Figure 5: Secretion of cytoplasmic effectors involves exocyst components Exo70 and Sec5. | Nature Communications

Figure 5: Secretion of cytoplasmic effectors involves exocyst components Exo70 and Sec5.

From: Two distinct secretion systems facilitate tissue invasion by the rice blast fungus Magnaporthe oryzae

Figure 5

(a–c) Wild-type strain Guy11 and corresponding mutants expressed Pwl2:mCherry:NLS and Bas4:GFP. Images clockwise from the upper left: bright-field; merged mCherry (red) and GFP (green); fluorescence intensity linescans for mCherry (red) and GFP (green) along the path of the white arrow; single channel GFP or mCherry fluorescence shown as black and white inverse images, respectively. (a) Pwl2:mCherry:NLS in wild type shows preferential BIC localization, translocation and accumulation in the rice nucleus (white asterisk) with no fluorescence observed in the BIC-associated IH cell. Bas4:GFP localizes to the EIHM compartment. (b) Δexo70 mutant shows partial retention of Pwl2:mCherry:NLS, predominantly in the BIC-associated IH cell. Bas4:GFP secretion appears normal. Internal red fluorescence in the BIC-associated IH cell is further visualized by a black and white inverse image (lower left) and by fluorescence intensity scans, which show red fluorescence between the peaks of green fluorescence marking the boundary of the EIHM. Faint red fluorescence in the rice nucleus (white asterisk) is consistent with effector secretion being only partially blocked in the mutants. (c) Δsec5 mutant showing partial retention of Pwl2:mCherry:NLS inside the BIC-associated IH cell, but no retention of Bas4:GFP, as further visualized by black and white inverse images and fluorescence intensity scans. (d–f) Images left to right: merged bright-field and mRFP; single channel mRFP fluorescence as a black and white inverse image; and corresponding fluorescence intensity linescan. (d) Wild type Guy11 expressing cytoplasmic effector Bas1:mRFP. Bas1:mRFP fluorescence is not observed inside wild type IH cells. (e) Bas1:mRFP expressed by an Δexo70 mutant shows significant retention inside the IH, predominantly in the BIC-associated cells. (f) Bas1:mRFP expressed by an Δsec5 mutant strain shows significant retention of Bas1:mRFP inside the IH, predominantly in the BIC-associated cells. Linescans shown are representative of wild type (n=20) and knockout mutants (n=20 for each). Scale bars, 10 μm.

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