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Physcomitrella patens, Marchantia polymorpha, Glomus intraradices, infection
Different forms of mycorrhiza are being distinguished based on the type of fungus and whether it surrounds or penetrates the tissue, typically the root. The most abundant form of mycorrhiza are AM (arbuscular mycorrhiza), formed by non-septate fungi belonging to the taxon Glomeromycota, exhibiting a broad host specifity. The success of mycorrhiza since the Ordovician, 450 MYA, result from their role in capturing nutrients from the soil and providing them to the plant and their contribution to plant fitness and tolerance against biotic and abiotic stresses. Mycorrhiza therefore are present in almost all ecosystems and plant taxa, play an important role in plant biodiversity and receive increasing attention in agricultural practice. Besides seed plants, associations with Glomales have been described in all three divisions comprising the bryophytes (mosses, hornworts and liverworts). Due to their early branching phylogenetic position and the ease of axenic cultivation under controlled conditions, bryophytes are ideal candidates to elucidate the mechanisms underlying the initial establishment of mycorrhizal symbiosis concomitant with the water-to-land-transition of plant life. The moss Physcomitrella patens, with its completely sequenced genome and a broad palette of available molecular tools, provides a tractable system to study nearly all aspects of AM. Similarly, the liverwort Marchantia polymorpha, the genome of which is currently being sequenced, offers a large selection of genetic tools. This technical note provides protocols for axenic in vitro establishment and evaluation of non-seed plant association with the AM fungus Glomus intraradices, including media preparation, cultivation, infection strategies and staining procedures.