Leaf senescence plays an important role in crop developmental processes that dramatically affect crop yield and grain quality. The genetic regulation of leaf senescence is complex, involving many metabolic and signaling pathways.Here, we identified a rapid leaf senescence 3(rls3) mutant that displayed accelerated leaf senescence, shorter plant height and panicle length, and lower seed set rate than the wild type. Map-based cloning revealed that RLS3 encodes a protein with AAA+ domain, localizing it to chloroplasts. Sequence analysis found that the rls3 gene had a single-nucleotide substitution(G→A) at the splice site of the 10~(th)intron/11~(th) exon, resulting in the cleavage of the first nucleotide in 11 ~(th) exon and premature termination of RLS3 protein translation.Using transmission electron microscope, the chloroplasts of the rls3 mutant were observed to degrade much faster than those of the wild type. The investigation of the leaf senescence process under dark incubation conditions furtherrevealed that the rls3 mutant displayed rapid leaf senescence.Thus, the RLS3 gene plays key roles in sustaining the normal growth of rice, while loss of function in RLS3 leads to rapid leaf senescence. The identification of RLS3 will be helpful to elucidate the mechanisms involved in leaf senescence in rice. The genetic regulation of leaf senescence is complex, involving many metabolic and signaling pathways. Here, we identified a rapid leaf senescence 3 (rls3) mutant that displayed accelerated leaf senescence, shorter plant height and panicle length, and lower seed set rate than the wild type. Map-based cloning revealed that RLS3 encodes a protein with AAA + domain, localizing it to chloroplasts. Sequence analysis found that the rls3 gene had a single -nucleotide substitution (G → A) at the splice site of the 10 ~ (th) intron / 11 ~ (th) exon, resulting in the cleavage of the first nucleotide in 11 ~ (th) exon and premature termination of RLS3 protein translation .Using transmission electron microscope, the chloroplasts of the rls3 mutant were observed to degrade much faster than those of the wild type. The investigation of the leaf senescence process under dark incubation cond itions furtherrevealed that the rls3 mutant displayed rapid leaf senescence.Thus, the RLS3 gene plays a key role in sustaining the normal growth of rice, while loss of function in RLS3 leads to rapid leaf senescence. The identification of RLS3 will be helpful to elucidate the mechanisms involved in leaf senescence in rice.