Free Standard AU & NZ Shipping For All Book Orders Over $80!
Register      Login
Functional Plant Biology Functional Plant Biology Society
Plant function and evolutionary biology
RESEARCH ARTICLE

The role of low molecular weight ligands in nickel hyperaccumulation in Hybanthus floribundus subspecies floribundus

Anthony G. Kachenko A C D , Balwant Singh A and Naveen Bhatia B
+ Author Affiliations
- Author Affiliations

A Faculty of Agriculture, Food and Natural Resources, John Woolley Building A20, The University of Sydney, Sydney, NSW 2006, Australia.

B Institute for Environmental Research, Australian Nuclear Science and Technology Organisation (ANSTO), Lucas Heights, NSW 2234, Australia.

C Present address: Nursery and Garden Industry Australia (NGIA), PO Box 907, Epping, NSW 1710, Australia.

D Corresponding author. Email: anthony.kachenko@ngia.com.au

Functional Plant Biology 37(12) 1143-1150 https://doi.org/10.1071/FP10080
Submitted: 11 April 2010  Accepted: 21 July 2010   Published: 17 November 2010

Abstract

The mechanisms responsible for nickel (Ni) hyperaccumulation in Hybanthus floribundus (Lindl.) F.Muell. subspecies floribundus are obscure. In this study, organic acids and free amino acids (AAs) were quantified in 0.025 M HCl H. floribundus subsp. floribundus shoot extracts using HPLC and ultra performance liquid chromatography (UPLC). In a 20 week pot experiment, plants exposed to five levels of Ni (0–3000 mg kg–1 Ni) accumulated up to 3200 mg Ni kg–1 dry weight in shoots, and the shoot : root Ni concentration ratios were >1.4. Concentration of organic acids followed the order malic acid > citric acid > oxalic acid. Citric acid concentration significantly increased upon Ni exposure, with concentrations between 2.3- and 5.9-fold higher in Ni treated plants that in control plants. Molar ratios of Ni to citric acid ranged from 1.3 : 1 to 1.7 : 1 equivalent to >60% of the accumulated Ni. Malic acid concentration also increased upon exposure to applied Ni. However, concentrations were statistically at par across 0–3000 mg kg–1 Ni treatments, suggesting that the production of malic acid is a constitutive property of the subspecies. Total AA concentrations were stimulated upon exposure to external Ni treatment, with glutamine, alanine and aspartic acids being the predominant acids. These AAs accounted for up to 64% of the total free AA concentration in control plants and up to 75% for the 2000 mg kg–1 Ni treatment plants. These results suggest that citric acid in addition to the aforementioned AAs are synthesised in H. floribundus subsp. floribundus plants following exposure to elevated concentrations of Ni and may act as potential ligands for detoxification and possibly storage of accumulated Ni.

Additional keywords: amino acids, citric acid, compartmentation, hyperaccumulator, shrub violet, organic acids.


Acknowledgements

We thank Malcolm Nobel (UNSW, Sydney) for organic acid analyses, and Bernie McInerney and Leon McQuade for AA analyses conducted at the Australian Proteome Analysis Facility (Macquarie University, Sydney) established under the Australian Government’s Major National Research Facilities program. Anthony G Kachenko acknowledges financial support from the University of Sydney and the Australian Commonwealth Government through an Australian Postgraduate Award scholarship. Thanks are also extended to Marilyn Sprague for providing plant material.


References


Bhatia NP, Walsh KB, Baker AJM (2005) Detection and quantification of ligands involved in nickel detoxification in a herbaceous Ni hyperaccumulator Stackhousia tryonii Bailey. Journal of Experimental Botany 56, 1343–1349.
Crossref | GoogleScholarGoogle Scholar | CAS | PubMed | open url image1

Bidwell SD (2001) Hyperaccumulation of metals in Australian native plants. PhD Thesis, School of Botany, The University of Melbourne.

