作物学报2016，Vol.42Issue(12)：1743-1753,11.DOI:10.3724/SP.J.1006.2016.01743
烟草ATP合酶F0部分4个亚基基因转录本编辑位点分析
Editing Sites in Transcript of Four F0-ATPase Subunit Gene in Tobacco
摘要
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 ste-rility (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 genesatp6, atp9, orf25,andorfB 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,andorfB 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. Theorf25andorfB gene transcripts had the same RNA editing sites between male sterile and fertile lines. Foratp6 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. Theatp9 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 ofatp9 gene resulted in pro-ducing 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.关键词
烟草/细胞质雄性不育/ATP合酶/亚基基因/RNA编辑Key words
Tobacco/Cytoplasmic male sterility/ATPase/Subunit gene/RNA editing引用本文复制引用
陶瑶,王瑜,钟思荣,吴凌敏,谢丽娟,聂亚平,周玮,王建革,刘齐元..烟草ATP合酶F0部分4个亚基基因转录本编辑位点分析[J].作物学报,2016,42(12):1743-1753,11.基金项目
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。This study was supported by the National Natural Science Foundation of China (31260350,31301388), China Postdoctoral Science Founda-tion (2015T80870,2014M562109), and Science and Technology Plan Projects of Jiangxi Province Education Department (GJJ13275) （31260350,31301388）
