title: Two distinct families of protein kinases are required for plant growth under high external Mg2+ concentrations in Arabidopsis.
abstract: Protein phosphorylation events play key roles in maintaining cellular ion homeostasis in higher plants, and the regulatory roles of these events in Na(+) and K(+) transport have been studied extensively. However, the regulatory mechanisms governing Mg(2+) transport and homeostasis in higher plants remain poorly understood, despite the vital roles of Mg(2+) in cellular function. A member of subclass III sucrose nonfermenting-1-related protein kinase2 (SnRK2), SRK2D/SnRK2.2, functions as a key positive regulator of abscisic acid (ABA)-mediated signaling in response to water deficit stresses in Arabidopsis (Arabidopsis thaliana). Here, we used immunoprecipitation coupled with liquid chromatography-tandem mass spectrometry analyses to identify Calcineurin B-like-interacting protein kinase26 (CIPK26) as a novel protein that physically interacts with SRK2D. In addition to CIPK26, three additional CIPKs (CIPK3, CIPK9, and CIPK23) can physically interact with SRK2D in planta. The srk2d/e/i triple mutant lacking all three members of subclass III SnRK2 and the cipk26/3/9/23 quadruple mutant lacking CIPK26, CIPK3, CIPK9, and CIPK23 showed reduced shoot growth under high external Mg(2+) concentrations. Similarly, several ABA biosynthesis-deficient mutants, including aba2-1, were susceptible to high external Mg(2+) concentrations. Taken together, our findings provided genetic evidence that SRK2D/E/I and CIPK26/3/9/23 are required for plant growth under high external Mg(2+) concentrations in Arabidopsis. Furthermore, we showed that ABA, a key molecule in water deficit stress signaling, also serves as a signaling molecule in plant growth under high external Mg(2+) concentrations. These results suggested that SRK2D/E/I- and CIPK26/3/9/23-mediated phosphorylation signaling pathways maintain cellular Mg(2+) homeostasis.
authors.affiliation: Laboratories of Plant Molecular Physiology (J. Mo., T.Y., Y.T., K.M., S.K., J.Mi., K.Y.-S.) andPlant Nutrition and Fertilizers (T.F., S.N.), Graduate School of Agricultural and Life Sciences andBiotechnology Research Center (S.Y., T.I.), University of Tokyo, Bunkyo-ku, Tokyo 113-8657, Japan;Biological Resources and Post-Harvest Division, Japan International Research Center for Agricultural Sciences, Tsukuba, Ibaraki 305-8686, Japan (Y.F.);Laboratory of Plant Stress Biology, Graduate School of Life and Environmental Sciences, University of Tsukuba, Ibaraki 305-8577, Japan (Y.F.); andRIKEN Center for Sustainable Resource Science, Yokohama, Kanagawa 230-0045, Japan (H.N., Y.N., F.T., K.S.).
authors.forename: Junro
authors.initials: J
authors.lastname: Mogami
authors.affiliation: Laboratories of Plant Molecular Physiology (J. Mo., T.Y., Y.T., K.M., S.K., J.Mi., K.Y.-S.) andPlant Nutrition and Fertilizers (T.F., S.N.), Graduate School of Agricultural and Life Sciences andBiotechnology Research Center (S.Y., T.I.), University of Tokyo, Bunkyo-ku, Tokyo 113-8657, Japan;Biological Resources and Post-Harvest Division, Japan International Research Center for Agricultural Sciences, Tsukuba, Ibaraki 305-8686, Japan (Y.F.);Laboratory of Plant Stress Biology, Graduate School of Life and Environmental Sciences, University of Tsukuba, Ibaraki 305-8577, Japan (Y.F.); andRIKEN Center for Sustainable Resource Science, Yokohama, Kanagawa 230-0045, Japan (H.N., Y.N., F.T., K.S.).
authors.forename: Yasunari
authors.initials: Y
authors.lastname: Fujita
authors.affiliation: Laboratories of Plant Molecular Physiology (J. Mo., T.Y., Y.T., K.M., S.K., J.Mi., K.Y.-S.) andPlant Nutrition and Fertilizers (T.F., S.N.), Graduate School of Agricultural and Life Sciences andBiotechnology Research Center (S.Y., T.I.), University of Tokyo, Bunkyo-ku, Tokyo 113-8657, Japan;Biological Resources and Post-Harvest Division, Japan International Research Center for Agricultural Sciences, Tsukuba, Ibaraki 305-8686, Japan (Y.F.);Laboratory of Plant Stress Biology, Graduate School of Life and Environmental Sciences, University of Tsukuba, Ibaraki 305-8577, Japan (Y.F.); andRIKEN Center for Sustainable Resource Science, Yokohama, Kanagawa 230-0045, Japan (H.N., Y.N., F.T., K.S.).
