Flavonoids play a key role in plant defense. This study utilizes a fluorescent biosensor to track flavonoid synthesis in maize without extensive chemical analysis. E. coli expressing the biosensor is exposed to flavonoid standards to establish fluorescence baselines. Spectrophotometry quantifies flavonoid levels, enabling efficient and non-destructive detection. This approach aids in developing high-flavonoid maize germplasm for pest and disease resistance, demonstrating the potential of biosensor technology for high-throughput metabolic screening and agricultural sustainability.