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Related Experiment Videos

Glutamine utilization by Rhizobium etli.

Rosarita Tatè1, Simona Ferraioli, Stefania Filosa

  • 1Institute of Genetics and Biophysics A. Buzzati-Traverso, CNR, Via G. Marconi 10, 80125 Naples, Italy.

Molecular Plant-Microbe Interactions : MPMI
|July 10, 2004
PubMed
Summary

Rhizobium etli utilizes glutamine (Gln) for energy via the citric acid cycle. Gln metabolism impacts cell shape and stress resistance, with key genes dispensable for nodulation but essential for growth.

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Area of Science:

  • Microbiology
  • Biochemistry
  • Bacterial Physiology

Background:

  • Rhizobium etli are nitrogen-fixing bacteria crucial for legume symbiosis.
  • Understanding nutrient utilization is key to optimizing bacterial growth and function.
  • Glutamine (Gln) is a potential carbon and energy source for bacteria.

Purpose of the Study:

  • To investigate the utilization of glutamine (Gln) as a carbon and energy source by Rhizobium etli.
  • To identify genes involved in Gln metabolism and their physiological consequences.
  • To determine the role of Gln utilization genes in bacterial nodulation ability.

Main Methods:

  • Tn5-induced mutagenesis to isolate mutants with defects in Gln utilization.
  • Growth assays on various media.

Related Experiment Videos

  • Glutamine uptake assays.
  • Measurement of CO2 production from Gln.
  • Analysis of bacterial cell morphology and oxidative stress susceptibility.
  • Nodulation assays on host plants.

    • Main Results:

    • Identified genes essential for Gln utilization, including amino acid transporters and glutamate dehydrogenase.
    • Demonstrated Gln degradation via the citric acid cycle.
    • Observed morphological changes (bacillary to coccoid) and increased oxidative stress susceptibility in Gln-grown cells.
    • Showed that glucose addition reversed these effects, indicating carbon starvation conditions.
    • Found Gln transport and degradation genes dispensable for nodulation, unlike gluconeogenesis and nucleotide biosynthesis genes.

    Conclusions:

    • Rhizobium etli can utilize glutamine as a sole carbon and energy source, involving specific transporters and metabolic pathways.
    • Gln metabolism induces physiological changes indicative of carbon starvation and oxidative stress.
    • Genes for Gln transport and degradation are not essential for symbiosis, highlighting distinct requirements for free-living growth versus nodulation.