Abstract: 【Objective】To perform transcriptome sequencing on the wild-type strain of Fusarium oxysporum f. sp. cubense（Foc）and the cystathionine γ-synthase gene（FoMETB）knockout mutant（ΔFoMETB），and to analyze the functions of differentially expressed genes（DEGs）enriched in the amino acid synthesis pathway，which could provide reference for exploring the virulence mechanism of this fungus and green control of banana fusarium wilt disease.【Method】 Transcriptome sequencing technology was used to conduct transcriptome sequencing on Foc wild type strain and the ΔFoMETB strain after 18 h of shaking culture in liquid PDB medium. The DEGs were annotated for GO functional categories and analyzed for KEGG signaling pathway enrichment. In silico technology was employed to analyze the functions of DEGs enriched in the amino acid synthesis pathway. Real-time fluorescence quantitative PCR was used to validate the results of the transcriptomic sequencing.【Result】Transcriptome sequencing of wild type strain and ΔFoMETB strain identified a total of 10140 genes，among which 923 genes were specifically expressed in wild type strain，706 genes were specifically expressed in ΔFoMETB strain，and 8511 genes were expressed in both wild type strain and ΔFoMETB strain. Compared with wild type strain，there were 3077 DEGs in ΔFoMETB strain，including 1598 up-regulated genes and 1479 down-regulated genes. GO function annotation analysis showed that 2465 DEGs were enriched in 529 GO terms，with the top 4 significantly enriched GO terms related to molecular functions such as coenzyme binding and biological processes including organic acid metabolic process，oxygen acid metabolic process and carboxylic acid metabolic process. KEGG signaling pathway enrichment analysis revealed that 8 out of the top 20 pathways with DEGs enrichment number were associated with amino acid synthesis and metabolism. In silico analysis of 51 DEGs in the amino acid synthesis pathway found that 22 genes have been identified in different types of fungi，involved in regulating fungal pathogenicity，growth and development，nitrogen source utilization，amino acid synthesis and secondary metabolite synthesis. Real-time fluorescence quantitative PCR was used to detect the expression of 10 DEGs，and the results were consistent with the transcriptomic sequencing data.【Conclusion】The FoMETB gene affects the physiological characteristics and pathogenicity of Fusarium oxysporum f. sp. cubense by regulating the key DEGs in the amino acid synthesis pathway.