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Vesicle trafficking via the Spitzenkörper during hyphal tip growth in Rhizoctonia solani

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Abstract

Growing hyphae of Rhizoctonia solani were stained with the endocytic marker dye FM4-64 and imaged by confocal microscopy. Staining of the plasma membrane was followed by labeling of organelles in the cytoplasm (after ~1 min) and of the Spitzenkörper (Spk; after ~2 min). Fluorescence recovery after photobleaching (FRAP) of the stained Spk demonstrated the vectorial flow of secretory vesicles from the apical cytoplasm to the Spk. This flux was modelled in a two-compartment model. The turnover time of the vesicles of the Spk was estimated to be 1.3–2.5 min. These results are roughly consistent with the expected flux of vesicles through the Spk based on the number of secretory vesicles within the Spk and the number of secretory vesicles that would be necessary to fuse with the apical plasma membrane to maintain hyphal extension rates. These results suggest that membrane retrieval via endocytosis is not as significant as previously suggested.

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Acknowledgments

JD was funded by a Leverhulme Grant. The authors are very much indebted to Nick Read, University of Edinburgh, for in depth discussions and suggestions during writing of the manuscript. The authors thank N. Christine Øien for assistance with some of the experiments. The authors are also indebted to Patrick Hickey and Sabine Fisher-Parton for helpful suggestions and technical advice and Rosamaria López-Franco and Charles Bracker for useful discussions.

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  1. Jan Dijksterhuis

    Present address: Applied and Industrial Mycology, CBS Fungal Biodiversity Centre, Uppsalalaan 8, 3584 CT, Utrecht, The Netherlands

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  1. Fungal Cell Biology Group, University of Edinburgh, Daniel Rutherford Building, Edinburgh, EH9 3JH, UK

    Jan Dijksterhuis

  2. Molecular Cell Physiology, VU University Amsterdam, De Boelelaan 1085, 1081 HV, Amsterdam, The Netherlands

    Douwe Molenaar

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Correspondence to Jan Dijksterhuis.

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Dijksterhuis, J., Molenaar, D. Vesicle trafficking via the Spitzenkörper during hyphal tip growth in Rhizoctonia solani . Antonie van Leeuwenhoek 103, 921–931 (2013). https://doi.org/10.1007/s10482-012-9873-1

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