Zinc (Zn) is one of the most essential micronutrients for plant growth and metabolism, but Zn excess can impair many basic metabolic processes in plant cells. In agriculture, crops often experience low phosphate (Pi) and high Zn double nutrient stresses because of inordinate agro-industrial activities, while the dual benefit of arbuscular mycorrhizal (AM) fungi protects plants from experiencing both deficient and toxic nutrient stresses. Although crosstalk between Pi and Zn nutrients in plants have been extensively studied at the physiological level, the molecular basis of how Pi starvation triggers Zn over-accumulation in plants and how AM plants coordinately modulate the Pi and Zn nutrient homeostasis remains to be elucidated. Here, we report that a novel AsZIP2 gene, a Chinese milk vetch (Astragalus sinicus) member of the ZIP gene family, participates in the interaction between Pi and Zn nutrient homeostasis in plants. Phylogenetic analysis revealed that this AsZIP2 protein was closely related to the orthologous Medicago MtZIP2 and Arabidopsis AtZIP2 transporters. Gene expression analysis indicated that AsZIP2 was highly induced in roots by Pi starvation or Zn excess yet attenuated by arbuscular mycorrhization in a Pi-dependent manner. Subcellular localization and heterologous expression experiments further showed that AsZIP2 encoded a functional plasma membrane-localized transporter that mediated Zn uptake in yeast. Moreover, overexpression of AsZIP2 in A. sinicus resulted in the over-accumulation of Zn concentration in roots at low Pi or excessive Zn concentrations, whereas AsZIP2 silencing lines displayed an even more reduced Zn concentration than control lines under such conditions. Our results reveal that the AsZIP2 transporter functioned in Zn over-accumulation in roots during Pi starvation or high Zn supply but was repressed by AM symbiosis in a Pi-dependent manner. These findings also provide new insights into the AsZIP2 gene acting in the regulation of Zn homeostasis in mycorrhizal plants through Pi signal.

Arbuscular mycorrhizal (AM) fungi can improve the lead (Pb) tolerance of host plants and accumulate intensive Pb in mycorrhizal roots. However, the detailed contribution of AM fungal extraradical hyphae to the plants' Pb uptake remains unknown. In this study, mulberry (Morus alba) colonized by the AM fungus (Rhizophagus irregularis) with light treatments were linked by fungal extraradical hyphae using a three-compartment system (pot test), and their differences in responding to Pb application were compared. Shading inhibited mulberry photosynthesis and the growth of mulberry. In this study, Pb application did not affect the colonization of R. irregularis when symbiosis had already formed as the root was not exposed to Pb during the colonization and formation of the AM fungal hyphae network. The R. irregularis preferred to transfer more Pb to the unshaded mulberry than to the shaded mulberry, a condition capable of providing more C supply for fungal survival than to low-light mulberry. The Pb transferred through the mycorrhizal pathway to mulberry had low mobility and might be compartmented in the root by R. irregularis until exceeding a threshold. The relatively high expressions of MaABCG16 with high Pb concentrations in plants suggest that MaABCG16 might play an important role in Pb translocation.

Eucalyptus grandis (E. grandis) has been reported to form a symbiosis with arbuscular mycorrhizal fungi (AMF), which plays an important role in improving plant tolerance of heavy metal. However, the mechanism of how AMF intercept and transport cadmium (Cd) at the subcellular level in E. grandis still remains to be researched. In this study, a pot experiment was conducted to investigate the growth performance of E. grandis under Cd stress and Cd absorption resistance of AMF and explored the Cd localization in the root by using transmission electron microscopy and energy dispersive X-ray spectroscopy. The results showed that AMF colonization could enhance plant growth and photosynthetic efficiency of E. grandis and reduce the translocation factor of Cd under Cd stress. After being treated with 50, 150, 300, and 500 μM Cd, the translocation factor of Cd in E. grandis with AMF colonization decreased by 56.41%, 62.89%, 66.67%, and 42.79%, respectively. However, the mycorrhizal efficiency was significant only at low Cd concentrations (50, 150, and 300 μM). Under 500 μM Cd concentration condition, the colonization of AMF in roots decreased, and the alleviating effect of AMF was not significant. Ultrastructural observations showed that Cd is abundant in regular lumps and strips in the cross-section of E. grandis root cell. AMF protected plant cells by retaining Cd in the fungal structure. Our results suggested that AMF alleviated Cd toxicity by regulating plant physiology and altering the distribution of Cd in different cell sites.