Identification of Zn-Nicotianamine and Fe-2'-Deoxymugineic Acid in the Phloem Sap from Rice Plants (Oryza sativa L.)

Department of Applied Biological Chemistry, Graduate School of Agricultural and Life Sciences, the University of Tokyo, Yayoi 1-1-1, Bunkyo-ku, Tokyo, 113-8657 Japan.
Plant and Cell Physiology (Impact Factor: 4.93). 01/2012; 53(2):381-90. DOI: 10.1093/pcp/pcr188
Source: PubMed


In higher plants, the supply of metals such as Zn and Fe via phloem is important for the growth and physiology of young organs. However, little information is available on the speciation (chemical forms) of these metals in the phloem fluids. Because the pH of phloem fluids is slightly alkaline and the concentration of phosphate, which may bind to metals, is high, Zn and Fe in phloem fluids could be precipitated if these metals do not form complexes with some ligand compounds. In the present experiment, we examined the chemical forms of Zn and Fe in phloem sap collected from rice (Oryza sativa L.) by separating the phloem sap using size-exclusion and anion-exchange chromatography, and identifying the contents using electrospray ionization time-of-flight mass spectrometry. The low molecular weight chemical forms of Zn and Fe were identified as Zn-nicotianamine and Fe(III)-2'-deoxymugineic acid complexes, respectively. This report is the first to identify metal-chelate complexes in rice phloem sap.

23 Reads
  • Source
    • "This was much higher than the NA values reported of 66 μMol L-1[39] and 76 μMol L-1[8]. In rice it has been found that the main Zn complex ligand was NA whilst for Fe, DMA was the main metabolite responsible for complexing this metal [8]. Glutathione has been found to play a role in Cd transportation as part of the detoxification process [40]. "
    [Show abstract] [Hide abstract]
    ABSTRACT: Background Biofortification of staple crops with essential micronutrients relies on the efficient, long distance transport of nutrients to the developing seed. The main route of this transport in common wheat (Triticum aestivum) is via the phloem, but due to the reactive nature of some essential micronutrients (specifically Fe and Zn), they need to form ligands with metabolites for transport within the phloem. Current methods available in collecting phloem exudate allows for small volumes (μL or nL) to be collected which limits the breadth of metabolite analysis. We present a technical advance in the measurement of 79 metabolites in as little as 19.5 nL of phloem exudate. This was achieved by using mass spectrometry based, metabolomic techniques. Results Using gas chromatography–mass spectrometry (GC-MS), 79 metabolites were detected in wheat phloem. Of these, 53 were identified with respect to their chemistry and 26 were classified as unknowns. Using the ratio of ion area for each metabolite to the total ion area for all metabolites, 39 showed significant changes in metabolite profile with a change in wheat reproductive maturity, from 8–12 to 17–21 days after anthesis. Of these, 21 were shown to increase and 18 decreased as the plant matured. An amine group derivitisation method coupled with liquid chromatography MS (LC-MS) based metabolomics was able to quantify 26 metabolites and semi-quantitative data was available for a further 3 metabolites. Conclusions This study demonstrates that it is possible to determine metabolite profiles from extremely small volumes of phloem exudate and that this method can be used to determine variability within the metabolite profile of phloem that has occurred with changes in maturity. This is also believed to be the first report of the presence of the important metal complexing metabolite, nicotianamine in the phloem of wheat.
    Plant Methods 08/2014; 10(1):27. DOI:10.1186/1746-4811-10-27 · 3.10 Impact Factor
  • Source
    • "AtYSL4 and AtYSL6 have been described as iron transporters localized to organelles, either the chloroplast (Divol et al., 2013) or other endomembrane systems (Conte et al., 2013). The pH of the phloem is slightly alkaline (Dinant et al., 2010), and therefore zinc has to travel in the phloem bound to a metal chelator, which often appear to be nicotianamine (Curie et al., 2009; Nishiyama et al., 2012). "
    [Show abstract] [Hide abstract]
    ABSTRACT: An important goal of micronutrient biofortification is to enhance the amount of bioavailable zinc in the edible seed of cereals and more specifically in the endosperm. The picture is starting to emerge for how zinc is translocated from the soil through the mother plant to the developing seed. On this journey, zinc is transported from symplast to symplast via multiple apoplastic spaces. During each step, zinc is imported into a symplast before it is exported again. Cellular import and export of zinc requires passage through biological membranes, which makes membrane-bound transporters of zinc especially interesting as potential transport bottlenecks. Inside the cell, zinc can be imported into or exported out of organelles by other transporters. The function of several membrane proteins involved in the transport of zinc across the tonoplast, chloroplast or plasma membranes are currently known. These include members of the ZIP (ZRT-IRT-like Protein), and MTP (Metal Tolerance Protein) and heavy metal ATPase (HMA) families. An important player in the transport process is the ligand nicotianamine that binds zinc to increase its solubility in living cells and in this way buffers the intracellular zinc concentration.
    Frontiers in Plant Science 02/2014; 5:30. DOI:10.3389/fpls.2014.00030 · 3.95 Impact Factor
  • Source
    • "The first study was conducted on Ricinus communis, revealing the presence of an Fe transport protein (RcITP; Krüger et al., 2002). A later work conducted in rice showed that DMA is the major Fe chelator in the phloem of this species, whereas NA was in fact mainly bound to zinc (Nishiyama et al., 2011). "
    [Show abstract] [Hide abstract]
    ABSTRACT: Iron (Fe) is one of the most abundant elements on earth, but its limited bioavailability poses a major constraint for agriculture and constitutes a serious problem in human health. Due to an improved understanding of the mechanisms that control Fe homeostasis in plants, major advances toward engineering biofortified crops have been made during the past decade. Examples of successful biofortification strategies are, however, still scarce and the process of Fe loading into seeds is far from being well understood in most crop species. In particular in grains where the embryo represents the main storage compartment such as legumes, increasing the seed Fe content remains a challenging task. This review aims at placing the recently identified actors in Fe transport into the unsolved puzzle of grain filling, taking the differences of Fe distribution between various species into consideration. We summarize the current knowledge on Fe transport between symplasmic and apoplasmic compartments, and provide models for Fe trafficking and localization in different seed types that may help to develop high seed Fe germplasms.
    Frontiers in Plant Science 01/2014; 4:535. DOI:10.3389/fpls.2013.00535 · 3.95 Impact Factor
Show more