Extended Data Fig. 8: Chitin deposition in the appressorium cell wall is impaired in a Δsln1 mutant.
From: A sensor kinase controls turgor-driven plant infection by the rice blast fungus
a, Conidia were collected from Guy11 and the Δsln1 mutant and inoculated on glass coverslips to form appressoria. At 24 h.p.i., appressoria were stained with 50 µM calcofluor white for 5 min in the dark, washed and images captured by epifluorescence microscopy. Line-scan graphs represent calcofluor white fluorescence in a transverse section of an individual appressorium. Images are representative of n = 3 independent biological replicates. Scale bar, 10 µm. b, The Sln1 kinase interacts with Sum1, Pkc1 and Mps1 in a yeast two-hybrid assay. Simultaneous co-transformation of pGAD-Sln1 (prey vector) with pGBK-Mps1, pGBK-PKC and pGBK-Sum1 (bait vectors) and pGBKT7-53 and pGADT7-T (positive-control vectors) into the Y2H Gold strain resulted in the activation of three reporter genes and growth on medium-stringency medium (−Ade, −Leu, –Trp, +X-α-gal). Co-transformation also activates the expression of MEL1, which results in the secretion of α–galactosidase and the hydrolysis of X-α-gal in the medium, turning the yeast colonies blue. Images are representative of n = 2 biological replications of the experiment.
