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References

  1. Audus, L. J. 1972. Plant Growth Substances, Vol. 1. Leonard Hill, London.
  2. Brelles-Marino, C. and E. J. Bedmar. 2001. Detection, purification and characterization of quorum-sensing signal molecules in plant-associated bacteria. J. Biotechnol. 91: 197-209.
    CrossRef
  3. Brinton, W. F. 1997. Dynamic chemical processes underlying BD horn manure (500) preparation. J. Biodynamics 214: 1-8.
  4. Canellas, L. P., L. B. Dobbss, A. L. Oliveira, J. G. Chagasa, N. O. Aguiar, V. M. Rumjanek, et al. 2012. Chemical properties of humic matter as related to induction of plant lateral roots. Eur. J. Soil Sci. 63: 315-324.
    CrossRef
  5. Canellas, L. P., A. Piccolo, L. B. Dobbss, R. Spaccini, F. L. Olivares, D. B. Zandonadi, and A. R. Façanha. 2010. Chemical composition and bioactivity properties of size-fractions separated from a vermicompost humic acid. Chemosphere 78: 457-466.
    Pubmed CrossRef
  6. Carpenter-Boggs, L., J. P. Reganold, and A. C. Kennedy. 2000. Biodynamic preparations: Short-term effects on crops, soils, and weed populations. Am. J. Altern. Agric. 15: 96-114.
    CrossRef
  7. Carpenter-Boggs, L., J. P. Reganold, and A. C. Kennedy. 2000. Organic and biodynamic management: Effects on soil biology. Soil Sci. Soc. Am. J. 64: 1651-1659.
    CrossRef
  8. Davidson, E. A., L. F. Galloway, and M. K. Strand. 1987. Assessing available carbon: Comparison of techniques across selected forest soils. Commun. Soil Sci. Plant Anal. 18: 45-64.
    CrossRef
  9. Deffune, G. and A. M. Scolfied. 1995. Effects of humic acids and three bio-dynamic preparations on the growth of wheat seedlings. Proc. 3rd ESA Congress, Abano-Podova, Paper Ref No. 3-56.
  10. D’Haeze, W. P., P. Mergaert, J. C. Promé, and M. Holsters. 2000. Nod factor requirements for efficient stem and root nodulation of the tropical legume Sesbania rostrata. J. Biol. Chem. 275: 15676-15684.
    Pubmed CrossRef
  11. Dobbs, L. B., L. P. Canellas, F. L. Olivares, N. O. Aguiar, L. E. Pereira Peres, M. Azevedo, et al. 2010. Bioactivity of chemically transformed humic matter from vermicompost on plant root growth. J. Chem. 58: 3681-3688.
  12. Ertani, A., M. Schiavon, A. Muscolo, and S. Nardi. 2013. Alfalfa plant-derived biostimulant stimulate short-term growth of salt stressed Zea mays L. plants. Plant Soil 364: 145.
    CrossRef
  13. Ertani, A., O. Francioso, V. Tugnoli, V. Righi, and S. Nardi. 2011. Effect of commercial lignosulfonate-humate on Zea mays L. metabolism. J. Agric. Food Chem. 59: 11940-11948.
    Pubmed CrossRef
  14. Fornasier, F. and A. Margon. 2007. Bovine serum albumin and Triton X-100 greatly increase phosphomonoesterases and arylsulphatase extraction yield from soil. Soil Biol. Biochem. 39: 2682-2684.
    CrossRef
  15. Koepf, H. H., B. B. Pettersson, and W. Schaumann. 1976. Biodynamic Agriculture. The Anthroposophic Press, Spring Valley, New York.
  16. Mäder, P., L. Pfiffner, A. Fließbach, and U. Niggli. 1995. Biodiversity of soil biota in biodynamic, organic and conventional farming systems, pp. 45-57. In J. Isart and J. J. Llerena (eds.). Biodiversity and Land Use: The Role of Organic Farming. Bonn.
  17. Mader, P., A. Fliessbach, D. Dubois, L. Gunst, P. Fried, and U. Niggli. 2002. Soil fertility and biodiversity in organic farming. Science 296: 1694-1697.
    Pubmed CrossRef
  18. Marchesi, J. R., T. Sato, A. J. Weightman, T. A. Martin, J. C. Fry, S. J. Hiom, and W. G. Wade. 1998. Design and evaluation of useful bacterium-specific PCR primers that amplify genes coding for bacterial 16S rRNA. Appl. Environ. Microbiol. 64:795-799.
    Pubmed PMC
  19. Miller, M. B. and B. L. Bassler. 2001. Quorum sensing in bacteria. Annu. Rev. Microbiol. 55: 165-199.
    Pubmed CrossRef
  20. Nannipieri, P., J. Ascher, M. T. Ceccherini, L. Landi, G. Pietramellara, and G. Renella. 2003. Microbial diversity and soil functions. Eur. J. Soil Sci. 54: 655-670.
    CrossRef
  21. Nardi, S., G. Concheri, and G. Dell’Agnola. 1996. Biological activity of humus, pp. 361-406. In A. Piccolo (ed.). Humic Substances in Terrestrial Ecosystems. Elsevier, New York.
    CrossRef
  22. Osborn, A. M., E. R. B. Moore, and K. N. Timmis. 2000. An evaluation of terminal-restriction fragment length polymorphism (T-RFLP) analysis for the study of microbial community structure and dynamics. Environ. Microbiol. 2: 39-50.
    Pubmed CrossRef
  23. Rossen, L., C. A. Shearman, A. W. B. Johnston, and J. A. Downie. 1985. The nodD gene of Rhizobium leguminosarum is autoregulatory and in the presence of plant exudate induces the nodABC genes. EMBO J. 4: 3369-3373.
    Pubmed PMC
  24. Russo, R. O. and G. P. Berlyn. 1990. The use of organic biostimulants to help low input sustainable agriculture. J. Sustain. Agric. 1: 19-42.
    CrossRef
  25. Shaw, P. D., P. Gao, S. L. Daly, C. Cha, J. E. Cronan Jr., K. L. Rinehart, and S. K. Farrand. 1997. Detecting and characterizing N-acyl-homoserine lactone signal molecules by thin-layer chromatography Proc. Natl. Acad. Sci. USA 94: 6036-6041.
    Pubmed PMC CrossRef
  26. Spaccini, R., P. Mazzei, A. Squartini, M. Giannattasio, and A. Piccolo. 2012. Molecular properties of a fermented manure preparation used as field spray in biodynamic agriculture. Environ. Sci. Pollut. Res. Int. 19: 4214-4225.
    Pubmed CrossRef
  27. Stearn, W. C. 1976. Effectiveness of two biodynamic preparations on higher plants and possible mechanism s for the observed response. M.S. Thesis, Ohio State Univ., Columbus, OH.
  28. Tejada, M., C. Benítez, I. Gómez, and J. Parrado. 2011. Use of biostimulants on soil restoration: Effects on soil biochemical properties and microbial community. Appl. Soil Ecol. 49: 11- 17.
    CrossRef
  29. Tsavkelova, E. A., T. A. Cherdyntseva, S. Y. Klimova, A. I. Shestakov, S. G. Botina, and A. I. Netrusov. 2007. Orchidassociated bacteria produce indole-3-acetic acid, promote seed germination, and increase their microbial yield in response to exogenous auxin. Arch. Microbiol. 188: 655-664.
    Pubmed CrossRef
  30. Turinek, M., S. Grobelnik-Mlakar, M. Bavec, and F. Bavec. 2009. Biodynamic agriculture research progress and priorities. Renew. Agricult. Food Syst. 24: 146-154.
    CrossRef
  31. Vessey, K. J. 2003. Plant growh promoting rhizobacteria as biofertilizers. Plant Soil 255: 571-586.
    CrossRef

