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Subcellular localization screening of Colletotrichum higginsianum effector proteins

Robin's article in Frontiers in Plant Science
BIOGER postdoc Guillaume Robin and collaborators published an article in which they determined the localization of 61 Colletotrichum higginsianum effector candidates (ChECs) inside plant cells by transiently expressing the proteins fused to GFP. Plant peroxisomes, Golgi bodies and microtubules were revealed as novel targets for fungal effectors.

The genome of the crucifer anthracnose fungus, C. higginsianum, encodes ~350 ChECs that are expressed at particular stages of host infection (appressoria, biotrophy and necrotrophy). However, almost nothing is known about where they go inside the plant after secretion by the fungus. Here, we set out to analyze the subcellular localization of 61 biotrophy-associated ChECs based on their transient expression as GFP-tagged fusion proteins in Nicotiana benthamiana, without their signal peptides. Although most of the fusion proteins showed a nucleo-cytoplasmic distribution similar to GFP expressed alone, 16 ChECs were addressed to specific plant compartments, including nine that were imported into plant nuclei. The remaining proteins were targeted to plant compartments that have not previously been reported for the effectors of any other filamentous pathogens. Thus, three were imported into the matrix of plant peroxisomes, three decorated plant microtubule arrays and one labelled plant Golgi bodies. Two of the peroxisome-targeted ChECs have C-terminal tripeptide signals for peroxisome import via the PTS1 pathway, and we verified that these signals are required for peroxisome localization.  Our finding that multiple Colletotrichum effectors converge on plant peroxisomes and microtubules suggests that manipulation of host functions associated with these structures could be critical for fungal pathogenesis. Overall, the information obtained from this medium-thoughput screen has allowed us to prioritize effector candidates for future functional analysis and provides clues to potential plant interacting proteins.


Reference: Robin GP, Kleemann J, Neumann U, Cabre L, Dallery J-F, Lapalu N, O’Connell RJ. 2018. Subcellular localization screening of Colletotrichum higginsianum effector candidates identifies fungal proteins targeted to plant peroxisomes, Golgi bodies and microtubules. Frontiers in Plant Science, 02 May 2018.