Pamamycin, produced by Streptomyces alboflavus TD-1, showed good application prospects in the prevention and treatment of Aspergillus flavus due to its significant biological activity. In order to break through the bottleneck of low pamamycin production, the molecular regulation mechanism of low-phosphate conditions to promote the efficient synthesis of pamamycin was analyzed by transcriptome sequencing and RT-qPCR technology with Streptomyces alboflavus TD-1 as the research object. The results showed that compared with the control group, the low phosphorus condition significantly increased the level of pamamycin synthesis by Streptomyces alboflavus TD-1 (2.01-fold increase in pamamycin production at 120 μmol/L phosphate concentration). Transcriptome analysis showed that 655 differentially expressed genes (DEGs) were detected after 48h of culture under low phosphorus conditions. At 96 h, the number of DEGs increased to 9 802, including 1 777 up-regulated genes and 8 025 down-regulated genes. GO and KEGG analyses showed that DEGs were significantly enriched in GO terms such as translation and primary metabolic processes, as well as KEGG pathways such as ribosome synthesis and TCA cycle. Metabolic pathway analysis further found that low phosphorus stress activated the expression of genes in the pamamycin synthesis pathway genes, the central carbon metabolism pathway genes, and the SenX3-RegX3 two-component system genes. At the same time, it inhibited the expression of genes in the precursor competitive pathway of pamamycin, such as biotin synthesis genes (bioA, bioB) and fatty acid synthesis genes (fabF, fabH). The results showed that under low phosphorus conditions, Streptomyces albus TD-1 sensed phosphorus signals through the SenX3-RegX3 two-component system, activated central carbon metabolism and inhibited fatty acid synthesis, so as to accumulate the precursor of pamamycin synthesis and activate the expression of pamamycin synthesis genes to promote its efficient synthesis. The aim of this study was to provide a basis for the efficient biosynthesis of pamamycin and promote its application in the biological control of Aspergillus flavus. 〖