Acta Univ. Agric. Silvic. Mendelianae Brun. 2011, 59(3), 241-246 | DOI: 10.11118/actaun201159030241

Methods of plant root exudates analysis: a review

Peter Dundek, Ladislav Holík, Tomáš Rohlík, Ladislav Hromádko, Valerie Vranová, Klement Rejšek, Pavel Formánek
Ústav geologie a pedologie, Mendelova univerzita v Brně, Zemědělská 3, 613 00 Brno, Česká republika

The aim of this review is to summarise current knowledge on methods being used to determine individual compounds and properties of water-soluble plant root exudates. These compounds include amino acids, organic acids and simple sugars, as well as polysaccharides, proteins and organic substances. Qualitative composition of water-soluble root exudates and exudation rate are commonly measured with the aim of consequent synthetic preparation of plant root exudates to be supplied to soil to create artificial rhizosphere for different experimental purposes. Root exudates collection usually requires consequent filtration or centrifugation to remove solids, root detritus and microbial cell debris, and consequent concentration using an evaporator, lyophilizator or ultrafiltration. Methods used for analysis of total groups of compounds (total proteins and total carbohydrates) and total organic carbon are simple. On the other hand, HPLC or GS/MS are commonly used to analyse individual low molecular weight organic molecules (sugars, organic acids and amino acids) with separation using different columns. Other properties such as pH, conductivity or activity of different enzymes as well as gel electrophoresis of proteins are sometimes assessed. All of these methods are discussed in this work.

Keywords: amino acids, HPLC, GC/MS, organic acids, root exudates, sugars
Grants and funding:

The study was supported by the grant MSM6215648902 / Forest and Wood: the support of functionally integrated forest management and use of wood as a renewable raw material phase 4/2/3, part II "The management strategy of nature conservation areas", and by project "Extension of current knowledge on bioavailable amino acids in soil and their utilization by soil microorganisms and plant roots" (IGA FFWT MENDELU No. 47/2010-2012).

Received: January 13, 2011; Published: June 14, 2014  Show citation

ACS AIP APA ASA Harvard Chicago IEEE ISO690 MLA NLM Turabian Vancouver
Dundek, P., Holík, L., Rohlík, T., Hromádko, L., Vranová, V., Rejšek, K., & Formánek, P. (2011). Methods of plant root exudates analysis: a review. Acta Universitatis Agriculturae et Silviculturae Mendelianae Brunensis, 59(3), 241-246. doi: 10.11118/actaun201159030241
Share...
Download citation
  • BibTeX (.bib)
  • Bookends (.ris)
  • EasyBib (.ris)
  • EndNote (.enw)
  • EndNote 8 (.xml)
  • ISI WoS (.isi)
  • Medlars (.medlars)
  • Mendeley (.ris)
  • MODS (.xml)
  • Papers (.ris)
  • RefWorks (.txt)
  • RefManager (.ris)
  • RIS (.ris)
  • MS Word (.xml)
  • Zotero (.ris)
    • Open full article

