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Abstract：
BACKGROUND: Cytoplasmic male sterility (CMS), which naturally exists in higher plants, is a useful mechanism for analyzing nuclear and mitochondrial genome functions and identifying the role of mitochondrial genes in the plant growth and development. Polima (pol) CMS is the most universally valued male sterility type in oil-seed rape. Previous studies have described the pol CMS restorer gene Rfp and the sterility-inducing gene orf224 in oil-seed rape, located in mitochondria. However, the mechanism of fertility restoration and infertility remains unknown. Moreover, it is still unknown how the fecundity restorer gene interferes with the sterility gene, provokes the sterility gene to lose its function, and leads to fertility restoration.
RESULTS: In this study, we used multi-omics joint analysis to discover candidate genes that interact with the sterility gene orf224 and the restorer gene Rfp of pol CMS to provide theoretical support for the occurrence and restoration mechanisms of sterility. Via multi-omics analysis, we screened 24 differential genes encoding proteins related to RNA editing, respiratory electron transport chain, anther development, energy transport, tapetum development, and oxidative phosphorylation. Using a yeast two-hybrid assay, we obtained a total of seven Rfp interaction proteins, with orf224 protein covering five interaction proteins.
CONCLUSIONS: We propose that Rfp and its interacting protein cleave the transcript of atp6/orf224, causing the infertility gene to lose its function and restore fertility. When Rfp is not cleaved, orf224 poisons the tapetum cells and anther development-related proteins, resulting in pol CMS mitochondrial dysfunction and male infertility. The data from the joint analysis of multiple omics provided information on pol CMS\'s potential molecular mechanism and will help breed B. napus hybrids.
RESULTS: In this study, we used multi-omics joint analysis to discover candidate genes that interact with the sterility gene orf224 and the restorer gene Rfp of pol CMS to provide theoretical support for the occurrence and restoration mechanisms of sterility. Via multi-omics analysis, we screened 24 differential genes encoding proteins related to RNA editing, respiratory electron transport chain, anther development, energy transport, tapetum development, and oxidative phosphorylation. Using a yeast two-hybrid assay, we obtained a total of seven Rfp interaction proteins, with orf224 protein covering five interaction proteins.
CONCLUSIONS: We propose that Rfp and its interacting protein cleave the transcript of atp6/orf224, causing the infertility gene to lose its function and restore fertility. When Rfp is not cleaved, orf224 poisons the tapetum cells and anther development-related proteins, resulting in pol CMS mitochondrial dysfunction and male infertility. The data from the joint analysis of multiple omics provided information on pol CMS\'s potential molecular mechanism and will help breed B. napus hybrids.
摘要：
背景：细胞质雄性不育（CMS），天然存在于高等植物中，是分析核和线粒体基因组功能以及识别线粒体基因在植物生长和发育中的作用的有用机制。Polima（pol）CMS是油菜中最普遍重视的雄性不育类型。先前的研究已经描述了油菜中的polCMS恢复基因Rfp和不育诱导基因orf224，位于线粒体。然而,生育恢复和不孕症的机制仍然未知。此外，目前尚不清楚繁殖力恢复基因如何干扰不育基因，导致不育基因失去其功能，并导致生育能力恢复。
结果：在这项研究中，我们利用多组学联合分析发现了与不育基因orf224和polCMS修复基因Rfp相互作用的候选基因，为不育的发生和恢复机制提供了理论支持。通过多组学分析，我们筛选了24个编码与RNA编辑相关的蛋白质的差异基因，呼吸电子传递链,花药发育，能源运输,绒毡层发育，和氧化磷酸化。用酵母双杂交法,我们总共获得了七个Rfp相互作用蛋白，用orf224蛋白覆盖五个相互作用蛋白。
结论：我们建议Rfp及其相互作用蛋白切割atp6/orf224的转录本，导致不育基因失去功能并恢复生育能力。当Rfp没有裂开时，orf224毒害绒毡层细胞和花药发育相关蛋白，导致polCMS线粒体功能障碍和男性不育。来自多个组学联合分析的数据提供了有关polCMS潜在分子机制的信息，并将有助于培育欧洲油菜杂种。
结果：在这项研究中，我们利用多组学联合分析发现了与不育基因orf224和polCMS修复基因Rfp相互作用的候选基因，为不育的发生和恢复机制提供了理论支持。通过多组学分析，我们筛选了24个编码与RNA编辑相关的蛋白质的差异基因，呼吸电子传递链,花药发育，能源运输,绒毡层发育，和氧化磷酸化。用酵母双杂交法,我们总共获得了七个Rfp相互作用蛋白，用orf224蛋白覆盖五个相互作用蛋白。
结论：我们建议Rfp及其相互作用蛋白切割atp6/orf224的转录本，导致不育基因失去功能并恢复生育能力。当Rfp没有裂开时，orf224毒害绒毡层细胞和花药发育相关蛋白，导致polCMS线粒体功能障碍和男性不育。来自多个组学联合分析的数据提供了有关polCMS潜在分子机制的信息，并将有助于培育欧洲油菜杂种。
