In the study, the RbohC and RbohF gene sequences of rapeseed (Brassica rapa L.) were cloned from ‘Long you 6’ by RTPCR. And realtime quantitative PCR was used to analyze the expression of RbohC and RbohF genes under different tissues and abiotic stresses, which provided a basis for further study on the biological functions of RbohC and RbohF genes in rapeseed. The results indicated: (1) two fulllength 3 050 bp and 2 995 bp rapeseed RbohC (GenBank accession number: XM_009134386) and RbohF (GenBank accession number: XM_009114548) gene sequences were successfully cloned. (2) Bioinformatic analysis showed that RbohC and RbohF contained 2 733 bp and 2 847 bp ORFs, encoding 910 and 948 amino acid residues, respectively. It is predicted that the molecular mass of RbohC and RbohF proteins were 103 kDa and 108 kDa which isoelectric points were 9.47 and 9.21. The amino acid sequences encoding RbohC and RbohF of rapeseed have high homology with the amino acid sequences of various plant corresponding proteins such as radish. These sequences are highly conserved and contain a typical conserved domain of NADPH oxidase, including two the EF chiral motif structures, which can bind to Ca²⁺, the six transmembrane domains, the flavin adenine dinucleotide binding domain, the NAD pyrophosphate binding domain, and the NADP ribose conserved binding site in the Cterminal region. (3) RTPCR analysis showed that RbohC and RbohF were expressed in root, stem, leaf and hypocotyl of rapeseed with nontissue specificity. However, RbohC had the highest level expression in roots, while RbohF gene in hypocotyl. (4) Low temperature, drought, salt, ABA and H₂O₂ all induced the expression of RbohC and RbohF, indicating that RbohC and RbohF play an important role in rapeseed adaptation to stress. However, the RbohC and RbohF genes of ‘Long you 6’, which is highly resistant to cold, are more sensitive to stress, and the expression of RbohC gene is higher than that of RbohF gene. (5) After treatment with H₂O₂ scavenger DMTU, NADPH oxidase inhibitor DPI and IMD, MAPKK inhibitor U0126, the expression of RbohC and RbohF in rapeseed decreased, indicating that U0126 and DMTU can inhibit the expression of RbohC and RbohF. Studies have shown that RbohC and RbohF genes have an important role in adapting to stress in rapeseed. The expression of both genes is regulated by MAPK kinase signaling pathway, and is regulated by H₂O₂ feedback. The RbohC and RbohF genes in the cultivar, the coldresistant ‘Long you 6’, are more sensitive to the H₂O₂ and MAPK kinase signaling pathways.