TIME RESOLVED FLUORESCENCE SPECT OF RED SHIFTED GREEN FLUORESCENT PROTEIN MUTANT

Cornell University Ithaca, Ithaca NY

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Abstract

Mycorrhizal fungi form mutualistic symbiotic associations with plant roots and are of primary importance in nearly all terrestrial ecosystems. However, very little is known about the cytology of this association, more precisely the cytology of the fungus. AMF produce long hyphae which are not septate (they have no walls between cells in the hyphae). AMF are also multinucleate (they have 2000-5000 nuclei per spore) and when the AMF spores germinate (that is, produce new hyphae) the nuclei concentrated in them wander along the hyphe. It is thought that the "wandering" nuclei are essential to the establishment of the symbiosis (involved in plant-fungus recognition processes), however nothing is known about their behaviour and life cycle before and after meeting the host plant. Techniques have been developed that allow for the establishment of Mycorrhizal plants in tissue culture. The mycorrhizae fungal hyphae are stained with DAPI while growing in agar based growth medium. Using multi photon microscopy we have been able to visualize the plant root-fungi contact areas and study the movement of DAPI-labeled nuclei in fungal hyphae.