Bidwell SD, Crawford S, Woodrow I, Sommer-Knudsen J, Marshall A (2004) Subcellular localization of Ni in the hyperaccumulator, Hybanthus floribundus (Lindley) F.Muell. Plant, Cell & Environment 27, 705–716.
Crossref | GoogleScholarGoogle Scholar | CAS | open url image1

Brooks R, Shaw S, Asensi Marfil A (1981) The chemical form and physiological function of nickel in some Iberian Alyssum species. Plant Physiology 51, 167–170.
Crossref | GoogleScholarGoogle Scholar | CAS | open url image1

Callahan DL, Baker AJM, Kolev SD, Wedd AG (2006) Metal ion ligands in hyperaccumulating plants. Journal of Biological Inorganic Chemistry 11, 2–12.
Crossref | GoogleScholarGoogle Scholar | CAS | PubMed | open url image1

Callahan DL, Kolev SD, O’Hair RAJ, Salt DE, Baker AJM (2007) Relationships of nicotianamine and other amino acids with nickel, zinc and iron in Thlaspi hyperaccumulators. New Phytologist 176, 836–848.
Crossref | GoogleScholarGoogle Scholar | CAS | PubMed | open url image1

Callahan DL, Roessner U, Dumontet V, Perrier N, Wedd AG, O’Hair RAJ, Baker AJM, Kolev SD (2008) LC-MS and GC-MS metabolite profiling of nickel(II) complexes in the latex of the nickel-hyperaccumulating tree Sebertia acuminata and identification of methylated aldaric acid as a new nickel(II) ligand. Phytochemistry 69, 240–251.
Crossref | GoogleScholarGoogle Scholar | CAS | PubMed | open url image1

Clemens S, Palmgren MG, Kramer U (2002) A long way ahead: understanding and engineering plant metal accumulation. Trends in Plant Science 7, 309–315.
Crossref | GoogleScholarGoogle Scholar | CAS | PubMed | open url image1

Cohen SA (2001) Amino acid analysis using precolumn derivatisation with 6-aminoquinolyl-N-hydroxysuccinimidyl carbamate. In ‘Methods in molecular biology’. (Eds C Cooper, N Packer, K Williams) pp. 39–47. (Humana Press: Totowa, NJ)

Cohen SA, Michaud DP (1993) Synthesis of a fluorescent derivatizing reagent, 6-aminoquinolyl-N-hydroxysuccinimidyl carbamate, and its application for the analysis of hydrolysate amino acids via high performance liquid chromatography. Analytical Biochemistry 211, 279–287.
Crossref | GoogleScholarGoogle Scholar | CAS | PubMed | open url image1

Farago ME, Mahmoud IEDAW (1983) Plants that accumulate metals (Part VI): further studies of an Australian nickel accumulating plant. Environmental Geochemistry and Health 5, 113–121.
CAS |
open url image1

Farago ME, Mahmoud I, Clark AJ (1980) The amino acid content of Hybanthus floribundus, a nickel accumulating plant and the difficulty of detecting nickel amino acid complexes by chromatographic methods. Inorganic and Nuclear Chemistry Letters 16, 481–484.
Crossref | GoogleScholarGoogle Scholar | CAS | open url image1

Gerendás J, Polacco JC, Freyermuth SK, Sattelmacher B (1999) Significance of nickel for plant growth and metabolism. Journal of Plant Nutrition and Soil Science 162, 241–256.
Crossref | GoogleScholarGoogle Scholar | open url image1

Homer FA, Reeves RD, Brooks RR, Baker AJM (1991) Characterization of the nickel-rich extract from the nickel hyperaccumulator Dichapetalum gelonioides. Phytochemistry 30, 2141–2145.
Crossref | GoogleScholarGoogle Scholar | CAS | open url image1

Homer FA, Reeves RD, Brooks RR (1995) The possible involvement of amino acids in nickel chelation in nickel chelation in some nickel-accumulating plants. Current Topics in Phytochemistry 14, 31–33. open url image1

Ingle RA, Mugford ST, Rees JD, Campbell MM, Smith JAC (2005) Constitutively high expression of the histidine biosynthetic pathway contributes to nickel tolerance in hyperaccumulator plants. The Plant Cell 17, 2089–2106.
Crossref | GoogleScholarGoogle Scholar | CAS | PubMed | open url image1

