Journal of Integrative Agriculture ›› 2025, Vol. 24 ›› Issue (12): 4484-4495.DOI: 10.1016/j.jia.2024.02.022

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P型三角状五肽重复蛋白YGS对水稻叶绿体的发育至关重要

  

  • 收稿日期:2023-11-24 修回日期:2024-02-29 接受日期:2024-01-03 出版日期:2025-12-20 发布日期:2025-11-13

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. 1 College of Life Sciences, Dezhou University, Dezhou 253023, China

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

  • Received:2023-11-24 Revised:2024-02-29 Accepted:2024-01-03 Online:2025-12-20 Published:2025-11-13
  • 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 study.

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摘要：

三角状五肽重复序列（Pentatricopeptide repeat，PPR）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 perform essential functions in post-transcriptional regulation of gene expression, particularly RNA editing and RNA splicing, in plant organelles.  Although research on chloroplast biogenesis and development has been extensive, the functions of most PPR genes in this process in rice (Oryza sativa) remain incompletely understood.  This study identifies a novel P-type PPR protein, YELLOW-GREEN LEAF AND SEEDLING LETHALITY (YGS), which localizes to rice chloroplasts. YGS shows predominant expression in leaves.  The ygs mutants, generated through CRISPR/Cas9-mediated genome editing of the YGS gene, displayed yellow-green leaves and seedling lethality.  These phenotypes corresponded with reduced pigment levels and disrupted chloroplast ultrastructure compared to wild-type plants.  Furthermore, the expression of genes associated with chloroplast development and chlorophyll biosynthesis showed significant alterations in the ygs mutants.  The absence of YGS function affected RNA editing of rpl2 and intron splicing of ycf3-1 in the plastid genome.  Additionally, YGS demonstrated interaction with the chloroplast signal recognition particle protein OscpSRP54b in yeast two-hybrid and bimolecular fluorescence complementation analyses.  These results indicate that YGS participates in RNA editing and RNA splicing in chloroplasts, thus serving a vital role in rice chloroplast development

Key words: rice , YGS ,  PPR protein ,  chloroplast development