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15 - In Vitro Methods for Plant–Microbe Interaction and Biocontrol Studies in European Ash (Fraxinus excelsior L.)

from Part IV - Endophytes for Novel Biomolecules and In Vitro Methods

Published online by Cambridge University Press:  01 April 2019

By
Anindita Lahiri ,
Gerry C. Douglas ,
Brian R. Murphy  and
Trevor R. Hodkinson
Edited by
Trevor R. Hodkinson ,
Fiona M. Doohan ,
Matthew J. Saunders  and
Brian R. Murphy
Trevor R. Hodkinson
Affiliation:
Trinity College Dublin
Fiona M. Doohan
Affiliation:
University College Dublin
Matthew J. Saunders
Affiliation:
Trinity College Dublin
Brian R. Murphy
Affiliation:
Trinity College Dublin
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Summary

In vitro tissue culture systems are required for plant–microbe interaction studies on European ash, Fraxinus excelsior. Methods are needed for plant micropropagation and for physiological experimentation including pathogen/resistance testing and biocontrol studies. For example, systems are required for experiments on ash dieback disease, caused by the ascomycete fungus Hymenoscyphus fraxineus, that is killing ash plantations and natural populations across its native range. Methods are also needed to optimise the number of endophytes cultured from ash tissue and to taxonomically identify them. We present endophyte isolation protocols and media for ash, provide an optimised DNA barcoding procedure for endophyte identification and describe in vitro tissue culture methods suitable for ash–microbe interaction studies in both roots and shoots. Methods for both embryo culture and seed culture (with precutting) and for the bulking up of genotypes via single node culture are outlined. We also discuss the potential of tissue culture for establishing microbe/endophyte-free cultures.

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Endophytes for a Growing World , pp. 328 - 340
Publisher: Cambridge University Press
Print publication year: 2019

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