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Viability and physiological state transitions of Rhizopus oligosporus sporangiospores in tempe starter culture

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Abstract

The viability and various physiological characteristics of individual sporangiospores of Rhizopus oligosporus in tempe starter cultures that had been stored for 8, 10, 16 and 30 months were examined by flow cytometry in combination with fluorescent dyes. Besides live, dead, and dormant spores we distinguished a category of sublethally damaged spores. Results indicated that the shelf-life of tempe starters was not limited by the death of spores, but by sublethal damage to spores as well as by dormancy which can be overcome by resuscitation, respiratory activation. During storage, the number of dormant and sublethally damaged spores increased: the longer the starter cultures were stored, the less dormant spores could still be activated. In contrast, the transition from sublethally damaged (spores that are not able to transform cFDA and emit green fluorescence except by activation treatment) to activated spores did not decrease with longer storage. However, after very long (30 months) storage, sublethally damaged spores could still be activated but could not germinate anymore. The shelf-life of spores in tempe starter is related to the physiological state of spores being sublethally damaged; a mechanism of physiological state transitions of R. oligosporus sporangiospores is proposed.

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Abbreviations

cFDA:

carboxyfluoresceindiacetate

FCM:

flow cytometry

PI:

propidium iodide

TOTO-1:

1-(4,4,7,7-tetramethyl-4,7-diazaundecamethylene)-bis-4-[3-methyl-2,3dihydro (benzo-1,3-oxazole)-2-methylidene]-1-(3-trimethyl ammonium propyl)-pyridinium tetraiodide

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Acknowledgements

This work was supported in part by IFS Grant No. C/3399-1. N.V. Thanh gratefully acknowledges receipt of the sandwich fellowship from the Netherlands organization for international cooperation in higher education (NUFFIC). The authors thank Patrick Verbaarschot (Laboratory of Entomology, Wageningen University) for his assistance with FCM analyses.

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Authors and Affiliations

  1. Biotechnology Research and Development Institute, Can Tho University, Can Tho, Vietnam

    Nguyen V. Thanh

  2. Laboratory of Food Microbiology, Wageningen University, P.O. Box 8129, 6700 EV, Wageningen, The Netherlands

    Nguyen V. Thanh, Frans M. Rombouts & M. J. Rob Nout

Authors
  1. Nguyen V. Thanh
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  2. Frans M. Rombouts
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  3. M. J. Rob Nout
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Correspondence to M. J. Rob Nout.

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Thanh, N.V., Rombouts, F.M. & Nout, M.R. Viability and physiological state transitions of Rhizopus oligosporus sporangiospores in tempe starter culture. Antonie van Leeuwenhoek 91, 35–44 (2007). https://doi.org/10.1007/s10482-006-9093-7

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  • DOI: https://doi.org/10.1007/s10482-006-9093-7

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