Elicitation of galanthamine and lycorine biosynthesis by Leucojum aestivum L. and L. aestivum ‘Gravety Giant’ plants cultured in bioreactor RITA®

Ptak, Agata; Morańska, Emilia; Saliba, Sahar; Zieliński, Andrzej; Simlat, Magdalena; Laurain-Mattar, Dominique

doi:10.1007/s11240-016-1113-3

Cited by 23 publications

(14 citation statements)

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“…For each a RITA Ò vessels 200 mL of medium and 5 g of plants were used. The physical parameters used for the culture were previously described (Ptak et al, 2017). The experiment was established in three replications.…”

Section: Treatment and Culture Conditionsmentioning

confidence: 99%

“…In the studies on biosynthesis of Amaryllidaceae alkaloids in in vitro cultures of L. aestivum the influence of mineral nutrients, auxins, cytokinins, ethylene and melatonin was tested (Laurain-Mattar & Ptak, 2018). Elicitation with methyl jasmonate and salicylic acid, and biotransformation using precursor 4′-O-methylnorbelladine were also studied (Saliba et al, 2016;Ptak et al, 2017). Additionally, the effects of physical factors, such as temperature and physical state of the medium, on the production of galanthamine and lycorine, in particular, were studied (Laurain-Mattar & Ptak, 2018).…”

Section: Introductionmentioning

confidence: 99%

“…To date, studies have been conducted only on the effect of sucrose concentration in shaken shoot cultures of N. confusus and L. aestivum (Sellés et al, 1997;Schumann et al, 2012). Studies on the adaptation of the temporary immersion system, RITA Ò , for growing L. aestivum shoot and plant cultures were carried out by Ivanov et al (2011) and Ptak et al (2017). However, the results of the research conducted by Ivanov et al (2012) and Georgiev et al (2014) on shoot cultures of L. aestivum, as well as Pancratium maritimum, with the use of standard amount of sucrose in the medium, showed that the optimal conditions for growth in the RITA Ò bioreactor (immersion frequency and stand-by periods) provided both the optimal biomass accumulation and the best sugar utilization from the medium.…”

Section: Introductionmentioning

confidence: 99%

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Carbohydrates stimulated Amaryllidaceae alkaloids biosynthesis inLeucojum aestivumL. plants cultured in RITA^®bioreactor

Ptak

Morańska

Skrzypek

et al. 2020

PeerJ

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Background Leucojum aestivum L. is an important medicinal plant which produces Amaryllidaceae alkaloids, especially galanthamine and lycorine. Research is currently exploring the possibility of producing these alkaloids using biotechnological methods, including in vitro cultures. The biosynthesis of alkaloids may be affected by the types and concentrations of carbohydrate sources used in the medium. In the present investigation we performed such studies on in vitro cultures of L. aestivum with a view to obtaining plant material of good quality, characterized, in particular, by a high content of valuable Amaryllidaceae alkaloids. Methods We examined the effects of various types of carbohydrate sources—sucrose, glucose, fructose and maltose—at different concentrations (30, 60 and 90 g/L)—on the quality of L. aestivum plants grown in the RITA® bioreactor. The plants’ quality was assessed by their biomass increments, as well by as analysing photosynthetic pigments, endogenous sugar, phenolics and Amaryllidaceae alkaloid content. We also investigated the effect of sugars on the activity of the antioxidant enzymes catalase (CAT), peroxidase (POD) and superoxide dismutase (SOD). Results The highest biomass increments were observed in plants cultivated in the medium containing 90 g/L sucrose. The highest CAT activity was noted in cultures growing in the medium supplemented with 90 g/L maltose, while the highest POD activity was observed in the presence of 90 g/L fructose and 60 g/L maltose. No differences in SOD activity were observed. Moreover, the sugars did not affect the contents of chlorophyll a and carotenoids, whereas the highest amount of chlorophyll b was recorded in plants growing in the medium with 60 g/L maltose. No statistically significant differences were observed in the contents of endogenous sugars and phenolics in any in vitro conditions. However, the addition of sugar had a decisive effect on the biosynthesis of the Amaryllidaceae alkaloids. The highest distribution of alkaloids occurred in plants cultured in the medium containing 60 g/L sucrose. Six Amaryllidaceae alkaloids were detected in the plant tissue. The addition of 30 g/L fructose in the medium resulted in the accumulation of five alkaloids, including ismine, which was not identified in other analysed tissues. The highest concentration of galanthamine was observed in plants cultured in the presence of 30 g/L fructose and 60 g/L sucrose (39.2 and 37.5 µg/g of dry weight (DW), respectively). The plants grown in the medium containing 60 g/L sucrose exhibited the highest lycorine content (1048 µg/g of DW). Conclusions The type and concentration of sugar used in the medium have an essential influence on the biosynthesis of Amaryllidaceae alkaloids in L. aestivum plants cultured in a RITA® bioreactor. The results point to an interesting approach for commercial production of galanthamine and lycorine.

