Yang Hang, Wang Yunlong, Tian Yunlu, Teng Xuan, Lv Zehui, Lei Jie, Duan Erchao, Dong Hui, Yang Xue, Zhang Yuanyan, Sun Yinglun, Chen Xiaoli, Bao Xiuhao, Chen Rongbo, Gu Chuanwei, Zhang Yipeng, Jiang Xiaokang, Ma Wenyu, Zhang Pengcheng, Ji Yi, Zhang Yu, Wang Yihua, Wan Jianmin
State Key Laboratory for Crop Genetics and Germplasm Enhancement, Jiangsu Plant Gene Engineering Research Center, Nanjing Agricultural University, Nanjing, 210095, China.
National Key Facility for Crop Resources and Genetic Improvement, Institute of Crop Sciences, Chinese Academy of Agricultural Sciences, Beijing, 100081, China.
J Integr Plant Biol. 2023 Mar;65(3):755-771. doi: 10.1111/jipb.13402. Epub 2023 Jan 5.
Most of the reported P-type pentatricopeptide repeat (PPR) proteins play roles in organelle RNA stabilization and splicing. However, P-type PPRs involved in both RNA splicing and editing have rarely been reported, and their underlying mechanism remains largely unknown. Here, we report a rice floury endosperm22 (flo22) mutant with delayed amyloplast development in endosperm cells. Map-based cloning and complementation tests demonstrated that FLO22 encodes a mitochondrion-localized P-type PPR protein. Mutation of FLO22 resulting in defective trans-splicing of mitochondrial nad1 intron 1 and perhaps causing instability of mature transcripts affected assembly and activity of complex Ⅰ, and mitochondrial morphology and function. RNA-seq analysis showed that expression levels of many genes involved in starch and sucrose metabolism were significantly down-regulated in the flo22 mutant compared with the wild type, whereas genes related to oxidative phosphorylation and the tricarboxylic acid cycle were significantly up-regulated. In addition to involvement in splicing as a P-type PPR protein, we found that FLO22 interacted with DYW3, a DYW-type PPR protein, and they may function synergistically in mitochondrial RNA editing. The present work indicated that FLO22 plays an important role in endosperm development and plant growth by participating in nad1 maturation and multi-site editing of mitochondrial messager RNA.
大多数已报道的P型五肽重复(PPR)蛋白在细胞器RNA稳定和剪接中发挥作用。然而,参与RNA剪接和编辑的P型PPR却鲜有报道,其潜在机制也 largely未知。在此,我们报道了一个水稻粉质胚乳22(flo22)突变体,其胚乳细胞中造粉体发育延迟。基于图谱的克隆和互补试验表明,FLO22编码一种定位于线粒体的P型PPR蛋白。FLO22的突变导致线粒体nad1内含子1的反式剪接缺陷,可能导致成熟转录本不稳定,影响了复合体Ⅰ的组装和活性,以及线粒体的形态和功能。RNA测序分析表明,与野生型相比,flo22突变体中许多参与淀粉和蔗糖代谢的基因表达水平显著下调,而与氧化磷酸化和三羧酸循环相关的基因则显著上调。除了作为P型PPR蛋白参与剪接外,我们还发现FLO22与DYW型PPR蛋白DYW3相互作用,它们可能在线粒体RNA编辑中协同发挥作用。目前的研究表明,FLO22通过参与nad1成熟和线粒体信使RNA的多位点编辑,在胚乳发育和植物生长中发挥重要作用。