## 2 Materials and Experimental Methods
### Soft Plasma Jet Device, Plasma-Activated Water Treatments, and Spore Viability  
A nonthermal atmospheric pressure air plasma jet (simply called a soft plasma jet) device consists of a power supply, a powered needle electrode, a quartz tube, and a grounded tube electrode. The quartz tube served as a dielectric barrier between the electrodes to induce an electrical discharge. Plasma radicals were generated by electrodes separated by a dielectric barrier. Electric power for the plasma device was supplied by an inverter that converts DC input to a 67 kHz AC output. A detailed description of the soft plasma jet system was presented in a previous paper [22]. The optical emission spectrum of the soft plasma jet exhibited emission lines corresponding to \(N_{2}\), atomic nitrogen, atomic oxygen, and hydroxyl radicals, as reported previously [22]. The electrical signal supplied to the soft plasma jet device showed a peak voltage of 1.24 kV (rms voltage 0.54 kV) and a peak current of 2.96 \(A\) (rms current, 97 mA). The period of the applied voltage/current was 12 us, and the duty ratio of pulse-on and pulse-off times was 18.4%. The soft plasma jet could transfer 3.9 \(J\) per second to the sample. To find the level of RONS in the PAW generated by the soft plasma jet, \(H_{2}O_{2}\) and \(NO_{x}\) concentrations in the PAW were investigated for the plasma activation times of 1, 3, 6, and 10 min. To study the influence of PAW treatment on \(A\). _trasiliensis_ spores, PAW was prepared by activating sterile de-ionized (DI) water with a soft plasma jet. DI water (3 mL) was dispensed into each well of six-well culture plates (SPL Life Science Co., Pocheon-si, Gyeonggi-do, Korea). The water in each well was activated by the soft plasma jet for 1 min, 3 min, and 6 min, and was coded as PAW-1, PAW-3, and PAW-6, respectively. Because the level of RONS in the PAW is saturated for plasma activation times longer than 6 min, the effects of PAW treatment on the viability of \(A\). _trasiliensis_ spores were examined using PAW-1, PAW-3, and PAW-6. The PAW treatment against fungal spores was performed right after the generation of the PAW. For the PAW treatment, the prepared spores were suspended in the PAW in the well of each plate at a concentration of \(5\times 10^{7}\) spores/mL, and incubated at room temperature for 30 min. After PAW treatment, the viability of PAW-treated spores was measured with the PAW-untreated spores as the control. To examine the effect of specific reactive species on the \(A\). _trasiliensis_ spores, \(H_{2}O_{2}\) and \(HNO_{3}\) solutions were prepared in five different concentrations (0.001%, 0.01%, 0.1%, 0.3%, and 1.0%). The fungal spores were treated for 30 min with each concentration of \(H_{2}O_{2}\) and \(HNO_{3}\) solutions as with the PAW treatment. To measure spore viability, all the spores treated with the PAW, plasma, and \(H_{2}O_{2}\) and \(HNO_{3}\) solutions were diluted with sterile water up to 10-fold, spread on PDA media, and kept at 25 \({}^{\circ}\)C for 7 days, which is long enough to observe visible mycelia of fungal colonies from the survived spores. The live fungal colonies were counted and the number of colony-forming units (CFUs) was calculated to measure viability. Morphology and Cell Wall Structure Analysis of \(A\). brasiliensis Spores Treated with Plasma-Activated Water  
The PAW-treated \(A\). _trasiliensis_ spores were subjected to flow cytometry measurements and scanning electron microscope (SEM) imaging analysis to investigate the effects of PAW treatment on the structure, integrity, and morphology of the cell wall. Field emission SEM (Hitachi, S-4300, Tokyo, Japan) was operated according to the protocol of [PERSON] et al. [26] to examine microstructures of the PAW-treated spores, as reported previously [22]. For morphological analysis, more than 200 individual spores of each treatment were observed at different magnifications.