The ability to solubilize insoluble inorganic pho- sphate compounds by Gluconacetobacter diazotrophicus was studied using different cul-ture approaches. Qualitative plate assays using tricalcium phosphate as the sole P-source showed that G. diazotrophicus produced solu-bilization only when aldoses were used as the C-source. Extracellular aldose oxidation via a pyrroloquinoline quinone-linked glucose dehy-drogenase (PQQ-GDH) is the main pathway for glucose metabolism in G. diazotrophicus. In batch cultures with 5 g l-1 of hydroxyapatite as the P-source and glucose as the C-source, more than 98% of insoluble P was solubilized. No solubilization was observed neither using glyc-erol nor culturing a PQQ-GDH mutant of G. di-azotrophicus. Solubilizaton was not affected by adding 100 mmol l-1 of MES buffer. Continuous cultures of G. diazotrophicus showed significant activities of PQQ-GDH either under C or P limi-tation. An intense acidification in the root envi-ronment of tomato and wheat seedlings inocu-lated with a G. diazotrophicus PAL5 was ob-served. Seedlings inoculated with a PQQ-GDH mutant strain of G. diazotrophicus showed no acidification. Our results suggest that G. di-azotrophicus is an excellent candidate to be used as biofertilizer because in addition to the already described plant growth-promoting abili-ties of this organism, it shows a significant mineral phosphate solubilization capacity.
Goldstein, A.H. (1986) Bacterial mineral phosphate solubilization: Historical perspective and future prospects. American Journal of Alternative Agriculture, 1, 57-65.
Sashidhar, B. and Podile, A.R. (2010) Mineral phosphate solubilization by rhizosphere bacteria and scope for manipulation of the direct oxidation pathway involving glucose dehydrogenase. Journal of Applied Microbiology, 109, 1-12.
Rodriguez, H. and Fraga, R. (1999) Phosphate solubilizing bacteria and their role in plant growth promotion. Biotechnology Advances, 17, 319-339. doi:10.1016/S0734-9750(99)00014-2
Gyaneshwar, P., Kumar, G.N., Parekh, L.J. and Poole, P.S. (2002) Role of soil microorganisms in improving P nutrition of plants. Plant and Soil, 245, 83-93. doi:10.1023/A:1020663916259
Intorne, A.C., de Oliveira, M.V., Lima, M.L., da Silva, J.F., Olivares, F.L. and de Souza Filho, G.A. (2009) Identifcation and characterization of Gluconacetobacter diazotrophicus mutants defective in the solubilization of phosphorus and zinc. Archives of Microbiology, 191, 477-483. doi:10.1007/s00203-009-0472-0
Pedraza, R.O. (2008) Recent advances in nitrogen-fixing acetic acid bacteria. International Journal of Food Microbiology, 125, 25-35. doi:10.1016/j.ijfoodmicro.2007.11.079
Luna, M.F., Galar, M.L., Aprea, J., Molinari M.L. and Boiardi, J.L. (2010) Colonization of sorghum and wheat by seed inoculation with Gluconacetobacter diazotrophicus. Biotechnology Letters, 32, 1071-1076. doi:10.1007/s10529-010-0256-2
Maheshkumar, K.S., Krishnaraj, P.U. and Alagwadi, A.R. (1999) Mineral solubilising activity of Acetobacter diazotrophicus, a bacterium associated with sugarcane. Current Science, 76, 874-875.
