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Cell re-entry assays do not support models of pathogen- independent translocation of AvrM and AVR3a effectors into plant cells

By Benjamin Petre, Michaela Kopischke, Alexandre Evrard, Silke Robatzek, Sophien Kamoun

Posted 29 Jan 2016
bioRxiv DOI: 10.1101/038232

The cell re-entry assay is widely used to evaluate pathogen effector protein uptake into plant cells. The assay is based on the premise that effector proteins secreted out of a leaf cell would translocate back into the cytosol of the same cell via a yet unknown host-derived uptake mechanism. Here, we critically assess this assay by expressing domains of the effector proteins AvrM-A of Melampsora lini and AVR3a of Phytophthora infestans fused to a signal peptide and fluorescent proteins in Nicotiana benthamiana. We found that the secreted fusion proteins do not re-enter plant cells from the apoplast and that the assay is prone to false-positives. We therefore emit a cautionary note on the use of the cell re-entry assay for protein trafficking studies.

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