## 4 Discussion
\begin{table}
\begin{tabular}{c c c c c c c c c} \hline \hline  & \multicolumn{2}{c}{**Mutations (\%)**} & \multicolumn{2}{c}{**Mutations**} & \multicolumn{2}{c}{**Missense**} & \multicolumn{2}{c}{**Missense Mutations**} \\  & & \multicolumn{2}{c}{**in Exons (\%)**} & \multicolumn{2}{c}{**Mutations (\%)**} & \multicolumn{2}{c}{**Genes (\%)**} \\ \hline
**Treatment Time (min)** & 5 & 20 & 5 & 20 & 5 & 20 & 5 & 20 \\ \hline
**Chromosome 1** & 0.00186 & 0.00160 & 0.00005 & 0.00000 & 0.00002 & 0.00000 & 0.04808 & 0.00000 \\ \hline
**Chromosome 2** & 0.00168 & 0.00115 & 0.00000 & 0.00000 & 0.00000 & 0.00000 & 0.00000 & 0.00000 \\ \hline
**Chromosome 3** & 0.00125 & 0.00142 & 0.00000 & 0.00000 & 0.00000 & 0.00000 & 0.00000 & 0.00000 \\ \hline
**Chromosome 4** & 0.00262 & 0.00309 & 0.00027 & 0.00012 & 0.00016 & 0.00010 & 0.19280 & 0.25707 \\ \hline
**Chromosome 5** & 0.00126 & 0.00128 & 0.00007 & 0.00013 & 0.00007 & 0.00011 & 0.13193 & 0.19789 \\ \hline
**Chromosome 6** & 0.00131 & 0.00138 & 0.00017 & 0.00007 & 0.00010 & 0.00002 & 0.22539 & 0.07513 \\ \hline
**Chromosome 7** & 0.00150 & 0.00112 & 0.00051 & 0.00024 & 0.00027 & 0.0014 & 0.31847 & 0.10616 \\ \hline
**Mitochondria** & 0.00000 & 0.00000 & 0.00000 & 0.00000 & 0.00000 & 0.00000 & 0.00000 \\ \hline \hline \end{tabular}
\end{table} 
Table 3: The percentage of mutations in each chromosome and mitochondrial genome of _A. oryzae_ after treatment with plasma for 5 min and 20 min. a plasma jet is likely to promote the secretion of \(\alpha\)-amylase and the activity of secreted \(\alpha\)-amylase in _A. oryzae_. The oxidative state of the PDB media seems to be closely associated with fungal activation. The highest elevation of \(\alpha\)-amylase activity was observed in plasma-treated samples, where PDB media exhibited ORP values of \(\sim\)310-320 mV. This value was higher than that of the control (0 min, 290-305 mV). H\({}_{2}\)O\({}_{2}\) and other reactive oxygen species generated upon plasma treatment may have contributed to the elevation in the ORP level in the PDB media, because H\({}_{2}\)O\({}_{2}\) can behave as an oxidant or a reductant in redox reactions [30]. Our results suggest that a certain level of oxidizing conditions outside the cells (310-320 mV in our case) may be needed for activation of the enzyme or enzyme secretion. Studies have shown that increased ORP values in plasma-treated water have a high impact on the oxidizing capacity and anti-microbial potential [31; 32; 33; 34]. However, our study demonstrated that microbial activation can also be induced under certain levels of the ORP. As observed in our previous study, NOx was stably detected in the PDB media during incubation post plasma treatment in this study as well. This indicates that NOx may be crucial for enhancing enzyme activity. In addition, the long-lived species of NO\({}_{2}\)\({}^{-}\) and NO\({}_{3}\)\({}^{-}\) were detected in the PDB media upon plasma treatment, indicating that these could be major players in the plasma-mediated activation of the enzyme or enzyme secretion. Although NOx levels were higher in the plasma-treated samples, no significant changes were observed in the pH levels, which were close to the optimal pH range of 5.0-6.0, which is suitable for \(\alpha\)-amylase enzyme production by _Aspergillus sp._ in a submerged fermentation culture [1]. Treatment with a plasma jet caused a small portion of variations in the genomic DNA of _A. oryzae_ in our study. Some of these variations occurred in the protein coding region. The plasma treatment time did not seem to affect the extent of genetic variations because similar levels of variation were observed upon plasma treatment for 5 min and 20 min. The soft rays and free radicals generated from NTAPP are known to damage DNA and RNA with increasing treatment times [35; 36]. This may lead to a mutational change in the sequence. In addition, recent studies using _A. oryzae_ and _C.