Summary
A chemostat was used as a model system to study competitive interactions of diazotrophic microorganisms. Enrichment experiments were carried out under microaerobic conditions (8.7 μmol O2/l) with malate as the sole carbon source. The starting material was a Korean rice soil including intact root pieces. The enrichment process was governed by the dilution rate. High dilution rates resulted in the enrichment ofAzospirillum lipoferum, whereas low dilution rates led to the predominance of an unidentified organism, named Isolate R. Dilution rates were set in the range from D=0.005 to D=0.1 h−1. The growth kinetics of both organisms followed Monod's model in the enrichment culture. From the experiments, the maximum specific growth rate ofA. lipoferum and Isolate R were 0.069 h−1 and 0.025 h−1, respectively. The corresponding Ks-values were 8.4 and 0.9 (mg. 1−1). The point of theoretical coexistence of both organisms was calculated to occur at a substrate concentration of s=3.0 (mg.l−1) with a growth of rate μ=0.018 h−1. Hence the preset nutritional niches occupied by at least two organisms.Azospirillum lipoferum seems to represent the copiotroph microflora and Isolate R is of the oligotroph type. In addition to its high substrate affinity Isolate R liberatedca. 75% of the fixed nitrogen into the medium, which indicates its potential role for mutualistic interactions in the rhizosphere.
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References
Aragno M and Schlegel H G 1981 The hydrogen oxidizing bacteria.In The Prokaryotes. Eds. M P Starr, H Stolp, H G Trüper, A Balows and H G Schlegel. pp 865–893, Springer, Berlin.
Armstrong W 1969 Rhizosphere oxidation in rice: An analysis of intervarietal differences in oxygen flux from the roots. Physiol. Plant. 22, 296–303.
Balandreau J 1983 Microbiology of the association. Can. J. Microbiol. 29, 851–859.
Bally R D, Thomas-Bauzon D, Heulin T, Balandreau J, Richard C and De Ley J 1983 Determination of the most frequent N2-fixing bacteria in the rice rhizosphere. Can. J. Microbiol. 29, 881–887.
Bergersen F J 1980 Measurement of nitrogen fixation by direct means.In Methods for Evaluating Biological Nitrogen Fixation. Ed F J Bergersen. pp 65–110, John Wiley and Sons, Chichester.
Boureau M 1977 Application de la chromatographie en phase gazeuse a l'etude de l' exudation racinaire du riz. Cah. ORSTOM, ser Biol. 7, 75–81.
Buchanan R E and Gibbons N E 1974 Bergey's Manual of Determinative Bacteriology, 8th ed. Williams and Wilkins Company, Baltimore.
Caceres E A R 1982 Improved medium for isolation of Azospirillum spp. Appl. Environ. Microbiol. 44, 990–991.
Coleman D C, Cole C V, Hunt H W and Klein D A 1978 Trophic interactions in soils as they affect energy and nutrient dynamics. I. Introduction. Microb. Ecol. 4, 345–349.
Dalton H 1980 The cultivation of diazotrophic microorganisms.In. Methods for Evaluating Biological Nitrogen Fixation. Ed. F J Bergersen. pp. 13–64, John Wiley and Sons, Chichester.
Döbereiner J and De Polli H 1980 Diazotrophic rhizocoenosis.In Nitrogen Fixation. Eds W D P Stewart and J R Gallon. pp 301–333, Academic Press, London.
Fredrickson A G and Tsichuya H M 1977 Microbial kinetics and dynamics.In Chemical Reactor Theory. Eds L Lapidus and N R Amundson. pp 405–483. Prentice-Hall, Englewood Cliffs.
Gerhardt P, Murray R G E, Costilow R N, Nester E W, Wood W A, Kreig N R and Phillips G B 1981 Manual of Methods for General Bacteriology. ASM, Washington.
Hartmann A, Kleiner D and Klingmüller W 1984 Ammonium uptake and release by Azospirillum.In Advances in Nitrogen fixation Research. Eds. C Veeger and W E Newton. pp 227, Nijhoff/Junk, The Hague.
Herbert D, Elsworth R and Telling R C 1956 The continuous culture of bacteria: A theoretical and experimental study. J. Gen. Microbiol. 14, 601–622.
Herzberg M A, Klein D A and Coleman D C 1978 Trophic interactions in soils as they affect energy and nutrient dynamics II. Physiological responses of selected rhizosphere bacteria. Microb., Ecol. 4, 351–359.
Hohorst H J 1970 L-(−)-Malat, Bestimmung mit Malatdehydrogenase und NAD.In Methoden der Enzymatischen Analyse. Ed. H U Bergmeyer. pp 1548, Verlag Chemie, Weinheim.
Jannasch H W 1967 Enrichment of aquatic bacteria in continuous culture. Arch. Microbiol. 59, 165–173.
