Abu-Darwish Doaa, Shibli Rida, Al-Abdallat Ayed M
Department of Horticulture and Crop Science, School of Agriculture, The University of Jordan, Amman 11942, Jordan.
Department of Agricultural Biotechnology and Genetic Engineering, Faculty of Agriculture Technology, Al-Ahliyya Amman University, Amman 19328, Jordan.
Plants (Basel). 2022 May 17;11(10):1326. doi: 10.3390/plants11101326.
Callus and microshoot cultures were established for (Vhal.) Brullo. (Asteraceae), a medicinal plant known for producing volatile organic compounds (VOCs). Callus induction was achieved successfully by culturing leaf explants on full-strength Murashige and Skoog medium (MS) supplemented with 2.2 µM 2, 4-dichlorophenoxy acetic acid (2,4-D) and 6.9 µM kinetin (Kin). Successful direct shoot regeneration was achieved using nodal explants cultured onto half-strength MS media supplemented with 1.4 μM Gibberellic Acid (GA) and 4.4 μM 6-Benzylaminopurine (BAP). Indirect microshoots were successfully regenerated using callus cultured on MS media supplemented with 8.8 μM BAP, 2.2 μM Zeatin, and 1.4 μM GA followed by culturing on MS media supplemented with 8.8 μM BAP and 0.5 μM naphthalene acetic acid (NAA). Using wild plant aerial parts, callus and microshoots samples, VOCs were extracted successfully using Headspace Solid-Phase Micro-Extraction (HS-SPME) and analyzed by gas chromatography-mass spectrometry (GC-MS). In wild plant extracts, sesquiterpene hydrocarbons were found to be predominant with the following principal components: Alloaromadendrene (11.92%), trans-Cadina-1(6),4-diene (7.54%), and α-caryophyllene (6.77%). The analysis of in vitro microshoots revealed high levels of oxygenated monoterpenes with -Myrtanol (16.62%), and β-Cyclocitral (14.3%) as the main components. Callus extract was dominated by monoterpene hydrocarbons and the main compounds identified were (Z)-β-Ocimene (22.27%), p-Cymene (15.13%), and α-pinene (13.78%). In conclusion, an efficient in vitro production system of VOCs in was established that can be used in the future for boosting their production without endangering wild plants.
为（Vhal.）布鲁洛（菊科）建立了愈伤组织和微芽培养体系，该植物是一种以产生挥发性有机化合物（VOCs）而闻名的药用植物。通过在添加了2.2 μM 2,4-二氯苯氧乙酸（2,4-D）和6.9 μM激动素（Kin）的全强度Murashige和Skoog培养基（MS）上培养叶片外植体，成功实现了愈伤组织诱导。使用培养在添加了1.4 μM赤霉素（GA）和4.4 μM 6-苄基腺嘌呤（BAP）的半强度MS培养基上的节外植体，成功实现了直接芽再生。使用培养在添加了8.8 μM BAP、2.2 μM玉米素和1.4 μM GA的MS培养基上的愈伤组织，然后在添加了8.8 μM BAP和0.5 μM萘乙酸（NAA）的MS培养基上培养成功再生了间接微芽。使用野生植物地上部分、愈伤组织和微芽样品，通过顶空固相微萃取（HS-SPME）成功提取了VOCs，并通过气相色谱-质谱联用（GC-MS）进行了分析。在野生植物提取物中，倍半萜烃类占主导地位，主要成分如下：别香橙烯（11.92%）、反式-杜松-1(6),4-二烯（7.54%）和α-石竹烯（6.77%）。对离体微芽的分析显示，氧化单萜含量较高，主要成分是桃金娘醇（16.62%）和β-环柠檬醛（14.3%）。愈伤组织提取物以单萜烃类为主，鉴定出的主要化合物为（Z）-β-罗勒烯（22.27%）、对伞花烃（15.13%）和α-蒎烯（13.78%）。总之，建立了一种有效的体外VOCs生产系统，该系统未来可用于提高其产量，而不会危及野生植物。
