title: Grapevine and Arabidopsis Cation-Chloride Cotransporters Localize to the Golgi and Trans-Golgi Network and Indirectly Influence Long-Distance Ion Transport and Plant Salt Tolerance.
abstract: Plant cation-chloride cotransporters (CCCs) have been implicated in conferring salt tolerance. They are predicted to improve shoot salt exclusion by directly catalyzing the retrieval of sodium (Na(+)) and chloride (Cl(-)) ions from the root xylem. We investigated whether grapevine (Vitis vinifera [Vvi]) CCC has a role in salt tolerance by cloning and functionally characterizing the gene from the cultivar Cabernet Sauvignon. Amino acid sequence analysis revealed that VviCCC shares a high degree of similarity with other plant CCCs. A VviCCC-yellow fluorescent protein translational fusion protein localized to the Golgi and the trans-Golgi network and not the plasma membrane when expressed transiently in tobacco (Nicotiana benthamiana) leaves and Arabidopsis (Arabidopsis thaliana) mesophyll protoplasts. AtCCC-green fluorescent protein from Arabidopsis also localized to the Golgi and the trans-Golgi network. In Xenopus laevis oocytes, VviCCC targeted to the plasma membrane, where it catalyzed bumetanide-sensitive (36)Cl(-), (22)Na(+), and (86)Rb(+) uptake, suggesting that VviCCC (like AtCCC) belongs to the Na(+)-K(+)-2Cl(-) cotransporter class of CCCs. Expression of VviCCC in an Arabidopsis ccc knockout mutant abolished the mutant's stunted growth phenotypes and reduced shoot Cl(-) and Na(+) content to wild-type levels after growing plants in 50 mm NaCl. In grapevine roots, VviCCC transcript abundance was not regulated by Cl(-) treatment and was present at similar levels in both the root stele and cortex of three Vitis spp. genotypes that exhibit differential shoot salt exclusion. Our findings indicate that CCC function is conserved between grapevine and Arabidopsis, but neither protein is likely to directly mediate ion transfer with the xylem or have a direct role in salt tolerance.
authors.affiliation: Australian Research Council Centre of Excellence in Plant Energy Biology and the University of Adelaide School of Agriculture, Food and Wine (S.W.H., S.W., J.Q., S.D.T., M.G.) andCommonwealth Scientific and Industrial Research Organization Agriculture (D.H.B., A.R.W., R.R.W.), Waite Research Precinct, Glen Osmond, South Australia 5064, Australia.
authors.forename: Sam W
authors.identifier: http://orcid.org/0000-0003-3019-1891
authors.initials: SW
authors.lastname: Henderson
authors.affiliation: Australian Research Council Centre of Excellence in Plant Energy Biology and the University of Adelaide School of Agriculture, Food and Wine (S.W.H., S.W., J.Q., S.D.T., M.G.) andCommonwealth Scientific and Industrial Research Organization Agriculture (D.H.B., A.R.W., R.R.W.), Waite Research Precinct, Glen Osmond, South Australia 5064, Australia.
authors.forename: Stefanie
authors.identifier: http://orcid.org/0000-0002-7232-5889
authors.initials: S
authors.lastname: Wege
authors.affiliation: Australian Research Council Centre of Excellence in Plant Energy Biology and the University of Adelaide School of Agriculture, Food and Wine (S.W.H., S.W., J.Q., S.D.T., M.G.) andCommonwealth Scientific and Industrial Research Organization Agriculture (D.H.B., A.R.W., R.R.W.), Waite Research Precinct, Glen Osmond, South Australia 5064, Australia.
authors.forename: Jiaen
authors.initials: J
authors.lastname: Qiu
authors.affiliation: Australian Research Council Centre of Excellence in Plant Energy Biology and the University of Adelaide School of Agriculture, Food and Wine (S.W.H., S.W., J.Q., S.D.T., M.G.) andCommonwealth Scientific and Industrial Research Organization Agriculture (D.H.B., A.R.W., R.R.W.), Waite Research Precinct, Glen Osmond, South Australia 5064, Australia.
authors.forename: Deidre H
authors.identifier: http://orcid.org/0000-0002-2008-493X
authors.initials: DH
authors.lastname: Blackmore
authors.affiliation: Australian Research Council Centre of Excellence in Plant Energy Biology and the University of Adelaide School of Agriculture, Food and Wine (S.W.H., S.W., J.Q., S.D.T., M.G.) andCommonwealth Scientific and Industrial Research Organization Agriculture (D.H.B., A.R.W., R.R.W.), Waite Research Precinct, Glen Osmond, South Australia 5064, Australia.
authors.forename: Amanda R
authors.identifier: http://orcid.org/0000-0002-7596-8484
authors.initials: AR
authors.lastname: Walker
authors.affiliation: Australian Research Council Centre of Excellence in Plant Energy Biology and the University of Adelaide School of Agriculture, Food and Wine (S.W.H., S.W., J.Q., S.D.T., M.G.) andCommonwealth Scientific and Industrial Research Organization Agriculture (D.H.B., A.R.W., R.R.W.), Waite Research Precinct, Glen Osmond, South Australia 5064, Australia.
authors.forename: Stephen D
authors.identifier: http://orcid.org/0000-0003-2455-1643
authors.initials: SD
authors.lastname: Tyerman
authors.affiliation: Australian Research Council Centre of Excellence in Plant Energy Biology and the University of Adelaide School of Agriculture, Food and Wine (S.W.H., S.W., J.Q., S.D.T., M.G.) andCommonwealth Scientific and Industrial Research Organization Agriculture (D.H.B., A.R.W., R.R.W.), Waite Research Precinct, Glen Osmond, South Australia 5064, Australia.
authors.forename: Rob R
authors.initials: RR
authors.lastname: Walker
authors.affiliation: Australian Research Council Centre of Excellence in Plant Energy Biology and the University of Adelaide School of Agriculture, Food and Wine (S.W.H., S.W., J.Q., S.D.T., M.G.) andCommonwealth Scientific and Industrial Research Organization Agriculture (D.H.B., A.R.W., R.R.W.), Waite Research Precinct, Glen Osmond, South Australia 5064, Australia matthew.gilliham@adelaide.edu.au.
authors.forename: Matthew
authors.identifier: http://orcid.org/0000-0003-0666-3078
authors.initials: M
authors.lastname: Gilliham
mesh_terms: D000818:Animals; D017360:Arabidopsis / Q000235:genetics / Q000502:physiology*; D027682:Cation Transport Proteins / Q000235:genetics / Q000378:metabolism*; D002712:Chlorides / Q000378:metabolism; D006056:Golgi Apparatus / Q000378:metabolism; D017136:Ion Transport; D009154:Mutation; D009865:Oocytes; D010641:Phenotype; D018515:Plant Leaves / Q000235:genetics / Q000502:physiology; D010940:Plant Proteins / Q000235:genetics / Q000378:metabolism; D018517:Plant Roots / Q000235:genetics / Q000502:physiology; D030821:Plants, Genetically Modified; D011523:Protoplasts; D055049:Salt Tolerance; D012965:Sodium Chloride / Q000378:metabolism*; D014026:Nicotiana / Q000235:genetics / Q000502:physiology; D027843:Vitis / Q000235:genetics / Q000502:physiology*; D014981:Xenopus; D052584:Xylem / Q000235:genetics / Q000502:physiology; D021601:trans-Golgi Network / Q000378:metabolism
chemical_list: D027682:Cation Transport Proteins; D002712:Chlorides; D010940:Plant Proteins; D012965:Sodium Chloride