In vitro Cultures Open New Prospects for Basic Research in Arbuscular Mycorrhizas
It is now 20 years since the publication of Bécard and Fortin’s article (1988) in which the modern basis for arbuscular mycorrhizal (AM) monoxenic cultures was established. Since then, many research projects have been carried out using in vitro systems, a
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Introduction Arbuscular Mycorrhizas In Vitro Cultures: The Starting Material
It is now 20 years since the publication of Bécard and Fortin’s article (1988) in which the modern basis for arbuscular mycorrhizal (AM) monoxenic cultures was established. Since then, many research projects have been carried out using in vitro systems, and new prospects have been opened up by utilizing the amazing research material provided by monoxenic plates. More and more researchers now acknowledge that in vitro AM production is the sole way of getting large amounts of clean, clonal, contamination-free AM fungal material. This has opened the doors for molecular biology and biochemical techniques to be applied to mycorrhizal research, and the direct consequence of this is an exponential increase in our knowledge in the basic biology of this mutualistic symbiosis over the last 10 years. However, monoxenic cultures have far more to offer than just being AM tissue factories; fields of AM research as different (yet interlinked) as colony architecture and dynamics, intra- and extraradical fungal morphology, mycorrhizal physiology, biotic and abiotic stress responses, microbial interactions and even the production of ultrapure, mycorrhizabased biofertilizers have benefited from this ‘in vitro revolution’. This chapter aims to summarize the opportunities offered by using in vitro culture technology, and to encourage researchers to (1) utilize existing, yet little known monoxenic culture techniques, and (2) improve them and design new in vitro experimental systems. It should not be regarded as a compilation of methods, but rather as a brainstorming exercise designed to create new avenues of mycorrhizal research.
Alberto Bago Departamento de Microbiología del Suelo y Sistemas Simbióticos, Estación Experimental del Zaidín (CSIC), calle Profesor Albareda 1, 18008-Granada, Spain e-mail: [email protected]
A. Varma (ed.) Mycorrhiza, © Springer-Verlag Berlin Heidelberg 2008
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C. Cano et al.
Starting a New Arbuscular Mycorrhizal Monoxenic Culture (AMMC): The Real Challenge
Research using monoxenic cultures in the late 1980s began mainly with using Gigaspora margarita DAOM 194757 (Bécard and Fortin 1988; Bécard and Piché 1989a, 1989b; Chabot et al. 1992a). From the 1990s, the AM fungal isolate, Glomus intraradices DAOM 197198 (Chabot et al. 1992b), was mostly used. This was mainly due to the fact that this AMF isolate is well-adapted to the monoxenic culture conditions, producing important amounts of tissue and being quite easy to replicate. As a consequence of this, G. intraradices DAOM 197198 has become a kind of ‘model AMF species’, comparable to the model plant Arabidopsis thaliana or bacteria Escherichia coli. It was plain from the beginning, however, that diversification using and comparing different AMF isolates/species/genera was mandatory in order to have a global and realistic view of mycorrhizal symbiosis. The difficulties of introducing some AMF isolates/species into monoxenics lead some researchers to claim
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