Figure 7

SsCVNH was secreted from the hyphal tips during infection.
(a) Western blot analysis showed that SsCVNH-FLAG could be detected in the engineered strains. SDS-polyacrylamide gel electrophoresis showed the equal loading amounts of total proteins used for the western blot analysis. Alkaline phosphatase-conjugated secondary antibody detected an approximately 17 kDa band in the SsCVNH-FLAG-engineered strains but not in the wild-type strain. (b) Immunofluorescence detection of SsCVNH-FLAG during infection. Onion bulb epidermis was inoculated with SsCVNH-FLAG-engineered strains for 36 h at 20 °C. Two different secondary antibodies conjugated with RRX or FITC respectively, were used independently to exhibit the specificity of fluorescence signal. The fluorescence of both RRX (red) and FITC (green) was detected in the SsCVNH-FLAG-engineered strains, but not in the wild-type strain. Arrows show the concentrated distribution of SsCVNH in the hyphal tips during infection. Scale bar = 50 μm.