Kachenko AG, Siegele R, Bhatia NP, Singh B, Ionescu M (2008a) Evaluation of specimen preparation techniques for micro-PIXE localisation of elements in hyperaccumulating plants. Nuclear Instruments & Methods in Physics Research. Section B, Beam Interactions with Materials and Atoms 266, 1598–1604.
Crossref | GoogleScholarGoogle Scholar | CAS | open url image1

Kachenko AG, Singh B, Bhatia NP, Siegele R (2008b) Quantitative elemental localisation in leaves and stems of nickel hyperaccumulating shrub Hybanthus floribundus subsp. floribundus using micro-PIXE spectroscopy. Nuclear Instruments & Methods in Physics Research. Section B, Beam Interactions with Materials and Atoms 266, 667–676.
Crossref | GoogleScholarGoogle Scholar | CAS | open url image1

Kerkeb L, Krämer U (2003) The role of free histidine in xylem loading of nickel in Alyssum lesbiacum and Brassica juncea. Plant Physiology 131, 716–724.
Crossref | GoogleScholarGoogle Scholar | CAS | PubMed | open url image1

Kersten WJ, Brooks RR, Reeves RD, Jaffré A (1980) Nature of nickel complexes in Psychotria douarrei and other nickel-accumulating plants. Phytochemistry 19, 1963–1965.
Crossref | GoogleScholarGoogle Scholar | CAS | open url image1

Krämer U, Cotter-Howells JD, Charnock JM, Baker AJM, Smith JAC (1996) Free histidine as a metal chelator in plants that accumulate nickel. Nature 379, 635–638.
Crossref | GoogleScholarGoogle Scholar | open url image1

Krämer U, Pickering IJ, Prince RC, Raskin I, Salt DE (2000) Subcellular localization and speciation of nickel in hyperaccumulator and non-accumulator Thlaspi species. Plant Physiology 122, 1343–1354.
Crossref | GoogleScholarGoogle Scholar | PubMed | open url image1

Lee J, Brooks RR, Reeves RD, Boswell CR, Jaffré T (1977) Plant-soil relationships in a New Caledonian serpentine flora. Plant and Soil 46, 675–680.
Crossref | GoogleScholarGoogle Scholar | CAS | open url image1

Lee J, Reeves RD, Brooks RR (1978) The relationship between nickel and citric acid in some nickel-accumulating plants. Phytochemistry 17, 1033–1035.
Crossref | GoogleScholarGoogle Scholar | CAS | open url image1

Martell EA , Smith RM (1974) ‘Critical stability constants.’ (Plenum Press: New York)

McIntyre T (2003) Phytoremediation of heavy metals from soils. In ‘Advances in biochemical engineering biotechnology’. (Ed. D Tsao) pp. 97–123. (Springer-Verlag: Berlin)

McNear DH, Chaney RL, Sparks DL (2010) The hyperaccumulator Alyssum murale uses complexation with nitrogen and oxygen donor ligands for Ni transport and storage. Phytochemistry 71, 188–200.
Crossref | GoogleScholarGoogle Scholar | CAS | PubMed | open url image1

Meagher RB (2000) Phytoremediation of toxic elemental and organic pollutants. Current Opinion in Plant Biology 3, 153–162.
Crossref | GoogleScholarGoogle Scholar | CAS | PubMed | open url image1

Miller RO (1998) Nitric-perchloric acid wet digestion in an open vessel. In ‘Handbook of reference methods for plant analysis’. (Ed. Y Kalra) pp. 57–61. (CRC Press: Boca Raton, FL)

Montargès-Pelletier E, Chardot V, Echevarria G, Michot LJ, Bauer A, Morel J-L (2008) Identification of nickel chelators in three hyperaccumulating plants: an X-ray spectroscopic study. Phytochemistry 69, 1695–1709.
Crossref | GoogleScholarGoogle Scholar | PubMed | open url image1

Pelosi P, Fiorentini R, Galoppini C (1976) On the nature of nickel compunds in Alyssum bertolonii Desv. II. Agricultural and Biological Chemistry 40, 1641–1642.
CAS |
open url image1

Pence NS, Larsen PB, Ebbs SD, Letham DLD, Lasat MM, Garvin DF, Eide D, Kochian LV (2000) The molecular physiology of heavy metal transport in the Zn/Cd hyperaccumulator Thlaspi caerulescens. Proceedings of the National Academy of Sciences of the United States of America 97, 4956–4960.
Crossref | GoogleScholarGoogle Scholar | CAS | PubMed | open url image1

