BackgroundRatoon sugarcane (Saccharum officinarum) is susceptible to chlorosis, significantly reducing production. The molecular mechanism underlying this phenomenon remains unknown. We analyzed the transcriptome and metabolome of chlorotic and non-chlorotic sugarcane leaves from the same field to gain insight into the symptom. ResultsThe agronomic traits, like plant height, leaf number, stalk nod number, and tiller number, declined in chlorotic sugarcane. The chlorophyll content in chlorosis leaves was significantly lower than non-chlorotic leaves. A total of 11,776 differentially expressed genes (DEGs) were discovered in transcriptome analysis. In the KEGG enriched chlorophyll metabolism pathway, sixteen DEGs were found, eleven of which were down-regulated. Two photosynthesis pathways were also enriched, with 32 genes downregulated and four genes upregulated. Among the 81 enriched GO biological processes, there were four categories related to metal ion homeostasis and three related to metal ion transport. Approximately 400 metabolites were identified in metabolome analysis. The thirteen classified differentially expressed metabolites (DEMs) were found all down-regulated. The phenylpropanoid biosynthesis pathway was enriched in DEGs and DEMs, indicating phenylpropanoids' vital role in chlorosis. ConclusionsAccording to our study, chlorophyll production, metal ion metabolism, photosynthesis, and some secondary metabolites of the phenylpropanoid biosynthesis pathway, were considerably altered in chlorotic ratoon sugarcane. Our finding revealed the relation between chlorosis and these pathways, which would further the understanding of the mechanism of ratoon sugarcane chlorosis.