    References

    1. ASHWELL, G., 1966: New colorimetric methods of sugar analysis. In Methods in enzymology, Vol. VIII, E. F. Neufeld, V. Ginsburg (Eds). pp.85-95. Academic Press, New York. ISBN 978-0-12-181808-1. Go to original source...
    2. AULAKH, M. S., WASSMANN, R., BUENO, C., KREUZWIESER, J., RENNENBERG, H., 2001: Characterization of root exudates at different growth stages of ten rice (Oryza sativa L.) cultivars. Plant Biology 3, 2: 139-148. ISSN 1438-8677. DOI: 10.1055/s-2001-12905 Go to original source...
    3. BACILIO-JIMENEZ, M., AGUILAR-FLORES, S., VENTURA-ZAPATA, E., PÉREZ-CAMPOS, E., BOUQUELET, S., ZENTENO, E., 2003: Chemical characterization of root exudates from rice (Oryza sativa) and their effect on the chemotactic response of endophytic bacteria. Plant and Soil 249, 2: 271-277. ISSN 0032-079X. DOI: 10.1023/A:1022888900465 Go to original source...
    4. BADOUD, R., PRATZ, G., 1986: Improved high-performance liquid chromatographic analysis of some carboxylic acids in food and beverages as their p-nitrobenzyl esters. Journal of Chromatography 360, 119-136. ISSN 0021-9673. DOI: 10.1016/S0021-9673(00)91656-X Go to original source...
    5. BRADFORD, M. M., 1976: A rapid and sensitive method for the quantitation of microgram quantities of protein utilising the principle of protein-dye binding. Analytical Biochemistry 72, 2: 248-254. ISSN 0003-2697. DOI: 10.1016/0003-2697(76)90527-3 Go to original source...
    6. BRINK, R. H. J., DUBACH, P., LYNCH, D. L., 1960: Measurement of carbohydrates in soil hydrolyzates with anthrone. Soil Science 89, 3: 157-166. ISSN 0038-075X. DOI: 10.1097/00010694-196003000-00006 Go to original source...
    7. COHEN, S. A., MICHAUD, D. P., 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, 2: 279-287. ISSN 0003-2697. DOI: 10.1006/abio.1993.1270 Go to original source...
    8. CURL, E. A., TRUELOVE, B., 1986: The Rhizosphere. Springer-Verlag, New York. Springer-Verlag, Berlin-Heidelberg-New York-Tokyo, 288 p. ISBN 3-540-15803-0.
    9. DANIELS, L., HANSON, R. S., PHILIPS, J. A., 1994: Chemical analysis. In: Gerhard, P. et al. (Eds). Method for general and molecular bacteriology. American Society for Microbiology, Wahington, DC, 512-554.
    10. DENVOR, A. W., 1950: Carbohydrate tests using sulphonated α-naphthol. Journal of the American Chemical Society 72, 2008-2012. DOI: 10.1021/ja01161a037 Go to original source...
    11. DOUSSET, S., MOREL, J. L., JACOBSON, A., BITTON, G., 2001: Copper binding capacity of root exudates of cultivated plants and associated weeds. Biology and Fertility of Soils 34, 4: 230-234. ISSN 0178-2762. DOI: 10.1007/s003740100404 Go to original source...
    12. DUBOIS, M., GILLES, K. A., HAMILTON, J. K., REBERS, P. A., SMITH, F., 1956: Colorimetric method for determination of sugars and related substances. Analytical Chemistry 28, 3: 350-356. ISSN 0003-2700. DOI: 10.1021/ac60111a017 Go to original source...
    13. ERICKSON, J., SCHOTT, D., REVERRI, T., MUHSIN, W., RUTTLEDGE, T., 2001: GC-MS analysis of hydrophobic root exudates of Sorghum and implications on the parasitic plant Striga asiatica. Journal of Agricultural and Food Chemistry 49, 5537-5542. ISSN 0021-8561. DOI: 10.1021/jf0111099 Go to original source...
    14. FORMÁNEK, P., KLEJDUS, B., VRANOVÁ, V., 2005: Bio-available amino acids extraction from soil by demineralized water and 0.5 M ammonium acetate. Amino Acids 28, 4: 427-429. ISSN 0939-4451. DOI: 10.1007/s00726-005-0190-0 Go to original source...
    15. GEORGE, T. S., GREGORY, P. J., WOOD, M., READ, D., BURESH, R. J., 2002: Phosphatase activity and organic acids in the rhizosphere of potential agroforestry species and maize. Soil Biology and Biochemistry 34, 10: 1487-1494. ISSN 0038-0717. DOI: 10.1016/S0038-0717(02)00093-7 Go to original source...
    16. GERKE, J., RÖMER, W., JUNK, A., 1994: The excretion of citric and malic acid by proteoid roots of Lupinus albus L.: effect on soil solution concentrations of phosphate, iron and alluminium in the proteoid rhizosphere sample of an oxisol and a luvisol. Journal of Plant Nutrition and Soil Science 157, 4: 289-294. ISSN 1436-8730. Go to original source...