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Article

Research article

J. Microbiol. Biotechnol. 2013; 23(5): 644-651

Published online May 25, 2013 https://doi.org/10.4014/jmb.1212.12004

Copyright © The Korean Society for Microbiology and Biotechnology.

Microbiological Features and Bioactivity of a Fermented Manure Product (Preparation 500) Used in Biodynamic Agriculture

Matteo Giannattasio 1, Elena Vendramin 1, Flavio Fornasier 2, Sara Alberghini 1, Marina Zanardo 1, Fabio Stellin 1, Giuseppe Concheri 1, Piergiorgio Stevanato 1, Andrea Ertani 1, Serenella Nardi 1, Valeria Rizzi 3, Pietro Piffanelli 3, Riccardo Spaccini 4, Pierluigi Mazzei 5, Alessandro Piccolo 4 and Andrea Squartini 1*

1Department of Agronomy, Animals, Natural Resources and Environment - DAFNAE. University of Padua, Viale dell’Universita '16, 35020 Legnaro (Padova), Italy, 2C.R.A.-R.P.S., Council for Research in Agriculture, Center for the Study of the Relationships between Plant and Soil, Via Trieste 23, 34170 Gorizia, Italy, 3Parco Tecnologico Padano Foundation, Via Einstein 1, Cascina Codazza - 26900 Lodi, Italy, 4Department of Soil, Plant, Environment and Animal Production (DiSSPAPA), University of Naples Federico II, Via Universita 100, 80055 Portici, Italy, 5Interdepartmental Research Centre for Nuclear Magnetic Resonance Spectroscopy (CERMANU), Via Universita 100, 80055 Portici, Italy

