The HvNramp5 Transporter Mediates Uptake of Cadmium and Manganese, But Not Iron.
The Natural Resistance Associated Macrophage Protein (Nramp) represents a transporter family for metal ions in all organisms. Here, we functionally characterized a member of Nramp family in barley (Hordeum vulgare), HvNramp5. This member showed different expression patterns, transport substrate specificity, and cellular localization from its close homolog in rice (Oryza sativa), OsNramp5, although HvNramp5 was also localized to the plasma membrane. HvNramp5 was mainly expressed in the roots and its expression was not affected by Cd and deficiency of Zn, Cu, and Mn, but slightly up-regulated by Fe deficiency. Spatial expression analysis showed that the expression of HvNramp5 was higher in the root tips than that in the basal root regions. Furthermore, analysis with laser microdissection revealed higher expression of HvNramp5 in the outer root cell layers. HvNramp5 showed transport activity for both Mn2+and Cd2+, but not for Fe2+when expressed in yeast. Immunostaining with a HvNramp5 antibody showed that this protein was localized in the root epidermal cells without polarity. Knockdown of HvNramp5 in barley resulted in a significant reduction in the seedling growth at low Mn supply, but this reduction was rescued at high Mn supply. The concentration of Mn and Cd, but not other metals including Cu, Zn, and Fe, was decreased in both the roots and shoots of knockdown lines compared with the wild-type barley. These results indicate that HvNramp5 is a transporter required for uptake of Mn and Cd, but not for Fe, and that barley has a distinct uptake system from rice.
Institute of Plant Science and Resources, Okayama University, Kurashiki 710-0046, Japan (D.W., N.Y., M.Y., M.K., K.S., J.F.M.); and.|Department of Agronomy, Zhejiang University, Hangzhou 310058, China (D.W.).
Dezhi
D
Wu
Institute of Plant Science and Resources, Okayama University, Kurashiki 710-0046, Japan (D.W., N.Y., M.Y., M.K., K.S., J.F.M.); and.|Department of Agronomy, Zhejiang University, Hangzhou 310058, China (D.W.).
Naoki
N
Yamaji
Institute of Plant Science and Resources, Okayama University, Kurashiki 710-0046, Japan (D.W., N.Y., M.Y., M.K., K.S., J.F.M.); and.|Department of Agronomy, Zhejiang University, Hangzhou 310058, China (D.W.).
Miki
M
Yamane
Institute of Plant Science and Resources, Okayama University, Kurashiki 710-0046, Japan (D.W., N.Y., M.Y., M.K., K.S., J.F.M.); and.|Department of Agronomy, Zhejiang University, Hangzhou 310058, China (D.W.).
Miho
M
Kashino-Fujii
Institute of Plant Science and Resources, Okayama University, Kurashiki 710-0046, Japan (D.W., N.Y., M.Y., M.K., K.S., J.F.M.); and.|Department of Agronomy, Zhejiang University, Hangzhou 310058, China (D.W.).
Kazuhiro
0000-0001-8818-5203
K
Sato
Institute of Plant Science and Resources, Okayama University, Kurashiki 710-0046, Japan (D.W., N.Y., M.Y., M.K., K.S., J.F.M.); and maj@rib.okayama-u.ac.jp.|Department of Agronomy, Zhejiang University, Hangzhou 310058, China (D.W.) maj@rib.okayama-u.ac.jp.
Jian
0000-0003-3411-827X
J
Feng Ma
D001692:Biological Transport; D002104:Cadmium / Q000378:metabolism*; D018506:Gene Expression Regulation, Plant; D055785:Gene Knockdown Techniques; D001467:Hordeum / Q000235:genetics / Q000378:metabolism*; D007501:Iron / Q000378:metabolism*; D008345:Manganese / Q000378:metabolism*; D026901:Membrane Transport Proteins / Q000235:genetics / Q000378:metabolism*; D012275:Oryza / Q000378:metabolism; D010641:Phenotype; D010802:Phylogeny; D010940:Plant Proteins / Q000235:genetics / Q000378:metabolism*; D030821:Plants, Genetically Modified; D034622:RNA Interference; D012441:Saccharomyces cerevisiae / Q000378:metabolism; D013347:Subcellular Fractions / Q000378:metabolism
D026901:Membrane Transport Proteins; D010940:Plant Proteins; D002104:Cadmium; D008345:Manganese; D007501:Iron