洋葱细胞质雄性不育基因分子标记的开发.docVIP

洋葱细胞质雄性不育基因分子标记的开发 　　摘 要： 　　根据细香葱orfA501开发的特异引物，以洋葱细胞质雄性不育系为试材进行PCR扩增，对获得的片段进行回收测序，根据测序结果设计引物进行染色体步移，获得部分侧翼序列，开发了鉴定洋葱细胞质基因型的SCAR标记，命名为OC2175。该标记在洋葱不育系中扩增出一条2 175 bp的特异条带和1 053 bp的条带，而保持系中仅扩增出1 053 bp的条带。对17组不同遗传背景的不育系及其相应的保持系进行验证，该SCAR标记的鉴定结果与田间表型判断结果完全吻合。 　　关键词：洋葱;细胞质雄性不育;分子标记 　　中图分类号：S633.203.6 文献标识号：A 文章编号：1001-4942（2015）06-0001-04 　　Development of a Molecular Marker Identifying 　　Cytoplasmic Male Sterility Gene in Onion（Allium cepa L.） 　　Li Yongbo1，2，Liu Bingjiang2，Huo Yumeng2，Miao Jun2，Yang Jianping1，Wu Xiong2，Yang Yanyan2* 　　（1. College of Horticulture Science and Engineering of Shandong Agricultural University， Taian 271018， China; 　　2. Vegetable Research Institute of Shandong Academy of Agricultural Sciences，Jinan 250100，China） 　　Abstract The specific primers of orfA501 in chives and cytoplasmic male sterile line of onion were used to PCR amplification. The amplified fragment was extracted and sequenced. The primers were designed according to the obtained sequence for genome walking， and partial flanking sequence was obtained. Then a SCAR marker was developed and named as OC2175. With this marker， a special fragment of 2 175 bp could be obtained from the male sterile lines and a 1 053 bp fragment was amplified from both （s）-cytoplasm and （N）-cytoplasm. Seventeen pairs of male sterile lines and their corresponding maintainer lines with different genetic backgrounds were used to verify this SCAR marker. The results indicated that the SCAR marker was perfectly suitable for distinguishing the cytoplasm types S or N in onion. 　　Key words Onion; Cytoplasmic male sterility; Molecular marker 　　 洋葱（Allium cepa L.），又称圆葱、葱头，百合科葱属，已有5 000多年的栽培历史。1936年美国的Jones和Emsweller 首次在“Italian Red”品种中发现雄性不育株[1]，1943年Jones和Clarke又发现了雄性不育系[2]。随后洋葱成为世界上最早利用杂种优势的蔬菜作物之一。洋葱是两年生植物，其育种年限是白菜、萝卜等蔬菜的2～4倍，与传统的测交-回交、自交等方法相比，分子标记技术直接以DNA形式出现，在植物体的各个组织、各发育时期均可检测，不受季节、环境的限制，可大大缩短不育系和保持系的选育年限，减少工作量，加快育种进程。 　　国内外关于洋葱细胞质雄性不育的研究报道较少，Havey[3]利用RFLP分子标记手段找出了洋葱雄性保持系和不育系线粒体基因组之间的差异。Engel