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Schematic diagram of a concentric-tube airlift bioreactor (with working volume of 1.0 l). 1 Reactor column; 2 inner draft tube (with its height of 15 cm); 3 sparger; B bottom clearance (2.5 cm); d diameter of draft tube (4.5 cm); D reactor diameter (7.0 cm); H column height (40 cm)
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Plant cell culture has recently received much attention as a useful technology for the production of valuable plant-derived secondary metabolites such as paclitaxel and ginseng saponin. The numerous problems that yet bewilder the optimization and scale-up of this process have not been over emphasized. In spite of the great progress recorded in rece...
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... feeding strategy was studied in an airlift bioreactor (Fig. 4) in order to en- hance the productivities of both ginseng saponin and ginseng polysaccharide on a reactor scale. It was found that sucrose should be fed before the specific oxygen uptake rate (SOUR) experiences a sharp decrease (at day 13) during the cultivation, i.e., the SOUR drop rate should not be larger than 0.5 mg O 2 (g DW) -1 h -1 d -1 . The added sugar could not be efficiently used for cell growth and metabolite synthesis if it was fed when the SOUR of the cells had already un- dergone a sharp drop and was very low (Table 6). Figure 5 shows the kinetics of cell growth and sugar consumption in the air- lift fed-batch cultivation (AFB). It indicates that sugar feeding did promote the final cell density, and a very high cell concentration of 29.7 g DW l -1 was suc- cessfully achieved on day 17. A very high biomass productivity of 1518 mg DW l -1 d -1 was also obtained in this case. The cell density was comparable to a previous record in flask cultures [45], while the productivity obtained here was much higher than that case, where it was 1188 mg DW l -1 d -1 [75]. In high-den- sity cultures of carrot cells in a 1-l airlift reactor, the highest cell density obtain- ed was 15.2 g l -1 (1.09 ¥ 10 7 cells ml -1 ) [77]. In the case of Catharanthus roseus suspension cultures, the maximum biomass accumulation obtained in an airlift reactor was 23.4 g l -1 , although a higher cell density over 30 g l -1 was achieved in a shake flask control ...
Citations
... Research on plants has a long-standing tradition in cell culture systems. The systems offer a promising resource for precious secondary metabolites of industrial importance, as well as drugs and food additives (Zhong 2001). In addition, it is also possible to extend our approach to produce antioxidants in medicine, contributing to the sustainable production of industrially important compounds and preserving natural sources (Pyo et al. 2004). ...
... However, the effectiveness of elicitation depends on several parameters such as concentration of biotic and abiotic elicitors, the time of exposure, and the type of plant material (Patel and Krishnamurthy 2013). In several instances, the choice of option for the enhanced production of secondary metabolites would be elicitors such as chitosan, polyamines, abscisic acid, methyl jasmonate, salicylic acid, nitric oxide, and heavy metals (Zhong 2001;Ramirez-Estrada et al. 2016). Khojasteh et al. (2016) reported the use of methyl jasmonate in Satureja khuzistanica improved the production of rosmarinic acid. ...
An understanding of plant secondary metabolites may prove to be important for novel drug development. The objective of this investigation was to enhance the biosynthesis of alkaloid compounds aervine and methylaervine in callus-derived cell suspension cultures of Aerva javanica using biotic and abiotic elicitors. The effect of five different elicitors on the biosynthesis of aervine (AE) and methylaervine (MAE) contents was studied for 20 days to determine the concentrations suitable for their accumulation in A. javanica. The callus obtained from the shoots of A. javanica on Murashige and Skoog (MS) medium containing 2.0 mg L−1 2,4-dichlorophenoxy acetic acid (2,4-D) and 0.5 mg L−1 α-naphthalene acetic acid (NAA), and their cell suspension cultures were used for the elicitation purposes. The result proved the maximum accumulation of AE and MAE contents in cell suspension culture. It was found to be 72.26 ± 0.30 mg g−1 DW of MAE, showing an increase of 8.66-fold, and AE content (7.48 ± 0.39 mg g−1 DW) in sodium carbonate (Na2CO3), whereas AE (34.10 ± 0.84 mg g−1 DW) and MAE (9.69 ± 0.04 mg g−1 DW) showed a 2.51-fold increase in salicylic acid. It was observed that the production of hairy root using Agrobacterium rhizogenes (MTCC 532) helps in improving aervine (42.22 ± 1.04 mg g−1 DW) and methylaervine (8.30 ± 0.09 mg g−1 DW) accumulation. This study will serve as an alternate protocol to improve alkaloid quantity as well as quality by elicitor stimulation. Furthermore, it may help in the sustainable production of A. javanica taxon and thereby helping in rescuing the natural sources recommended to cure several ailments.
