Isolation and Characterization of SPOTTED LEAF42 Encoding a Porphobilinogen Deaminase in Rice

The formation and development of chloroplasts play a vital role in the breeding of high-yield rice (Oryza sativa L.). Porphobilinogen deaminases (PBGDs) act in the early stage of chlorophyll and heme biosynthesis. However, the role of PBGDs in chloroplast development and chlorophyll production remains elusive in rice. Here, we identified the spotted leaf 42 (spl42) mutant, which exhibited a reddish-brown spotted leaf phenotype. The mutant showed a significantly lower chlorophyll content, abnormal thylakoid morphology, and elevated activities of reactive oxygen species (ROS)-scavenging enzymes. Consistently, multiple genes related to chloroplast development and chlorophyll biosynthesis were significantly down-regulated, whereas many genes involved in leaf senescence, ROS production, and defense responses were upregulated in the spl42 mutant. Map-based cloning revealed that SPL42 encodes a PBGD. A C-to-T base substitution occurred in spl42, resulting in an amino acid change and significantly reduced PBGD enzyme activity. SPL42 targets to the chloroplast and interacts with the multiple organelle RNA editing factors (MORFs) OsMORF8-1 and OsMORF8-2 to affect RNA editing. The identification and characterization of spl42 helps in elucidating the molecular mechanisms associated with chlorophyll synthesis and RNA editing in rice.

Automated summary

The mutant spl42 exhibits a reddish-brown spotted leaf phenotype, with significantly lower chlorophyll content and abnormal thylakoid morphology. Reactive oxygen species (ROS)-scavenging enzyme activities are elevated in the mutant. Genes related to chloroplast development and chlorophyll biosynthesis are down-regulated, while genes involved in leaf senescence, ROS production, and defense responses are upregulated in the spl42 mutant. Map-based cloning revealed that SPL42 encodes a porphobilinogen deaminase (PBGD), with significantly reduced enzyme activity due to a base substitution.