Novel method for identifying bacterial mutants with reduced epiphytic fitness

  • Lindow S
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In order to identify novel traits involved in epiphytic colonization, a technique for the rapid identification of bacterial mutants with quantitatively different population sizes in a natural habitat based on measurements of ice nucleation activity was developed. The threshold freezing temperatures of leaves harboring different numbers of cells of ice nucleation-active Pseudomonas syringae B728a differed substantially. While few leaves containing less than about 106 cells per g (fresh weight) froze at assay temperatures of -2.75{degrees}C or higher, nearly all leaves froze at these temperatures when population sizes of this strain increased to about 107 cells per g (fresh weight). Presumptive epiphytic fitness mutants could readily be identified as strains which initiated freezing in fewer leaves than did other strains within a given experiment. Most Tn5-induced mutants of strain B728a which conferred a low frequency of ice nucleation on inoculated bean leaves generally had a smaller population size than the parental strain at the time of the leaf freezing assay. The leaf freezing assay was capable of differentiating samples which varied by approximately three- to fivefold in mean bacterial population size.

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  • ISSN: 00992240
  • SCOPUS: 2-s2.0-0027289358
  • SGR: 0027289358
  • PUI: 23141175
  • PMID: 16348938
  • ISBN: 0099-2240


  • S. E. Lindow

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