... Extracts derived from plant cell cultures, in the form of fractions and isolated compounds, have garnered significant attention as potential bioactive agents. 13,14 The induction of callus through in vitro cultivation has been observed to yield a diverse array of active compounds, including ferulic acid, catechin, rutin, coumarins, quercetin, caffeic acid, and gallic acid. The modulation of secondary metabolite production is affected by external stimuli such as nanoelicitors and light stress. ...
In vitro plant cultures have emerged as a viable source, holding auspicious reservoirs for medicinal applications. This study aims to delineate the antioxidant and hypoglycemic potential of phytosynthesized selenium nanoparticle (SeNP)- and light stress-mediated in vitro callus cultures of Caralluma tuberculata extract. The morphophysicochemical characteristics of biogenic SeNPs were assessed through a combination of analytical techniques, including UV–visible spectrophotometry, scanning electron microscopy, energy-dispersive X-rays, Fourier transform infrared spectrometry, and zeta potential spectroscopy. The antioxidative potential of the callus extract 200 and 800 μg/mL concentrations was assessed through various tests and exhibited pronounced scavenging potential in reducing power (26.29%), ABTS + scavenging (42.51%), hydrogen peroxide inhibition (37.26%), hydroxyl radical scavenging (40.23%), and phosphomolybdate (71.66%), respectively. To inspect the hypoglycemic capacity of the callus extract, various assays consistently demonstrated a dosage-dependent relationship, with higher concentrations of the callus extract exerting a potent inhibitory impact on the catalytic sites of the alpha-amylase (78.24%), alpha-glucosidase (71.55%), antisucrase (59.24%), and antilipase (74.26%) enzyme activities, glucose uptake by yeast cells at 5, 10, and 25 mmol/L glucose solution (72.18, 60.58 and 69.33%), and glucose adsorption capacity at 5, 10, and 25 mmol/L glucose solution (74.37, 83.55, and 86.49%), respectively. The findings of this study propose selenium NPs and light-stress-mediated in vitro callus cultures of C. tuberculata potentially operating as competitive inhibitors. The outcomes of the study were exceptional and hold promising implications for future medicinal applications.
... Optimization of tissue culture media is useful in applied research and engineering of secondary metabolite biosynthetic pathways (Heidargholinezhad et al. 2023) to produce biomass and secondary metabolite production on a large scale (Bienaimé et al. 2015). The culture media are easy to manipulate by changing the sugar composition, type, and concentration of plant growth hormones, phosphate, and nitrogen levels (Zhong 2001). Increasing knowledge about economically important plant secondary metabolite pathways has implications for increasing the use of plant cultures to produce bioactive compounds (Vanisree et al. 2004). ...
The use of medicinal plants (MPs) that produce a secondary metabolite continues to grow along with population growth. As a result, overexploitation occurs and causes species extinction and genetic erosion. This article aims to answer the question about the role of biotechnology in increasing the production of secondary metabolite and their conservation. The biotechnologies that can be used to increase secondary metabolite are tissue culture and bioreactor. Secondary metabolites of tissue culture MPs improved through methods such as such as (1) selection of high-yield lines; (2) optimization of culture conditions (nutrient media composition, inoculum density, temperature, light, agitation, and aeration); (3) elicitation; (4) addition of precursors; and (5) immobilization. Meanwhile, the conservation of MPs can be carried out by utilizing in vitro technology, especially in vitro cold storage and cryopreservation. This shows that humans, biotechnology, and plants have a close relationship and can be optimized to meet the MPs needs and their conservation. Nevertheless, the social impact of the use of biotechnology still needs to be studied.
... Cell viability, total sugar content, and growth index measurements were carried out as described by Estrada-Zúñiga et al. [19]. Different system responses in plant cell suspension cultures (e.g., growth) can be used to study whether the plant cells are subjected to factors that cause stress [54]. Additionally, the tolerance of the cells was calculated based on the growth tolerance index using Equation (1) according to Rout et al. [36], with some modifications: ...
