Abstract BACKGROUND Chenopodium glaucum is one of the most serious weeds in Medicago sativa fields in northern China, and its control relies primarily on chemical herbicides. The excessive use of chemical herbicides has led to issues such as environmental pollution. Microbial herbicides are regarded as a promising alternative for sustainable weed management. RESULTS Strain MXBP304 was identified as a new species based on morphological and molecular analyses. Its spore suspension showed great potential as a pre‐emergence bioherbicide for M. sativa fields, evaluated by determining germination rates. Eighty‐eight percent of C . glaucum failed to germinate at 1.19 × 10 6 spores mL −1 , and the inhibitory effect did not differ from that of commercial pendimethalin. Meanwhile, the suspension at this concentration was safe for nine crops, including M. sativa . Whole‐genome sequence of MXBP304 showed that the genome size was 37 809 574 bp, with a GC content of 52.13% and encoding 10 277 protein‐coding genes. Comparative genomic analysis was further conducted between MXBP304 and 11 related species via phylogenetic and genomic evolutionary analyses. The antiSMASH predicted 10 known secondary metabolite biosynthetic gene clusters in the genome of MXBP304, which may play key roles in its herbicidal activity. CONCLUSION MXBP304 strain, isolated from the rhizosphere of M. sativa , was identified as a new species of the genus Mortierella and exhibited great potential as a pre‐emergence bioherbicide for multiple crops. Whole‐genome sequence of MXBP304 has provided a basis for exploring its herbicidal mechanism, which may be crucial for its commercial application. © 2026 Society of Chemical Industry.
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