Abstract
Reversible transition of the phytopathogenic gram-negative bacterium Erwinia carotovora, subsp. atroseptica, strain SCRI1043, to a dormant state was demonstrated; it was associated with a complete loss of cell ability to form colonies on the standard medium, i.e., with acquiring “non-culturability”. Entering of Erwinia cells to a nonculturable state occurred after long-term incubation (100–150 days) of the stationary-phase cell suspensions in either a fresh complete medium or in the carbon-free mineral medium or treatment with a chemical analogue of microbial anabiosis autoinducers (4 × 10−4 M of C12-alkylhydroxybenzene, AHB). However, confocal laser microscopy of the cells stained with the Live/Dead BacLight kit revealed that the majority of E. carotovora cells (90%) from long-incubated suspensions retained membrane integrity. In these suspensions, round cells of smaller size prevailed, with the envelope, containing an electron-dense outer layer and an underlying layer of lower density; the cytoplasm was coarse-granulated. Detection of “nonculturable” E. carotovora cells by quantitative real-time PCR analysis (Q-PCR) with specific primers by using standard procedures of sample preparation was shown to be inefficient. A special procedure including cell washing from the incubation medium in the absence of growth stimulation was developed, which promoted recovery of the colony-forming ability of the cells (up to 10% of the initial CFU number) and improved cell detection by Q-PCR from the number of genomic copies. The results provided further insight into the ways of long-term survival of phytopathogenic bacteria under environmental changes and carbon starvation.
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Original Russian Text © V.Yu. Gorshkov, O.E. Petrova, N.E. Mukhametshina, M.V. Ageeva, A.L. Mulyukin, Yu.V. Gogolev, 2009, published in Mikrobiologiya, 2009, Vol. 78, No. 5, pp. 647–655.
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Gorshkov, V.Y., Petrova, O.E., Mukhametshina, N.E. et al. Formation of “Nonculturable” dormant forms of the phytopathogenic enterobacterium Erwinia carotovora . Microbiology 78, 585–592 (2009). https://doi.org/10.1134/S0026261709050099
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DOI: https://doi.org/10.1134/S0026261709050099