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Comparison of the development in planta of a pyrrolnitrin-resistant mutant of Botrytis cinerea and its sensitive wild-type parent isolate

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Abstract

Botrytis cinerea is able to build-up resistance to pyrrolnitrin, an antibiotic produced by diverse biocontrol agents, possibly compromising the durability of this method of disease control. The development of two near-isogenic lines of B. cinerea differing in their level of resistance to pyrrolnitrin was compared in tomato plants and on PDA medium. In tomato plants, significant differences in the percentage of infected petioles 1 day after inoculation and in symptom progression on petioles and stems were observed between the resistant mutant and the sensitive wild-type parent, suggesting a difference in their level of aggressiveness. Cytohistological investigations revealed that conidia of both near-isogenic lines germinated 6 h after inoculation and mycelium developed within petiole tissues 12 h after inoculation. However, while the wild-type parent isolate spread throughout the petiole and rapidly invaded the stem tissues via the leaf-abscission zone 72 h after inoculation, the pyrrolnitrin-resistant mutant failed to extend beyond petiole tissues to invade the stem. Moreover, 72 h after inoculation, the mycelial development of the pyrrolnitrin-resistant mutant was accompanied by abnormal glycogen accumulation and chlamydospore-like cell formation. In contrast, wild-type parent mycelium was normally structured with intensive colonization of stem tissues. Additionally, on PDA medium the mycelium of the pyrrolnitrin-resistant mutant was less vigorous than the wild-type isolate. These results suggest that the acquisition of pyrrolnitrin-resistance in B. cinerea is accompanied by changes in mycelial structure and reduction in mycelial growth, leading to a noticeable loss of aggressiveness on tomato plants.

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Acknowledgements

The authors gratefully acknowledge Claire Troulet and Gisèle Riqueau for excellent technical assistance in the plant tests. They also acknowledge Isabelle Bornard for the additional microscopical observations of the hyphal structure done in vitro and Veronique Decognet for critically reading the manuscript. This work was supported in part by the Agence Nationale de la Recherche (ANR-Ecoserre project) and by INRA (SPE project). A grant for studies was provided by the Syrian government for Sakhr Ajouz.

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Authors and Affiliations

  1. INRA, UR407, Plant Pathology Unit, Domaine St Maurice, 84140, Montfavet, France

    Sakhr Ajouz, Marc Bardin & Philippe C. Nicot

  2. Université d’Avignon, BP 21239, 84916, Avignon Cedex 9, France

    Mohamed El Maâtaoui

  3. INRA, UMR 406 UAPV/INRA, Avignon, 84914, France

    Mohamed El Maâtaoui

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  1. Sakhr Ajouz
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  2. Marc Bardin
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  3. Philippe C. Nicot
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  4. Mohamed El Maâtaoui
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Correspondence to Marc Bardin.

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Ajouz, S., Bardin, M., Nicot, P.C. et al. Comparison of the development in planta of a pyrrolnitrin-resistant mutant of Botrytis cinerea and its sensitive wild-type parent isolate. Eur J Plant Pathol 129, 31–42 (2011). https://doi.org/10.1007/s10658-010-9638-5

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