Received: December 3, 2012; Accepted: January 14, 2013

Abstract

The fermented manure derivative known as Preparation
500 is traditionally used as a field spray in biodynamic
agriculture for maintaining and increasing soil fertility.
This work aimed at characterizing the product from a
microbiological standpoint and at assaying its bioactive
properties. The approach involved molecular taxonomical
characterization of the culturable microbial community;
ARISA fingerprints of the total bacteria and fungal
communities; chemical elemental macronutrient analysis
via a combustion analyzer; activity assays for six key
enzymes; bioassays for bacterial quorum sensing and
chitolipooligosaccharide production; and plant hormonelike
activity. The material was found to harbor a bacterial
community of 2.38 × 108 CFU/g dw dominated by Grampositives
with minor instances of Actinobacteria and
Gammaproteobacteria. ARISA showed a coherence of
bacterial assemblages in different preparation lots of the
same year in spite of geographic origin. Enzymatic
activities showed elevated values of β-glucosidase, alkaline
phosphatase, chitinase, and esterase. The preparation had
no quorum sensing-detectable signal, and no rhizobial nod
gene-inducing properties, but displayed a strong auxin-like
effect on plants. Enzymatic analyses indicated a bioactive
potential in the fertility and nutrient cycling contexts. The
IAA activity and microbial degradation products qualify
for a possible activity as soil biostimulants. Quantitative
details and possible modes of action are discussed.

Keywords: Preparation 500, biodynamic agriculture, auxin-like activity, hornmanure, biostimulants

