Visualizing the Movement of Magnaporthe oryzae Effector Proteins in Rice Cells During Infection
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During biotrophy, filamentous pathogens such as the rice blast fungus Magnaporthe oryzae deliver effector proteins into live host cells to facilitate colonization. We describe three complementary assays for visualizing M. oryzae effector translocation into the rice cytoplasm and cell-to-cell movement during infection. Our assays make use of live-cell confocal imaging of optically clear rice sheath cells infected with transgenic strains of M. oryzae that express the fluorescent protein-tagged effector known as PWL2. We highlight several important considerations for the analysis of effector translocation and movement dynamics during infection of host plants.
Key wordsBiotrophic interfacial complex Confocal imaging Effector proteins Fluorescent proteins Host translocation assay Live-cell imaging Magnaporthe oryzae Oryza sativa Plant–pathogen interactions Rice blast
We acknowledge the assistance of the Biomedical Microscopy Core at the University of Georgia with imaging using a Zeiss LSM 710 confocal microscope. This work was supported by the Agriculture and Food Research Initiative competitive grants program, Award number 2014-67013-21717 from the USDA National Institute of Food and Agriculture.
- 4.Giraldo MC, Dagdas YF, Gupta YK, Mentlak TA, Yi M, Martinez-Rocha AL, Saitoh H, Terauchi R, Talbot NJ, Valent B (2013) Two distinct secretion systems facilitate tissue invasion by the rice blast fungus Magnaporthe oryzae. Nat Commun 4:1996. https://doi.org/10.1038/ncomms2996CrossRefPubMedPubMedCentralGoogle Scholar
- 5.Mentlak TA, Kombrink A, Shinya T, Ryder LS, Otomo I, Saitoh H, Terauchi R, Nishizawa Y, Shibuya N, Thomma BP, Talbot NJ (2012) Effector-mediated suppression of chitin-triggered immunity by Magnaporthe oryzae is necessary for rice blast disease. Plant Cell 24:322–335CrossRefPubMedPubMedCentralGoogle Scholar
- 7.Park C-H, Chen S, Shirsekar G, Zhou B, Khang CH, Songkumarn P, Afzal AJ, Ning Y, Wang R, Bellizzi M, Valent B, Wang G-L (2012) The Magnaporthe oryzae effector AvrPiz-t targets the RING E3 ubiquitin ligase APIP6 to suppress pathogen-associated molecular pattern–triggered immunity in rice. Plant Cell 24:4748–4762CrossRefPubMedPubMedCentralGoogle Scholar
- 9.Kleemann J, Rincon-Rivera LJ, Takahara H, Neumann U, Ver Loren van Themaat E et al (2012) Sequential delivery of host-induced virulence effectors by appressoria and intracellular hyphae of the phytopathogen Colletotrichum higginsianum. PLoS Pathog 8:e1002643. https://doi.org/10.1371/journal.ppat.1002643CrossRefPubMedPubMedCentralGoogle Scholar
- 10.Jones K, Zhu J, Jenkinson CB, Kim DW, Khang CH (2017) Disruption of the interfacial membrane leads to Magnaporthe oryzae effector re-location and lifestyle switch during rice blast disease. bioRxiv. https://doi.org/10.1101/177147
- 17.Shipman EN, Jones K, Jenkinson CB, Kim DW, Zhu J, Khang CH (2017) Nuclear and structural dynamics during the establishment of a specialized effector-secreting cell by Magnaporthe oryzae in living rice cells. BMC Cell Biol 18:11. https://doi.org/10.1186/s12860-017-0126-zCrossRefPubMedPubMedCentralGoogle Scholar