Relative expression of putative genes involved in galanthamine and other Amaryllidaceae alkaloids biosynthesis in Narcissus field and in vitro tissues

Aleya, Ferdusi, Chang, Xianmin, Hall, Anthony and Jones, Meriel (2021) Relative expression of putative genes involved in galanthamine and other Amaryllidaceae alkaloids biosynthesis in Narcissus field and in vitro tissues. Gene, 774. ISSN 0378-1119

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Abstract

The Narcissus pseudonarcissus cv. Carlton contains Amaryllidaceae alkaloids namely galanthamine, lycorine, homolycorine, narciclasine, which are noted for their pharmaceutical properties such as for the treatment of early to mid-stage Alzheimer’s diseases, cancer, tumor etc. Alkaloid biosynthesis using plant in vitro systems has been considered as a tool for drug discovery and the pathways are starting to be understood but still far from complete. Therefore, the study was emphasized to observe the relative expressions of putative genes involved in the biosynthetic pathway leading to the Amaryllidaceae alkaloids in field grown bulbs and developing cell culture systems in Narcissus. MS media fortified with growth regulators were used for the development of tissue culture from Carlton twinscale explants. MS medium with high auxin, 20 mg/l NAA was the best medium for callus growth and maintenance while media with low auxin, 4 mg/l NAA and MS basal media gave the maximum bulblets. Field tissues showed a higher amount of galanthamine content; i.e. basal plate (1050–1310 μg Gal/g FW) and bulb(980–1150 μg Gal/g FW) than the culture derived samples; callus (1.0–7.0 μg Gal/g FW) and bulblets (12–215 μg Gal/g FW) on a fresh weight (FW) basis. GC–MS chromatograms of samples under study also showed the presence of other important alkaloids i.e. lycorine, homolycorine, lycorenine, haemanthamine, crinamine, lycoramine and tazettine. RNA extracted from in vitro callus, bulblets and field grown bulb, basal plate were used for PCR to detect the relative expression of putative genes; P450, PAL, TYDC and NpO4OMT normalized to actin. The selected transcripts for P450s and TYDC were expressed in both field and in vitro tissues. Higher expressions of PAL were observed in calli than field samples. The expression of NpN4OMT was notably higher in field samples than in vitro tissues. Therefore, in vitro tissues could be a good source for the reproducible and easy extraction of alkaloids from plants.

Item Type: Article
Keywords: Narcissus Galanthamine Alkaloids In vitro P450 (Cytochrome P450) PAL (Phenyl ammonia lyase) TYDC (Tyrosine decarboxylase) and NpO4OMT (norbelladine 4′-O-methyltransferase)
Divisions: Agriculture, Food and Environment
Depositing User: Professor Xianmin Chang
Date Deposited: 03 Feb 2021 10:01
Last Modified: 19 Oct 2022 14:45
URI: https://rau.repository.guildhe.ac.uk/id/eprint/16440

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