King E O, Ward M K and Raney D E 1954 Two simple media for demonstration of pyocyanin and fluorescin. J. Lab. Cli. Med. 44, 301–307.
Kloss M., Iwannek K-H and Fendrik I 1983 Physiological properties ofAzospirillum brasilense Sp7 in a malate limited chemostat. J. Gen. Appl. Microbiol (Tokyo) 29, 447–457.
Kloss M, Iwannek K-H and Fendrik I 1984 Malate limited growth ofAzospirillum brasilense Sp7 in a chemostat.In Advances in Nitrogen Fixation Research. Eds C Veeger and W E Newton. p 231 Nijhoff/Junk, The Hague.
Kloss M, Iwannek K-H, Fendrik I and Niemann E-G 1984 Organic acids in the root exudates ofDiplachne fusca (Linn.) Beauv. Exp. Environ. Bot 24, 179–188.
Kuenen J G and Harder W 1982 Microbial competition in continuous culture.In Experimental Microbial Ecology. Eds. R G Burns and J H Slater. pp 342–367. Backwell Scientific Publications, Oxford.
Ladha J K, Barraquio W L and Watanabe I 1982 Immunological techniques to identify Azospirillum associated with wetland rice. Can. J. Microbiol. 28, 478–485.
Loh W H T, Randles C, Sharp W R and Millers R H 1984 Intermediary carbon metabolism ofAzospirillum brasilense. J. Bacteriol. 158, 264–268.
Lynch J M 1982 The rhizosphere.In Experimental Microbial Ecology. Eds. R G Burns and J H Slater. pp 395–411, Blackwell Scientific Publications, Oxford.
Monod J 1949 The growth of bacterial cultures. Annu. Rev. Microbiol. 3, 371–394.
Müller-Beissenhirz W and Keller H 1965 Zur Bestimmung des Blutammoniaks. Klin. Woschr. 43 43–49.
Newman E I and Watson A 1977 Microbial abundance in the rhizosphere: A computer model. Plant and Soil 48, 17–56.
Parkes R J 1982 Methods for enriching, isolating, and analysing microbial communities in laboratory systems.In Microbial Interactions and Communities. eds. A T Bull and J H Slater. pp 45–102, Academic Press, London.
Patriquin D G, Döbereiner J and Jain D K 1983 Sites and processes of the association between diazotrophs and grasses. Can. J Microbiol. 29, 900–915.
Pfennig N and Jannasch H W 1962 Biologische Grundfragen bei der homokontinuierlichen Kultur von Mikroorganismen. Ergeb. Biol. 25, 93–135.
Pirt S J 1975 The Principles of Microbe and Cell Cultivation. Blackwell Scientific Publications, Oxford.
Powell E O 1958 Criteria for growth of contaminants and mutants in continuous culture. J. Gen. Microbiol. 18, 259–268.
Slater J H and Bull A T 1978 Interactions between microbial populations.In Companion to Microbiology. Eds A T Bull and P M Meadow. 181–206, Longman, London.
Slater J H and Hardmann D J 1982 Microbiol ecology in the laboratory: Experimental systems.In Experimental Microbial Ecology. Eds. R G Burns and J H Slater. pp. 255–274, Blackwell Scientific Publications Oxford.
Tarrand J J, Krieg N R and Döbereiner J 1978 A taxonomic study of theSpirillum lipoferum group, with descriptions of a new genus,Azospirillum gen nov. and two new species,Azospirillum lipoferum (Beijerinck) comb. Nov. andAzospirillum brasilense sp. nov. Can. J. Microbiol. 24, 967–980.
Tempest D W and Neijssel O M 1978 Ecophysiological aspects of microbial growth in aerobic nutrient-limited environments. Adv. Microbial Ecol. 2, 105–153.
Thomas-Bauzon D P, Weinhard P, Villecourt P and Balandreau J 1982 The spermosphere model I. Its use in growing, counting, and isolating N2-fixing bacteria from the rhizosphere of rice. Can. J. Microbiol. 28, 922–928.
Trolldenier G 1977 Influence of some environmental factors on nitrogen fixation in the rhizosphere of rice. Plant and Soil 47, 203–217.
Vancura V 1964 Root exudates of plants. I. Analysis of root exudates of barley and wheat in their initial phases of growth. Plant and Soil 21, 231–248.
Veldkamp H and Jannasch H W 1972 Mixed culture studies with the chemostat. J. Appl. Chem. Biotechnol. 22 105–123.
Warembourg F R and Billies G 1979 Estimating the carbon transfer in the rhizosphere.In The Soil-Root Interface. Eds. J L Harley and R Scott-Russel. pp 181–196, Academic Press, London.
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Kloss, M., Iwannek, K.H., Fendrik, I. et al. Enrichment of diazotrophic bacteria from rice soil in continuous culture. Plant Soil 90, 151–164 (1986). https://doi.org/10.1007/BF02277394
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DOI: https://doi.org/10.1007/BF02277394