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作物学报 ›› 2012, Vol. 38 ›› Issue (07): 1212-1220.doi: 10.3724/SP.J.1006.2012.01212

• 作物遗传育种·种质资源·分子遗传学 • 上一篇    下一篇

油菜Nsa CMS候选恢复基因的来源及表达

张洪，付丽，李云昌，刘佳，梅德圣，彭鹏飞，陈玉峰，胡琼*   

  1. 中国农业科学院油料作物研究所国家油料作物改良中心 / 农业部油料作物生物学与遗传育种重点开放实验室，湖北武汉430062
  • 收稿日期:2011-11-15 修回日期:2012-02-22 出版日期:2012-07-12 网络出版日期:2012-04-23
  • 通讯作者: 胡琼, E-mail: huqiong@oiolcrops.cn, Tel: 027-86711556
  • 基金资助:

    本研究由国家高技术发展研究计划(863计划)项目(2011AA10A104), 国家自然科学基金项目(30871553, 31000725)和武汉市学科带头人项目(210151730564)资助。

Origin and Expression of Nsa CMS Candidate Restorer Gene

ZHANG Hong,FU Li,LI Yun-Chang,LIU Jia,MEI De-Sheng,PENG Peng-Fei,CHEN Yu-Feng,HU Qiong*   

  1. Oil Crops Research Institute, Chinese Academy of Agricultural Sciences, National Center for Oil Crops Improvement / Key Laboratory for Biological Sciences and Genetic Improvement of Oil Crops, Ministry of Agriculture, Wuhan 430062, China
  • Received:2011-11-15 Revised:2012-02-22 Published:2012-07-12 Published online:2012-04-23
  • Contact: 胡琼, E-mail: huqiong@oiolcrops.cn, Tel: 027-86711556

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摘要： 根据同源序列法克隆的Nsa CMS候选恢复基因PPR618的序列，采用PCR方法在Nsa CMS不育系、恢复系、原始体细胞杂交亲本以及其他甘蓝型油菜品种中共克隆出22个同源序列。序列分析表明，野芥亲本野油18、甘蓝型油菜亲本中双4号各含有2个同源序列，而Nsa CMS 4个恢复系则分别含有3、1、1、1个同源序列。恢复系中候选恢复基因的序列与野芥亲本野油18的同源序列的一致性在93%以上，其中恢1、恢3和恢4中至少有一个同源序列与野油18的第一个同源序列完全相同，恢2中的同源序列与野油18的第2个同源序列完全相同；但与甘蓝型油菜的同源序列一致性均低于80%，说明候选恢复基因来源于新疆野芥，而不是甘蓝型油菜。除了原来的PPR618外，获得了3个来源于恢复系的新候选恢复基因。候选恢复基因在序列上与萝卜和矮牵牛的恢复基因一致性较高。半定量RT-PCR分析表明，候选恢复基因在恢复系中多个组织都有表达，但在根、茎中表达量特别少。随着营养器官到生殖器官的发育逐渐升高，候选恢复基因在恢复系中表达量最高的是雄性败育关键时期, 即1.5~2.5 mm的花蕾中，但不育系中的同源基因在茎中的表达量则相对高于其他组织。

关键词: Nsa CMS, 恢复基因, 同源序列, 表达

Abstract:  Nsa cytoplasmic male sterility (CMS) is a novel CMS system develpoed by somatic hybridization between Brassica napus and Sinapis arvensis. The cloning the restorer genes for Nsa CMS is important for both the development of better restorers and the mechanism understanding of fertility restoration. A candidate restorer gene named PPR618 of Nsa CMS was cloned based on the homologue sequencing strategy previously. In this study, based on sequence information of PPR618, 22 homologous sequences were identified in Nsa CMS male sterile line, Nsa CMS restorer lines, the original fusion parents which gave rise to Nsa CMS and several other B. napus lines as well as one B. oleracea and one B. rapa accessions. Sequence analysis showed that there are two PPR618 homologues in each of the fusion parental lines, S. arvensis var. Yeyou 18 and B. napus var. Zhongshuang 4, whereas there were three, one, one, and one PPR618 homologues in the four restorers, Hui 1, Hui 2, Hui 3, and Hui 4, respectively. The identity of the homologous genes in restorers was above 93% to those in Yeyou 18, but less than 80% to those in Zhongshuang 4, implicating the candidate restorer genes of Nsa CMS originated from S. arvensis. Both homologues in S. arvensis were found in the restorers, one in Hui1, Hui 3, and Hui 4, the other in Hui 2. Besides PPR618, three new candidate restoring genes were identified. The homologous genes from restorers was also found to have relations with the restoring genes for CMS system of radish and petunia. Semi-quantitative RT-PCR results showed that the candidate restoring gene expressed in all tested organs. The gene expression gradually increased along with the developmental process from vegetative to reproductive stages, peaked in the bud of 1.5–2.5 mm in diameter, and slumped in roots and stems. In contrast, the highest expression of homologous gene in male sterile line was detected in stems.

Key words: Nsa CMS, Restorer gene, Homologous sequence, Expression

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