Chakraborty Anirban, Mitra Joy, Bhattacharyya Jagannath, Pradhan Subrata, Sikdar Narattam, Das Srirupa, Chakraborty Saikat, Kumar Sachin, Lakhanpaul Suman, Sen Soumitra K
Advanced Laboratory for Plant Genetic Engineering, Indian Institute of Technology, Kharagpur, 721302, India.
Planta. 2015 Jun;241(6):1463-79. doi: 10.1007/s00425-015-2269-5. Epub 2015 Mar 10.
Over-expression of the unedited mitochondrial orfB gene product generates male sterility in fertile indica rice lines in a dose-dependent manner. Cytoplasmic male sterility (CMS) and nuclear-controlled fertility restoration are widespread developmental features in plant reproductive systems. In self-pollinated crop plants, these processes often provide useful tools to exploit hybrid vigour. The wild abortive CMS has been employed in the majority of the "three-line" hybrid rice production since 1970s. In the present study, we provide experimental evidence for a positive functional relationship between the 1.1-kb unedited orfB gene transcript, and its translated product in the mitochondria with male sterility. The generation of the 1.1-kb unedited orfB gene transcripts increased during flowering, resulting in low ATP synthase activity in sterile plants. Following insertion of the unedited orfB gene into the genome of male-fertile plants, the plants became male sterile in a dose-dependent manner with concomitant reduction of ATPase activity of F1F0-ATP synthase (complex V). Fertility of the transgenic lines and normal activity of ATP synthase were restored by down-regulation of the unedited orfB gene expression through RNAi-mediated silencing. The genetic elements deciphered in this study could further be tested for their use in hybrid rice development.
未编辑的线粒体orfB基因产物的过表达以剂量依赖的方式在可育籼稻品系中产生雄性不育。细胞质雄性不育(CMS)和核控制的育性恢复是植物生殖系统中广泛存在的发育特征。在自花授粉作物中,这些过程通常为利用杂种优势提供有用的工具。自20世纪70年代以来,野败型CMS已被用于大多数“三系”杂交水稻生产。在本研究中,我们提供了实验证据,证明1.1kb未编辑的orfB基因转录本与其在线粒体中的翻译产物与雄性不育之间存在正功能关系。1.1kb未编辑的orfB基因转录本的产生在开花期间增加,导致不育植株中ATP合酶活性降低。将未编辑的orfB基因插入雄性可育植物的基因组后,植株以剂量依赖的方式变为雄性不育,同时F1F0-ATP合酶(复合体V)的ATP酶活性降低。通过RNAi介导的沉默下调未编辑的orfB基因表达,可恢复转基因系的育性和ATP合酶的正常活性。本研究中解析的遗传元件可进一步测试其在杂交水稻开发中的用途。