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作物学报 ›› 2016, Vol. 42 ›› Issue (12): 1743-1753.doi: 10.3724/SP.J.1006.2016.01743

• 作物遗传育种·种质资源·分子遗传学 • 上一篇    下一篇

烟草ATP合酶F0部分4个亚基基因转录本编辑位点分析

陶瑶1，王瑜1,3，钟思荣1，吴凌敏1，谢丽娟1，聂亚平1，周玮2，王建革4，刘齐元1,?*   

  1. 1江西农业大学农学院/作物生理生态与遗传育种教育部重点实验室/江西省作物生理生态与遗传育种重点实验室,江西南昌330045;2湖南农业大学植物保护学院/植物病虫害生物学与防控湖南省重点实验室,湖南长沙410128;3贵州省黔西南州农业委员会,贵州兴义562400;4江西农业大学园林与艺术学院,江西南昌330045
  • 收稿日期:2016-04-18 修回日期:2016-07-11 出版日期:2016-12-12 网络出版日期:2016-07-28
  • 通讯作者: 刘齐元,E-mail:qiyuanl@126.com
  • 基金资助:

    本研究由国家自然科学基金项目(31260350,31301388)，中国博士后科学基金项目(2015T80870,2014M562109)和江西省教育厅科技计划项目(GJJ13275)资助。

Editing Sites in Transcript of Four F0-ATPase Subunit Genein Tobacco

TAO Yao1,WANG Yu1,3,ZHONG Si-Rong1,WU Lin-Min1,XIE Li-Juan1,NIE Ya-Ping1,ZHOU Wei2,WANG Jian-Ge4,LIU Qi-Yuan1,*   

  1. 1 Key Laboratory of Crop Physiology, Ecology and Genetic Breeding, Ministry of Education/Key Laboratory of Crop Physiology,Ecology and Genetic Breeding of Jiangxi Province/College of Agronomy, Jiangxi Agricultural University, Nanchang 330045,China;2 Hunan Provincial Key Laboratory for Biology and Control of Plant Diseases and Insect Pests, College of Plant Protection, Hunan Agricultural University,Changsha 410128,China;3 Agricultural committee of Guizhou province Qianxinanzhou, Xingyi 562400,China; 4 College of Landscape Architecture and Art, Jiangxi Agricultural University, Nanchang 330045,China
  • Received:2016-04-18 Revised:2016-07-11 Published:2016-12-12 Published online:2016-07-28
  • Contact: Liu Qiyuan,E-mail:qiyuanl@126.com
  • Supported by:
    This study was supported by the National Natural Science Foundation of China (31260350, 31301388), China Postdoctoral Science Foundation (2015T80870, 2014M562109), and Science and Technology Plan Projects of Jiangxi Province Education Department (GJJ13275).

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摘要：

RNA编辑是高等植物线粒体基因转录后表达调控的一种重要方式。为探究ATP合酶F0部分的4个亚基基因与植物雄性不育性的关系, 本研究以3个烟草雄性不育系(MS中烟90、MS云烟85和MS K326)及其同型保持系为供试材料, 比较分析atp6、atp9、orf25和orfB线粒体基因转录本的编辑位点。结果表明, orf25和orfB基因转录本在不育系和保持系中发生的编辑位点是一致的。atp6基因在不育系中未发生编辑, 在保持系中共有6处发生了RNA编辑。与保持系相比, atp9基因在不育系中除8处共同的C→T编辑外, 还缺少2个C→T的特异编辑位点, 其中1个导致氨基酸类型的改变。推测不育胞质因缺少特异的线粒体基因转录本编辑而导致烟草的细胞质雄性不育。

关键词: 烟草, 细胞质雄性不育, ATP合酶, 亚基基因, RNA编辑

Abstract:

RNA editing exits extensively in mitochondria of higher plants and is one of the most important post-transcriptional regulation methods of gene expression in mitochondrial genomes of higher plants. At the same time, it is an essential process for forming function proteins. RNA editing can induce mutations in mitochondrial genes including nucleotide insertion, substitution, or deletion, which further affects the splicing and processing of primary transcripts, ultimately resulting in cytoplasmic male sterility (CMS). The results of research using multiple species showed that there is an obvious relationship between the four subunit genesof F0-ATPase and CMS. To explore the relationship, we studied RNA editing status of four mitochondrial genes atp6, atp9, orf25,and orfB from three tobacco male sterility lines (MS Zhongyan 90, MS Yunyan 85, MS K326) and their corresponding fertile lines . The four mitochondrial genes atp6, atp9, orf25,and orfB and their cDNA were distinctively amplified by PCR from six tobacco lines. After that, by means of making a comparison between the DNA sequences and the cDNA sequences of target genes to find RNA editing sites. The orf25 and orfB gene transcripts had the same RNA editing sites between male sterile and fertile lines. For atp6 gene, RNA editing didn't occur in male sterile lines, while there were six RNA editing sites in fertile lines, which all caused changes in the type of amino acids and there were four editing sites enhancing hydrophobicity of the amino acids. It inferred that the difference of protein's hydrophobicity was most likely to cause CMS. The atp9 gene had ten RNA editing sites in fertile lines, eight of which were the same as those in male sterile lines, while two C→T unique editing sites were absent in male sterile lines, of which one caused changes in amino acid. The nucleotide variations at 223 site of atp9 gene resulted in producing a termination code, which might be the necessary RNA editing to produce normal functional protein. These results suggest that lacking of the unique RNA editing sites might contribute to CMS property in tobacco.

Key words: Tobacco, Cytoplasmicmalesterility, ATPase, Subunitgene, RNAediting

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