title: Multi-Element Bioimaging of Arabidopsis thaliana Roots.
abstract: Better understanding of root function is central for the development of plants with more efficient nutrient uptake and translocation. We here present a method for multielement bioimaging at the cellular level in roots of the genetic model system Arabidopsis (Arabidopsis thaliana). Using conventional protocols for microscopy, we observed that diffusible ions such as potassium and sodium were lost during sample dehydration. Thus, we developed a protocol that preserves ions in their native, cellular environment. Briefly, fresh roots are encapsulated in paraffin, cryo-sectioned, and freeze dried. Samples are finally analyzed by laser ablation-inductively coupled plasma-mass spectrometry, utilizing a specially designed internal standard procedure. The method can be further developed to maintain the native composition of proteins, enzymes, RNA, and DNA, making it attractive in combination with other omics techniques. To demonstrate the potential of the method, we analyzed a mutant of Arabidopsis unable to synthesize the metal chelator nicotianamine. The mutant accumulated substantially more zinc and manganese than the wild type in the tissues surrounding the vascular cylinder. For iron, the images looked completely different, with iron bound mainly in the epidermis of the wild-type plants but confined to the cortical cell walls of the mutant. The method offers the power of inductively coupled plasma-mass spectrometry to be fully employed, thereby providing a basis for detailed studies of ion transport in roots. Being applicable to Arabidopsis, the molecular and genetic approaches available in this system can now be fully exploited in order to gain a better mechanistic understanding of these processes.
authors: Department of Plant and Environmental Sciences and Copenhagen Plant Science Center, Faculty of Science, University of Copenhagen, DK-1871 Frederiksberg C, Denmark (D.P.P., A.C., J.K.S., S.H.);Laboratory of Genetics, Wageningen University, 6708 PB Wageningen, The Netherlands (M.G.M.A.); andCentre for Plant Integrative Biology, University of Nottingham, Nottingham LE12 5RD, United Kingdom (D.E.S.) dap@plen.ku.dk.
authors: Daniel Pergament
authors: 0000-0003-3976-190X
authors: DP
authors: Persson
authors: Department of Plant and Environmental Sciences and Copenhagen Plant Science Center, Faculty of Science, University of Copenhagen, DK-1871 Frederiksberg C, Denmark (D.P.P., A.C., J.K.S., S.H.);Laboratory of Genetics, Wageningen University, 6708 PB Wageningen, The Netherlands (M.G.M.A.); andCentre for Plant Integrative Biology, University of Nottingham, Nottingham LE12 5RD, United Kingdom (D.E.S.).
authors: Anle
authors: 0000-0001-8439-5721
authors: A
authors: Chen
authors: Department of Plant and Environmental Sciences and Copenhagen Plant Science Center, Faculty of Science, University of Copenhagen, DK-1871 Frederiksberg C, Denmark (D.P.P., A.C., J.K.S., S.H.);Laboratory of Genetics, Wageningen University, 6708 PB Wageningen, The Netherlands (M.G.M.A.); andCentre for Plant Integrative Biology, University of Nottingham, Nottingham LE12 5RD, United Kingdom (D.E.S.).
authors: Mark G M
authors: 0000-0001-5257-0740
authors: MG
authors: Aarts
authors: Department of Plant and Environmental Sciences and Copenhagen Plant Science Center, Faculty of Science, University of Copenhagen, DK-1871 Frederiksberg C, Denmark (D.P.P., A.C., J.K.S., S.H.);Laboratory of Genetics, Wageningen University, 6708 PB Wageningen, The Netherlands (M.G.M.A.); andCentre for Plant Integrative Biology, University of Nottingham, Nottingham LE12 5RD, United Kingdom (D.E.S.).
authors: David E
authors: DE
authors: Salt
authors: Department of Plant and Environmental Sciences and Copenhagen Plant Science Center, Faculty of Science, University of Copenhagen, DK-1871 Frederiksberg C, Denmark (D.P.P., A.C., J.K.S., S.H.);Laboratory of Genetics, Wageningen University, 6708 PB Wageningen, The Netherlands (M.G.M.A.); andCentre for Plant Integrative Biology, University of Nottingham, Nottingham LE12 5RD, United Kingdom (D.E.S.).
authors: Jan K
authors: 0000-0002-2852-3298
authors: JK
authors: Schjoerring
authors: Department of Plant and Environmental Sciences and Copenhagen Plant Science Center, Faculty of Science, University of Copenhagen, DK-1871 Frederiksberg C, Denmark (D.P.P., A.C., J.K.S., S.H.);Laboratory of Genetics, Wageningen University, 6708 PB Wageningen, The Netherlands (M.G.M.A.); andCentre for Plant Integrative Biology, University of Nottingham, Nottingham LE12 5RD, United Kingdom (D.E.S.).
authors: Søren
authors: 0000-0003-2020-1902
authors: S
authors: Husted
mesh_terms: D017360:Arabidopsis / Q000235:genetics / Q000378:metabolism*; D029681:Arabidopsis Proteins / Q000378:metabolism; D017136:Ion Transport / Q000235:genetics; D007501:Iron / Q000378:metabolism; D008345:Manganese / Q000378:metabolism; D013058:Mass Spectrometry / Q000379:methods*; D009154:Mutation; D009696:Nucleic Acids / Q000378:metabolism; D018517:Plant Roots / Q000235:genetics / Q000378:metabolism*; D015203:Reproducibility of Results; D052584:Xylem / Q000235:genetics / Q000378:metabolism*; D015032:Zinc / Q000378:metabolism
chemical_list: D029681:Arabidopsis Proteins; D009696:Nucleic Acids; D008345:Manganese; D007501:Iron; D015032:Zinc