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. 1985 Aug;50(2):398–405. doi: 10.1128/aem.50.2.398-405.1985

Inoculant Production with Diluted Liquid Cultures of Rhizobium spp. and Autoclaved Peat: Evaluation of Diluents, Rhizobium spp., Peats, Sterility Requirements, Storage, and Plant Effectiveness

P Somasegaran 1
PMCID: PMC238633  PMID: 16346860

Abstract

Fully grown broth cultures of various fast- and slow-growing rhizobia were deliberately diluted with various diluents before their aseptic incorporation into autoclaved peat in polypropylene bags (aseptic method) or mixed with the peat autoclaved in trays (tray method). In a factorial experiment with the aseptic method, autoclaved and irradiated peat samples from five countries were used to prepare inoculants with water-diluted cultures of three Rhizobium spp. When distilled water was used as the diluent, the multiplication and survival of rhizobia in the peat was similar to that with diluents having a high nutrient status when the aseptic method was used. In the factorial experiment, the mean viable counts per gram of inoculant were log 9.23 (strain TAL 102) > log 8.92 (strain TAL 82) > log 7.89 (strain TAL 182) after 24 weeks of storage at 28°C. The peat from Argentina was the most superior for the three Rhizobium spp., with a mean viable count of log 9.0 per g at the end of the storage period. The quality of inoculants produced with diluted cultures was significantly (P = 0.05) better with irradiated than with autoclaved peat, as shown from the factorial experiment. With the tray method, rhizobia in cultures diluted 1,000-fold or less multiplied and stored satisfactorily in the presence of postinoculation contaminants, as determined by plate counts, membrane filter immunofluorescence, and plant infection procedures. All strains of rhizobia used in both the methods showed various degrees of population decline in the inoculants when stored at 28°C. Fast- and slow-growing rhizobia in matured inoculants produced by the two methods showed significant (P < 0.01) decline in viability when stored at 4°C, whereas the viability of some strains increased significantly (P < 0.01) at the same temperature. The plant effectiveness of inoculants produced with diluted cultures and autoclaved peat did not differ significantly from that of inoculants produced with undiluted cultures and gamma-irradiated peat.

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Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Bohlool B. B., Schmidt E. L. Nonspecific staining: its control in immunofluorescence examination of soil. Science. 1968 Nov 29;162(3857):1012–1014. doi: 10.1126/science.162.3857.1012. [DOI] [PubMed] [Google Scholar]
  2. Crist D. K., Wyza R. E., Mills K. K., Bauer W. D., Evans W. R. Preservation of Rhizobium viability and symbiotic infectivity by suspension in water. Appl Environ Microbiol. 1984 May;47(5):895–900. doi: 10.1128/aem.47.5.895-900.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Hobbie J. E., Daley R. J., Jasper S. Use of nuclepore filters for counting bacteria by fluorescence microscopy. Appl Environ Microbiol. 1977 May;33(5):1225–1228. doi: 10.1128/aem.33.5.1225-1228.1977. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Hoben H. J., Somasegaran P. Comparison of the Pour, Spread, and Drop Plate Methods for Enumeration of Rhizobium spp. in Inoculants Made from Presterilized Peat. Appl Environ Microbiol. 1982 Nov;44(5):1246–1247. doi: 10.1128/aem.44.5.1246-1247.1982. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Schmidt E. L., Bakole R. O., Bohlool B. B. Fluorescent-antibody approach to study of rhizobia in soil. J Bacteriol. 1968 Jun;95(6):1987–1992. doi: 10.1128/jb.95.6.1987-1992.1968. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Somasegaran P., Halliday J. Dilution of liquid Rhizobium cultures to increase production capacity of inoculant plants. Appl Environ Microbiol. 1982 Aug;44(2):330–333. doi: 10.1128/aem.44.2.330-333.1982. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Strijdom B. W., van Rensburg H. J. Effect of steam sterilization and gamma irradiation of peat on quality of Rhizobium inoculants. Appl Environ Microbiol. 1981 Jun;41(6):1344–1347. doi: 10.1128/aem.41.6.1344-1347.1981. [DOI] [PMC free article] [PubMed] [Google Scholar]

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