    17. GRAMSS, G., RIDESCHKO, O., 1998: Activities of oxidoreductase enzymes in tissue extracts and sterile root exudates of three crop plants, and some properties of the peroxidase component. New Phytologists 138, 3: 401-409. ISSN 0028-646X. DOI: 10.1046/j.1469-8137.1998.00128.x Go to original source...
    18. GRANSEE, A., 2001: Effects of root exudates on nutrient availability in the rhizosphere. In: Horst, W. J. et al. 2001. Plant nutrition-food security and sustainability of agro-ecosystems, 626-627. Food security and sustainability and sustainability of agro-ecosystems through basic and applied research. ISBN 0792371054.
    19. GRANSEE, A., WITTENMAYER, L., 2000: Qualitative and quantitative analysis of water-soluble root exudates in relation to plant species and development. Journal of Plant Nutrition and Soil Science 163, 4: 381-385. ISSN 1436-8730. DOI: 10.1002/1522-2624(200008)163:43.0.CO;2-7 Go to original source...
    20. GRAYSTON, S. J., VAUGHAN, D., JONES, D., 1996: Rhizosphere carbon flow in trees, in comparison with annual plants: the importance of root exudation and its impact on microbial activity and nutrient availability. Applied Soil Ecology 5, 1: 29-56. ISSN 0929-1393. DOI: 10.1016/S0929-1393(96)00126-6 Go to original source...
    21. JOHNSON, J. F., ALLAN, D. L., VANCE, C. P., WEIBLEN, G., 1996: Root Carbon Dioxide Fixation by Phosphorus-Deficient Lupinus albus (Contribution to Organic Acid Exudation by Proteoid Roots). Plant Physiology 112, 1: 19-30. ISSN 0032-0889. DOI: 10.1104/pp.112.1.31 Go to original source...
    22. KRAVCHENKO, L. V., AZAROVA, T. S., MAKAROVA, N. M., TIKHONOVICH, I. A., 2004: The effect of tryptophan present in plant roots exudates on the phytostimulating activity of rhizobacteria. Microbiology 73, 2: 156-158. ISSN 0026-2617. DOI: 10.1023/B:MICI.0000023982.76684.9d Go to original source...
    23. KRAVCHENKO, L. V., LEONOVA, E. I., 1993: The use of the root exometabolite tryptophan by plant-associated bacteria for the biosynthesis of indole-3-acetic acid. Microbiologiya 62, 3: 453-459. ISSN 0026-3656.
    24. LI, Y. G., LI, L. H., JIANG, G. M., NIU, S. L., LIU, M. Z., GAO, L. M., PENG, Y., JIANG, C. D., 2004: Traits of chlorophyll fluorescence in 99 plant species from the sparse-elm grassland in Hunshandak Sandland. Photosynthetica 42, 2: 243-249. ISSN 0300-3604. DOI: 10.1023/B:PHOT.0000040596.39460.6f Go to original source...
    25. LIN, C. H., LERCH, R. N., GARRETT, H. E., GEORGE, M. F., 2004: Incorporating forage grasses in riparian buffers for bioremediation of atrazine, isoxaflutole and nitrate in Missouri. Agroforestry Systems 63, 1: 91-99. ISSN 0167-4366. DOI: 10.1023/B:AGFO.0000049437.70313.ef Go to original source...
    26. MA, Z., MIYASAKA, S. C., 1998: Oxalate exudation by taro in respone to Al. Plant Physiology 118, 5: 861-865. ISSN 0032-0889. DOI: 10.1104/pp.118.3.861 Go to original source...
    27. MENCH, M., 1985: Influence des exudats racinaires solubles sur la dynamique des métaux dans la rhizosphére du maďs (Zea mays L.). Thése de doctorat, University of Nancy. FRBNF 37594962.
    28. MIYA, R. K., FIRESTONE, M. K., 2001: Enhanced phenanthrene biodegradation in soil by slender oat root exudates and root debris. Journal of Environmental Quality 30, 6: 1911-1918. ISSN 0047-2425. DOI: 10.2134/jeq2001.1911 Go to original source...
    29. NARDI, S., SESSI, E., PIZZEGHELLO, D., STURARO, A., RELLA, R., PARVOLI, G., 2002: Biological activity of soil organic matter mobilized by root exudates. Chemosphere 46, 7: 1075-1081. ISSN 0045-6535. DOI: 10.1016/S0045-6535(01)00160-6 Go to original source...
    30. NARDI, S., TOSONI, M., PIZZEGHELLO, D., PROVENZANO, M. R., CILENTI, A., STURARO, A., RELLA, R., VIANELLO, A., 2005: Chemical characteristics and biological activity of organic substances extracted from soils by root exudates. Soil Science Society of America Journal 69, 2012-2019. ISSN 0361-5995. DOI: 10.2136/sssaj2004.0401 Go to original source...
