Abstract
The chloroplast serves as the primary site of photosynthesis, and its development plays a crucial role in regulating plant growth and morphogenesis. The Pentatricopeptide Repeat Sequence (PPR) proteins constitute a vast protein family that function in the post-transcriptional modification of RNA within plant organelles. In this study, we characterized mutant of rice with pale green leaves (pgl3a). The chlorophyll content of pgl3a at the seedling stage was significantly reduced compared to the wild type (WT). Transmission electron microscopy (TEM) and quantitative PCR analysis revealed that pgl3a exhibited aberrant chloroplast development compared to the wild type (WT), accompanied by significant alterations in gene expression levels associated with chloroplast development and photosynthesis. The Mutmap analysis revealed that a single base deletionin the coding region of Os03g0136700 in pgl3a. By employing CRISPR/Cas9 mediated gene editing, two homozygous cr-pgl3a mutants were generated and exhibited a similar phenotype to pgl3a, thereby confirming that Os03g0136700 was responsible for pgl3a. Consequently, it was designated as OsPGL3A. OsPGL3A belongs to the DYW-type PPR protein family and is localized in chloroplasts. Furthermore, we demonstrated that the RNA editing efficiency of rps8-182 and rpoC2-4106, and the splicing efficiency of ycf3-1 were significantly decreased in pgl3a mutants compared to WT. Collectively, these results indicate that OsPGL3A plays a crucial role in chloroplast development by regulating the editing and splicing of chloroplast genes in rice.
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Acknowledgements
We thank Prof. Chunming Wang for the assistance in Mutmap data analysis.
Funding
This study was supported by the National Natural Science Foundation of China (32171989 and U20A2025), Heilongjiang Key Research and Development Program (2022ZX02B03).
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Min Xu: performed the subcellular localization, expression analysis and the RNA editing. Xinying Zhang: performed the RNA splicing. Jinzhe Cao and Jiali Liu: analyzed the phenotypes and cloned PGL3. Deyong Ren: examined the ultrastructure of chloroplasts. Yiyuan He: performed some of the experiments and provided technical assistance. Qingjie Guan, Xiaojie Tian, Jiaqi Tang, and Xiufeng Li: helped with the discussion of the work. Zhenyu Wang: conceived and supervised the entire project, analyzed the data, and wrote the original draft. Qingyun Bu: conceived and supervised the entire project, refined preliminary manuscripts. All authors commented on previous versions of the manuscript, and all authors read and approved the final manuscript.
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Xu, M., Zhang, X., Cao, J. et al. OsPGL3A encodes a DYW-type pentatricopeptide repeat protein involved in chloroplast RNA processing and regulated chloroplast development. Mol Breeding 44, 29 (2024). https://doi.org/10.1007/s11032-024-01468-7
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DOI: https://doi.org/10.1007/s11032-024-01468-7