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(a) Schematic view of the Ar plasma jet device. The distance between the tip of the needle (inner electrode) and the surface of the microtiter plate was 17 mm, and the condition for plasma breakdown was a voltage of 4 kV, a current of 13 mA, a repetition rate of 22 kHz, and an Ar gas flow rate of 150 sccm controlled by a mass flow controller (MFC). (b) Optical emission spectrum of the Ar plasma jet.
(Color online) (a) Germination of spores treated in water and VM with plasma. Open and closed columns represent the control (treated with Ar gas only) and the plasma-treated samples, respectively. Spores were germinated on FGS (Fructose, Glucose, Sucrose) agar plates after 1 day of incubation at 30 °C. Each value was an average of three replicate counts. (b) Spore morphology observed under an optical microscope at 400× magnification. Spores treated in water and VM by plasma for 180 s. were photographed using a Samsung CCD SCB-2000 camera (Samsung, Seoul, Korea). (c) pH of water and VM treated with Ar gas only (open column) and plasma (closed column). Each value was an average of three replicate measurements. (d) To further examine plasma or pH effect, spores (2 × 107) were placed in water and then exposed to plasma (left graph) and were incubated in plasma treated (middle graph) or acidic (right graph) water. Then, germination of spores was assessed. Direct plasma treatment of spores in water (Ar gas treatment as control) was performed as described in the text. Spores were exposed to plasma treated or acidic water as follows: Deionized water was treated with plasma or only Ar gas (control) for 10, 30, 60, and 180 s and then spores were incubated in the plasma or Ar gas treated water for 1 h. Acidic water of pH 4 and 2 was made by deionized water and nitric acid, and spores were incubated in the acidic water for 1 h. In the left and middle graphs, open and closed column represent Ar gas only and plasma treatment, respectively.
Agarose (1%) gel electrophoresis of genomic DNA extracted from N. crassa tissues (16 h culture) treated with Ar gas only (designated as A) and plasma (designated as P). A 1 kb DNA ladder was loaded in the left lane.
(Color online) Responses of the tah-3 deletion mutant to plasma. (a) Flask culture of wild-type and the tah-3 deletion mutant under normal conditions. (b) Vegetative growth on the VM agar plate of the wild-type and the tah-3 deletion mutant exposed to Ar gas only (left panel) and plasma (right panel) for 30 s. Pictures were taken after incubation at 30 °C for 2 days. (c) The spore germination percentage of the wild-type (diamond bullet line) and the tah-3 deletion mutant (square bullet line). The spore germination percentage was calculated as follows: (germinated number of spores exposed to plasma/germinated number of spores exposed to only Ar gas) × 100.
Ion analysisa in water exposed to plasma.
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