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| OsPPR9 encodes a DYW-type PPR protein that affects editing efficiency of multiple RNA editing sites and is essential for chloroplast development |
| CHEN Chang-zhao1*, WANG Ya-liang1*, HE Meng-xing1, 2, LI Zhi-wen1, 3, SHEN Lan1, LI Qing1, REN De-yong1, HU Jiang1, ZHU Li1, ZHANG Guang-heng1, GAO Zhen-yu1, ZENG Da-li1, GUO Long-biao1, QIAN Qian1#, ZHANG Qiang1# |
1 State Key Lab of Rice Biology, China National Rice Research Institute, Hangzhou 310006, P.R.China
2 College of Chemistry and Life Sciences, Zhejiang Normal University, Jinhua 321004, P.R.China
3 College of Life and Environmental Sciences, Hangzhou Normal University, Hangzhou 310006, P.R.China
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摘要
叶片的光合作用主要发生在叶绿体中，叶绿体的发育受核基因编码蛋白调控。其中，PPR蛋白参与细胞器RNA编辑。在水稻中虽然鉴定出了约450个PPR蛋白家族成员，但只有少数被证明影响水稻叶绿体RNA编辑。利用基因编辑技术创造了新的水稻种质资源和突变体，能够用于水稻育种和基因功能研究。本研究鉴定了一个DYW类型PPR蛋白OsPPR9在水稻叶绿体RNA编辑中的功能。通过CRISPR/Cas9基因编辑技术获得了Osppr9突变体，该突变体叶片黄化和致死表型；在突变体中，叶绿体发育相关基因表达量降低，光合作用相关蛋白的积累减少。此外，OsPPR9蛋白功能的缺失降低了叶绿体中rps8-C182, rpoC2-C4106, rps14-C80和ndhB-C611 RNA编辑位点的编辑效率，影响水稻叶绿体的生长发育。OsPPR9在水稻叶片中表达量最高和编码一个定位于叶绿体的PPR蛋白。此外，通过酵母双杂验证OsPPR9与OsMORF2和OsMORF9相互作用。总之，我们的研究为探明PPR蛋白在水稻叶绿体发育中的作用提供了线索。
Abstract
Photosynthesis occurs mainly in chloroplasts, whose development is regulated by proteins encoded by nuclear genes. Among them, pentapeptide repeat (PPR) proteins participate in organelle RNA editing. Although there are more than 450 members of the PPR protein family in rice, only a few affect RNA editing in rice chloroplasts. Gene editing technology has created new rice germplasm and mutants, which could be used for rice breeding and gene function study. This study evaluated the functions of OsPPR9 in chloroplast RNA editing in rice. The osppr9 mutants were obtained by CRISPR/Cas9, which showed yellowing leaves and a lethal phenotype, with suppressed expression of genes associated with chloroplast development and accumulation of photosynthetic-related proteins. In addition, loss of OsPPR9 protein function reduces the editing efficiency of rps8-C182, rpoC2-C4106, rps14-C80, and ndhB-C611 RNA editing sites, which affects chloroplast growth and development in rice. Our data showed that OsPPR9 is highly expressed in rice leaves and encodes a DYW-PPR protein localized in chloroplasts. Besides, the OsPPR9 protein was shown to interact with OsMORF2 and OsMORF9. Together, our findings provide insights into the role of the PPR protein in regulating chloroplast development in rice.
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Received: 11 February 2022
Accepted: 12 April 2022
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| Fund:
This research was funded by the Central Public-Interest Scientific Institution Basal Research Fund, China (CPSIBRF-CNRRI-202111 and CPSIBRF-CNRRI-202110), the Agricultural Science and Technology Innovation Program, Chinese Academy of Agricultural Sciences (ASTIP), the Project of State Key Laboratory of Rice Biology, China (2020ZZKT10205), and the Key Research and Development Project of China Rice Research Institute (CNRRI-2020-01).
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| About author: CHEN Chang-zhao, E-mail: chenchangzhao97@163.com; WANG Ya-liang, E-mail: wangyaliang@caas.cn; #Correspondence QIAN qian, Tel: +86-571-63370378, E-mail: qianqian188@hotmail.com; ZHANG Qiang, Tel: +86-571-63370378, E-mail: Zhangqiang9024@126.com
* These authors contributed equally to this study.
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Cite this article:
CHEN Chang-zhao, WANG Ya-Liang, HE Meng-xing, LI Zhi-wen, SHEN Lan, LI Qing, RE De-yong, HU Jiang, ZHU Li, ZHANG Guang-heng, GAO Zhen-yu, ZENG Da-li, GUO Long-biao, QIAN Qian, ZHANG Qiang.
2023.
OsPPR9 encodes a DYW-type PPR protein that affects editing efficiency of multiple RNA editing sites and is essential for chloroplast development. Journal of Integrative Agriculture, 22(4): 972-980.
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Chen G L, Zou Y, Hu J H, Ding Y. 2018. Genome-wide analysis of the rice PPR gene family and their expression profiles under different stress treatments. BMC Genomics, 19, 720.
He P, Wu S Y, Jiang Y L, Zhang L H, Tang M J, Xiao G H, Yu J N. 2019. GhYGL1d, a pentatricopeptide repeat protein, is required for chloroplast development in cotton. BMC Plant Biology, 19, 350.
Huang C, Li Z R, Yu Q B, Ye L S, Cui Y L, Molloy D P, Yang Z N. 2019. MORF2 tightly associates with MORF9 to regulate chloroplast RNA editing in Arabidopsis. Plant Science, 278, 64–69.
