Chloroplasts are essential sites for plant photosynthesis, and the biogenesis of the photosynthetic complexes involves the interaction of nuclear genes and chloroplast genes. In this study, we identified a rice pale green leaf mutant, crs2. The crs2 mutant showed different degrees of low chlorophyll phenotypes at different growth stages, especially at the seedling stage. Fine mapping and DNA sequencing of crs2 revealed a single nucleotide substitution (G4120A) in the eighth exons of CRS2, causing a G-to-R mutation of the 229th amino acid of CRS2 (G229R). The results of complementation experiments confirmed that this single-base mutation in crs2 is responsible for the phenotype of the crs2 mutant. CRS2 encodes a chloroplast RNA splicing 2 protein localized in the chloroplast. Western blot results revealed an abnormality in the abundance of the photosynthesis-related protein in crs2. However, the mutation of CRS2 leads to the enhancement of antioxidant enzyme activity, which could reduce ROS levels. Meanwhile, with the release of Rubisco activity, the photosynthetic performance of crs2 was improved. In summary, the G229R mutation in CRS2 causes chloroplast protein abnormalities and affects photosystem performance in rice; the above findings facilitate the elucidation of the physiological mechanism of chloroplast proteins affecting photosynthesis.
Leaves are the primary photosynthetic organs, and alterations in leaf color can affect photosynthesis and plant biomass. In an EMS-mutagenized SN9816 population, we identified two white-striped mutants, ws21-1 and ws21-2. Both mutants showed severely reduced pigment content, defective chloroplasts, and elevated reactive oxygen species. The ws21-2 allele caused a near-complete albino phenotype, while ws21-1 resulted in milder striping. Genetic mapping and cloning identified causal mutations in OsRNRS1, encoding the small subunit of ribonucleotide reductase. The G583R (ws21-1) and Y365F (ws21-2) mutations likely impair enzyme activity, disrupting the dNTP pool for plastid genome replication and causing aberrant chloroplast development. Correspondingly, the expression of genes for chlorophyll synthesis, photosynthesis, and ROS metabolism was altered. Our findings directly link nuclear-encoded nucleotide metabolism to chloroplast biogenesis and demonstrate that dNTP homeostasis is critical for maintaining photosynthetic capacity and redox balance in plants.