Gel-filtration analysis to detect Al-binding ligands in Eucalyptus camaldulensis

Published Date
August 2015, Volume 20, Issue 4, pp 411–414

Title 

Gel-filtration analysis to detect Al-binding ligands in Eucalyptus camaldulensis

• Author 

• Ko Tahara, 
• Katsumi Kojima, 
• Kenji Shinohara

• Abstract 

• Aluminum (Al) toxicity is a major factor limiting plant growth on acid soils; inhibited root elongation is a characteristic symptom. To elucidate mechanisms of Al resistance in plants, we developed a simple method to separate Al-binding ligands from plant roots by gel-filtration chromatography. In this method, an eluent consisting of Homopipes buffer flows through a Sephadex G-25 column. Free Al ions do not pass through the column, but Al-ligand complexes can be fractionated according to differences in their molecular sizes. We applied the method to Eucalyptus camaldulensis (Myrtaceae) and successfully separated Al-binding ligands from roots. One peak of Al-ligand complex increased with Al treatment of seedlings, which can be explained by Al-induced accumulation of citrate in roots.

• References 

• 1. Brunner I, Sperisen C (2013) Aluminum exclusion and aluminum tolerance in woody plants. Front Plant Sci 4:172PubMedCentralPubMedCrossRef
  2. Delhaize E, Ma JF, Ryan PR (2012) Transcriptional regulation of aluminium tolerance genes. Trends Plant Sci 17:341–348PubMedCrossRef
  3. Ikka T, Ogawa T, Li DH, Hiradate S, Morita A (2013) Effect of aluminum on metabolism of organic acids and chemical forms of aluminum in root tips of Eucalyptus camaldulensisDehnh. Phytochemistry 94:142–147PubMedCrossRef
  4. Kinraide TB, Sweeney BK (2001) Buffered, phosphate-containing media suitable for aluminum toxicity studies. Plant Soil 235:75–83CrossRef
  5. Kochian LV (1995) Cellular mechanisms of aluminum toxicity and resistance in plants. Annu Rev Plant Physiol Plant Mol Biol 46:237–260CrossRef
  6. Kochian LV, Hoekenga OA, Piñeros MA (2004) How do crop plants tolerate acid soils? Mechanisms of aluminum tolerance and phosphorous efficiency. Annu Rev Plant Biol 55:459–493PubMedCrossRef
  7. Ma JF (2007) Syndrome of aluminum toxicity and diversity of aluminum resistance in higher plants. Int Rev Cytol 264:225–252PubMedCrossRef
  8. Öhman LO (1988) Equilibrium and structural studies of silicon(IV) and aluminum(III) in aqueous solution. 17. Stable and metastable complexes in the system H+-Al3+-citric acid. Inorg Chem 27:2565–2570CrossRef
  9. Osawa H, Endo I, Hara Y, Matsushima Y, Tange T (2011) Transient proliferation of proanthocyanidin-accumulating cells on the epidermal apex contributes to highly aluminum-resistant root elongation in camphor tree. Plant Physiol 155:433–446PubMedCentralPubMedCrossRef
  10. Ryan PR, Tyerman SD, Sasaki T, Furuichi T, Yamamoto Y, Zhang WH, Delhaize E (2011) The identification of aluminium-resistance genes provides opportunities for enhancing crop production on acid soils. J Exp Bot 62:9–20PubMedCrossRef
  11. Sjöberg S, Öhman LO (1985) Equilibrium and structural studies of silicon(IV) and aluminium(III) in aqueous solution. Part 13. A potentiometric and 27Al nuclear magnetic resonance study of speciation and equilibria in the aluminium(III)-oxalic acid-hydroxide system. J Chem Soc Dalton Trans (12):2665–2669
  12. Tahara K, Norisada M, Hogetsu T, Kojima K (2005a) Aluminum tolerance and aluminum-induced deposition of callose and lignin in the root tips of Melaleuca and Eucalyptus species. J For Res 10:325–333CrossRef
  13. Tahara K, Norisada M, Tange T, Yagi H, Kojima K (2005b) Ectomycorrhizal association enhances Al tolerance by inducing citrate secretion in Pinus densiflora. Soil Sci Plant Nutr 51:397–403CrossRef
  14. Tahara K, Norisada M, Yamanoshita T, Kojima K (2008) Role of aluminum-binding ligands in aluminum resistance of Eucalyptus camaldulensis and Melaleuca cajuputi. Plant Soil 302:175–187CrossRef
  15. Tahara K, Hashida K, Otsuka Y, Ohara S, Kojima K, Shinohara K (2014) Identification of a hydrolyzable tannin, oenothein B, as an aluminum-detoxifying ligand in a highly aluminum-resistant tree, Eucalyptus camaldulensis. Plant Physiol 164:683–693PubMedCentralPubMedCrossRef
  16. von Uexküll HR, Mutert E (1995) Global extent, development and economic impact of acid soils. Plant Soil 171:1–15CrossRef
  17. Wenzl P, Patiño GM, Chaves AL, Mayer JE, Rao IM (2001) The high level of aluminum resistance in signalgrass is not associated with known mechanisms of external aluminum detoxification in root apices. Plant Physiol 125:1473–1484PubMedCentralPubMedCrossRef

• For further details log on website :

• http://link.springer.com/article/10.1007/s10310-015-0487-0