Allelopathic effects of artemisinin on ectomycorrhizal fungal isolates in vitro

• Artemisinin inhibited significantly the growth and nutrient uptake (nitrogen, phosphorus and potassium) of all studied ectomycorrhizal fungi in vitro.
• Artemisinin increased the rate of fungal oxalate efflux, presumably suggesting membrane damage and abnormal opening of anion channels on hyphae cell membranes.
• The growth, nutrient uptake and oxalate efflux of ectomycorrhizal fungi varied greatly in response to artemisinin.

The anti-malarial drug artemisinin is extracted from the leaves of Artemisia annua L. The release of artemisinin into forest soils could produce a potential risk for forest ecosystems, including effects on ectomycorrhizal fungal nutrient uptake, in areas where commercial and continual cultivation of the medicinal plant A. annua L. is practiced. Therefore, growth, proton and oxalate efflux, and nutrient uptake (nitrogen, phosphorus and potassium) of three isolates of Suillus luteus (S. luteus 1, S. luteus 13, and S. luteus 11) and of one isolate of Suillus subluteus (S. subluteus 12) were compared in culture solutions with different nominal artemisinin concentrations. The results showed that artemisinin inhibited significantly the growth of all studied fungi. With 25 mg artemisinin L−1 added, fungal biomass was decreased by 78.6% (S. luteus 1), 96.7% (S. luteus 13), 77.8% (S. luteus 11) and 86.8% (S. subluteus 12) compared with the control (without artemisinin). This could explain, at least in part, why ectomycorrhizal fungal sporocarps in forests are consistently not found near cultivated A. annua L. fields. The amount of proton efflux by the fungal isolates also decreased as nominal artemisinin concentrations increased, indicating the limited ability of ectomycorrhizal fungi to mobilize nutrients from soil minerals. However, nominal artemisinin significantly increased the rate of fungal oxalate efflux, suggesting membrane damage and the abnormal opening of anion channels on hyphae cell membranes. Nominal artemisinin also decreased the uptake of nitrogen, phosphorus and potassium by the fungal isolates, which may not benefit from the nutrient uptake by ectomycorrhizae. Therefore, artemisinin released from large A. annua L. plantations may inhibit ectomycorrhizal fungal growth, nutrition and functions in forest ecosystems in Southwest China.