Figure 1.

Necrosis-inducing activity of the B. cinerea secretome. A, Bean leaves were inoculated with a suspension of B. cinerea spores. At the designated time points post inoculation, the suspension was collected and centrifuged, the spores and mycelia fragments were removed by filtering through 45-μm filters, and the clean suspension was infiltrated into N. benthamiana leaves. Images show leaves 3 d after treatment. B, Clean spore suspension was boiled and mixed with loading buffer, 20 μL was loaded onto an SDS-PAGE gel and separated by electrophoresis, and the gel was stained with silver reagent. C, Suspension was collected from inoculated bean leaves or from a fungus that was grown in Gamborg’s B5 on a glass slide. Images were taken 3 d after infiltration of leaves with the clean suspension. D, Clean suspension was treated by boiling for 10 min, precipitation of proteins with ammonium sulfate, or dialysis through a 10-kD membrane. Images were taken 3 d after infiltration of leaves with boiled suspension (Boiled), suspension after protein precipitation [(NH₄)₂SO₄], dialyzed suspension (10 kDa), or the flow through after dialysis (Flow through).