mSphere00936-19 Supplementary Fig. S4

Supplementary Fig. S4. MoLAEA shows no role in pathogenicity. (A) Conidial germination of the wild type and mutant strains were studied as the ratio of germinating conidia on hydrophobic surfaces to total conidia. (B) The average rate of appressorium formation by the over-expression and knock-down strains compared to the wild type was determined. 100µl of spore suspension containing 1 × 105/ml spore suspension was inoculated on hydrophobic surfaces. After 24 h, the number of appressoria formed by 50 conidia was counted and a mean value generated. (C) Rate of appressorium penetration. The number of appressoria that had penetrated the leaf cuticle and developed visible IH, from a total of 50 appressoria observed was determined. The average was determined from three independent replicates. (D) Invasive hyphae growth was determined for each strain, at 48 hpi, using the four-point scale. According to this scale, 1 = IH length shorter than 10 mm with no branching; 2 = IH length is 10–20 mm with 0–2 branches; 3 = IH length is longer than 20 mm and/or with more than 2 branches within one cell; 4 = IH has spread to adjacent cells. Three leaf sheaths were inoculated with each strain, and 50 IH growth rates were determined for each replicate to generate a value for the mean IH growth rate. (E) Spores isolated from the lesions of the leaves infected by the punch-inoculation method and counted on a hemocytometer under a microscope. (F) Quantification of lesion density in the leaves inoculated with wild type nad each strains. Disease lesions were counted in over 20 diseased leaves and the average lesion number was then calculated. (G) The quantitative analysis of the lesion area in 20 diseased leaves and the average lesion area was calculated. Bars with the same letter are not significantly different at p≤0.05