Molybdenum (Mo) is required by the enzymes involved in many metabolic processes related to plant growth and development. However, the effects of Mo addition on plant growth and beneficial microorganisms in mixed grasslands are unclear. We conducted a greenhouse experiment to examine the effects of different Mo addition levels (10 and 20 mg Mo kg−1 soil in the form of Na2MoO4) on the growth of perennial ryegrass–white clover in two low-Mo soils, as well as their symbiotic microorganisms. Our results showed that the addition of Mo had a significant impact on plant growth in limestone soil but not in yellow loam soil (p < 0.05). Compared with no addition of Mo fertilizer in limestone soil, an addition of 10 mg Mo kg−1 significantly increased the plant community shoot and root biomass (p < 0.05). However, this improvement was not observed with an addition of 20 mg Mo kg−1. The shoot nitrogen and phosphorus content in both soil types was unaffected by the Mo addition (p > 0.05), whereas the 10 mg Mo kg−1 addition significantly increased the shoot nitrogen and phosphorus uptake in limestone soil (p < 0.05). This increase in plant community productivity was primarily due to the increased growth of both species, caused by the enhanced activation of the symbiotic rhizobium. We conclude that Mo supply may promote N utilization and uptake in mixed grassland by increasing the activity of symbiotic rhizobium, resulting in a higher yield of mixed grassland, which is critical for sustainable agricultural development in low-Mo soils.
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