Buddleja cordata cell suspension cultures could be used as a tool for investigating the capabilities of this species to tolerate heavy metals (HMs) and for assessing the effects of HMs on the accumulation of phenolic compounds in this species. It grows in a wide range of habitats in Mexico, including ultramafic soils, and mobilizes some HMs in the soil. The mobilization of these HMs has been associated with phenolic substances. In addition, this species is used in Mexican traditional medicine. In the present study, a B. cordata cell suspension culture was grown for 18 days in a culture medium enriched with Cu (0.03–0.25 mM), Fe (0.25–1.5 mM), Mn (0.5–3.0 mM), or Zn (0.5–2.0 mM) to determine the effects of these HMs on growth and HM accumulation. We also assessed the effects of the HMs on phenolic compound accumulation after 1 and 18 days of HM exposure. Cells were able to grow at almost all tested HM concentrations and accumulated significant amounts of each HM. The highest accumulation levels were as follows: 1160 mg Cu kg−1, 6845 mg Fe kg−1, 3770 mg Mn kg−1, and 6581 mg Zn kg−1. Phenolic compound accumulation was affected by the HM exposure time and corresponded to each HM and its concentration. Future research should analyze whole plants to determine the capabilities of Buddleja cordata to accumulate abnormally high amounts of HM and to evaluate the physiological impact of changes in the accumulation of phenolic compounds.
... Such strategies have been reported to effectively increase the metabolite production to greater extent both under in-vitro as well as in-vivo conditions. The various aspects focussed in the recent past were based on the idea of manipulating signal transduction pathways in one way or the other viz., the moderation in plant nutrient medium for boosting production of SMs mainly through chemical processing or medium provided with common elicitors which can enhance the overall growth of plant (Zhong 2001;Marchev et al. 2020). Genetic manipulation of regulatory genes including the study of transcription factors for improving targeted SMs (Memelink et al. 2001;Wang et al. 2016). ...
Elicitation, the process of stimulating plants to enhance the production of valuable secondary metabolites, has emerged as a significant avenue in the field of plant biotechnology. This review paper provides a comprehensive examination of elicitation, encompassing its mechanisms, the imperative for its utilization, and the diverse array of elicitors employed, including abiotic, biotic, and nanoparticle-based agents. By exploring the intricate signaling pathways and molecular responses triggered by external stimuli in this review, we gain insights into how plants tailor their metabolite production in reaction to their environment. Intricately intertwined with the mechanisms, the types of elicitors utilized are extensively discussed. Abiotic elicitors, encompassing physical factors and chemicals, biotic elicitors involving microbe-derived molecules, plant hormones and symbiotic microorganisms, and the emerging application of nanoparticles as elicitation agents are explored. The review further examines the application of these elicitors in both in-vitro and in-vivo cultures, showcasing their practical utility in controlled laboratory settings and real-world environments. By assessing the efficacy of diverse elicitors, this paper provides valuable insights into tailoring elicitation strategies to enhance secondary metabolite yields for various applications. Addressing the limitations and leveraging emerging technologies will undoubtedly steer elicitation research towards innovative breakthroughs, further enhancing our capacity to harness plants for sustainable and efficient secondary metabolite production.
... In general, it should be noted once again that the response of plant cell cultures to hydrodynamic stress is individual and depends both on the nature and intensity of the stress and on the physiological state of the cell culture, the age of the population (cells are most susceptible to stress in the lag phase and stationary phase of the growth cycle), the concentration of components in the nutrient medium, the presence of inhibitory metabolites, etc. The hydrodynamic effects of mixing should be particularly considered in large-scale cultivation processes [19,128,138,139]. ...
The cultivation of plant cells in large-scale bioreactor systems has long been considered a promising alternative for the overexploitation of wild plants as a source of bioactive phytochemicals. This idea, however, faced multiple constraints upon realization, resulting in very few examples of technologically feasible and economically effective biotechnological companies. The bioreactor cultivation of plant cells is challenging. Even well-growing and highly biosynthetically potent cell lines require a thorough optimization of cultivation parameters when upscaling the cultivation process from laboratory to industrial volumes. The optimization includes, but is not limited to, the bioreactor’s shape and design, cultivation regime (batch, fed-batch, continuous, semi-continuous), aeration, homogenization, anti-foaming measures, etc., while maintaining a high biomass and metabolite production. Based on the literature data and our experience, the cell cultures often demonstrate cell line- or species-specific responses to parameter changes, with the dissolved oxygen concentration (pO2) and shear stress caused by stirring being frequent growth-limiting factors. The mass transfer coefficient also plays a vital role in upscaling the cultivation process from smaller to larger volumes. The Experimental Biotechnological Facility at the K.A. Timiryazev Institute of Plant Physiology has operated since the 1970s and currently hosts a cascade of bioreactors from the laboratory (20 L) to the pilot (75 L) and a semi-industrial volume (630 L) adapted for the cultivation of plant cells. In this review, we discuss the most appealing cases of the cell cultivation process’s adaptation to bioreactor conditions featuring the cell cultures of medicinal plants Dioscorea deltoidea Wall. ex Griseb., Taxus wallichiana Zucc., Stephania glabra (Roxb.) Miers, Panax japonicus (T. Nees) C.A.Mey., Polyscias filicifolia (C. Moore ex E. Fourn.) L.H. Bailey, and P. fruticosa L. Harms. The results of cell cultivation in bioreactors of different types and designs using various cultivation regimes are covered and compared with the literature data. We also discuss the role of the critical factors affecting cell behavior in bioreactors with large volumes.
