型表达,可能与洋葱的花粉发育存在某种关联。利用orf393在S型细胞质雄性不育系和保持系之间的序列差异,开发了一个稳定可靠的鉴别洋葱细胞质类型的SCAR分子标记GXX393。The differential expression of 58 open reading frames（ORFs）identified in the intergenic sequence of the mitochondrial genome was analyzed using a male sterile line and a maintainer line of onion（Allium cepa L.）,through which,a differentially expressed fragment was obtained,designated as orf393. By quantitative real-time PCR,the temporal-spatial expression of orf393 was carried out in different tissues and organs,including roots,leaves,stalks,and the buds of different developmental stages（pollen mother cells,tetrad,mononuclear,and matured pollen grains）. The results showed that orf393 was only expressed in the samples of the male sterile line;its expression could be detected in all samples of the male sterile line,especially the buds during the tetrad stage,which had the highest expression of orf393. The expression level of orf393 returned to the same level as that during the vegetative stage. Our results suggest that orf393 is constitutively expressed and may associate with the male sterility of onion. Based on the sequence difference of orf393 between the male sterile line and its maintainer line,we developed a reliable molecular marker,namely GXX393,for the identification of cytoplasm types of onion.

关 键 词：洋葱 细胞质雄性不育 orf393 基因表达 分子标记 

分 类 号：S633.2[农业科学—蔬菜学]