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. 2020 Jul 25;6(3):118. doi: 10.3390/jof6030118

Figure 9.

Figure 9

Capture of volatiles. (A). Trapping materials. Pa10 was grown on TSA for 72 h at 37 °C in the double plate method, with a second plate (trapping phase) as indicated. With a delay of <3 sec, the Pa plate was replaced with a plate of TSA with 10AF inoculum (exposure phase), incubated 48 or 72 h, and the growth was measured. The 48 h exposure data is shown. Unexposed plates (controls) are compared to exposed plates. (B). No inhibition of growth from an uninoculated plate. In a double plate experiment similar to that in Figure 9A, 40 μL from a Pa10 109/mL suspension on TSA, or an uninoculated TSA plate, were paired with 15 mL of deionized water in the second plate, and incubated for 72 h at 37 °C. The 2 water plates were each then sealed in a second double plate setup with a TSA plate inoculated with 10 μL of 5 × 104/mL conidial suspension of 10AF, incubated for 48 h at 37 °C, and the fungal growth measured. Three asterisks = p < 0.001. (C). Effect of time of exposure. Pa10 (40 μL from a 109/mL suspension) inoculated on TSA plates, or uninoculated TSA plates (control), were incubated for 48 h at 37 °C in the double plate setup with 15 mL of water in the second plate (trapping phase). In the second part of the experiment, the double plate setups contained TSA plates inoculated with 10 μL of 5 × 104/mL conidial suspension of 10AF, with the second plate containing the water that had been exposed to the Pa volatiles. This was incubated for 24 h at 37°C. Then, 24 and 48 h later, the water was replaced with water that had just trapped the Pa volatiles for 48 h. After 48 and 72 h of total co-incubation, the fungal growth was measured. “Interaction” refers to the cumulative time of 24 h of co-incubations of Af with the water that had trapped the Pa volatiles. Three asterisks = p < 0.001 compare the left 2 bars, and 3 daggers = p < 0.001 compare the right 2 bars. Refreshing the exposure of Af with fresh water that had trapped Pa volatiles maintains the inhibition over a 72 h period. (D). Optimal trapping duration. In the double plate method, a plate of deionized water was combined with a plate of Pa10 on TSA for 24, 48, or 72 h, and each water plate was then tested in exposure to 10AF on TSA.