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Hydrolysis of organic phosphorus in soil water suspensions after addition of phosphatase enzymes

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Abstract

Additions of enzymes involved in organic phosphorus (P) hydrolysis can be used to characterize the hydrolyzability of molybdate-unreactive P (MUP) in soil water extracts. Our aim was to test the feasibility of enzyme additions to soil water suspensions with respect to (1) suitable enzyme preparations and (2) recovery of molybdate-reactive P (MRP). To this end, we compared the substrate specificity of seven commercially available enzyme preparations (acid and alkaline phosphomonoesterase, phytase, and nuclease preparations) and optimized the assay conditions in microplates. We then measured MRP release after the addition of the enzymes to soil water suspensions and filtrates of two Swiss grassland soils (midland and alpine). In some cases, commercial preparations of the same enzyme differed in their specificity, presumably due to contamination with other enzymes, and also in their efficiency in soil suspensions. Addition of EDTA to the buffer was required to decrease sorption of released P in soil suspensions. Enzymatic release of P was consistently equal or higher in soil suspensions than in soil filtrates. However, also more dissolved MUP was present in soil suspensions than in filtrates, since the buffer interacted with the solid phase. Of the total dissolved MUP in soil suspensions, 94 and 61 % were hydrolyzable in midland and alpine soil, respectively. More specifically, 60 and 17 % of MUP were in nucleic acids, 6 and 39 % in simple monoesters, and 28 and 5 % in inositol hexakisphosphate in midland and alpine soil, respectively. Thus, we show that the characterization of hydrolyzable organic P in soil suspensions with hydrolytic enzyme preparations may be useful to better understand the availability of soil organic P to enzymatic hydrolysis, but that it requires soil-specific adaptation for optimum P recovery.

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References

  • Anderson JM, Ingram JSI (1993) Tropical soil biology and fertility. A handbook of methods, 2nd edn. CAB International, Wallingford, p 221 pp

    Google Scholar 

  • Bernasconi SM, Bauder A, Bourdon B, Brunner I, Bünemann E, Christl I, Derungs N, Edwards P, Farinotti D, Frey B, Frossard E, Furrer G, Gierga M, Göransson H, Gulland K, Hagedorn F, Hajdas I, Hindshaw R, Ivy-Ochs S, Jansa J, Jonas T, Kiczka M, Kretzschmar R, Lemarchand E, Luster J, Magnusson J, Mitchell EAD, Venterink HO, Plotze M, Reynolds B, Smittenberg RH, Stahli M, Tamburini F, Tipper ET, Wacker L, Welc M, Wiederhold JG, Zeyer J, Zimmermann S, Zumsteg A (2011) Chemical and biological gradients along the Damma glacier soil chronosequence. Switzerland Vadose Zone J 10:867–883

    Article  CAS  Google Scholar 

  • Bünemann EK (2008) Enzyme additions as a tool to assess the potential bioavailability of organically bound nutrients. Soil Biol Biochem 40:2116–2129

    Article  Google Scholar 

  • Bünemann EK, Keller B, Hoop D, Jud K, Boivin P, Frossard E (2013) Increased availability of phosphorus after drying and rewetting of a grassland soil: processes and plant use. Plant Soil. doi 10.1007/s11104-11013-11651-y.

  • Bünemann EK, Oberson A, Liebisch F, Keller F, Annaheim KE, Huguenin-Elie O, Frossard E (2012) Rapid microbial phosphorus immobilization dominates gross phosphorus fluxes in a grassland soil with low inorganic phosphorus availability. Soil Biol Biochem 51:84–95

    Article  Google Scholar 

  • Celi L, Barberis E (2005) Abiotic stabilization of organic phosphorus in the environment. In: Turner BL, Frossard E, Baldwin D (eds) Organic phosphorus in the environment. CABI, Wallingford, pp 113–132

    Chapter  Google Scholar 

  • Chen CR, Condron LM, Davis MR, Sherlock RR (2002) Phosphorus dynamics in the rhizosphere of perennial ryegrass (Lolium perenne L.) and radiata pine (Pinus radiata D. Don.). Soil Biol. Biochem 34:487–499

