Fig 6. Ca²⁺ detection in hyphopodia and vegetative hyphal tips of wild-type and mutant strains.

A. Detection of intracellular Ca²⁺ elevation in wild-type and mutant strains. V592, mutant strains VdΔnoxb and VdΔpls1, and complemented strains VdΔnoxb/VdNoxB and VdΔpls1/VdPls1 were grown on MM medium overlaid with the cellophane membrane for 2 days, and intracellular Ca²⁺ was detected with Fluo-4 AM. A tip-high gradient Ca²⁺ was observed in hyphopodia of V592 and complemented strains but not in the defective hyphopodia of VdΔnoxb or VdΔpls1. Bar = 5μm. B. Detection of cytoplasmic Ca²⁺ in vegetative hyphal tips in V592, VdΔnoxb and VdΔpls1. Cytoplasmic Ca²⁺ was detected in wild-type V592 and both mutant strains upon Fluo-4 AM treatment. Bar = 5μm. C. Elevation of Ca²⁺ in VdΔnoxb and VdΔpls1 upon application of OH·. VdΔnoxb and VdΔpls1 grown on cellophane were treated by OH· and cytoplasmic Ca²⁺ was detected with Fluo-4AM. Bar = 5μm.