# In Vitro Cultures and Volatile Organic Compound Production in _Chiliadenus montanus_ ([PERSON].) [PERSON]
## 4 Conclusions  
In this study, efficient micropropagation and in vitro culture systems for _C. montanus_ were successfully established using different types of explants and media compositions. Callus culture was successfully established using leaf explants cultured on full-strength MS media supplemented with 2.2 uM 2,4-D and 6.9 uM kinetin Kin. Microshoots were successfully regenerated directly from nodal explants cultured onto half-strength MS media supplemented with 1.4 uM GA\({}_{3}\) and 4.4 uM BAP, while indirect microshoots were regenerated using callus explant cultured on MS media supplemented with 8.8 uM BAP, 2.2 uM Zeatin, and 1.4 uM GA\({}_{3}\). The VOCs from in vivo and in vitro samples were extracted successfully using HS-SPME and analyzed using GC-MS. Clear variations in the qualitative and quantitative aspects of VOCs between the wild plant and in vitro cultures of _C. montanus_ were observed. Callus cultures were found to be rich in monoterpeak hydrocarbons, while microshoot cultures were rich in oxygenated monoterpeak when compared to the wild plant extract that was rich in sesquiterpen. In conclusion, the in vitro cultures of _C. montanus_ reported in this study can be used to boost the production of VOCs under aseptic conditions without affecting wild-grown plants.  
\begin{table}
\begin{tabular}{c c c c c c c} \hline \hline \multirow{2}{*}{**Compound Name**} & \multirow{2}{*}{**Classification**} & \multirow{2}{*}{**RI-Exp 2**} & \multirow{2}{*}{**RI-Lit 3**} & \multicolumn{3}{c}{**Content (\%) 1**} \\ \cline{3-6}  & & & & **Wild** & **Microshoot** & **Callus** \\ \hline \multirow{3}{*}{Gleenol} & Oxygenated & 1589 & 1587 & 1.59 & - & - \\  & sesquiterpenes & & & & & \\  & Oxygenated & 1634 & 1631 & 22.19 & - & - \\  & sesquiterpenes & & & & & \\  & Sesquiterpene & 1649 & 1640 & - & 1.14 & - \\ \hline \end{tabular}  
* The contents percentage is based on the GC peak; 2 RI-Exp: experimental retention index related to (C8–C20) n-alkanes; 3 RI-Lit: reported retention index.  
\end{table}
Table 4: _Cont._  
**Supplementary Materials:** The following supporting information can be downloaded at: [[https://www.mdpi.com/article/10.3390/plants11101326/s1](https://www.mdpi.com/article/10.3390/plants11101326/s1)]([https://www.mdpi.com/article/10.3390/plants11101326/s1](https://www.mdpi.com/article/10.3390/plants11101326/s1)), Table S1: Effect of different concentrations of auxins and cytokins on callus induction using leaf explants of _C. montanus_.
* [PERSON] and [PERSON] conceived and designed the experiments and wrote the manuscript. [PERSON] conceived the in vitro culture work and VOCs extraction and analysis. [PERSON] and [PERSON] helped with data analysis. All authors edited and provided a critical review of the manuscript. All authors have read and agreed to the published version of the manuscript.
* The Deanship of Scientific research\(\backslash\)The University of Jordan to DAD (Grant number: 19-2020-99).
* Not Applicable.
* The data sets supporting the results of this article will be freely available upon reasonable request from A.M.A.
* We sincerely acknowledge [PERSON] for his valuable help.
* The authors declare no conflict of interest.