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Hereditary properties of strong growth and huge accumulation of biomass in Oryza officinalis exhibit a great potential; however, the genes that code for its high photosynthesis performance are not established. This study screened eight hybrids, using biomass accumulation and photosynthesis analysis, based on the introgression lines constructed by analyzing distant hybridization patterns between Oryza officinalis and cultivars HY-8. We designed 23 primer pairs using transcriptome sequencing of Oryza officinalis and screened two types of photosynthetic enzymes: phosphoenolpyruvate carboxylase (PEPC) and pyruvate orthophosphate dikinase (PPDK), which are two related proteins of ribulose-1,5-biphosphate carboxylase/oxygenase (Rubisco), its binding protein (rubis-subs-bind), and a small subunit (rbcS). Results showed that C4 photosynthetic pathway enzymes, PEPC and PPDK, were highly expressed in hybrids and the source plant, Oryza officinalis. Homology analysis also indicated that the sequences of those two genes were different from those of the C3 and C4 plants investigated. Therefore, a better understanding of the photosynthetic characteristics of Oryza officinalis would provide clues for further isolation of valuable genes from this plant.