authors.forename: Takuya
authors.initials: T
authors.lastname: Yoshida
authors.affiliation: Laboratories of Plant Molecular Physiology (J. Mo., T.Y., Y.T., K.M., S.K., J.Mi., K.Y.-S.) andPlant Nutrition and Fertilizers (T.F., S.N.), Graduate School of Agricultural and Life Sciences andBiotechnology Research Center (S.Y., T.I.), University of Tokyo, Bunkyo-ku, Tokyo 113-8657, Japan;Biological Resources and Post-Harvest Division, Japan International Research Center for Agricultural Sciences, Tsukuba, Ibaraki 305-8686, Japan (Y.F.);Laboratory of Plant Stress Biology, Graduate School of Life and Environmental Sciences, University of Tsukuba, Ibaraki 305-8577, Japan (Y.F.); andRIKEN Center for Sustainable Resource Science, Yokohama, Kanagawa 230-0045, Japan (H.N., Y.N., F.T., K.S.).
authors.forename: Yoshifumi
authors.initials: Y
authors.lastname: Tsukiori
authors.affiliation: Laboratories of Plant Molecular Physiology (J. Mo., T.Y., Y.T., K.M., S.K., J.Mi., K.Y.-S.) andPlant Nutrition and Fertilizers (T.F., S.N.), Graduate School of Agricultural and Life Sciences andBiotechnology Research Center (S.Y., T.I.), University of Tokyo, Bunkyo-ku, Tokyo 113-8657, Japan;Biological Resources and Post-Harvest Division, Japan International Research Center for Agricultural Sciences, Tsukuba, Ibaraki 305-8686, Japan (Y.F.);Laboratory of Plant Stress Biology, Graduate School of Life and Environmental Sciences, University of Tsukuba, Ibaraki 305-8577, Japan (Y.F.); andRIKEN Center for Sustainable Resource Science, Yokohama, Kanagawa 230-0045, Japan (H.N., Y.N., F.T., K.S.).
authors.forename: Hirofumi
authors.initials: H
authors.lastname: Nakagami
authors.affiliation: Laboratories of Plant Molecular Physiology (J. Mo., T.Y., Y.T., K.M., S.K., J.Mi., K.Y.-S.) andPlant Nutrition and Fertilizers (T.F., S.N.), Graduate School of Agricultural and Life Sciences andBiotechnology Research Center (S.Y., T.I.), University of Tokyo, Bunkyo-ku, Tokyo 113-8657, Japan;Biological Resources and Post-Harvest Division, Japan International Research Center for Agricultural Sciences, Tsukuba, Ibaraki 305-8686, Japan (Y.F.);Laboratory of Plant Stress Biology, Graduate School of Life and Environmental Sciences, University of Tsukuba, Ibaraki 305-8577, Japan (Y.F.); andRIKEN Center for Sustainable Resource Science, Yokohama, Kanagawa 230-0045, Japan (H.N., Y.N., F.T., K.S.).
authors.forename: Yuko
authors.initials: Y
authors.lastname: Nomura
authors.affiliation: Laboratories of Plant Molecular Physiology (J. Mo., T.Y., Y.T., K.M., S.K., J.Mi., K.Y.-S.) andPlant Nutrition and Fertilizers (T.F., S.N.), Graduate School of Agricultural and Life Sciences andBiotechnology Research Center (S.Y., T.I.), University of Tokyo, Bunkyo-ku, Tokyo 113-8657, Japan;Biological Resources and Post-Harvest Division, Japan International Research Center for Agricultural Sciences, Tsukuba, Ibaraki 305-8686, Japan (Y.F.);Laboratory of Plant Stress Biology, Graduate School of Life and Environmental Sciences, University of Tsukuba, Ibaraki 305-8577, Japan (Y.F.); andRIKEN Center for Sustainable Resource Science, Yokohama, Kanagawa 230-0045, Japan (H.N., Y.N., F.T., K.S.).
authors.forename: Toru
authors.initials: T
authors.lastname: Fujiwara
authors.affiliation: Laboratories of Plant Molecular Physiology (J. Mo., T.Y., Y.T., K.M., S.K., J.Mi., K.Y.-S.) andPlant Nutrition and Fertilizers (T.F., S.N.), Graduate School of Agricultural and Life Sciences andBiotechnology Research Center (S.Y., T.I.), University of Tokyo, Bunkyo-ku, Tokyo 113-8657, Japan;Biological Resources and Post-Harvest Division, Japan International Research Center for Agricultural Sciences, Tsukuba, Ibaraki 305-8686, Japan (Y.F.);Laboratory of Plant Stress Biology, Graduate School of Life and Environmental Sciences, University of Tsukuba, Ibaraki 305-8577, Japan (Y.F.); andRIKEN Center for Sustainable Resource Science, Yokohama, Kanagawa 230-0045, Japan (H.N., Y.N., F.T., K.S.).
