aquaporin PvTIP4;1 from Pteris vittata may mediate arsenite uptake

• Published in: The New phytologist 2016-01, Vol.209 (2), p.746-761
• Main Authors: He, Zhenyan, Yan, Huili, Chen, Yanshan, Shen, Hongling, Xu, Wenxiu, Zhang, Haiyan, Shi, Lei, Zhu, Yong‐Guan, Ma, Mi
• Format: Article
• Language: English
• Subjects: Accumulation; aquaporin; Aquaporins; Aquaporins - chemistry; Aquaporins - genetics; Aquaporins - metabolism; Arabidopsis; Arabidopsis - drug effects; Arabidopsis - genetics; Arabidopsis - metabolism; Arginine; Aromatic compounds; aromatic/arginine (Ar/R); Arsenic; arsenic (As); Arsenic - toxicity; Arsenite; arsenite (As(III)); arsenites; Arsenites - metabolism; Arsenites - pharmacokinetics; Biological Transport; Callus; cDNA libraries; Complementary DNA; Cysteine; DNA; Domains; Ferns; ferns and fern allies; Fusion protein; Gene Expression Regulation, Plant; Genes; heterologous gene expression; hyperaccumulators; Localization; Membrane Proteins - genetics; Mutagenesis; Mutation; Nucleotide sequence; PCR; Plant Proteins - chemistry; Plant Proteins - genetics; Plant Proteins - metabolism; Plants, Genetically Modified; plasma membrane; Proteins; Protoplasts; Pteris - genetics; Pteris - metabolism; Pteris vittata; PvTIP4; 1; quantitative polymerase chain reaction; roots; Saccharomyces cerevisiae; Saccharomyces cerevisiae - drug effects; Saccharomyces cerevisiae - genetics; Saccharomyces cerevisiae Proteins - genetics; Site-directed mutagenesis; Tissue; tonoplast; Tonoplast intrinsic protein; Toxicity; transgenic plants; Uptake; Yeast; Yeasts
• ISSN: 0028-646X; 1469-8137
• DOI: 10.1111/nph.13637

The fern Pteris vittata is an arsenic hyperaccumulator. The genes involved in arsenite (As(III)) transport are not yet clear. Here, we describe the isolation and characterization of a new P. vittata aquaporin gene, PvTIP4;1, which may mediate As(III) uptake. PvTIP4;1 was identified from yeast functional complement cDNA library of P. vittata. Arsenic toxicity and accumulating activities of PvTIP4;1 were analyzed in Saccharomyces cerevisiae and Arabidopsis. Subcellular localization of PvTIP4;1–GFP fusion protein in P. vittata protoplast and callus was conducted. The tissue expression of PvTIP4;1 was investigated by quantitative real‐time PCR. Site‐directed mutagenesis of the PvTIP4;1 aromatic/arginine (Ar/R) domain was studied. Heterologous expression in yeast demonstrates that PvTIP4;1 was able to facilitate As(III) diffusion. Transgenic Arabidopsis showed that PvTIP4;1 increases arsenic accumulation and induces arsenic sensitivity. Images and FM4‐64 staining suggest that PvTIP4;1 localizes to the plasma membrane in P. vittata cells. A tissue location study shows that PvTIP4;1 transcripts are mainly expressed in roots. Site‐directed mutation in yeast further proved that the cysteine at the LE1 position of PvTIP4;1 Ar/R domain is a functional site. PvTIP4;1 is a new represented tonoplast intrinsic protein (TIP) aquaporin from P. vittata and the function and location results imply that PvTIP4;1 may be involved in As(III) uptake.