scholarly journals Optimization of Biomass Accumulation and Production of Phenolic Compounds in Callus Cultures of Rhodiola rosea L. Using Design of Experiments

Plants ◽  
2022 ◽  
Vol 11 (1) ◽  
pp. 124
Author(s):  
Anna A. Erst ◽  
Anastasia A. Petruk ◽  
Andrey S. Erst ◽  
Denis A. Krivenko ◽  
Nadezhda V. Filinova ◽  
...  

Rhodiola rosea L. is a valuable medicinal plant with adaptogenic, neuroprotective, antitumor, cardioprotective, and antidepressant effects. In this study, design of experiments methodology was employed to analyze and optimize the interacting effects of mineral compounds (concentration of NO3− and the ratio of NH4+ to K+) and two plant growth regulators [total 6-benzylaminopurine (BAP) and α-naphthylacetic acid (NAA) concentration and the ratio of BAP to NAA] on the growth and the production of total phenolic compounds (TPCs) in R. rosea calluses. The overall effect of the model was highly significant (p < 0.0001), indicating that NH4+, K+, NO3−, BAP, and NAA significantly affected growth. The best callus growth (703%) and the highest production of TPCs (75.17 mg/g) were achieved at an NH4+/K+ ratio of 0.33 and BAP/NAA of 0.33, provided that the concentration of plant growth regulators was 30 μM and that of NO3− was ≤40 mM. According to high-performance liquid chromatography analyses of aerial parts (leaves and stems), in vitro seedlings and callus cultures of R. rosea contain no detectable rosarin, rosavin, rosin, and cinnamyl alcohol. This is the first report on the creation of an experiment for the significant improvement of biomass accumulation and TPC production in callus cultures of R. rosea.

Molecules ◽  
2019 ◽  
Vol 24 (11) ◽  
pp. 2112 ◽  
Author(s):  
Mariella Bonello ◽  
Uroš Gašić ◽  
Živoslav Tešić ◽  
Everaldo Attard

The production of secondary metabolites in tissue culture has been considered as an alternative to the cultivation and harvesting of crops intended for this purpose. The present study was aimed at the growth of callus and production of polyphenolic compound of callus derived from a Maltese indigenous grapevine variety, Ġellewża. Callus was inoculated onto plant growth regulators-enriched Murashige Skoog media (MSm) to determine whether polyphenols are produced in vitro as well as to determine the best combination of plant growth regulators needed for the production of these metabolites. From results obtained, it was observed that the best callus production was obtained by auxin-enriched MSm. In fact, indole acetic acid and indole acetic acid /6-benzyl aminopurine enhanced biomass accumulation (3.04 g and 3.39 g) as opposed to the others (<1.97 g). On the other hand, parameters showing the presence of flavonoids (tonality, 3.80), particularly anthocyanins (24.09 mg/kg) and total polyphenols (1.42 mg/g), were optimum in the presence of cytokinins, particularly 6-benzyl aminopurine. Analysis for single polyphenols revealed a high amount a particular stilbene: polydatin (glucoside of resveratrol). Resveratrol and other typical polyphenols, found in mature berries, were also found in significant quantities, while the other polyphenolic compounds were found in minimal quantities. This is the first study to describe the production and composition of polyphenols in Ġellewża callus cultures. From the results obtained, it can be seen that this grape tissue is an excellent alternative for the production of polyphenols from the stilbene group, which can be upscaled and exploited commercially.


2020 ◽  
Vol 44 (2) ◽  
pp. 163-173 ◽  
Author(s):  
Niloofar Hemmati ◽  
Monireh Cheniany ◽  
Ali Ganjeali

This study was undertaken to investigate the effect of different plant growth regulators (PGRs) on callus induction in Salvia tebesana explants grown in vitro and to evaluate the content of secondary phenolic compounds and their antioxidant potential. The explants (shoot apical meristem, leaf and petiole) were dissected from an 8-week-old plant of S. tebesana growing in vitro and cultured on MS media containing different concentrations of 2,4-D (0, 0.5, 1, 1.5 and 2 mg L-1), NAA (0, 0.5 and 1 mg L-1) and BAP (0, 0.5 and 1 mg L-1), either alone or in a blend with each other. Morphological characteristics of the callus (consistency and colour), biomass increase based on fresh and dry weight and the percentage of induction were recorded after 56 days. Levels of total phenols, ortho-diphenols, phenolic acids, flavonoids, proanthocyanidins and flavonols of callus, as well as antioxidant activities, were evaluated in vitro. The maximum callus formation (100%) was obtained from shoot apical meristem on MS medium supplemented with 0.5 and 1.5 mg L-1 2,4-D + 1 mg L-1 BAP and with 1 and 1.5 mg L-1 2,4-D + 0.5 mg L-1 BAP, whereas the highest fresh (15.06 ? 0.88 g) and dry (0.33 ? 0.02 g) weights of call were observed in a medium containing1.5 mg L-1 2,4-D + 0.5 mg L-1 NAA. It was noted that MS media augmented with combined PGRs had the highest accumulation of polyphenols, phenolic acids and flavonoid compounds, with levels of content varying in the following order: 2,4-D + BAP > NAA + BAP > 2,4-D + NAA. Strong linear correlations were established between total phenolic content of callus extracts and results of the DPPH and FRAP assays (r2 = 0.896 and r2 = 0.946, p < 0.01, respectively). The obtained results suggest that the described method could be utilised as a tool for large-scale production of medicinal metabolites of S. tebesana by tissue culture.


