Fig. 3.

Z. tritici is easily dislodged from the wheat leaf. Leaves were inoculated with Z. tritici at 10⁷ cfu/ml and allowed to dry at RT for 1 h. Leaves were either: sampled immediately (controls); gently misted with water to runoff; rinsed by placing in a 15 ml tube half filled with water and inverting ten times; or vigorously shaken in a half-filled 15 ml tube of water or 0.1% (v/v) Tween. Inoculated leaves were visualised using confocal microscopy and all individuals in a given area enumerated and their location scored. Values are means of 2 or 3 independent experiments, each consisting of 5 biological replicates each including 3 samples; control n = 30–200. Error bars represent SE. Asterisks indicate significant differences from control (Bonferroni corrected t-tests). Fungi were scored as encountering a stoma if any part of the fungus touched or crossed the guard cells. Penetration was scored if fungus was visible in the substomatal aperture. Trichome association indicates that some part of the fungus grew across a trichome. Cfu counts are given as a percentage of the number seen on control (unwashed) samples (first set of bars). Other values represent the percentage of fungi within each treatment in the indicated location.