    CAS  Google Scholar 

  • Condron LM, Turner BL, Cade-Menun BJ (2005) Chemistry and dynamics of soil organic phosphorus. In: Sims JT, Sharpley AN (eds) Phosphorus: agriculture and the environment. ASA, CSSA and SSSA, Madison, pp 87–122

    Google Scholar 

  • Dao TH (2004) Ligands and phytase hydrolysis of organic phosphorus in soils amended with dairy manure. Agron J 96:1188–1195

    Article  Google Scholar 

  • Firsching BM, Claassen N (1996) Root phosphatase activity and soil organic phosphorus utilization by Norway spruce Picea abies (L) Karst. Soil Biol Biochem 28:1417–1424

    Article  CAS  Google Scholar 

  • Frossard E, Achat DL, Bernasconi SM, Bünemann EK, Fardeau J-C, Jansa J, Morel C, Rabeharisoa L, Randriamanantsoa L, Sinaj S, Tamburini F, Oberson A (2011) The use of tracers to investigate phosphate cycling in soil–plant systems. In: Bünemann EK, Oberson A, Frossard E (eds) Phosphorus in action, Biological processes in soil phosphorus cycling. Springer, Heidelberg, pp 59–91

    Chapter  Google Scholar 

  • Frossard E, Sinaj S (1998) The isotope exchange kinetic technique: a method to describe the availability of inorganic nutrients. Applications to K, P, S and Zn. Isot Environ Heal Stud 34:61–77

    Article  Google Scholar 

  • George TS, Richardson AE, Simpson RJ (2005) Behaviour of plant derived extracellular phytase upon addition to soil. Soil Biol Biochem 37:977–988

    Article  CAS  Google Scholar 

  • George TS, Simpson RJ, Gregory PJ, Richardson AE (2007a) Differential interaction of Aspergillus niger and Peniophora lycii phytases with soil particles affects the hydrolysis of inositol phosphates. Soil Biol Biochem 39:793–803

    Article  CAS  Google Scholar 

  • George TS, Simpson RJ, Hadobas PA, Marshall DJ, Richardson AE (2007b) Accumulation and phosphatase-lability of organic phosphorus in fertilised pasture soils. AusJAgricRes Aus.J.Agric.Res. 58:47–55

    Article  Google Scholar 

  • Giaveno C, Celi L, Richardson AE, Simpson RJ, Barberis E (2010) Interaction of phytases with minerals and availability of substrate affect the hydrolysis of inositol phosphates. Soil Biol Biochem 42:491–498

    Article  CAS  Google Scholar 

  • Greiner R (2007) Phytate-degrading enzymes: regulation of synthesis in microorganisms and plants. In: Turner BL, Richardson AE, Mullaney EJ (eds) Inositol phosphates: linking agriculture and the environment. CABI, Wallingford, pp 78–96

    Chapter  Google Scholar 

  • Harrison AF (1982) 32P-method to compare rates of mineralization of labile organic phosphorus in woodland soils. Soil Biol Biochem 14:337–341

    Article  CAS  Google Scholar 

  • Harrison AF (1987) Soil organic phosphorus, A review of world literature. CAB International, Wallingford

    Google Scholar 

  • Häussling M, Marschner H (1989) Organic and inorganic soil phosphates and acid phosphatase activity in the rhizosphere of 80-year-old Norway spruce [Picea abies (L) Karst] trees. Biol Fert Soils 8:128–133

    Article  Google Scholar 

  • He ZQ, Griffin TS, Honeycutt CW (2004) Enzymatic hydrolysis of organic phosphorus in swine manure and soil. J Environ Qual 33:367–372

    PubMed  CAS  Google Scholar 

  • Kuo S (1996) Phosphorus. In: Sparks DL (ed) Methods of soil analysis. Part 3. Chemical methods. SSSA/ASA, Madison, pp 869–919.

