Integrated GC-MS and UPLC-ESI-QTOF-MS based untargeted metabolomics analysis of in vitro raised tissues of Digitalis purpurea L

• Published in: Frontiers in plant science 2024-08, Vol.15, p.1433634
• Main Authors: Bansal, Yashika, Mujib, A, Mamgain, Jyoti, Syeed, Rukaya, Mohsin, Mohammad, Nafees, Afeefa, Dewir, Yaser Hassan, Mendler-Drienyovszki, Nóra
• Format: Article
• Language: English
• Subjects: biochemical; GC-MS; indirect organogenesis; metabolomics; Plant Science; POD; SOD
• ISSN: 1664-462X; 1664-462X
• DOI: 10.3389/fpls.2024.1433634

L. is one of the important plant species of Nilgiris, Kashmir and Darjeeling regions of India, belonging to the family Plantaginaceae, with well-known pharmacological applications. In the present investigation, an culture technique of indirect shoot organogenesis of is being explored; the biochemical attributes, the antioxidant activities and the metabolomic analyses were made by utilizing untargeted Gas Chromatography-Mass Spectrometry (GC-MS) and Ultra Performance Liquid Chromatography coupled with electronspray ionization/quadrupole-time-of-flight-mass spectrometry (UPLC-ESI-QTOF-MS) approaches. Initially, the leaf explants were used for callus induction and proliferation and maximum callusing frequency (94.44%) and fresh biomass (4.9 g) were obtained on MS, fortified with 8.8 µM BAP (6-benzyl amino purine) + 0.9 µM 2,4-D (2,4-dichlorophenoxyacetic acid), subsequently shoot formation (indirect organogenesis) was noted on the same MS medium with a shoot induction frequency of 83.33%. Later on, the biochemical and antioxidant potential of -, grown leaf and leaf derived callus were assessed. Significantly higher total phenol, flavonoid, DPPH (2,2-diphenyl-1-picrylhydrazyl), POD (peroxidase) and SOD (superoxide dismutase) activities were noticed in grown callus and leaf tissues compared with field grown leaf. The GC-MS analysis of each methanolic extract ( -, derived leaf and leaf derived callus) displayed the presence of more than 75 bioactive compounds viz loliolide, stigmasterin, alpha-tocopherol, squalene, palmitic acid, linoleic acid, beta-amyrin, campesterol etc. possessing immense therapeutic importance. The UPLC-MS based metabolite fingerprinting of each methanolic extracts were conducted in both positive and negative ionization mode. The obtained results revealed variation in phytochemical composition in field - and laboratory grown tissues, indicating the impact of culture conditions on plant tissues. The detected phytocompounds belongs to various classes such as flavonoids, steroids, terpenoids, carbohydrates, tannins, lignans etc. The medicinally important metabolites identified were 20, 22-dihydrodigoxigenin, digoxigenin monodigitoxoside, apigenin, luteolin, kaempferide, rosmarinic acid, nepitrin and others. The results of the present study suggest that culture of could successfully be utilized for the novel drug discovery by producing such important phytocompounds of commercial interest in shorter duration without harming the plants' natural p