Huang W F, Zhang Y, Shen L Q, Fang Q, Liu Q, Gong C B, Zhang C, Zhou Y, Mao C, Zhu Y L, Zhang J H, Chen H P, Zhang Y, Lin Y, Bock R, Zhou F. 2020. Accumulation of the RNA polymerase subunit RpoB depends on RNA editing by OsPPR16 and affects chloroplast development during early leaf development in rice. New Phytologist, 228, 1401–1416.
Jiang T, Zhang J, Rong L W, Feng Y J, Wang Q, Song Q L, Zhang L X, Ouyang M. 2018. ECD1 functions as an RNA-editing trans-factor of rps14-149 in plastids and is required for early chloroplast development in seedlings. Journal of Experimental Botany, 69, 3037–3051.
Li X L, Sun M D, Liu S J, Teng Q, Li S H, Jiang Y S. 2021. Functions of PPR proteins in plant growth and development. International Journal of Molecular Sciences, 22, 11274.
Liu X Y, Jiang R C, Wang Y, Tang J J, Sun F, Yang Y Z, Tan B C. 2021. ZmPPR26, a DYW-type pentatricopeptide repeat protein, is required for C-to-U RNA editing at atpA-1148 in maize chloroplasts. Journal of Experimental Botany, 72, 4809–4821.
Nakai M. 2018. New perspectives on chloroplast protein import. Plant and Cell Physiology, 59, 1111–1119.
Ramos-Vega M, Guevara-García A, Llamas E, Sánchez-León N, Olmedo-Monfil V, Vielle-Calzada J P, León P. 2015. Functional analysis of the Arabidopsis thaliana CHLOROPLAST BIOGENESIS 19 pentatricopeptide repeat editing protein. New Phytologist, 208, 430–441.
Small I D, Schallenberg-Rüdinger M, Takenaka M, Mireau H, Ostersetzer-Biran O. 2020. Plant organellar RNA editing: what 30 years of research has revealed. The Plant Journal, 101, 1040–1056.
Singh R, Singh S, Parihar P, Singh V P, Prasad S M. 2015. Retrograde signaling between plastid and nucleus: A review. Journal of Plant Physiology, 181, 55–66.
Tang J P, Zhang W W, Wen K, Chen G M, Sun J, Tian Y L, Tang W J, Yu J, An H Z, Wu T T, Kong F, Terzaghi W, Wang C M, Wan J M. 2017. OsPPR6, a pentatricopeptide repeat protein involved in editing and splicing chloroplast RNA, is required for chloroplast biogenesis in rice. Plant Molecular Biology, 95, 345–357.
Xiao H J, Xu Y H, Ni C Z, Zhang Q N, Zhong F Y, Huang J S, Liu W, Peng L L, Zhu Y G, Hu J. 2018. A rice dual-localized pentatricopeptide repeat protein is involved in organellar RNA editing together with OsMORFs. Journal of Experimental Botany, 69, 2923–2936.
Xin K X, Pan T, Gao S, Yan S P. 2021. A transcription factor regulates gene expression in chloroplasts. International Journal of Molecular Sciences, 22, 6769.
Yan J, Zhang Q, Yin P. 2018. RNA editing machinery in plant organelles. Science China Life Sciences, 61, 162–169.
Yu Q B, Jiang Y, Chong K, Yang Z N. 2009. AtECB2, a pentatricopeptide repeat protein, is required for chloroplast transcript accD RNA editing and early chloroplast biogenesis in Arabidopsis thaliana. The Plant Journal, 59, 1011–1023.
Yu X Q, Xie W, Liu H, Liu W, Zeng D L, Qian Q, Ren D Y. 2022. Characterization and fine mapping of a semi-rolled leaf mutant srl3 in rice. Journal of Integrative Agriculture, 21, 3103–3113.
Zeng Z H, Lin T Z, Zhao J Y, Zheng T H, Xu L F, Wang Y H, Liu L L, Jiang L, Cheng S H, Wan J M. 2020. OsHemA gene, encoding glutamyl-tRNA reductase (GluTR) is essential for chlorophyll biosynthesis in rice (Oryza sativa). Journal of Integrative Agriculture, 19, 612–623.
Zhang J H, Guo Y P, Fang Q, Zhu Y L, Zhang Y, Liu X J, Lin Y J, Barkan A, Zhou F. 2020. The PPR-SMR protein ATP4 is required for editing the chloroplast rps8 mRNA in rice and maize. Plant Physiology, 184, 2011–2021.
Zhang Q, Shen L, Wang Z W, Hu G L, Ren D Y, Hu J, Zhu L, Gao Z Y, Zhang G H, Guo L B, Zeng D L, Qian Q. 2019. OsCAF1, a CRM domain containing protein, influences chloroplast development. International Journal of Molecular Sciences, 20, 4386.
Zhang Q, Wang Y L, Xie W, Chen C Z, Ren D Y, Hu J, Zhu L, Zhang G H, Gao Z Y, Guo L B, Zeng D L, Shen L, Qian Q. 2021. OsMORF9 is necessary for chloroplast development and seedling survival in rice. Plant Science, 307, 110907.
Zhang Z G, Cui X A, Wang Y W, Wu J X, Gu X F, Lu T G. 2017. The RNA editing factor WSP1 is essential for chloroplast development in rice. Molecular Plant, 10, 86–98.
Zhou W, Cheng Y, Yap A, Chateigner-Boutin A L, Delannoy E, Hammani K, Small I, Huang J. 2009. The Arabidopsis gene YS1 encoding a DYW protein is required for editing of rpoB transcripts and the rapid development of chloroplasts during early growth. The Plant Journal, 58, 82–96.
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