  • Leprince F, Quiquampoix H (1996) Extracellular enzyme activity in soil: effect of pH and ionic strength on the interaction with montmorillonite of two acid phosphatases secreted by the ectomycorrhizal fungus Hebeloma cylindrosporum. Eur J Soil Sci 47:511–522

    Article  CAS  Google Scholar 

  • Lung SC, Leung A, Kuang R, Wang Y, Leung P, Lim BL (2008) Phytase activity in tobacco (Nicotiana tabacum) root exudates is exhibited by a purple acid phosphatase. Phytochemistry 69:365–373

    Article  PubMed  CAS  Google Scholar 

  • Macklon AES, Grayston SJ, Shand CA, Sim A, Sellars S, Ord BG (1997) Uptake and transport of phosphorus by Agrostis capillaris seedlings from rapidly hydrolysed organic sources extracted from 32P-labelled bacterial cultures. Plant Soil 190:163–167

    Article  CAS  Google Scholar 

  • Nadeau JA, Qualls RG, Nowak RS, Blank RR (2007) The potential bioavailability of organic C, N, and P through enzyme hydrolysis in soils of the Mojave Desert. Biogeochem 82:305–320

    Article  CAS  Google Scholar 

  • Nannipieri P, Giagnoni L, Renella G, Puglisi E, Ceccanti B, Masciandaro G, Fornasier F, Moscatelli MC, Marinari S (2012) Soil enzymology: classical and molecular approaches. Biol Fert Soils 48:743–762

    Article  Google Scholar 

  • Nannipieri P, Gianfreda L (1998) Kinetics of enzyme reactions in soil environments. In: Huang PM, Senesi N, Buffle J (eds) Structure and surface reactions of soil particles. Wiley, New York, pp 449–479

    Google Scholar 

  • Ohno R, Zibilske LM (1991) Determination of low concentrations of phosphorus in soil extracts using malachite green. Soil Sci Soc Am J 55:892–895

    Article  CAS  Google Scholar 

  • Olsson R, Giesler R, Loring JS, Persson P (2012) Enzymatic hydrolysis of organic phosphates adsorbed on mineral surfaces. ES&T 46:285–291

    Article  CAS  Google Scholar 

  • Philipp A, Huguenin-Elie O, Flisch R, Gago R, Stutz C, Kessler W (2004) Einfluss der Phosphordüngung auf eine Fromentalwiese. Agrarforschung 11:86–91

    Google Scholar 

  • Saunders WMH, Williams EG (1955) Observations on the determination of total organic phosphorus in soils. J Soil Sci 6:254–267

    Article  CAS  Google Scholar 

  • Staunton S, Razzouk R, Abadie J, Quiquampoix H (2012) Water-extractable soil organic matter inhibits phosphatase activity. Soil Biol Biochem 55:14–16

    Article  CAS  Google Scholar 

  • Tiessen H, Moir JO (1993) Characterisation of available P by sequential extraction. In: Carter MR (ed) Soil sampling and methods of analysis. CRC, Boca Raton, pp 75–86

    Google Scholar 

  • Turner BL, McKelvie ID, Haygarth PM (2002) Characterisation of water-extractable soil organic phosphorus by phosphatase hydrolysis. Soil Biol Biochem 34:27–35

    Article  CAS  Google Scholar 

Download references

Acknowledgments

We thank Arne Korsbak from Novozyme (DSM Nutritional Products) and Roland Betz from BASF (Ludwigshafen) for the supply of Phytase I and II, respectively. Olivier Huguenin-Elie (ART Reckenholz) kindly provided access to the Watt field site and Julia Coffin did language corrections on the manuscript. This study was conducted within a project funded by the Swiss National Science Foundation.

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Correspondence to E. K. Bünemann.

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Annaheim, K.E., Rufener, C.B., Frossard, E. et al. Hydrolysis of organic phosphorus in soil water suspensions after addition of phosphatase enzymes. Biol Fertil Soils 49, 1203–1213 (2013). https://doi.org/10.1007/s00374-013-0819-1

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  • DOI: https://doi.org/10.1007/s00374-013-0819-1

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