    31. NELSON, D. W., SOMMERS, L. E., 1996: Total carbon, organic carbon, and organic matter. In: Sparks, D. L. (Ed.), Methods of Soil Analysis, Chemical Methods, Vol. 5, part 3. Soil Science Society of America Inc., Madison, WI, USA, 961-1010. ISSN 0361-5995. Go to original source...
    32. NEUMAN, G., HÜLSTER, A., RÖMHELD, V., 1999: PCDD/PCDF-mobilizing compounds in root exudates of zucchini. Organohalogen Compounds 41: 331-334. ISSN 1026-4892.
    33. NEUMAN, G., RÖMHELD, V., 2007: The release of root exudates as affected by the plant physiological status. In: Pinton, R., Varanini, Z., Nannipieri, P. (Eds.). The rhizosphere: Biochemistry and organic substances at the soil-plant interface, 23-72. ISBN 9780849338557. Go to original source...
    34. NÓBREGA, F. M., SANTOS, I. S., DA DUNHA, M., CARVALHO, A. O., GOMES, V. M., 2005: Antimicrobial proteins from cowpea root exudates: inhibitory activity against Fusarium oxysporum and purification of chitinase-like proteins. Plant and Soil 272, 1-2: 223-232. ISSN 0032-079X. DOI: 10.1007/s11104-004-4954-1 Go to original source...
    35. PAYNEL, F., MURRAY, J., CLIQUET, J. B., 2001: Root exudates: a pathway for short-term N transfer from clover and ryegrass. Plant and Soil 229, 2: 235-243. ISSN 0032-079X. DOI: 10.1023/A:1004877214831 Go to original source...
    36. PERSONENI, E., NGUYEN, C., MARCHAL, P., PAGÉS, L., 2007: Experimental evaluation of an efflux-influx model of C exudation by individual apical root segments. Journal of Experimental Botany 58, 8: 2091-2099, ISSN 0022-0957. DOI: 10.1093/jxb/erm065 Go to original source...
    37. PHILIPS, D. A., FOX, T. C., KING, M. D., BHUVANESWARI, T. V., TEUBER, L. R., 2004: Microbial products trigger amino acids exudation from plant roots. Plant Physiology 136, 1: 2887-2894. ISSN 0032-0889. DOI: 10.1104/pp.104.044222 Go to original source...
    38. SAKAI, H., TADANO, T., 1993: Characteristics of response of acid phosphatase secreted by the roots of several crops to various conditions in the grow media. Soil Science and Plant Nutrition 39, 3: 437-444. ISSN 0038-0768. DOI: 10.1080/00380768.1993.10419784 Go to original source...
    39. SCHWAB, S. M., MENGE, J. A., LEONARD, R.T., 1983: Quantitative and qualitative effects of phosphorus on extracts and exudates of sudangrass roots in relation to vesicular-arbuscular mycorrhiza formation. Plant Physiology 73, 3: 761-765. ISSN 0032-0889. DOI: 10.1104/pp.73.3.761 Go to original source...
    40. SPIES, J. R., 1957: Colorimetric procedures for amino acids. In: Colowick, S. P., Kaplan, N. O. Methods in Enzymology, Vol. III. pp. 468-471. Academic Press, New York. Go to original source...
    41. UREN, N. C., 2007: Type, amounts, and possible functions of compounds released into the rhizosphere by soil-grown plants. In: Pinton, R., Varanini, Z., Nannipieri, P. (Eds.), The rhizosphere: Biochemistry and organic substances at the soil-plant interface, pp.1-21, CRC Press, Boca Raton, FL. Go to original source...
    42. VANČURA, V., 1988: Plant metabolites in soil. In: Vančura, V., Kunc, F. (Eds.). Soil microbial associations: control of structures and functions. Academia Praha, 57-144. Go to original source...
    43. VONDERHEIDE, A. P., MOUNICOU, S., MEIJA, J., HENRY, H. F., CARUSO, J. A., SHANN, J. R., 2006: Investigation of selenium-containing root exudates of Brassica juncea using HPLC-ICP-MS and ESI-qTOF-MS. Analyst 131, 33-40. ISSN 0003-2654. DOI: 10.1039/b510712a Go to original source...
    44. WANG, P., Bi, S., WANG, S., DING, Q., 2006: Variation of wheat root exudates under aluminium stress. Journal of Agricultural and Food Chemistry 54, 10040-10046. ISSN 0021-8561. DOI: 10.1021/jf061249o Go to original source...
    45. WHIPPS, J. M., LYNCH, J. M., 1990: Substrate flow in the rhizosphere. Plant and Soil 129, 1: 1-10. ISSN 0032-079X. Go to original source...

    This is an open access article distributed under the terms of the Creative Commons Attribution-NonCommercial-NoDerivatives 4.0 International License (CC BY NC ND 4.0), which permits non-comercial use, distribution, and reproduction in any medium, provided the original publication is properly cited. No use, distribution or reproduction is permitted which does not comply with these terms.


    Return to the content