ORIGINAL PAPER A novel phosphorus biofertilization strategy using cattle manure treated with phytase–nanoclay complexes Daniel Menezes-Blackburn & Milko A. Jorquera & Liliana Gianfreda & Ralf Greiner & María de la Luz Mora Received: 4 July 2013 /Revised: 2 October 2013 /Accepted: 4 October 2013 # Springer-Verlag Berlin Heidelberg 2013 Abstract The aim of this work was to evaluate the treatment of cattle manure with phytases stabilized in allophanic nanoclays as a potential novel phosphorus (P) biofertilization technology for crops grown in volcanic soils (Andisol). Furthermore, because the optimal pH for commercial phytase catalysis does not match the natural pH of manure, a comple- mentary experiment was set up to evaluate the effect of manure inoculation with an alkaline phytase-producing bac- terium. Finally, phytase-treated soil, manure, and soil–manure mixtures were evaluated for their P-supplying capacity to wheat plants grown under greenhouse conditions. Treating cattle manure with phytases stabilized in nanoclays resulted in a significant (P ≤ 0.05) increase of inorganic P in soil extracts (NaOH-EDTA and Olsen). The use of phytase- treated cattle manure increased dry weights by 10 % and the P concentration by 39 % in wheat plants grown under green- house conditions, which is equivalent to a P fertilizer rate of about 150 kg of P per hectare. The inoculation of cattle manure with β-propeller phytase-producing bacteria led to an ∼10 % increase in inorganic P in the manure extracts. However, applying inoculated manure to soil did not signifi- cantly increase wheat yield or P acquisition responses. Our results suggest that the novel approach of incubating cattle manure with phytases stabilized in nanoclay enhances the organic P cycling and P nutrition of plants grown in P- deficient soils. Keywords Phytase . Phytate . Phosphorus . Organic phosphorus . Biofertilization . Volcanic soils . Cattle manure . Wheat Introduction Soil organic phosphorus (Po) accounts for 40–80 % of total phosphorus (P t ), and its concentration in Chilean Andisols averages about 1,000 mg kg -1 (Pinochet et al. 2001; Borie and Rubio 2003; Briceño et al. 2004; Redel et al. 2007). Among all the Po forms, phytate is usually expected to be the most abundant form of P in soils (Turner et al. 2002; Borie and Rubio 2003). In addition to phytate, other forms of phytase-hydrolyzable Po are ubiquitous in soil and manure environments (Menezes-Blackburn et al. 2013). Soil phytase- labile organic P pools (P Phy-lab ) naturally accumulate in re- sponse to P fixation mechanisms. Nevertheless, the accumu- lation of these compounds is significantly enhanced by both inorganic and organic P fertilization (Turner et al. 2002; George et al. 2007b). Rock phosphate used to produce inorganic phosphate fer- tilizers are a nonrenewable-limited natural resource, and its imminent scarcity scenario has been recently speculated (Abelson 1999; Ashley et al. 2011). Due to this global P problem, renewed interest has recently been directed toward developing technologies and practices related to increasing P D. Menezes-Blackburn Chile–Italy International Doctorate in Environmental Resources Science, Universidad de La Frontera, Ave.nida Francisco Salazar, 01145 Temuco, Chile M. A. Jorquera : M. de la Luz Mora (*) Scientific and Technological Bioresource Nucleus (BIOREN), Universidad de La Frontera, Avenida Francisco Salazar, 01145 Temuco, Chile e-mail: mariluz.mora@ufrontera.cl L. Gianfreda Dipartimento di Scienze del Suolo, della Pianta e dell’Ambiente, e delle Produzioni Animali, Università di Napoli, Federico II, Portici, Italy D. Menezes-Blackburn : R. Greiner Department of Food Technology and Bioprocess Engineering, Max Rubner-Institut, Federal Research Institute of Nutrition and Food, Karlsruhe, Germany Biol Fertil Soils DOI 10.1007/s00374-013-0872-9