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. 2021 Jun 17;9:681734. doi: 10.3389/fcell.2021.681734

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Copyright © 2021 Jones, Zhu, Jenkinson, Kim, Pfeifer and Khang.

This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.

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The occurrence of EIHM disruption increases proportionally with nuclear stage. (A) Schematic diagram three-dimensionally depicting an epidermal rice cell invaded by M. oryzae expressing sec-GFP (green) and nuclear tdTomato (magenta). (B,C) Confocal images of rice cells invaded by M. oryzae CKF2187 expressing sec-GFP (green) and nuclear tdTomato (magenta). (B) Images of the same infection taken at different focal planes. The top three panels show merged fluorescence and bright-field from individual focal planes (indicated in the top right corner), while the last panel shows a merged fluorescence projection of all focal planes; 12 z-slices in total spanning 24 μm over the z-axis. Note that all five fungal nuclei are fully visible only in the projection view. Bar = 10 μm. (C) A merged fluorescence projection of infected rice cells at 30 hpi showing different patterns of sec-GFP localization at different IH growth stages determined by the nuclear number for each infection (white numbers). The infection to the far left shows EIHMx-localized sec-GFP (intact EIHM), while the other infections show host-localized sec-GFP (disrupted EIHM). Note the infected rice cell at the bottom right was invaded by two separate appressoria. Rice cell walls are indicated by white outlines. Bar = 50 μm. (D) A plot of the frequency of EIHM disruption according to fungal nuclear number for 390 infections of M. oryzae CKF2187 in the first-invaded host cell between 28 and 33 hpi. Sec-GFP localization patterns revealed the distribution of intact (n = 235) and disrupted (n = 155) EIHMs (y-axis) at each group of two nuclear stages (x-axis).