Abstract
We developed a method using nitrocellulose membranes and image analysis to localise and quantify acid phosphatase activity in the rhizosphere of two plant species, one with cluster roots (Dryandra sessilis (Knight) Domin) and another with ectomycorrhizal roots (Pinus taeda L.). Membranes were placed in contact with roots and then treated with a solution of x, α-naphthyl phosphate and Fast Red TR. Acid phosphatase activity was visualised as a red imprint on the membrane. We quantified acid phosphatase activity by image analysis of scanned imprints. The method was used to estimate the spatial distribution of acid phosphatase activity within particular root classes (lateral roots, mycorrhizal roots, root clusters). Over 95% of the acid phosphatase activity of the root system of D. sessilis was associated with cluster roots, and between 20 and 32% of the root surface active. About 26 % of the acid phosphatase activity of the root system of P. taeda was associated with mycorrhizal roots and unsuberised white root tips and less than 10% of the root surface was active, irrespective of root type. This non-destructive method can be used for rapid, semi-quantitative assessment of acid phosphatase activity in the laboratory and in situ.
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Antibus R K, Bower D and Dighton J 1997 Root surface phosphatase activities and uptake of 32P-labelled inositol phosphate in field-collected gray birch and red maple roots. Mycorrhiza 7, 39–46.
Bresson L-M and Guilloré P 1994 On direct digital image acquisition from thin sections. In Soil Micromorphology: Studies in Management and Genesis. Eds A J Ringrose-Voase and G S Humphreys. pp 515–520. Developments in Soil Science 22, Elsevier, Amsterdam.
Burns R G 1978 Enzyme activity in soil: some theoretical and practical considerations. In Soil Enzymes. Ed R G Burns. pp 259–340. Academic Press, New York.
Dighton J and Boddy L 1989 Role of fungi in nitrogen, phosphorus and sulphur cycling in temperate forest ecosystems. In Nitrogen Phosphorus and Sulphur Utilization by Fungi. Eds L Boddy, R Marchant and D Read. pp 269–298. Cambridge University Press, Cambridge.
Dinkelaker B and Marschner H 1992 In vivo demonstration of acid phosphatase activity in the rhizosphere of soil-grown plants. Plant Soil 144, 199–205.
Dodd J C, Burton C C, Burns R G and Jeffries P 1987 Phosphatase activity associated with the roots and the rhizosphere of plants infected with vesicular-aibuscular mycorrhizal fungi. New Phytol. 107, 163–172.
Dracup MN H, Barrett-Lennard E G, Greenway H and Robson AD 1984 Effect of phosphorus deficiency on phosphatase activity of cell walls from roots of subterranean clover. J. Exp. Bot. 35, 466–480.
Firsching B-M and Claassen N 1996 Root phosphatase activity and soil organic phosphorus utilization by Norway spruce [Picea abies (L.) Karst.]. Soil Biol Biochem. 28, 1417–1424.
Gahan P B 1984 Plant Histochemistry and Cytochemistry. Academic Press, London.
Gallagher S R and Leonard R T 1982 Effect of vanadate, molybdate, and azide on membrane-associated ATPase and soluble phosphatase activity of corn roots. Plant Physiol. 70, 1335–1340.
Gundlach G and Mühlhausen B 1980 Untersuchungen zur Kupplung des 1-Naphthols mit Fast Red TR. Untersuchungen zur Optimierung einer kontinuierlichen Bestimmung der sauren Phosphatase. 1. Mitteilung. J. Clin. Chem. Clin. Biochem. 18, 603–610.
Häussling M and 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. Fertil. Soils 8. 128–133.
Hayano K 1987 Characterization of a phosphodiesterase component in a forest soil extract. Biol. Fertil. Soils 3, 128–133.
Helal H M and Sauerbeck D R 1987 Phosphatase-Aktivitat von Planzenwurzeln und Böden in Abhängigkeit von der PVersorgung. VDLUFA-Schriftenreihe 23, 195–201.
Hendrick R L and Pregitzer K 5 1996 Applications of minirhizotrons to understanding root function in forests and other natural ecosystems. Plant Soil 185, 293–304.
Jacobsen J V and Knox R B 1973 Cytochemical localisation and antigenicity of α-amylase in barley aleurone tissue. Planta 112, 213–224.
Jaillard B, Ruiz L and Arvieu J C 1996 pH mapping in transparent gel using colour indicator videodensitometry. Plant Soil 183, 85–95.
Lambais M R and Mehdy M C 1998 Spatial distribution of chitinases and β-1,3-glucanase transcripts in bean arbuscular mycorrhizal roots under low and high soil phosphate conditions. New Phytol. 140, 33–42.
Leprince F and Quiquampoix H 1996 Extracellular enzyme activity in soil: effect of pH and ionic strength on the interaction with montruorillonite of two acid phosphatases secreted by the ectomycorrhizal fungus Hebeloma cylindrosporum. European J Soil Sci. 47, 511–522.
Lin N 5, Hsu Y H and Hsu H T 1990 Immunological detection of plant viruses and a mycoplasma-like organism by direct tissue blotting on nitrocellulose transfer membranes. Phytopathology 80, 824–828.
MacFall J, Slack SA and Iyer J 1991 Effects of Hebeloma arenosa and phosphorus fertility on root acid phosphatase activity of red pine (Pinus resinosa) seedlings. Can. J. Bot. 69. 380–383.
Magliano T M A and Casal J J 1998 In vitro cross-linking of extensin precursors by mustard extracellular isoforms of peroxidase that respond either to phvtochrome or to wounding. J. Exp. Bot. 49, 1491–1499.
Nannipieri P, Ceccanti B and Bianchi D 1988 Characterization of humus-phosphatase complexes extracted from soil. Soil Biol. Biochem. 20. 683–691.
Pan W L, Bolton R P Lundquist E J and Huller L K 1998 Portable rhizotron and colour scanner system for monitoring root development. Plant Soil 200, 107–412.
Shaykh M M and Roberts L W 1974. A histochemical study of phosphatases in root apical menstems. Ann. Bot. 38, 165–174.
Sinsabaugh R L 1994 Enzymic analysis of microbial pattern and process. Biol. Fertil. Soils 17, 69–74.
Skujins J 1978 History of abiontic soil enzyme research. In Soil Enzymes. Ed RG Burns. pp 1–50. Academic Press, New York.
Tabatabai M A and Bremner J M 1969 Use of p-nitrophenyl phosphate for assay of soil phosphatase activity. Soil Biol. Biochem. 1, 301–307.
Tarafdar J C and Claassen N 1988 Organic phosphorus compounds as a phosphorus source for higher plants through the activity of phosphatases produced by plant roots and microorganisms. Biol. Fertil. Soils 5, 308–312.
Tarafdar J C and Marschner H 1994 Phosphatase activity in the rhizosphere and hyphosphere of VA mycorrhizal wheat supplied with inorganic and organic phosphorus. Soil Biol. Biochem. 26, 387–395.
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Grierson, P.F., Comerford, N.B. Non-destructive measurement of acid phosphatase activity in the rhizosphere using nitrocellulose membranes and image analysis. Plant and Soil 218, 49–57 (2000). https://doi.org/10.1023/A:1014985327619
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DOI: https://doi.org/10.1023/A:1014985327619