2019 ◽  
Author(s):  
Tasiu Isah

Abstract Background: In vitro morphogenic response of mature seed embryo-derived callus cultures of Chonemorpha fragrans was studied using solid and liquid Murashige and Skoog medium amended with cytokinins or their combinations with naphthalene acetic acid at 0.5 mg L-1.Results: The tested plant growth regulators combination and concentrations could not stimulate organogenesis after three subcultivations of the callus cultures on the same PGRs-amended solid medium, and when cultivated in the liquid but, formation of morphogenic callus was observed. Evaluation of biomass and camptothecin production showed that the PGRs influenced biomass and CPT yield of the callus cultures. The alkaloid yield of various explants of 3–4 weeks old axenic seedlings was higher in roots (0.019 % CPT) followed by mature seed embryos (0.0053 %), cotyledons (0.0039 %), hypocotyls (0.0024 %) and leaves (0.0017 %). There was no significant difference in yield of CPT from callus induced from the various explants. Camptothecin yield of morphogenic callus cultures cultivated in liquid medium was lower than that of solid due to extracellular leaching effect of the alkaloid. Amount of synthesized CPT in the callus cultures also varied with PGR type and concentration amended in the cultivation medium, and showed association with biomass production.Conclusion: Results of the present study suggest that callus cultures offer alternative tissue source for in vitro CPT yield enhancement through biotechnological approaches, with application in the large-scale production of the alkaloid to conserve the ever decimated natural population of the medicinal woody climber for CPT.


2021 ◽  
Vol 13 (4) ◽  
pp. 11052
Author(s):  
Tasiu ISAH ◽  
Shruti SINGH

In vitro morphogenic response of mature seed embryo-derived callus cultures of Chonemorpha fragrans was studied using solid and liquid Murashige and Skoog medium amended with cytokinins or their combinations with naphthalene acetic acid at 0.5 mg L-1. The plant growth regulators (PGRs) combination and concentrations tested could not stimulate organogenesis after three subcultivations of the callus cultures on the same PGRs-amended solid medium and when cultivated in the liquid but, formation of morphogenic callus was observed. Evaluation of biomass and camptothecin production showed that the PGRs influenced biomass and CPT yield of the callus cultures. The alkaloid yield of various explants of 3-4 weeks old axenic seedlings was higher in roots (0.019% CPT) followed by mature seed embryos (0.0053%), cotyledons (0.0039%), hypocotyls (0.0024%) and leaves (0.0017%), and no significant difference was observed in yield of CPT from callus induced from the various explants. Camptothecin yield of morphogenic callus cultures cultivated in liquid medium was lower than that of solid due to extracellular leaching effect of the alkaloid. Amount of synthesized CPT in the callus cultures also varied with PGR type and concentration amended in the cultivation medium, and was association with biomass production. Results of the present study suggest that callus cultures offer alternative tissue source for in vitro CPT yield enhancement through biotechnological approaches, with application in the large-scale production of the alkaloid to conserve the ever-decimated natural population of the medicinal woody climber for CPT.


Plants ◽  
2021 ◽  
Vol 10 (3) ◽  
pp. 437
Author(s):  
Anza-Tshilidzi Ramabulana ◽  
Paul A. Steenkamp ◽  
Ntakadzeni E. Madala ◽  
Ian A. Dubery

Plant cell culture offers an alternative to whole plants for the production of biologically important specialised metabolites. In cultured plant cells, manipulation by auxin and cytokinin plant growth regulators (PGRs) may lead to in vitro organogenesis and metabolome changes. In this study, six different combination ratios of 2,4-dichlorophenoxyacetic acid (2,4-D) and benzylaminopurine (BAP) were investigated with the aim to induce indirect organogenesis from Bidens pilosa callus and to investigate the associated induced changes in the metabolomes of these calli. Phenotypic appearance of the calli and total phenolic contents of hydromethanolic extracts indicated underlying biochemical differences that were investigated using untargeted metabolomics, based on ultra-high-performance liquid chromatography quadrupole time-of-flight mass spectrometry (UHPLC–qTOF–MS), combined with multivariate data analysis. The concentration and combination ratios of PGRs were shown to induce differential metabolic responses and, thus, distinct metabolomic profiles, dominated by chlorogenic acids consisting of caffeoyl- and feruloyl-derivatives of quinic acid. Although organogenesis was not achieved, the results demonstrate that exogenous application PGRs can be used to manipulate the metabolome of B. pilosa for in vitro production of specialised metabolites with purported pharmacological properties.


