Enhancement of galanthamine production through elicitation and NMR-based metabolite profiling in Narcissus pseudonarcissus cv. Carlton in vitro callus cultures

Abstract

Plants are the unique sources of secondary metabolites (terpenes, phenolic and alkaloid) used as pharmaceuticals, food additives and industrial produces. However, very limited information is known about their biosynthesis in plants. Elicitors are chemicals often used to activate the accumulation of such metabolites in plant in vitro cultures. Since the ancient past, Narcissus is well-known for its ornamental purposes and medicinal versatility with diverse pharmaceutically important alkaloids including galanthamine, lycorine, haemanthamine and narciclasine. In this research, callus cultures induced from N. pseudonarcissus cv. Carlton bulb explants were subjected to MS media supplemented with growth regulators, yeast extract, methyl jasmonate, chitosan and trans-cinnamic acid for the determination of galanthamine content using GC-MS. Bulbs showed 538 to 1109 µg g -1 FW (fresh weight) and callus cultures subjected to MS medium without elicitors produced 7.88 µg g -1 FW of galanthamine. Whereas, callus cultures grown on MS with methyl jasmonate, chitosan and 25% sucrose showed approximately 5.6, 3 and 2 fold increased galanthamine production respectively. NMR (Nuclear Magnetic Resonance) depicted that the concentration of metabolites related to galanthamine production, tyrosine, tyramine, 3-chlorotyrosine, were higher in bulb tissue than callus.However, these metabolites were present in higher concentrations in elicitor-treated calluses, leading to the higher galanthamine accumulation than the non-treated calluses. Furthermore, the notable metabolites detected in elicitor-treated callus cultures were amino acids, phenols and sugar alcohols. The metabolites in media extracts were mainly related to sucrose and galactose metabolism. The results suggested that the application of elicitors could be a potential approach for enhanced production of valuable metabolites in Narcissus in vitro culture.

Comparative Analysis of Primary and Secondary Metabolites in Different In Vitro Tissues of Narcissus tazetta var. chinensis

“…Similar to our study, research on galantamine and metabolites using Narcissus pseudo narcissus , which is closely related to Narcissus gazzetta , also showed that the galantamine content is lowest in the callus. 37 However, unlike our data, this study was not able to detect a significant amount of galantamine in the root, which may be due to the different conditions for root induction. For example, unlike our tissue culture conditions, for root induction, we used cytokinin, which can act as a NAA antagonist.…”

“…This suggests an efficient and high-yielding method for the commercial-scale production of galantamine. Similar to our study, research on galantamine and metabolites using Narcissus pseudo narcissus, which is closely related to Narcissus gazzetta, also showed that the galantamine content is lowest in the callus . However, unlike our data, this study was not able to detect a significant amount of galantamine in the root, which may be due to the different conditions for root induction.…”

“…Another reason might be the time duration for the induction of the root tissue. For example, Ferdausi et al 37 analyzed the galantamine content in 14 week old root cultures, whereas in our study, we analyzed 6 week old cultures, which might explain the highest accumulation of galantamine in N. tazetta roots. Additionally, another study found that the timing of tissue harvest depends on the developmental stage, such as before and after flowering, which alters the galantamine content in a tissue-specific manner.…”

henolics profile and antioxidant activities of in vitro propagules and field-raised plant organs of Curculigo latifolia

“…The plantlets were cultured on MS basal medium with plant growth regulators. Plant callus as an alternative source of raw material can be induced to increase its metabolites content through several interventions, for example, by adding biotic or abiotic elicitors (Ferdausi et al, 2021), manipulating biosynthetic pathways (Inyai et al, 2021), or genetic engineering (Shi et al, 2021).…”

Elicitation Induced α-Amyrin Synthesis in Tylophora indica In Vitro Cultures and Comparative Phytochemical Analyses of In Vivo and Micropropagated Plants

“…Phytochemical profiling with gas chromatography–mass spectrometry (GC–MS) has recently emerged as one of the most valuable techniques to detect and identify bioactive compounds, like alkaloids, flavonoids, tannins, phenols, steroids, terpenoids and essential oils [ 8 , 9 ]. This technique requires a low extract level to analyze metabolites with precision and reproducibility [ 10 ]. There are reports of GC–MS-based profiling of in vitro regenerated plants, like Tecoma stans [ 11 ] and Curcuma zedoaria [ 12 ].…”