Background: Rose is an important economic horticultural crop. However, its field growth and quality are negatively affected by aphids. However, the defence mechanisms used by rose plants against aphids are unclear. A previous study showed that Macrosiphum rosivorum is the most common and harmful aphid species to Rosa plants and that Rosa longicuspis is highly resistant species to aphids. Therefore, to understand the defence mechanism of rose under aphid stress, we combined RNA sequencing and metabolomics techniques to investigate the changes in gene expression and metabolomic processes in R. longicuspis infected with M. rosivorum. Result: In our study, after inoculation with M. rosivorum, M. rosivorum quickly colonized. A total of 34202 genes and 758 metabolites were detected in all samples, with 2845, 2627 and 466 differentially expressed genes (DEGs) were found in CK-vs.-A3 d, CK-vs.-A5 d, and A3 d-vs.-A5 d, respectively. Among these metabolites, 65, 70 and 26 differentially expressed metabolites (DEMs) were found in CK-vs.-A3 d, CK-vs.-A5 d, and A3 d-vs.-A5 d, respectively. The combined omics approach revealed that M. rosivorum is perceived by effector-triggered immunity. Under M. rosivorum stress, R. longicuspis responded by signal transduction pathway activation, transcription factor expression, ROS production and hormone-mediated defence responses. Interestingly, the ‘brassinosteroid biosynthesis’ pathway was significantly enriched in A3 d-vs.-A5 d. Further analysis showed that M. rosivorum induced the transformation of starch and sucrose, the biosynthesis of terpenoids, tannins and phenolic acids and metabolism of cyanoamino acid. Importantly, the ‘tropane, piperidine and pyridine alkaloid biosynthesis’, ‘glutathione metabolic’ and ‘glucosinolate biosynthesis’ pathways were significantly enriched, which resulted in increased levels of metabolites that were involved in the plant defence response. Conclusion: Our study provides candidate genes and metabolites for Rosa defence against aphids. What’s more, this study provides a theoretical basis for further exploring the molecular regulation mechanism of rose aphid resistance and aphid resistance breeding in the future.