# In Vitro Cultures and Volatile Organic Compound Production in _Chiliadenus montanus_ ([PERSON].) [PERSON]
## 3 Results and Discussion
### Volatile Organic Compound Analysis  
The VOC components in wild, microshoot, and callus extracts of _C. montanus_ were identified using GC-MS analysis based on the comparison of the obtained RI and MS fragmentation patterns to standard compounds and computer matching with built-in libraries [33]. Accordingly, the wild plant extract was found to be richer in sesquiterpene hydrocarbons (47.97%) and oxygenated sesquiterpenes (35.32%), followed by oxygenated monoterpenes (12.22%) and monoterpene hydrocarbons (1.68%) (Table 3). On the other hand, the most abundant chemical groups in microshoot extracts were oxygenated monoterpenes (58.09%) and monoterpene hydrocarbons (23.68%), followed by sesquiterpene hydrocarbon (10.16%) and oxygenated sesquiterpenes (1.14%), while callus extracts contained only monoterpene hydrocarbons (63.35%) and oxygenated monoterpenes (35.64%) (Table 3). The occurrence of monoterpenes and sesquiterpenes in _C. montanus_ was reported previously [15; 41]. Ref. [41] reported that the predominant compounds identified in _C. montanus_ wild-growing plants were oxygenated monoterpenes, which was not in general agreement with [15]. In another study, clear variations in the composition of VOCs between wild populations of _C. ipbinoides_ were also observed [42]. In this study, microshoot and callus extracts produced higher amounts of monoterpenes when compared with the wild-type plant extract, which produced mainly sesquiterpenes compounds. In contrast to microshoot and wild plant extracts, callus culture extracts produced only monoterpenes compounds; such differences in VOC profiles between in vitro cultures and wild plant extracts were reported previously [43]. The analysis of VOCs obtained from the areal parts of wild _C. montanus_ resulted in the identification of 19 compounds representing 97.19% of the total oil content. The main VOCs found in wild plant extract included Alloaromadendrene (11.92%), trans-Cadina-1(6),4-diene (7.54%), and \(\alpha\)-caryophyllene (6.77%) from the sesquiterpene hydrocarbons group, while from the oxygenated sesquiterpenes group, Muurola-4,10(14)-dien-8.\(\alpha\)-ol (22.19%) and \(\delta\)-amorphene (9.32%) were the most abundant compounds (Table 4). For  
\begin{table}
\begin{tabular}{c c c c} \hline \hline
**Cytokinin Type** & **Concentration (\(\mu\)M)** & **Number of Leaves** & **Shoot Length (cm)** \\ \hline \multirow{4}{*}{BAP} & 0.0 & 10.70 \(\pm\) 0.74 \({}^{\mathrm{\,\text{\text{\text{\text{\textasci  
the monoterrepenoids group, Thymol methyl ether (9.17%) and \(\gamma\)-Terpineol (3.05%) were detected in wild plant extract (Table 4). In the microshoot extracts, 26 compounds were identified that accounted for 99.22% of the total VOCs with cis-Myrtanol (16.62%) and \(\beta\)-Cyclocitral (14.3%) as the main compounds from the oxygenated monoterrepenes group, and p-cymene (8.25%), (+)-2-Carene (6.43%), and (E)-\(\beta\)-ocimene as the main compounds from the monoterrepenos group (Table 4). For sesquiterpennes in microshoot extracts, \(\alpha\)-caryophyllene (2.80%) and \(\delta\)-Amorphene (2.35%) were identified as the main components. For callus extracts, the VOCs profile was dominated by monoterpenes, and 15 compounds were identified, representing 98.99% of the total content (Table 4). The main compounds of this fraction were (Z)-\(\beta\)-Ocimene (22.27%), p-Cymene (15.13%), and \(\alpha\)-Pinene (13.78%) from the monoterrepen hydrocarbons group, while Thymoquinone was as the main compound in the oxygenated monoterpenes group (13.25%).