... Plant cell culture provides an achievable basis for the large-scale production of secondary metabolites. Under suitable in vitro culture conditions, some secondary metabolites can be accumulated at a higher quantity in plant cultures than in whole plants, suggesting that the use of plant cell culture instead of whole plants for the production of specific secondary metabolites is promising [6]. Currently, C. bungei C. A. Mey cell engineering research is focused on the establishment of an in vitro fast propagation system [7] and the study of somatic cell embryogenesis [8]. ...
Catalpa bungei C. A. Mey is a unique and precious multi-purpose tree species that possesses great timber-related, ornamental and medicinal values. In this study, MS, N6 and DKW were used as basic media, and different concentrations of 6-BA and NAA were added for callus induction. The induction rate and total flavonoid content of callus tissue showed that the best callus induction medium was DKW + 2.0 mg·L−1 6-BA + 0.5–1.0 mg·L−1 NAA. The leaves of different bark phenotypes of C. bungei C. A. Mey and the callus tissue extracted from young leaves of C. bungei C. A. Mey were used as experimental materials to construct metabolomic profiles of widely targeted flavonoids. Based on the metabolomic databases, the predominant flavonoids were screened from the callus tissues. Eight flavonoid metabolites increased in callus, and diosmetin-7-O-rutinoside (diosmin) was the flavonoid constituent with the shortest retention time, most efficient detection and best medicinal functions among these 8. The optimal medium for callus induction was supplemented with different concentrations of elicitors (salicylic acid SA and yeast extract YE). The optimal elicitor and the amount to be added were determined by analyzing the induction rate of callus, as well as the total contents of flavonoids and diosmin. The addition of SA and YE in appropriate amounts increased the total flavonoid content in the callus, but only the addition of YE promoted the formation of diosmin in the callus. The optimal medium formulation to promote the formation of callus was DKW + 2.0 mg·L−1 6-BA + 1.0 mg·L−1 NAA + 30 g·L−1 sucrose + 6.8 g·L−1 agar + 10 μmol·L−1 SA. The medium formulation to optimally increase the content of geraniol glucoside was DKW + 2.0 mg·L−1 6-BA + 1.0 mg·L−1 NAA + 30 g·L−1 sucrose + 6.8 g·L−1 agar + 200 mg·L−1 YE. The results of the present study will provide a scientific basis for the subsequent increase in the content of the active components of the suspension cells via the addition of elicitors, and for the production of diosmin in factory settings.
... J.J. Zhong's (2001) research revealed that the accumulation of alkaloids in Holarrhena antidysenterica and shikonin in Lithospermum erythrorhizon suspension cell cultures were significantly affected by the carbon source. In 1980, Knobloch and Berlin examined the impact of saccharose concentration on the synthesis of indole alkaloids in Catharanthus roseus cell cultures, ranging between 4-12%. ...
... The induction of rapidly growing callus was observed during cultivation on MS solid medium with 4.0 mg/L -1 2,4-dichlorophenoxyacetic acid, but without additional nitrogen supply. According to J.J. Zhong (2001), raising the nitrogen content of the nutrient medium augments the anthocyanin composition in suspension cultures of Vitis vinifera. T.P. Magangana et al. (2018) demonstrated that the availability of nitrogen directly affects steviol concentration in microplants. ...
