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Thermo-stability, dose effects and shelf-life of antifungal metabolite-containing supernatants produced by Xenorhabdus szentirmaii

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Abstract

Entomopathogenic nematodes in the genus Steinernema are associated with Xenorhabdus spp. bacteria. When steinernematid colonise an insect host the nematode-bacterium association overcomes the insect immune system and kills the host within 48 h. Xenorhabdus spp. produce secondary metabolites that are antifungal to protect nematode-infected cadavers from fungal colonization. The concentrated, or cell-free metabolites of X. szentirmaii exhibit high toxicity against various fungal plant pathogens and show potential as natural bio-fungicides. In the current study, we determined 1) thermo-stability, 2) dose-response, and 3) shelf-life of antifungal metabolites of X. szentirmaii against Monilinia fructicola (cause of brown rot of peach and other stone fruit) and Glomerella cingulata (cause of antharacnose). Thermo-stability was determined by autoclaving bacterial culture broths (121 °C and 15 psi for 15 min) and measuring fungal growth on in potato dextrose agar (PDA) containing 10% of the supernatants. Autoclaving had no impact on the antifungal activity of the secondary metabolites. Over a test period of 9 months, the activity of both extract types did not decline when stored at 4 or 20 °C. A dose-response study (10, 20, 40, 60, 80 and 100% supernatant-containing metabolite) using both phytopathogens demonstrated that a greater dose of supernatant increased antifungal activity. The antifungal-metabolite containing supernatant of X. szentirmaii has potential as a bio-fungicide. These results demonstrate the metabolite(s) are thermo-stable, they have a long shelf-life and require no stabilizing formulation, even at room temperature.

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Acknowledgements

We thank to Technical and Research Council of Turkey (TUBITAK) for supporting S. Hazir within the 2219 fellowship program. D. I. Shapiro-Ilan and C.H. Bock acknowledge the research was partially supported by the USDA National Programs for Crop Protection & Quarantine, and Crop Production (Project Numbers: 6042-22000-023-00D and 6042-21220-012-00D, respectively). We also thank Stacy Byrd, Amy Euston, Benjamin Graham, Domonique Roth-Alday, Unicka Stokes, Wanda Evans, Amy Eubanks and Seth Richards for their valuable technical assistance. This article reports the results of research only. Mention of a proprietary product does not constitute an endorsement or recommendation by the USDA for its use.

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Authors and Affiliations

  1. Faculty of Arts and Sciences, Department of Biology, Adnan Menderes University, 09100, Aydin, Turkey

    Selcuk Hazir

  2. Southeastern Fruit and Tree Nut Research Laboratory, USDA-ARS, Byron, GA, 31008, USA

    David I. Shapiro-Ilan & Clive H. Bock

  3. L. G. Leite Instituto Biologico, APTA, CP 70, Campinas, SP, 13001-970, Brazil

    Luis G. Leite

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  1. Selcuk Hazir
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  2. David I. Shapiro-Ilan
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  3. Clive H. Bock
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  4. Luis G. Leite
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Correspondence to Selcuk Hazir or David I. Shapiro-Ilan.

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Hazir, S., Shapiro-Ilan, D.I., Bock, C.H. et al. Thermo-stability, dose effects and shelf-life of antifungal metabolite-containing supernatants produced by Xenorhabdus szentirmaii . Eur J Plant Pathol 150, 297–306 (2018). https://doi.org/10.1007/s10658-017-1277-7

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