References

  1. Audus, L. J. 1972. Plant Growth Substances, Vol. 1. Leonard Hill, London.
  2. Brelles-Marino, C. and E. J. Bedmar. 2001. Detection, purification and characterization of quorum-sensing signal molecules in plant-associated bacteria. J. Biotechnol. 91: 197-209.
    CrossRef
  3. Brinton, W. F. 1997. Dynamic chemical processes underlying BD horn manure (500) preparation. J. Biodynamics 214: 1-8.
  4. Canellas, L. P., L. B. Dobbss, A. L. Oliveira, J. G. Chagasa, N. O. Aguiar, V. M. Rumjanek, et al. 2012. Chemical properties of humic matter as related to induction of plant lateral roots. Eur. J. Soil Sci. 63: 315-324.
    CrossRef
  5. Canellas, L. P., A. Piccolo, L. B. Dobbss, R. Spaccini, F. L. Olivares, D. B. Zandonadi, and A. R. Façanha. 2010. Chemical composition and bioactivity properties of size-fractions separated from a vermicompost humic acid. Chemosphere 78: 457-466.
    Pubmed CrossRef
  6. Carpenter-Boggs, L., J. P. Reganold, and A. C. Kennedy. 2000. Biodynamic preparations: Short-term effects on crops, soils, and weed populations. Am. J. Altern. Agric. 15: 96-114.
    CrossRef
  7. Carpenter-Boggs, L., J. P. Reganold, and A. C. Kennedy. 2000. Organic and biodynamic management: Effects on soil biology. Soil Sci. Soc. Am. J. 64: 1651-1659.
    CrossRef
  8. Davidson, E. A., L. F. Galloway, and M. K. Strand. 1987. Assessing available carbon: Comparison of techniques across selected forest soils. Commun. Soil Sci. Plant Anal. 18: 45-64.
    CrossRef
  9. Deffune, G. and A. M. Scolfied. 1995. Effects of humic acids and three bio-dynamic preparations on the growth of wheat seedlings. Proc. 3rd ESA Congress, Abano-Podova, Paper Ref No. 3-56.
  10. D’Haeze, W. P., P. Mergaert, J. C. Promé, and M. Holsters. 2000. Nod factor requirements for efficient stem and root nodulation of the tropical legume Sesbania rostrata. J. Biol. Chem. 275: 15676-15684.
    Pubmed CrossRef
  11. Dobbs, L. B., L. P. Canellas, F. L. Olivares, N. O. Aguiar, L. E. Pereira Peres, M. Azevedo, et al. 2010. Bioactivity of chemically transformed humic matter from vermicompost on plant root growth. J. Chem. 58: 3681-3688.
  12. Ertani, A., M. Schiavon, A. Muscolo, and S. Nardi. 2013. Alfalfa plant-derived biostimulant stimulate short-term growth of salt stressed Zea mays L. plants. Plant Soil 364: 145.
    CrossRef
  13. Ertani, A., O. Francioso, V. Tugnoli, V. Righi, and S. Nardi. 2011. Effect of commercial lignosulfonate-humate on Zea mays L. metabolism. J. Agric. Food Chem. 59: 11940-11948.
    Pubmed CrossRef
  14. Fornasier, F. and A. Margon. 2007. Bovine serum albumin and Triton X-100 greatly increase phosphomonoesterases and arylsulphatase extraction yield from soil. Soil Biol. Biochem. 39: 2682-2684.
    CrossRef
  15. Koepf, H. H., B. B. Pettersson, and W. Schaumann. 1976. Biodynamic Agriculture. The Anthroposophic Press, Spring Valley, New York.
  16. Mäder, P., L. Pfiffner, A. Fließbach, and U. Niggli. 1995. Biodiversity of soil biota in biodynamic, organic and conventional farming systems, pp. 45-57. In J. Isart and J. J. Llerena (eds.). Biodiversity and Land Use: The Role of Organic Farming. Bonn.
  17. Mader, P., A. Fliessbach, D. Dubois, L. Gunst, P. Fried, and U. Niggli. 2002. Soil fertility and biodiversity in organic farming. Science 296: 1694-1697.
    Pubmed CrossRef
  18. Marchesi, J. R., T. Sato, A. J. Weightman, T. A. Martin, J. C. Fry, S. J. Hiom, and W. G. Wade. 1998. Design and evaluation of useful bacterium-specific PCR primers that amplify genes coding for bacterial 16S rRNA. Appl. Environ. Microbiol. 64:795-799.
    Pubmed KoreaMed
  19. Miller, M. B. and B. L. Bassler. 2001. Quorum sensing in bacteria. Annu. Rev. Microbiol. 55: 165-199.
    Pubmed CrossRef
  20. Nannipieri, P., J. Ascher, M. T. Ceccherini, L. Landi, G. Pietramellara, and G. Renella. 2003. Microbial diversity and soil functions. Eur. J. Soil Sci. 54: 655-670.
    CrossRef
  21. Nardi, S., G. Concheri, and G. Dell’Agnola. 1996. Biological activity of humus, pp. 361-406. In A. Piccolo (ed.). Humic Substances in Terrestrial Ecosystems. Elsevier, New York.
    CrossRef
  22. Osborn, A. M., E. R. B. Moore, and K. N. Timmis. 2000. An evaluation of terminal-restriction fragment length polymorphism (T-RFLP) analysis for the study of microbial community structure and dynamics. Environ. Microbiol. 2: 39-50.
    Pubmed CrossRef
  23. Rossen, L., C. A. Shearman, A. W. B. Johnston, and J. A. Downie. 1985. The nodD gene of Rhizobium leguminosarum is autoregulatory and in the presence of plant exudate induces the nodABC genes. EMBO J. 4: 3369-3373.
    Pubmed KoreaMed
  24. Russo, R. O. and G. P. Berlyn. 1990. The use of organic biostimulants to help low input sustainable agriculture. J. Sustain. Agric. 1: 19-42.
    CrossRef
  25. Shaw, P. D., P. Gao, S. L. Daly, C. Cha, J. E. Cronan Jr., K. L. Rinehart, and S. K. Farrand. 1997. Detecting and characterizing N-acyl-homoserine lactone signal molecules by thin-layer chromatography Proc. Natl. Acad. Sci. USA 94: 6036-6041.
    Pubmed KoreaMed CrossRef
  26. Spaccini, R., P. Mazzei, A. Squartini, M. Giannattasio, and A. Piccolo. 2012. Molecular properties of a fermented manure preparation used as field spray in biodynamic agriculture. Environ. Sci. Pollut. Res. Int. 19: 4214-4225.
    Pubmed CrossRef
  27. Stearn, W. C. 1976. Effectiveness of two biodynamic preparations on higher plants and possible mechanism s for the observed response. M.S. Thesis, Ohio State Univ., Columbus, OH.
  28. Tejada, M., C. Benítez, I. Gómez, and J. Parrado. 2011. Use of biostimulants on soil restoration: Effects on soil biochemical properties and microbial community. Appl. Soil Ecol. 49: 11- 17.
    CrossRef
  29. Tsavkelova, E. A., T. A. Cherdyntseva, S. Y. Klimova, A. I. Shestakov, S. G. Botina, and A. I. Netrusov. 2007. Orchidassociated bacteria produce indole-3-acetic acid, promote seed germination, and increase their microbial yield in response to exogenous auxin. Arch. Microbiol. 188: 655-664.
    Pubmed CrossRef
  30. Turinek, M., S. Grobelnik-Mlakar, M. Bavec, and F. Bavec. 2009. Biodynamic agriculture research progress and priorities. Renew. Agricult. Food Syst. 24: 146-154.
    CrossRef
  31. Vessey, K. J. 2003. Plant growh promoting rhizobacteria as biofertilizers. Plant Soil 255: 571-586.
    CrossRef