Functional Identification of 1-Deoxy-d-Xylulose-5-Phosphate Synthase (DXPS) as an Effective Herbicide Target for the Discovery of Novel Potent Inhibitors through Virtual Screening

1-Deoxy-d-xylulose-5-phosphate synthase (DXPS) is a key-limiting enzyme in the methylerythritol phosphate (MEP) pathway. With only two commercially available herbicides targeting DXPS, there is still a great amount of chemical space to explore new inhibitors targeting DXPS with different skeletons. In this work, we first used CRISPR/Cas9 and virus-induced gene silencing (VIGS) methods to identify the essential function of DXPS. Based on structure-based virtual screening and herbicidal activity assay, we screened two active inhibitors (19 and 24) with novel chemotypes. The analysis of chlorophyll content, surface plasmon resonance (SPR), and molecular docking revealed the mechanism of action of 19 and 24 targeting DXPS. In greenhouse tests, the postemergence and pre-emergence herbicidal activities of 24 (100 and 65%) and Bixlozone (100 and 61%) on BRACL (Brassica chinensis L.) at 750 g a.i./ha were comparable. This work provides a novel chemical framework and optimization basis for the development of herbicides based on DXPS.