Persans MW, Yan X, Patnoe J-MML, Krämer U, Salt DE (1999) Molecular dissection of the role of histidine in nickel hyperaccumulation in Thlaspi goesingense (Hálácsy). Plant Physiology 121, 1117–1126.
Crossref | GoogleScholarGoogle Scholar | CAS | PubMed | open url image1

Reuter DJ , Robinson JB , Peverill KI , Price GH (1988) Guidelines for collecting, handling and analysing plant material. In ‘Plant analysis: an interpretation manual’. (Eds DJ Reuter, JB Robinson) pp. 20–30. (Inkarta Press: Melbourne)

Sagner S, Kneer R, Wanner G, Cossons JP, Deus-Neumann B, Zenk MH (1998) Hyperaccumulation, complexation and distribution of nickel in Sebertia acuminata. Phytochemistry 47, 339–347.
Crossref | GoogleScholarGoogle Scholar | CAS | PubMed | open url image1

Salt DE, Smith RD, Raskin I (1998) Phytoremediation. Annual Review of Plant Physiology and Plant Molecular Biology 49, 643–668.
Crossref | GoogleScholarGoogle Scholar | CAS | PubMed | open url image1

Salt DE, Prince RC, Baker AJM, Raskin I, Pickering IJ (1999) Zinc ligands in the metal hyperaccumulator Thlaspi caerulescens as determined using X-ray absorption spectroscopy. Environmental Science & Technology 33, 713–717.
Crossref | GoogleScholarGoogle Scholar | CAS | open url image1

Schat H, Llugany M, Vooijs R, Hartley-Whitaker J, Bleeker PM (2002) The role of phytochelatins in constitutive and adaptive heavy metal tolerances in hyperaccumulator and non-hyperaccumulator metallophytes. Journal of Experimental Botany 53, 2381–2392.
Crossref | GoogleScholarGoogle Scholar | CAS | PubMed | open url image1

Schaumlöffel D, Ouerdane L, Bouyssiere B, Lobinski R (2003) Speciation analysis of nickel in the latex of a hyperaccumulating tree Sebertia acuminata by HPLC and CZE with ICP MS and electrospray MS-MS detection. Journal of Analytical Atomic Spectrometry 18, 120–127.
Crossref | GoogleScholarGoogle Scholar | open url image1

Seregin IV, Kozhevnikova A (2006) Physiological role of nickel and its toxic effects on higher plants. Russian Journal of Plant Physiology 53, 257–277.
Crossref | GoogleScholarGoogle Scholar | CAS | open url image1

Severne BC (1974) Nickel accumulation by Hybanthus floribundus. Nature 248, 807–808.
Crossref | GoogleScholarGoogle Scholar | CAS | PubMed | open url image1

Shen ZG, Zhao FJ, McGrath SP (1997) Uptake and transport of zinc in the hyperaccumulator Thlaspi caerulescens and the non-hyperaccumulator Thlaspi ochroleucum. Plant, Cell & Environment 20, 898–906.
Crossref | GoogleScholarGoogle Scholar | CAS | open url image1

Sun R-L, Zhou Q-X, Jin C-X (2006) Cadmium accumulation in relation to organic acids in leaves of Solanum nigrum L. as a newly found cadmium hyperaccumulator. Plant and Soil 285, 125–134.
Crossref | GoogleScholarGoogle Scholar | CAS | open url image1

Wójcik M, Skórzyn’ska-Polit E, Tukiendorf A (2006) Organic acids accumulation and antioxidant enzyme activities in Thlaspi caerulescens under Zn and Cd stress. Plant Growth Regulation 48, 145–155.
Crossref | GoogleScholarGoogle Scholar | open url image1

Yang X, Li T, Yang J, He Z, Lu L, Meng F (2006) Zinc compartmentation in root, transport into xylem, and absorption into leaf cells in the hyperaccumulating species of Sedum alfredii Hance. Planta 224, 185–195.
Crossref | GoogleScholarGoogle Scholar | CAS | PubMed | open url image1