title: The Arabidopsis nitrate transporter NRT1.8 functions in nitrate removal from the xylem sap and mediates cadmium tolerance.
abstract: Long-distance transport of nitrate requires xylem loading and unloading, a successive process that determines nitrate distribution and subsequent assimilation efficiency. Here, we report the functional characterization of NRT1.8, a member of the nitrate transporter (NRT1) family in Arabidopsis thaliana. NRT1.8 is upregulated by nitrate. Histochemical analysis using promoter-beta-glucuronidase fusions, as well as in situ hybridization, showed that NRT1.8 is expressed predominantly in xylem parenchyma cells within the vasculature. Transient expression of the NRT1.8:enhanced green fluorescent protein fusion in onion epidermal cells and Arabidopsis protoplasts indicated that NRT1.8 is plasma membrane localized. Electrophysiological and nitrate uptake analyses using Xenopus laevis oocytes showed that NRT1.8 mediates low-affinity nitrate uptake. Functional disruption of NRT1.8 significantly increased the nitrate concentration in xylem sap. These data together suggest that NRT1.8 functions to remove nitrate from xylem vessels. Interestingly, NRT1.8 was the only nitrate assimilatory pathway gene that was strongly upregulated by cadmium (Cd(2+)) stress in roots, and the nrt1.8-1 mutant showed a nitrate-dependent Cd(2+)-sensitive phenotype. Further analyses showed that Cd(2+) stress increases the proportion of nitrate allocated to wild-type roots compared with the nrt1.8-1 mutant. These data suggest that NRT1.8-regulated nitrate distribution plays an important role in Cd(2+) tolerance.
authors.affiliation: National Key Laboratory of Plant Molecular Genetics, Institute of Plant Physiology and Ecology, Shanghai Institutes for Biological Sciences, Chinese Academy of Sciences, Shanghai, People's Republic of China.
authors.forename: Jian-Yong
authors.initials: JY
authors.lastname: Li
authors.forename: Yan-Lei
authors.initials: YL
authors.lastname: Fu
authors.forename: Sharon M
authors.initials: SM
authors.lastname: Pike
authors.forename: Juan
authors.initials: J
authors.lastname: Bao
authors.forename: Wang
authors.initials: W
authors.lastname: Tian
authors.forename: Yu
authors.initials: Y
authors.lastname: Zhang
authors.forename: Chun-Zhu
authors.initials: CZ
authors.lastname: Chen
authors.forename: Yi
authors.initials: Y
authors.lastname: Zhang
authors.forename: Hong-Mei
authors.initials: HM
authors.lastname: Li
authors.forename: Jing
authors.initials: J
authors.lastname: Huang
authors.forename: Le-Gong
authors.initials: LG
authors.lastname: Li
authors.forename: Julian I
authors.initials: JI
authors.lastname: Schroeder
authors.forename: Walter
authors.initials: W
authors.lastname: Gassmann
authors.forename: Ji-Ming
authors.initials: JM
authors.lastname: Gong
mesh_terms: D000222:Adaptation, Physiological / Q000187:drug effects* / Q000235:genetics; D027321:Anion Transport Proteins / Q000235:genetics / Q000378:metabolism*; D017360:Arabidopsis / Q000166:cytology / Q000187:drug effects / Q000201:enzymology* / Q000235:genetics; D029681:Arabidopsis Proteins / Q000235:genetics / Q000378:metabolism*; D002104:Cadmium / Q000633:toxicity*; D002462:Cell Membrane / Q000187:drug effects / Q000201:enzymology; D020869:Gene Expression Profiling; D018506:Gene Expression Regulation, Plant / Q000187:drug effects; D006863:Hydrogen-Ion Concentration / Q000187:drug effects; D009154:Mutation / Q000235:genetics; D000091285:Nitrate Transporters; D009566:Nitrates / Q000378:metabolism*; D053147:Plant Exudates / Q000235:genetics / Q000378:metabolism*; D021381:Protein Transport / Q000187:drug effects; D013312:Stress, Physiological / Q000187:drug effects / Q000235:genetics; D013347:Subcellular Fractions / Q000187:drug effects / Q000201:enzymology; D015854:Up-Regulation / Q000187:drug effects; D052584:Xylem / Q000166:cytology / Q000187:drug effects / Q000201:enzymology* / Q000235:genetics
chemical_list: D027321:Anion Transport Proteins; D029681:Arabidopsis Proteins; C552677:NRT1.8 protein, Arabidopsis; D000091285:Nitrate Transporters; D009566:Nitrates; D053147:Plant Exudates; D002104:Cadmium