Rapid and Field-Deployable Detection of the S59G Mutation in Glutamine Synthetase Endowing Glufosinate Resistance in Eleusine indica

Glufosinate is a crucial nonselective herbicide used in both conventional and transgenic cropping systems. Its effectiveness is increasingly compromised by resistant weed species like Eleusine indica (L.) Gaertn. A single nucleotide mutation in the GS1–1 gene, specifically the S59G substitution in glutamine synthetase, is the only known target-site mutation conferring resistance to glufosinate in E. indica. This study presents a rapid and sensitive detection system for the GS1–1 S59G mutation, integrating recombinase-aided amplification (RAA) with CRISPR/Cas12a technology. We optimized key components to achieve robust performance, detecting target sequences at concentrations as low as approximately 104 aM. The system effectively differentiates between susceptible and S59G-mutated resistant E. indica using fluorescence readouts or lateral flow assays. This RAA-CRISPR/Cas12a detection system serves as an efficient molecular diagnostic tool for monitoring glufosinate resistance in E. indica, aiding in effective weed management strategies.