Arsenic-hyperaccumulator Pteris vittata is efficient in As uptake, probably through phosphate transporters (Pht). Here, for the first time, we cloned a new PvPht1;4 gene from P. vittata and investigated its role in arsenate (AsV) uptake and transport in yeast and transgenic tobacco plants. On the basis of quantitative real-time polymerase chain reaction (qRT-PCR), PvPht1;4 was abundantly expressed in P. vittata fronds and roots, with its transcripts in the roots being induced by both P deficiency and As exposure. PvPht1;4 was localized to the plasma membrane, which complemented a yeast-mutant defective in P uptake and showed higher P transport affinity than PvPht1;3. Under AsV exposure, PvPht1;4 yeast transformants showed comparable tolerance as PvPht1;3, but higher As accumulation than PvPht1;2 transformants, indicating that PvPht1;4 had considerable AsV and P transport activity. However, in soil and hydroponic experiments, PvPht1;4 expressing tobacco lines accumulated 26−44 and 37−55% lower As in the shoots than wild type plants, with lower root-to-shoot As translocation. In the roots of PvPht1;4 lines, higher glutathione (GSH) contents and expression levels of GSH synthetase gene NtGSH2 were observed. In addition, the transcripts of AsIII− GSH transporter NtABCC1 in PvPht1;4 lines were upregulated. The data suggested that PvPht1;4 lines probably detoxified As by reducing AsV to AsIII, which was then complexed with GSH and stored in the root vacuoles, thereby reducing As translocation in transgenic tobacco. Given its strong AsV transport capacity, expression of PvPht1;4 provides a new molecular approach to reduce As accumulation in plant shoots.

Arsenic-hyperaccumulator Pteris vittata is efficient in As accumulation and has been used in phytoremediation of As-contaminated soils. Arsenate (AsV) is the predominant As species in aerobic soils and is taken up by plants via phosphate transporters (Pht) including P. vittata. In this work, we cloned the PvPht1;3 full length coding sequence from P. vittata and investigated its role in As accumulation by yeast and plants. PvPht1;3 complemented a yeast P uptake mutant strain and showed a stronger affinity and transport capacity to AsV than PvPht1;2. In transgenic tobacco, PvPht1;3 enhanced AsV absorption and translocation, increasing As accumulation in the shoots under both hydroponic and soil experiments. On the basis of the expression patterns via qRT-PCR, PvPht1;3 was strongly induced by P deficiency but not As exposure. To further understand its expression pattern, transgenic Arabidopsis thaliana and soybean expressing the GUS reporter gene, driven by PvPht1;3 promoter, were produced. The GUS staining showed that the reporter gene was mainly expressed in the stele cells, indicating that PvPht1;3 was expressed in stele cells and was likely involved in P/As translocation. Taken together, the data suggested that PvPht1;3 was a high-affinity AsV transporter and was probably responsible for efficient As translocation in P. vittata. Our results suggest that expressing PvPht1;3 enhances As translocation and accumulation in plants, thereby improving phytoremediation of As-contaminated soils.

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