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Abstract
A suitable Agrobacterium-mediated transient expression assay was evaluated for rapid analysis of vacuole organisation in different cell types in vivo. By simple infiltration of Agrobacterium cells carrying appropriate plasmid constructs into Petunia hybrida leaves and petals, reproducible expression can be revealed by GFP fluorescence within one day without using expensive equipment (e.g. biolistic gun or electroporation apparatus) or complicated procedures (e.g. preparation of protoplasts). Different vacuolar markers for the neutral compartment (GFP-Chi) or the lytic one (Aleu-GFP), and an ER resident protein (GFP-KDEL) were used. Previously, it was shown that these markers could label different compartments but that such compartments are organised differently depending on plant species and tissues. Our results demonstrate that epidermal cells of petunia petals represent a case study that demands further investigation concerning vacuolar organisation, and that Agrobacterium-mediated transient expression is a simple and efficient method for in vivo assays of sub-cellular markers in this tissue. In the present study, this method revealed an unexpected difference between the anthocyan accumulating vacuole and the normal lytic vacuole labelled by Aleu-GFP.
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Acknowledgements
This work was made possible thanks to an EMBO Short Term Fellowship ASTF 350.00-05 and EU funding (MTKD-CT-2004-509253).