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Section: Treatment and Culture Conditionsmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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Carbohydrates stimulated Amaryllidaceae alkaloids biosynthesis inLeucojum aestivumL. plants cultured in RITA^®bioreactor

Ptak

Morańska

Skrzypek

et al. 2020

PeerJ

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“…It was established that shoot cultures had higher potential to synthesize this alkaloid as compared to hairy roots described by Diop and coworkers [11]. Yields of galanthamine, produced by shoot culture of L. aestivum, were strongly influenced from illumination [9], nutrient medium composition [12], and addition of elicitors [13][14][15][16][17]. Additionally, our group proposed a modified formulation of the nutrient medium composition for maximal biosynthesis of galanthamine by L. aestivum shoots cultivated under submerged conditions [9].…”

mentioning

confidence: 96%

In sito galanthamine extraction during the cultivation of Leucojum aestivum L. shoot culture in two‐phase bubble column cultivation system

Ivanov

Berkov

Pavlov

et al. 2019

Engineering in Life Sciences

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Two‐phase bioreactor cultivation system was developed and applied for in sito recovery of extracellular galanthamine during the cultivation of Leucojum aestivum L. shoot culture in a modified column bioreactor system. The inclusion of an external circulation column with adsorbent resin Amberlite XAD‐4 as a second phase, on the 21st day of the beginning of cultivation resulted in 1.25 folds increase in biomass accumulation and maximal amounts of accumulated galanthamine of 6 mg/L (3.1 mg/L intracellular and 2.9 mg/L extracellular). It was demonstrated that the inclusion of a second phase at the cultivation of the L. aestivum shoot culture in a bubble column bioreactor with internal sections redirected the alkaloid metabolism to galanthamine synthesis and inhibits the synthesis of hemanthamine and lycorine type alkaloids. Our research demonstrated that the application of the two‐phase cultivation systems could be an important tool to increase the yields of valuable secondary metabolites in plant tissue culture‐based bioprocess.

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“…Literature data have reported that contents of these alkaloids in these plants, particularly galanthamine, oscillate between 0.01 and 2.0% dry weight (DW). 10,11 To the earlier mentioned is added the fact that the multiplication of bulbous plants from seeds will show wide variability in morphological features, whereas separation of bulbs is a very slow process for obtaining a large number of plants in a short time. Normally, a plant produces two to three bulbs in a year of growth.…”

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confidence: 99%

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

Trujillo-Chacón

Pastene

Bustamante

et al. 2019

Phytochemical Analysis

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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.

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Elicitation of galanthamine and lycorine biosynthesis by Leucojum aestivum L. and L. aestivum ‘Gravety Giant’ plants cultured in bioreactor RITA®

Cited by 23 publications

References 34 publications

Carbohydrates stimulated Amaryllidaceae alkaloids biosynthesis inLeucojum aestivumL. plants cultured in RITA^®bioreactor

Carbohydrates stimulated Amaryllidaceae alkaloids biosynthesis inLeucojum aestivumL. plants cultured in RITA^®bioreactor

In sito galanthamine extraction during the cultivation of Leucojum aestivum L. shoot culture in two‐phase bubble column cultivation system

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

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Elicitation of galanthamine and lycorine biosynthesis by Leucojum aestivum L. and L. aestivum ‘Gravety Giant’ plants cultured in bioreactor RITA®

Cited by 23 publications

References 34 publications

Carbohydrates stimulated Amaryllidaceae alkaloids biosynthesis inLeucojum aestivumL. plants cultured in RITA®bioreactor

Carbohydrates stimulated Amaryllidaceae alkaloids biosynthesis inLeucojum aestivumL. plants cultured in RITA®bioreactor

In sito galanthamine extraction during the cultivation of Leucojum aestivum L. shoot culture in two‐phase bubble column cultivation system

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

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Product

Resources

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Carbohydrates stimulated Amaryllidaceae alkaloids biosynthesis inLeucojum aestivumL. plants cultured in RITA^®bioreactor

Carbohydrates stimulated Amaryllidaceae alkaloids biosynthesis inLeucojum aestivumL. plants cultured in RITA^®bioreactor