In vitro micropropagation and alkaloids analysis by GC–MS of Chilean Amaryllidaceae plants: Rhodophiala pratensis

Abstract

Introduction Plants from Amaryllidaceae family are of interest since they produce a particular type of alkaloid useful for the treatment of neurodegenerative diseases of the central nervous system, such as Galanthamine. Given the low content of these secondary metabolites in the plant, it is necessary to study mechanisms to increase the productivity of them. Objective To obtain fast qualitative and quantitative analysis of the alkaloids and extend the understanding of biosynthesis and metabolism in these kinds of plants. Furthermore, establish a reliable, simple and fast analytical method for the in vitro callus culture of vegetative organs for Rhodophiala pratensis species. Methods The alkaloids composition of the callus culture of R. pratensis were analysed by gas chromatography coupled with mass spectrometry (GC–MS). Results A methodology for the qualitative and quantitative analysis of the alkaloids present in fresh callus culture of this wild plant species was established. The analysis showed alternation in the alkaloids type ratio and number of compounds between wild bulbs, in vitro bulbs and callus. It was possible to identify 24 alkaloids from a pool of 60 signals whose fragmentation pattern corresponds to the alkaloids of Amaryllidaceae plants. Together with the aforementioned, the amount and type of alkaloid present in the plant material obtained by in vitro culture of R. pratensis was determined in the same way. The results show the high biosynthetic potential of in vitro grown bulbs and callus tissue that are able to produce significant amounts of pharmacologically relevant alkaloids from R. pratensis in various proportions that depend on the culture conditions such as supplementation with growth substances. The in vitro grown bulbs produce an alkaloidal extract that contain a 52.6% w/w of alkaloids. Conclusion This study allowed the alkaloid content in callus culture of R. pratensis to be found by means of GC–MS. These results allowed a relationship between the type of growth regulator and the type of alkaloids found to be established. Finally, we can say that the results achieved to state that the production of alkaloids using different combinations of growth regulators could be directed during in vitro micropropagation from provided plant material.

Alkaloids Analysis of Habranthus cardenasianus (Amaryllidaceae), Anti-Cholinesterase Activity and Biomass Production by Propagation Strategies

“…Thus, the extraction of bioactive metabolites from natural sources is currently not viable, and it is important to develop sustainable propagation methods for Amaryllidaceae domestication [17]. Such methods include seed propagation [18,19], cut-induced bulb division [20][21][22], and micropropagation [23][24][25]. Seed propagation is a simple and economical way to obtain seedlings [26] and allows a good multiplication rate, but it has the drawback of a high genetic drift [19].…”

“…Cut-induced bulb division consists of making incisions that induce hormonal mechanisms in the plant and promote the generation of new bulbils [20]. In-vitro culture is large scale micropropagation using tissue culture techniques [23]. Micropropagation is possible due to the totipotentiality of plant cells, which is the ability to regenerate a complete plant under the appropriate stimuli [17,24].…”

“…Micropropagation is possible due to the totipotentiality of plant cells, which is the ability to regenerate a complete plant under the appropriate stimuli [17,24]. Micropropagation is very useful in breeding programs because it produces a uniform quality of commercial plants, with a high multiplication rate, little genetic variability, and a stable production of the metabolites of interest [23]. To the best of our knowledge, this work represents the first chemical study of H. cardenasianus bulbs, the evaluation of their inhibitory activity against cholinesterases, and the analysis of biomass production by several propagation strategies to produce bioactive alkaloids.…”

Biological Investigation of Amaryllidaceae Alkaloid Extracts from the Bulbs of Pancratium trianthum Collected in the Senegalese Flora

“…However, identical fragmentations were reported in the literature and corresponded to 11,12-dehydroanhydrolycorine ( 2 ) and trisphaeridine ( 6 ) [ 13 ]. Hamayne ( 5 ), also not available in NIST 05 database, showed ions at m / z 287, 258, 242, 186, and 153 and was identified by similarity with the reported AA from Rhodophiala pratensis [ 36 ], while compound ( 4 ) displayed ions at m / z 273, 201, 175, 157, 141, and 128, identically to 8- O -demethylmaritidine from Amaryllis belladonna L. [ 35 ]. Additionally, an approximation of the relative proportion of identified AAs was estimated as a percentage of the total ion current (TIC) chromatogram.…”

Biotechnological Approaches to Optimize the Production of Amaryllidaceae Alkaloids

“…It can be applied to grow whole plants, plant parts, or undifferentiated tissues. Micropropagation, a technique that enables rapid vegetative/clonal multiplication of plants from limited or small size plants, has been successfully applied with Amaryllidaceae species such as Rhodophiala pratensis , Lapiedra martinezii , Eucrosia stricklandii , and Lycoris sprengeri , leading to plant development with similar morphometric traits [ 28 , 41 , 42 , 43 , 44 ]. Other cultivation methods of plant material in vitro (bulblets, seedlings, plantlets, shoots, roots, shoot–clump, callus) also provide an interesting opportunity to produce AAs, being effective for both conservation, long term growth, and industrial purpose.…”