authors.forename: Sho
authors.initials: S
authors.lastname: Nishida
authors.affiliation: Laboratories of Plant Molecular Physiology (J. Mo., T.Y., Y.T., K.M., S.K., J.Mi., K.Y.-S.) andPlant Nutrition and Fertilizers (T.F., S.N.), Graduate School of Agricultural and Life Sciences andBiotechnology Research Center (S.Y., T.I.), University of Tokyo, Bunkyo-ku, Tokyo 113-8657, Japan;Biological Resources and Post-Harvest Division, Japan International Research Center for Agricultural Sciences, Tsukuba, Ibaraki 305-8686, Japan (Y.F.);Laboratory of Plant Stress Biology, Graduate School of Life and Environmental Sciences, University of Tsukuba, Ibaraki 305-8577, Japan (Y.F.); andRIKEN Center for Sustainable Resource Science, Yokohama, Kanagawa 230-0045, Japan (H.N., Y.N., F.T., K.S.).
authors.forename: Shuichi
authors.initials: S
authors.lastname: Yanagisawa
authors.affiliation: Laboratories of Plant Molecular Physiology (J. Mo., T.Y., Y.T., K.M., S.K., J.Mi., K.Y.-S.) andPlant Nutrition and Fertilizers (T.F., S.N.), Graduate School of Agricultural and Life Sciences andBiotechnology Research Center (S.Y., T.I.), University of Tokyo, Bunkyo-ku, Tokyo 113-8657, Japan;Biological Resources and Post-Harvest Division, Japan International Research Center for Agricultural Sciences, Tsukuba, Ibaraki 305-8686, Japan (Y.F.);Laboratory of Plant Stress Biology, Graduate School of Life and Environmental Sciences, University of Tsukuba, Ibaraki 305-8577, Japan (Y.F.); andRIKEN Center for Sustainable Resource Science, Yokohama, Kanagawa 230-0045, Japan (H.N., Y.N., F.T., K.S.).
authors.forename: Tetsuya
authors.initials: T
authors.lastname: Ishida
authors.affiliation: Laboratories of Plant Molecular Physiology (J. Mo., T.Y., Y.T., K.M., S.K., J.Mi., K.Y.-S.) andPlant Nutrition and Fertilizers (T.F., S.N.), Graduate School of Agricultural and Life Sciences andBiotechnology Research Center (S.Y., T.I.), University of Tokyo, Bunkyo-ku, Tokyo 113-8657, Japan;Biological Resources and Post-Harvest Division, Japan International Research Center for Agricultural Sciences, Tsukuba, Ibaraki 305-8686, Japan (Y.F.);Laboratory of Plant Stress Biology, Graduate School of Life and Environmental Sciences, University of Tsukuba, Ibaraki 305-8577, Japan (Y.F.); andRIKEN Center for Sustainable Resource Science, Yokohama, Kanagawa 230-0045, Japan (H.N., Y.N., F.T., K.S.).
authors.forename: Fuminori
authors.initials: F
authors.lastname: Takahashi
authors.affiliation: Laboratories of Plant Molecular Physiology (J. Mo., T.Y., Y.T., K.M., S.K., J.Mi., K.Y.-S.) andPlant Nutrition and Fertilizers (T.F., S.N.), Graduate School of Agricultural and Life Sciences andBiotechnology Research Center (S.Y., T.I.), University of Tokyo, Bunkyo-ku, Tokyo 113-8657, Japan;Biological Resources and Post-Harvest Division, Japan International Research Center for Agricultural Sciences, Tsukuba, Ibaraki 305-8686, Japan (Y.F.);Laboratory of Plant Stress Biology, Graduate School of Life and Environmental Sciences, University of Tsukuba, Ibaraki 305-8577, Japan (Y.F.); andRIKEN Center for Sustainable Resource Science, Yokohama, Kanagawa 230-0045, Japan (H.N., Y.N., F.T., K.S.).
