Abstract
The chloroplast is a semiautonomous photosynthetic organelle that is essential for plant growth and development, particularly in crops via manipulation of its photosynthetic capacity and the biosynthesis of carbon skeletons. Plastid ribosomal proteins (PRPs) are crucial for the establishment of the transcription/translation apparatus during chloroplast differentiation. In this study, we isolated and characterized T-DNA-tagged rice mutants with defective chloroplasts, named prpl3, that exhibited a distinct albino seedling lethality. Transmission electronic microscopy (TEM) observations showed that the grana stacks in the mutant were not properly formed, with disrupted thylakoid structures in their chloroplasts. Chlorophyll content was also significantly reduced in the leaves of prpl3 mutant seedlings. PRPL3 contains nuclear genes encoding PRPs localized to the chloroplasts, and prpl3 represents a novel mutant presentation of an impaired PRPL3 gene. Our findings also demonstrated that PRPL3 is responsible for phenotypic alterations by generating additional mutant alleles thereof using CRISPR/Cas9 systems. Expression levels of genes involved in photosynthesis and chloroplast development, including plastidial transcription and translation and photosynthesis, were altered in the prpl3 mutant. These results collectively demonstrate that nuclear-encoded PRPL3 is indispensable for the proper development of chloroplasts in rice.
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Acknowledgements
The authors thank Junhyun Lim and Sang Dae Yun for taking care of the Arabidopsis plants. This work was supported by a grant from the Netx-Generation BioGreen 21 program (Project No. PJ01369001), Rural Development Administration, Republic of Korea.
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JL and SKP designed the research project; JL performed the research and wrote the manuscript; SR performed the TEM analysis; SL generated CRISPR/Cas9 plants; JP analyzed the chlorophyll contents; SJ, SL, and GA participated in the discussion; JL, SJ, and SKP wrote the manuscript. The authors declare no conflict of interest.
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Fig. S1. Sequence alignment of PRPL3 and PRPL31 protein.
Fig. S2. Comparing the amino acid sequences of PRPL3 between WT and KO mutants.
Fig. S3. Expression patterns of PRPL3 in different organs and developmental stages.
Table S1. List of primers used for qRT-PCR DNA constructs and genotyping in this study.
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Lee, J., Jang, S., Ryu, S. et al. Impaired Plastid Ribosomal Protein L3 Causes Albino Seedling Lethal Phenotype in Rice. J. Plant Biol. 62, 419–428 (2019). https://doi.org/10.1007/s12374-019-0380-z
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DOI: https://doi.org/10.1007/s12374-019-0380-z