Galar, M.L. and Boiardi, J.L. (1995) Evidence for a membrane-bound pyrroloquinoline quinone-linked glucose dehydrogenase in Acetobacter diazotrophicus. Applied Microbiology and Biotechnology, 43, 713-716. doi:10.1007/BF00164778
Luna, M.F., Bernardelli, C.E., Galar, M.L. and Boiardi, J.L. (2006) Glucose metabolism in batch and continuous cultures of Gluconacetobacter diazotrophicus PAL 3. Current Microbiology, 52, 163-168. doi:10.1007/s00284-005-4563-0
Luna, M.F., Mignone, C.F. and Boiardi, J.L. (2000) The carbon source influences the energetic efficiency of the respiratory chain of N2-fixing Acetobacter diazotrophicus. Applied Microbiology and Biotechnology, 54, 564-569. doi:10.1007/s002530000425
Stephan, M.P., Oliveira, M., Teixeira, K.R.S., Martínez-Drets, G. and D?bereiner, J. (1991) Physiology and dinitrogen fixation of Acetobacter diazotrophicus. FEMS Microbiology Letters, 77, 67-72. doi:10.1111/j.1574-6968.1991.tb04323.x
Nautiyal, C.S. (1999) An effcient microbiological growth medium for screening phosphate solubilizing microorganisms,” Microbiology Letters, 170, 265-270. doi:10.1111/j.1574-6968.1999.tb13383.x
Rodriguez, H., Gonzalez, T. and Selman, G. (2000) Expression of a mineral phosphate solubilizing gene from Erwinia herbicola in two rhizobacterial strains. Journal of Biotechnology, 84, 155-161. doi:10.1016/S0168-1656(00)00347-3
Clesscerl, L.S., Greenberg, A.E. and Eaton, A.D. (1998) Standard methods for the examination of water and wastewater. 20th Edition, APHA-AWWA-WEF, Washington, DC.
Matsushita, K. and Ameyama, M. (1982) D-Glucose dehydrogenase from Pseudomonas fluorescens, membrane-bound. Methods in Enzymology, 89, 149-155. doi:10.1016/S0076-6879(82)89026-5
Matsushita, K., Shinagawa, E. and Ameyama, M. (1982) D-gluconate dehydrogenase from bacteria, 2-keto-D-gluconate yielding, membrane bound. Methods in Enzymology, 89, 187-193. doi:10.1016/S0076-6879(82)89033-2
F?hraeus, G. (1957) The infection of clove root hairs by nodule bacteria studied by simple glass slide technique. Journal of General Microbiology, 16, 347-381
Mehta, S. and Nautiyal, C.S. (2001) An efficient method for qualitative screening of phosphate solubilizing bacteria. Current Microbiology, 43, 51-55. doi:10.1007/s002840010259
Olsthoorn, A.J. and Duine, J.A. (1998) On the mechanism and specificity of soluble, quinoprotein glucose dehydrogenase in the oxidation of aldose sugars. Biochemistry, 37, 13854-13861. doi:10.1021/bi9808868
Attwood, M.M., van Dijken, J.P. and Pronk, J.T. (1991) Glucose metabolism and gluconic acid production by Acetobacter diazotrophicus. Journal of Fermentation and Bioengineering, 72, 101-105. doi:10.1016/0922-338X(91)90317-A
Luna, M.F. and Boiardi, J.L. (2008) Growth yields and glucose metabolism of N2-fixing Gluconacetobacter diazotrophicus at different culture pH values. World Journal of Microbiology and Biotechnology, 24, 587-590. doi:10.1007/s11274-007-9507-3
Russell, J.B., and Cook, G.M. (1995) Energetics of bacterial growth: balance of anabolic and catabolic reactions. Microbiology and Molecular Biology Reviews, 59, 48-62.
Luna, M.F., Bernardelli, C.E., Mignone, C.F. and Boiardi, J.L. (2002) Energy generation by extracellular aldose oxidation in N2-fixing Gluconacetobacter diazotrophicus. Applied and Environmental Microbiology, 64, 2054-2056. doi:10.1128/AEM.68.4.2054-2056.2002
Lugtenberg, B.J.J. Kravchenko, L.V. and Simons, M. (1999) Tomato seed and root exudate sugars: composition, utilization by Pseudomonas biocontrol strains and role in rhizosphere colonization. Environmental Microbiology, 1, 439-446. doi:10.1046/j.1462-2920.1999.00054.x
Wang, P., Bi, S., Wang, S. and Ding, Q. (2006) Variation of wheat root exudates under aluminum stress. Journal of Agricultural and Food Chemistry, 54, 10040-10046. doi:10.1021/jf061249o