Stevia rebaudiana plants produce sweet tasting compounds, steviosides, which are 300 times sweeter as sugar and can be used as natural sweeteners. Moreover, stevia leaves contain high amount of antioxidants. The use of plant compounds as components of functional nutrition and nutraceuticals is extremely relevant today. The purpose of this study was to investigate the effect of increased nitrogen and saccharose supply on growth and accumulation of steviosides and anthocyanins as well as antioxidative activity in fast-growing shoot cultures of S. rebaudiana. The morphological changes of the leaves, the accumulation of steviosides and anthocyanins, and the antioxidant properties of the extracts were determined. It was shown that during three weeks of in vitro cultivation of shoots on MS medium with double concentration of nitrogen, the biomass increased by 71.0%, after application of double concentration of saccharose – by 133%, and after increasing nitrogen in combination with saccharose supply – by 162.0% compared to the control. Sucrose stimulated the accumulation of biomass. Anti-oxidative potential after nitrogen application was 2.4 times higher than the control, and with saccharose supply it increased 2.7 times. Shoots treated with increased nitrogen and saccharose concentration contained 50.7 and 57.8 mg/g-1 steviosides, respectively. However, the combination of nitrogen and saccharose led to accumulation of 73.4 1 mg/g-1 steviosides. Shoots grown on MS medium culture had 26.0 mg/g-1 of steviosides. The content of anthocyanins was 1.7 times greater under added nitrogen supply, and 2.3 times greater after the application of nitrogen and saccharose. Optimum cultivation media developed individually for each in vitro culture increase the production of valuable plant secondary metabolites up to 3 times
... On the other hand, there are promising efforts have recently been conducted in order to increase the discovery of plant SMs and their uses, focusing mainly on the following areas: (i) increasing the productivity of target single natural substance through the cell culturing process by altering the processing and bioreactor performance, or by using elicitors and/or other techniques, regardless of their underlying processes [27]; (ii) establishing the possible signaling pathways involving directly or indirectly the creation of some target SMs; (iii) investigating genetically the methods of regulating some transcription factors, such as altering the regulator genes to enhance the biosynthesis of target and desired SMs [28,29]; (iv) measuring and evaluating global gene expression under various settings to examine the gene transcripts for plant SMs in order to understand their mechanism of action better [30]. A detailed illustration of the various applications for plant SMs has been reported as follows. ...
Natural products are compounds produced by living organisms and can be divided into two main categories: primary (PMs) and secondary metabolites (SMs). Plant PMs are crucial for plant growth and reproduction since they are directly involved in living cell processes, whereas plant SMs are organic substances directly involved in plant defense and resistance. SMs are divided into three main groups: terpenoids, phenolics and nitrogen-containing compounds. The SMs contain a variety of biological capabilities that can be used as flavoring agents, food additives, plant-disease control, strengthen plant defenses against herbivores and, additionally, it can help plant cells to be better adapted to the physiological stress response. The current review is mainly focusing on certain key elements related to the significance, biosynthesis, classification, biochemical characterization and medical/pharmaceutical uses of the major categories of plant SMs. In addition, the usefulness of SMs in controlling plant diseases, boosting plant resistance and as potential natural, safe, eco-friendly substitutes for chemosynthetic pesticides were also reported in this review.
... Innovative approaches in biotechnology are corroborating the significance of in vitro plant tissue culture techniques in plant breeding and genetics for either biosynthesis of pharmaceutically novel metabolites or a model system to study plant physiology, biochemistry, and pathology [19,20]. The production of such novel metabolites in concentrated form through tissue culture techniques has been reported in the literature [21,22]. ...
Moringa oleifera, also called miracle tree, is a pharmaceutically important plant with a multitude of nutritional, medicinal, and therapeutic attributes. In the current study, an in-vitro-based elicitation approach was used to enhance the commercially viable bioactive compounds in an in vitro callus culture of M. oleifera. The callus culture was established and exposed to different monochromatic lights to assess the potentially interactive effects on biomass productions, biosynthesis of pharmaceutically valuable secondary metabolites, and antioxidant activity. Optimum biomass production (16.7 g/L dry weight), total phenolic contents (TPC: 18.03 mg/g), and flavonoid contents (TFC: 15.02 mg/g) were recorded in callus cultures placed under continuous white light (24 h), and of other light treatments. The highest antioxidant activity, i.e., ABTS (550.69 TEAC µM) and FRAP (365.37 TEAC µM), were also noted under white light (24 h). The analysis of phytochemicals confirmed the significant impact of white light exposures on the enhanced biosynthesis of plant secondary metabolites. The enhanced levels of secondary metabolites, i.e., kaempferol (1016.04 µg/g DW), neochlorogenic acid (998.38 µg/g DW), quercetin (959.92 µg/g DW), and minor compounds including luteolin, apigenin, and p-coumaric acid were observed as being highest in continuous white light (24 h with respect to the control (photoperiod). Similarly, blue light enhanced the chlorogenic acid accumulation. This study shows that differential spectral lights demonstrate a good approach for the enhancement of nutraceuticals along with novel pharmacologically important metabolites and antioxidants in the in vitro callus culture of M. oleifera.