authors.forename: Kyoko
authors.initials: K
authors.lastname: Morimoto
authors.affiliation: Laboratories of Plant Molecular Physiology (J. Mo., T.Y., Y.T., K.M., S.K., J.Mi., K.Y.-S.) andPlant Nutrition and Fertilizers (T.F., S.N.), Graduate School of Agricultural and Life Sciences andBiotechnology Research Center (S.Y., T.I.), University of Tokyo, Bunkyo-ku, Tokyo 113-8657, Japan;Biological Resources and Post-Harvest Division, Japan International Research Center for Agricultural Sciences, Tsukuba, Ibaraki 305-8686, Japan (Y.F.);Laboratory of Plant Stress Biology, Graduate School of Life and Environmental Sciences, University of Tsukuba, Ibaraki 305-8577, Japan (Y.F.); andRIKEN Center for Sustainable Resource Science, Yokohama, Kanagawa 230-0045, Japan (H.N., Y.N., F.T., K.S.).
authors.forename: Satoshi
authors.initials: S
authors.lastname: Kidokoro
authors.affiliation: Laboratories of Plant Molecular Physiology (J. Mo., T.Y., Y.T., K.M., S.K., J.Mi., K.Y.-S.) andPlant Nutrition and Fertilizers (T.F., S.N.), Graduate School of Agricultural and Life Sciences andBiotechnology Research Center (S.Y., T.I.), University of Tokyo, Bunkyo-ku, Tokyo 113-8657, Japan;Biological Resources and Post-Harvest Division, Japan International Research Center for Agricultural Sciences, Tsukuba, Ibaraki 305-8686, Japan (Y.F.);Laboratory of Plant Stress Biology, Graduate School of Life and Environmental Sciences, University of Tsukuba, Ibaraki 305-8577, Japan (Y.F.); andRIKEN Center for Sustainable Resource Science, Yokohama, Kanagawa 230-0045, Japan (H.N., Y.N., F.T., K.S.).
authors.forename: Junya
authors.initials: J
authors.lastname: Mizoi
authors.affiliation: Laboratories of Plant Molecular Physiology (J. Mo., T.Y., Y.T., K.M., S.K., J.Mi., K.Y.-S.) andPlant Nutrition and Fertilizers (T.F., S.N.), Graduate School of Agricultural and Life Sciences andBiotechnology Research Center (S.Y., T.I.), University of Tokyo, Bunkyo-ku, Tokyo 113-8657, Japan;Biological Resources and Post-Harvest Division, Japan International Research Center for Agricultural Sciences, Tsukuba, Ibaraki 305-8686, Japan (Y.F.);Laboratory of Plant Stress Biology, Graduate School of Life and Environmental Sciences, University of Tsukuba, Ibaraki 305-8577, Japan (Y.F.); andRIKEN Center for Sustainable Resource Science, Yokohama, Kanagawa 230-0045, Japan (H.N., Y.N., F.T., K.S.).
authors.forename: Kazuo
authors.initials: K
authors.lastname: Shinozaki
authors.affiliation: Laboratories of Plant Molecular Physiology (J. Mo., T.Y., Y.T., K.M., S.K., J.Mi., K.Y.-S.) andPlant Nutrition and Fertilizers (T.F., S.N.), Graduate School of Agricultural and Life Sciences andBiotechnology Research Center (S.Y., T.I.), University of Tokyo, Bunkyo-ku, Tokyo 113-8657, Japan;Biological Resources and Post-Harvest Division, Japan International Research Center for Agricultural Sciences, Tsukuba, Ibaraki 305-8686, Japan (Y.F.);Laboratory of Plant Stress Biology, Graduate School of Life and Environmental Sciences, University of Tsukuba, Ibaraki 305-8577, Japan (Y.F.); andRIKEN Center for Sustainable Resource Science, Yokohama, Kanagawa 230-0045, Japan (H.N., Y.N., F.T., K.S.) akys@mail.ecc.u-tokyo.ac.jp.
authors.forename: Kazuko
authors.identifier: http://orcid.org/0000-0002-0249-8258
authors.initials: K
authors.lastname: Yamaguchi-Shinozaki
mesh_terms: D000040:Abscisic Acid / Q000096:biosynthesis; D017360:Arabidopsis / Q000187:drug effects / Q000201:enzymology* / Q000254:growth & development*; D029681:Arabidopsis Proteins / Q000378:metabolism*; D002853:Chromatography, Liquid; D047468:Immunoprecipitation; D008274:Magnesium / Q000494:pharmacology*; D008954:Models, Biological; D005810:Multigene Family; D009154:Mutation / Q000235:genetics; D010641:Phenotype; D010766:Phosphorylation / Q000187:drug effects; D063245:Plant Development / Q000187:drug effects*; D011485:Protein Binding / Q000187:drug effects; D011494:Protein Kinases / Q000378:metabolism*; D053719:Tandem Mass Spectrometry
chemical_list: D029681:Arabidopsis Proteins; D000040:Abscisic Acid; D011494:Protein Kinases; D008274:Magnesium