Cytoplasmic effects (CEs) have been discovered to influence a diverse array of agronomic traits in crops,and understanding the underlying mechanisms can help accelerate breeding programs.Seed oil content (SOC) is of great agricultural,nutritional,and economic importance.However,the genetic basis of CEs on SOC (CE-SOC) remains enigmatic.In this study,we use an optimized approach to sequence the cytoplasmic (plastid and mitochondrial) genomes of allotetraploid oilseed rape (Brassica napus) cultivars,51218 and 56366,that bear contrasting CE-SOC.By combining comparative genomics and genome-wide transcriptome analysis,we identify mitochondria-encoded orf188 as a potential CE-SOC determinant gene.Functional analyses in the model system Arabidopsis thaliana and rapeseed demonstrated that orf188 goves CE-SOC and could significantly increase SOC,strikingly,through promoting the yield of ATP.Consistent with this finding,transcriptional profiling with microarray and RNA sequencing revealed that orf188 affects transcriptional reprogramming of mitochondrial energy metabolism to facilitate ATP production.Intriguingly,orf188 is a previously uncharacterized chimeric gene,and the presence of this genetic novelty endows rapeseed with positive CE-SOC.Our results shed light on the molecular basis of CEs on a key quantitative trait in polyploid crops and enrich the theory of mateal control of oil content,providing new scientific guidance for breeding high-oil rapeseed germplasms.