2019 ◽  
Author(s):  
Tasiu Isah ◽  
Shahid Umar

Abstract Background: In vitro morphogenic response of mature seed embryo-induced callus cultures of Chonemorpha fragrans was studied using solid and liquid Murashige and Skoog medium amended with cytokinins or their combinations with naphthalene acetic acid at 0.5 mg L-1. Results: The tested plant growth regulators combination and concentrations did not stimulate organogenesis after three subcultivations of the callus cultures on the same PGRs-amended solid medium, and when cultivated in the liquid but, formation of morphogenic callus was observed. Evaluation of biomass and camptothecin production showed that the PGRs influenced biomass and CPT yield of the callus cultures. The alkaloid yield of various explants of 3–4 weeks old axenic seedlings was higher in roots (0.019 % CPT) followed by mature seed embryos (0.0053 %), cotyledons (0.0039 %), hypocotyls (0.0024 %) and leaves (0.0017 %). There was no significant difference in the yield of CPT from callus induced from the various explants. Camptothecin yield of morphogenic callus cultures cultivated in liquid medium was lower than that of solid due to extracellular leaching effect of the alkaloid. Amount of synthesized CPT in the callus cultures also varied with the PGR type and concentration amended in the cultivation medium, and showed association with biomass production. Conclusion: Results of the present study suggest that callus cultures offer alternative tissue source for the in vitro CPT yield enhancement through biotechnological approaches, with application in the large-scale production of the alkaloid to conserve the ever decimated natural population of this medicinal woody climber for the alkaloid.


Agronomy ◽  
2021 ◽  
Vol 11 (2) ◽  
pp. 320
Author(s):  
Nisar Ahmad Zahid ◽  
Hawa Z.E. Jaafar ◽  
Mansor Hakiman

Ginger (Zingiber officinale Roscoe) var. Bentong is a monocotyledon plant that belongs to the Zingiberaceae family. Bentong ginger is the most popular cultivar of ginger in Malaysia, which is conventionally propagated by its rhizome. As its rhizomes are the economic part of the plant, the allocation of a large amount of rhizomes as planting materials increases agricultural input cost. Simultaneously, the rhizomes’ availability as planting materials is restricted due to the high demand for fresh rhizomes in the market. Moreover, ginger propagation using its rhizome is accompanied by several types of soil-borne diseases. Plant tissue culture techniques have been applied to produce disease-free planting materials of ginger to overcome these problems. Hence, the in vitro-induced microrhizomes are considered as alternative disease-free planting materials for ginger cultivation. On the other hand, Bentong ginger has not been studied for its microrhizome induction. Therefore, this study was conducted to optimize sucrose and plant growth regulators (PGRs) for its microrhizome induction. Microrhizomes were successfully induced in Murashige and Skoog (MS) medium supplemented with a high sucrose concentration (>45 g L−1). In addition, zeatin at 5–10 µM was found more effective for microrhizome induction than 6-benzylaminopurine (BAP) at a similar concentration. The addition of 7.5 µM 1-naphthaleneacetic acid (NAA) further enhanced microrhizome formation and reduced sucrose’s required dose that needs to be supplied for efficient microrhizome formation. MS medium supplemented with 60 g L−1 sucrose, 10 µM zeatin and 7.5 µM NAA was the optimum combination for the microrhizome induction of Bentong ginger. The in vitro-induced microrhizomes sprouted indoors in moist sand and all the sprouted microrhizomes were successfully established in field conditions. In conclusion, in vitro microrhizomes can be used as disease-free planting materials for the commercial cultivation of Bentong ginger.


2011 ◽  
Vol 3 (3) ◽  
pp. 97-100
Author(s):  
Naimeh SHARIFMOGHADAM ◽  
Abbas SAFARNEJAD ◽  
Sayed Mohammad TABATABAEI

The Almond (Amygdalus communis) is one of the most important and oldest commercial nut crops, belonging to the Rosaceae family. Almond has been used as base material in pharmaceutical, cosmetic, hygienically and food industry. Propagation by tissue culture technique is the most important one in woody plants. In the current research, in vitro optimization of tissue culture and mass production of almond was investigated. In this idea, explants of actively growing shoots were collected and sterilized, then transferred to MS medium with different concentrations and combinations of plant growth regulators. The experiment was done in completely randomized blocks design, with 7 treatment and 30 replications. After 4 weeks, calli induction, proliferation, shoot length and number of shoot per explants were measured. Results showed that the best medium for shoot initiation and proliferation was MS + 0.5 mg/l IAA (Indol-3-Acetic Acid) + 1 mg/l BA (Benzyl Adenine). Autumn was the best season for collecting explants. The shoots were transferred to root induction medium with different concentrations of plant growth regulators. The best root induction medium was MS + 0.5 mg/l IBA (Indol Butyric Acid).


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