Fig. 1: Mechanosensor N+-BDP reveals spatial variations in membrane tension of appressoria of M. oryzae.
a, Time course of infection-related development of M. oryzae. Images show developing appressoria of the wild-type strain Guy11 germinated on glass coverslips between 0 h and 24 h. Images are representative of n = 3 independent repeats of the experiment. Scale bar, 10 μm. b, Chemical structure of the N+-BDP rotor. c,d, Schematic illustration showing the molecular mechanism by which N+-BDP reports changes in membrane tension in 4 h and 24 h Guy11 wild-type appressoria. e, Representative FLIM images of 4 h and 24 h N+-BDP rotor-stained appressoria. The colour corresponds to the fluorescence lifetime values expressed in nanoseconds, as shown in the key 2–4 ns. The corresponding black and white images highlight spatial heterogeneity in membrane tension during turgor generation at 24 h. f, Dot plots showing the average fluorescence lifetime for 4 h and 24 h appressoria. Each dot corresponds to the average fluorescence lifetime obtained for an ROI drawn around the membrane of an individual appressorium in a 2D FLIM image. Total observations n = 45 appressoria examined in three biological replicates, typical range 21–24 appressoria; each biological replicate is colour coded (R1, replicate 1; R2, replicate 2; R3, replicate 3); data are presented as mean ± s.e.m. ****P < 0.0001, two-tailed unpaired Student’s t-test with Welch correction. g, Time-lapse FLIM images of appressorium development in Guy11 4.5–7 hpi (0–145 min, respectively). Images are representative of n = 3 independent repeats of the experiment. Scale bars, 1 μm. Image created with BioRender.com.
