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The chloroplast is a semiautonomous photosyntheticorganelle that is essential for plant growth and development,particularly in crops via manipulation of its photosyntheticcapacity and the biosynthesis of carbon skeletons. Plastidribosomal proteins (PRPs) are crucial for the establishmentof the transcription/translation apparatus during chloroplastdifferentiation. In this study, we isolated and characterized TDNA-tagged rice mutants with defective chloroplasts, namedprpl3, that exhibited a distinct albino seedling lethality. Transmission electronic microscopy (TEM) observationsshowed that the grana stacks in the mutant were not properlyformed, with disrupted thylakoid structures in their chloroplasts. Chlorophyll content was also significantly reduced in the leavesof prpl3 mutant seedlings. PRPL3 contains nuclear genesencoding PRPs localized to the chloroplasts, and prpl3represents a novel mutant presentation of an impaired PRPL3gene. Our findings also demonstrated that PRPL3 is responsiblefor phenotypic alterations by generating additional mutantalleles thereof using CRISPR/Cas9 systems. Expression levelsof genes involved in photosynthesis and chloroplastdevelopment, including plastidial transcription and translationand photosynthesis, were altered in the prpl3 mutant. Theseresults collectively demonstrate that nuclear-encoded PRPL3 isindispensable for the proper development of chloroplasts in rice.