title: Difference in the distribution and speciation of cellular nickel between nickel-tolerant and non-tolerant Nicotiana tabacum L. cv. BY-2 cells.
abstract: To evaluate Ni dynamics at the subcellular level, the distribution and speciation of Ni were determined in wild-type (WT) and Ni-tolerant (NIT) tobacco BY-2 cell lines. When exposed to low but toxic levels of Ni, NIT cells were found to contain 2.5-fold more Ni (14% of whole-cell Ni values) in their cell walls than WT cells (6% of whole-cell Ni values). In addition to higher levels of Ni in the apoplast, a higher proportion (94%) of symplastic Ni was localized in the vacuoles of NIT cells than in the vacuoles of WT cells (81%). The concentration of cytosolic Ni in the NIT cells was significantly lower (18 nmol g(-1) FW) than that in the WT cells (85 nmol g(-1) FW). In silico simulation showed that 95% of vacuolar Ni was in the form of Ni-citrate complexes, and that free Ni(2+) was virtually absent in the NIT cells. On the other hand, the amount of free metal ions was markedly increased in WT cells because free citrate was depleted by chelation of Ni. A protoplast viability assay using BCECF-AM further demonstrated that the main mechanism that confers strong Ni tolerance was present in the symplast as opposed to the cell wall.
authors.affiliation: Laboratory of Plant Production Chemistry, Department of Applied Biology and Chemistry, Tokyo University of Agriculture, Tokyo 156-8502, Japan.
authors.forename: Akihiro
authors.initials: A
authors.lastname: Saito
authors.forename: Misa
authors.initials: M
authors.lastname: Saito
authors.forename: Yusuke
authors.initials: Y
authors.lastname: Ichikawa
authors.forename: Masaaki
authors.initials: M
authors.lastname: Yoshiba
authors.forename: Toshiaki
authors.initials: T
authors.lastname: Tadano
authors.forename: Eitaro
authors.initials: E
authors.lastname: Miwa
authors.forename: Kyoko
authors.initials: K
authors.lastname: Higuchi
mesh_terms: D002460:Cell Line; D002473:Cell Wall / Q000378:metabolism*; D002951:Citrates / Q000378:metabolism; D008956:Models, Chemical; D009532:Nickel / Q000378:metabolism*; D014026:Nicotiana / Q000378:metabolism*; D014617:Vacuoles / Q000378:metabolism*
chemical_list: D002951:Citrates; D009532:Nickel