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Journal of Integrative Agriculture
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The P-type pentatricopeptide repeat protein YGS is essential for chloroplast development in rice
Zhennan Qiu1*, Dongdong Chen2*, Peiliang Zhang1*, Chunmiao Wang1, Guihong Liang1, Chunyang Jiao1, Shuo Han1, Cuiping Wen1, Xiliang Song1, Peiyan Guan1, Yan Li1, Shiyong Wen1#, Li Zhu2#

1 College of Life Science, Dezhou University, Dezhou 253023, China

2 State Key Laboratory of Rice Biology, China National Rice Research Institute, Hangzhou 310006, China

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摘要  三角状五肽重复序列(Pentatricopeptide repeatPPR)蛋白在植物细胞器基因表达的转录后调控发挥着至关重要的作用,特别是 RNA 编辑和 RNA 剪接中。尽管对叶绿体的研究由来已久,但大多数 PPR 基因在水稻(Oryza sativa)叶绿体生物发生和发育过程中的作用仍不清楚。在本研究中,我们鉴定了一种新的 P 型 PPR 蛋白 YELLOW-GREEN LEAF AND SEEDLING LETHAL (YGS),它定位于水稻叶绿体中。YGS基因在叶片中高表达。通过使用 CRISPR/Cas9 介导的基因组编辑技术敲除 YGS 基因,获得了 ygs 突变体;这些突变体表现出黄绿色叶片和幼苗致死表型。与野生型相比, ygs 突变体的色素含量较低,叶绿体超微结构异常,这与突变体的表型相符。此外,ygs 突变体中与叶绿体发育和叶绿素生物合成相关的基因的表达水平显著改变。另外,YGS 的功能丧失损害了叶绿体基因组中 rpl2 的 RNA 编辑和 ycf3-1 的内含子剪接。最后,酵母双杂交和双分子荧光互补试验证实YGS与叶绿体信号识别颗粒蛋白 OscpSRP54b 相互作用。这些发现表明 YGS 参与叶绿体中的 RNA 编辑和 RNA 剪接,从而在水稻叶绿体发育中起关键作用。

Abstract  Pentatricopeptide repeat (PPR) proteins play crucial roles in the post-transcriptional regulation of gene expression, specifically RNA editing and RNA splicing, in plant organelles. Despite longstanding research on chloroplast biogenesis and development, the roles of most PPR genes in this process in rice (Oryza sativa) remain unclear. In this study, we identified a novel P-type PPR protein, YELLOW-GREEN LEAF AND SEEDLING LETHAL (YGS), that is targeted to rice chloroplasts.  YGS is preferentially expressed in leaves.  The ygs mutants were obtained by knocking out YGS gene using CRISPR/Cas9-mediated genome editing; these mutants exhibited yellow-green leaves and a seedling-lethal phenotype.  Consistent with these phenotypes, the ygs mutants had lower levels of pigment contents and an abnormal chloroplast ultrastructure compared to the wild type.  Moreover, the expression levels of genes related to chloroplast development and chlorophyll biosynthesis were significantly altered in the ygs mutants.   In addition, loss of function of YGS impaired RNA editing of rpl2 and intron splicing of ycf3-1 in the plastid genome. Finally, YGS interacted with the chloroplast signal recognition particle protein OscpSRP54b in yeast two-hybrid and bimolecular fluorescence complementation assays.  These findings suggest that YGS is involved in RNA editing and RNA splicing in chloroplasts, thereby playing a crucial role in chloroplast development in rice.
Keywords:  rice       YGS              PPR protein              chloroplast development  
Online: 15 March 2024  
About author:  #Correspondence Li Zhu, Tel: +86-571-63371390, E-mail: zhuli05@caas.cn; Shiyong Wen, E-mail: wenshiyong434@163.com. * These authors contributed equally to this work.

Cite this article: 

Zhennan Qiu, Dongdong Chen, Peiliang Zhang, Chunmiao Wang, Guihong Liang, Chunyang Jiao, Shuo Han, Cuiping Wen, Xiliang Song, Peiyan Guan, Yan Li, Shiyong Wen, Li Zhu. 2024. The P-type pentatricopeptide repeat protein YGS is essential for chloroplast development in rice. Journal of Integrative Agriculture, Doi:10.1016/j.jia.2024.02.022

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