Skip to main content
PMC home page
Applied and Environmental Microbiology logoLink to Applied and Environmental Microbiology
. 1990 Mar;56(3):686–692. doi: 10.1128/aem.56.3.686-692.1990

Physiological and Morphological Changes Induced by Nutrient Limitation of Pseudomonas fluorescens 378 in Continuous Culture

Anders Persson 1, Göran Molin 1,*, Claes Weibull 1
PMCID: PMC183406  PMID: 16348142

Abstract

Pseudomonas fluorescens 378 was studied in continuous culture at a dilution rate of 0.05 or 0.15 h−1 and under a limitation of carbon/energy, nitrogen, phosphorus, iron(III), or oxygen. Cultures were examined for nutritional consumption, production of biosurfactant AP-6 and lipase, and electron microscopy morphology. Morphological features were lysis and plasmolysis of the cells, vacuoles in the cells, granules in cell nuclei, and DNA coagulation during transmission electron microscopy preparation. Biosurfactant and lipase production were lost after 8 to 15 retention times, but under iron limitation and at low dilution rate they were maintained for more than 30 retention times. Consumption of nutrients varied between different cultures. Between 2.4 and 6.0 g of succinic acid per g (dry weight) was consumed; the highest value was obtained under phosphorus limitation. The uptake of nitrogen was mostly about 0.16 g/g (dry weight), and that of phosphorus varied between 13 and 58 mg/g (dry weight). Phosphorus-limited cells reduced their phosphorus consumption by at least 50% compared with other limitations. Cell morphology varied among different cultures. Up to 25% cell lysis occurred at the higher dilution rate. The frequencies of plasmolysis varied between 0 and 85%. Granules in nuclei were found in 65 to 100% of the cells. Vacuoles appeared mostly in low numbers, but at the lower dilution rate under phosphorus or iron limitation the frequencies increased to between 25 and 85%. At high dilution rate, the DNA coagulated in 30 to 70% of the cells. Multivariate data analysis demonstrated a general difference between the two tested dilution rates; i.e., both nutritional and morphological features differed more between the two tested dilution rates than between the different limitations. Cultures at the lower dilution rate changed more with time; this was especially pronounced for phosphorus or iron limitation. The data analysis also showed a correlation between plasmolysis or vacuoles in the cells and an increased carbon uptake under phosphorus limitation.

Full text

PDF
686

Images in this article

689Image
on p.689
690Image
on p.690

Selected References

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

  1. Bjorn M. J., Sokol P. A., Iglewski B. H. Influence of iron on yields of extracellular products in Pseudomonas aeruginosa cultures. J Bacteriol. 1979 Apr;138(1):193–200. doi: 10.1128/jb.138.1.193-200.1979. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Blomquist G., Johansson E., Söderström B., Wold S. Classification of fungi by means of pyrolysis-gas chromatography-pattern recognition. J Chromatogr. 1979 May 11;173(1):19–32. doi: 10.1016/s0021-9673(01)80442-8. [DOI] [PubMed] [Google Scholar]
  3. CHANEY A. L., MARBACH E. P. Modified reagents for determination of urea and ammonia. Clin Chem. 1962 Apr;8:130–132. [PubMed] [Google Scholar]
  4. Dykhuizen D. E., Hartl D. L. Selection in chemostats. Microbiol Rev. 1983 Jun;47(2):150–168. doi: 10.1128/mr.47.2.150-168.1983. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Fernandez L., San José C., Cholette H., McKellar R. C. Characterization of a pyoverdine-deficient mutant of Pseudomonas fluorescens impaired in the secretion of extracellular lipase. Arch Microbiol. 1988;150(6):523–528. doi: 10.1007/BF00408243. [DOI] [PubMed] [Google Scholar]
  6. Guerra-Santos L., Käppeli O., Fiechter A. Pseudomonas aeruginosa biosurfactant production in continuous culture with glucose as carbon source. Appl Environ Microbiol. 1984 Aug;48(2):301–305. doi: 10.1128/aem.48.2.301-305.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Harder W., Dijkhuizen L. Physiological responses to nutrient limitation. Annu Rev Microbiol. 1983;37:1–23. doi: 10.1146/annurev.mi.37.100183.000245. [DOI] [PubMed] [Google Scholar]
  8. Höfle M. G. Long-Term Changes in Chemostat Cultures of Cytophaga johnsonae. Appl Environ Microbiol. 1983 Nov;46(5):1045–1053. doi: 10.1128/aem.46.5.1045-1053.1983. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Lee W. S., Cooper J. K., Lynch W. H. Membrane enzymes associated with the dissimilation of some citric acid cycle substrates and production of extracellular oxidation products in chemostat cultures of Pseudomonas fluorescens. Can J Microbiol. 1984 Mar;30(3):396–405. doi: 10.1139/m84-058. [DOI] [PubMed] [Google Scholar]
  10. Minnikin D. E., Abdolrahimzadeh H. The replacement of phosphatidylethanolamine and acidic phospholipids by an ornithine-amide lipid and a minor phosphorus-free lipid in Pseudomonas fluorescens NCMB 129. FEBS Lett. 1974 Aug 1;43(3):257–260. doi: 10.1016/0014-5793(74)80655-1. [DOI] [PubMed] [Google Scholar]
  11. NOVICK A., SZILARD L. Description of the chemostat. Science. 1950 Dec 15;112(2920):715–716. doi: 10.1126/science.112.2920.715. [DOI] [PubMed] [Google Scholar]
  12. NOVICK A., SZILARD L. Experiments with the Chemostat on spontaneous mutations of bacteria. Proc Natl Acad Sci U S A. 1950 Dec;36(12):708–719. doi: 10.1073/pnas.36.12.708. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. RYTER A., KELLENBERGER E., BIRCHANDERSEN A., MAALOE O. Etude au microscope électronique de plasmas contenant de l'acide désoxyribonucliéique. I. Les nucléoides des bactéries en croissance active. Z Naturforsch B. 1958 Sep;13B(9):597–605. [PubMed] [Google Scholar]
  14. Schwyn B., Neilands J. B. Universal chemical assay for the detection and determination of siderophores. Anal Biochem. 1987 Jan;160(1):47–56. doi: 10.1016/0003-2697(87)90612-9. [DOI] [PubMed] [Google Scholar]
  15. Stanier R. Y., Palleroni N. J., Doudoroff M. The aerobic pseudomonads: a taxonomic study. J Gen Microbiol. 1966 May;43(2):159–271. doi: 10.1099/00221287-43-2-159. [DOI] [PubMed] [Google Scholar]
  16. de Weger L. A., van Boxtel R., van der Burg B., Gruters R. A., Geels F. P., Schippers B., Lugtenberg B. Siderophores and outer membrane proteins of antagonistic, plant-growth-stimulating, root-colonizing Pseudomonas spp. J Bacteriol. 1986 Feb;165(2):585–594. doi: 10.1128/jb.165.2.585-594.1986. [DOI] [PMC free article] [PubMed] [Google Scholar]

Articles from Applied and Environmental Microbiology are provided here courtesy of American Society for Microbiology (ASM)

RESOURCES