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Clethra barbinervis  Sieb. et Zucc. is a tree species that grows naturally at several mine sites and seems to be tolerant of high concentrations of heavy metals, such as Cu, Zn, and Pb. The purpose of this study is to clarify the mechanism(s) underlying this species’ ability to tolerate the sites’ severe heavy-metal pollution by considering  C .  barbinervis  interaction with root fungal endophytes. We measured the heavy metal concentrations of root-zone soil, leaves, branches, and fine roots collected from mature  C .  barbinervis  at Hitachi mine. We isolated fungal endophytes from surface-sterilized root segments, and we examined the growth, and heavy metal and nutrient absorption of  C .  barbinervis  seedlings growing in sterilized mine soil with or without root fungal endophytes. Field analyses showed that  C .  barbinervis  contained considerably high amounts of Cu, Zn, and Pb in fine roots and Zn in leaves. The fungi,  Phialocephala fortinii ,  Rhizodermea veluwensis , and  Rhizoscyphus  sp. were frequently isolated as dominant fungal endophyte species. Inoculation of these root fungal endophytes to  C .  barbinervis  seedlings growing in sterilized mine soil indicated that these fungi significantly enhanced the growth of  C .  barbinervis  seedlings, increased K uptake in shoots and reduced the concentrations of Cu, Ni, Zn, Cd, and Pb in roots. Without root fungal endophytes,  C .  barbinervis  could hardly grow under the heavy-metal contaminated condition, showing chlorosis, a symptom of heavy-metal toxicity. Our results indicate that the tree  C .  barbinervis  can tolerate high heavy-metal concentrations due to the support of root fungal endophytes including  P .  fortinii ,  R .  veluwensis , and  Rhizoscyphus  sp. via growth enhancement, K uptake promotion and decrease of heavy metal concentrations.

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Root Fungal Endophytes Enhance Heavy-Metal Stress Tolerance of Clethra barbinervis Growing Naturally at Mining Sites via Growth Enhancement, Promotion of Nutrient Uptake and Decrease of Heavy-Metal Concentration