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Effects on plant growth and root-knot nematode infection of an endophytic GFP transformant of the nematophagous fungus Pochonia chlamydosporia

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Abstract

Pochonia chlamydosporia (Pc123) is a fungal parasite of nematode eggs which can colonize endophytically barley and tomato roots. In this paper we use culturing as well as quantitative PCR (qPCR) methods and a stable GFP transformant (Pc123gfp) to analyze the endophytic behavior of the fungus in tomato roots. We found no differences between virulence/root colonization of Pc123 and Pc123gfp on root-knot nematode Meloidogyne javanica eggs and tomato seedlings respectively. Confocal microscopy of Pc123gfp infecting M. javanica eggs revealed details of the process such as penetration hyphae in the egg shell or appressoria and associated post infection hyphae previously unseen. Pc123gfp colonization of tomato roots was low close to the root cap, but increased with the distance to form a patchy hyphal network. Pc123gfp colonized epidermal and cortex tomato root cells and induced plant defenses (papillae). qPCR unlike culturing revealed reduction in fungus root colonization (total and endophytic) with plant development. Pc123gfp was found by qPCR less rhizosphere competent than Pc123. Endophytic colonization by Pc123gfp promoted growth of both roots and shoots of tomato plants vs. uninoculated (control) plants. Tomato roots endophytically colonized by Pc123gfp and inoculated with M. javanica juveniles developed galls and egg masses which were colonized by the fungus. Our results suggest that endophytic colonization of tomato roots by P. chlamydosporia may be relevant for promoting plant growth and perhaps affect managing of root-knot nematode infestations.

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Acknowledgements

This research was funded by the Spanish Ministry of Science and Innovation Grants AGL 2008-00716/AGR and AGL 2011-29297. We thank Mr. E. Larriba (University of Alicante, Spain) for designing qPCR primers for VcP1 gene and Dr. F. J. Sorribas (Universitat Politècnica de Catalunya) for his help in the establishment of the nematode population. We also thank Professor Hans-Börje Jansson (University of Alicante) for critical comments on the manuscript.

In Memoriam

This paper is dedicated to the memory of the late Prof. Brian Kerry (Rothamsted Research, UK). He made key discoveries in the field of nematophagous fungi finding that soil suppression to plant parasitic nematodes was due to fungal parasites of nematode eggs. His work and scientific thought inspired the research of biologists all over the world including us. Many thanks Brian!

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  1. Laboratory of Plant Pathology, Department of Marine Sciences and Applied Biology, Multidisciplinary Institute for Environmental Studies (MIES) Ramon Margalef, University of Alicante, Apto 99, 03080, Alicante, Spain

    N. Escudero & L. V. Lopez-Llorca

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  2. L. V. Lopez-Llorca
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Correspondence to N. Escudero.

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Escudero, N., Lopez-Llorca, L.V. Effects on plant growth and root-knot nematode infection of an endophytic GFP transformant of the nematophagous fungus Pochonia chlamydosporia . Symbiosis 57, 33–42 (2012). https://doi.org/10.1007/s13199-012-0173-3

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