## 3 Results
### Effect of Salt Stress on the Relationship between Cell Vacuolization and the Provacuolar Index  
The increase in the vacuolization index was influenced by both the concentration of saline used and the exposure time, with a high degree of significance (\(p<0.001\)). The combined effect of the treatment and exposure time showed an increase in the vacuolization index. Thus, we can say that the vacuolization phenomenon is closely related to the cytotoxicity caused by the saline solutions used. Vacuolized cells are formed by the fusion of provacuoles in response to salt stress. The number of cells with multiple provacuoles increased considerably with increasing saline concentrations (\(p<0.001\)) (Table 5). Another important effect of salt toxicity is the increase in the provacuolar index (PVI). At the cellular level, under the influence of salt stress, a cytological process is observed consisting in the formation of provacuoles, which fuse to form an enlarged vacuole in which the genetic material is arranged around it. The vacuolization phenomenon develops when the biological material is exposed to salt stress with an increasing frequency as a function of the NaCl concentration. The vacuolization phenomenon showed an increasing frequency concomitant with the duration of exposure to the salt solutions, whereby for the 24-h exposure duration we recorded the lowest values for the salt concentrations, while the highest values were recorded for the 72-h exposure duration, culminating in a maximum value of more than 36% of cells vacuolized under a salt concentration of 150 mM (Table 6). Therefore, a correlation was found between the increases in the provacuolar index and vacuolization index (\(p<0.05\)). \begin{table}
\begin{tabular}{c c c c c c} \hline \hline  & \multicolumn{6}{c}{**Vacuolization Index (VI)**} \\ \hline
**Variance Source** & **DF*** & **SS*** & **Mean Sq** & **F Value** & **Pr (\(p\)-F)** \\ \hline Treatment & 3 & 37544 & 12515 & 2092.87 & \(2\times 10^{-16}\)*** \\ \hline Exposure time & 2 & 1396 & 698 & 116.73 & \(2\times 10^{-16}\)*** \\ \hline Treatment \(\times\) exposure time & 6 & 640 & 107 & 17.84 & \(5.16\times 10^{-5}\)*** \\ \hline Residual & 288 & 1722 & 6 & & \\ \hline \multicolumn{6}{c}{**Provacuolar Index (PVI)**} \\ \hline
**Variance Source** & **DF*** & **SS*** & **Mean Sq** & **F Value** & **Pr (\(p\)-F)** \\ \hline Treatment & 3 & 1307.1 & 435.7 & 184.728 & \(2\times 10^{-16}\)*** \\ \hline Exposure time & 2 & 7.1 & 3.5 & 1.502 & 0.2244 \\ \hline Treatment \(\times\) exposure time & 6 & 44.2 & 7.4 & 3.121 & 0.0056 *** \\ \hline Residual & 288 & 679.3 & 2.4 & & \\ \hline \hline \end{tabular}  
* DF—degrees of freedom; SS—sum of squares. The results were analyzed at significance levels of **
* \(p<0.001\), **
* \(p<0.01\) and  
\end{table} 
Table 5: Analysis of variance of the combined effect of different salt concentrations and exposure times on the vacuolization and provacuolar indices. Cell vacuolization occurs under stress conditions of different salt concentrations. Vacuolized cells show a distribution of genetic material around the vacuole, and the tonoplast of the vacuole is well defined. Vacuolization is a very important topic in salt stress determination, which has applications in the selection of salt-stress-resistant genotypes in plant research and development programs. Vacuolization also plays a role in the diagnosis of cell stress, as the vacuolization index is significantly higher in salt treatments than in salt-free treatments (\(p<0.05\)). Therefore, we can identify how at a physiological level, cells are stressed due to salinity or other stressors in both human and plant cells. The effect of salt stress was also observed in our analyses based on the appearance of cell vacuolization. According to our studies, we observed an increase in the rate of cells with provacuoles, ranging from two to seven provacuoles in some cases (Figure 4B). The provacuoles fused (Figure 4 AC), forming an enlarged vacuole with genetic material arranged around it (Figure 4F).