Search Results (17)

Ruta corsica is a rare and endemic plant native to Corsica. Due to its limited distribution and the priority to preserve natural sites, has been insufficiently studied. In vitro cultures provide an opportunity to research R. corsica under controlled conditions. In the present study, in vitro cultures of R. corsica were conducted in Plantform^TM bioreactors. The study aimed to assess the effects of growth cycle length (5 and 6 weeks) and different concentrations of plant growth regulators (NAA and BAP) at 0.1/0.1, 0.1/0.5, 0.5/0.5, 0.5/1.0, and 1.0/1.0 mg/L on biomass growth and secondary metabolite accumulation. HPLC analysis identified compounds in the furanocoumarin and furoquinoline alkaloid groups, with furanocoumarins being the primary secondary metabolites (maximum total content: 1571.5 mg/100 g DW). Among them, xanthotoxin, psoralen, and bergapten were dominant, with maximum concentrations of 588.1, 426.6, and 325.2 mg/100 g DW, respectively. The maximum total content of furoquinoline alkaloids was 661 mg/100 g DW, with γ-fagarine as the primary metabolite, reaching 448 mg/100 g DW. The optimal conditions for secondary metabolite accumulation in R. corsica cultures were a 5-week growth cycle and the LS 0.1/0.1 medium variant. Full article

Sundews (Drosera sp.) are the source of biologically active secondary metabolites: phenolic acids, flavonoids, and 1,4-naphtoquinones. Because obtaining them from the natural environment is impossible (rare and endangered species), in this study modifications of traditional tissue cultures grown in solid medium (SM), such as agitated cultures (ACs) (cultures in liquid medium with rotary shaking) and temporary immersion bioreactors Plantform^TM (TIB), were used for multiplication of four sundew species: Drosera peltata, Drosera indica, Drosera regia, and Drosera binata, with simultaneously effective synthesis of biologically active phenolic compounds. Each species cultivated on SM, AC, and TIB was tested for biomass accumulation, the content of total phenols and selected phenolic derivative concentrations (DAD-HPLC), the productivity on of phenolic compounds, as well as its antibacterial activity against two human pathogens: Staphylococcus aureus and Escherichia coli. The results showed that the type of culture should be selected for each species separately. Phytochemical analyses showed that the synthesis of secondary metabolites from the groups of phenolic acids, flavonoids, and 1,4-naphthoquinones can be increased by modifying the cultivation conditions. D. regia turned out to be the richest in phenolic compounds, including 1,4-naphtoquinones: plumbagin and ramentaceone. Extracts from D. indica and D. regia tissue showed strong antibacterial activity against both pathogens. It has also been shown that the growth conditions of sundews can modify the level of secondary metabolites, and thus, their biological activity. Full article

The propagation of Crocus sativus L. relies exclusively on corm multiplication. As underground storage organs, corms are susceptible to a wide range of pathogens, environmental stresses, and diseases, making traditional propagation methods often ineffective with the loss of valuable material. In vitro propagation offers an alternative for the saffron culture under controlled conditions. In particular, the innovative application of the Temporary Immersion System (TIS) represents a technological advancement for enhancing biomass production with a reduction in operational costs. The current study utilized the Plantform™ bioreactor to propagate in vitro saffron corms from the ‘Abruzzo’ region (Italy), integrating machine learning models to assess its performance. The evaluation of saffron explants after 30, 60, and 90 days of culture showed a marked improvement in growth and microcorm production compared to conventional in vitro culture on semisolid medium, supported by the machine learning analysis. Indeed, the Random Forest algorithm revealed a predictive accuracy with an R² value of 0.81 for microcorm number, showcasing the capability of machine learning models to forecast propagation outcomes effectively. These results confirm that applying TIS in saffron culture could lead to economically viable, large biomass production within a controlled environment, irrespective of seasonality. This study represents the first endeavor to use TIS technology to enhance the in vitro propagation of saffron in conjunction with machine learning, suggesting an innovative approach for cultivating high-value crops like saffron. Full article

This research’s scope encompassed biotechnological, phytochemical, and biological studies of Schisandra henryi, including investigations into its in vitro microshoot culture grown in PlantForm bioreactors (temporary immersion systems, TISs), as well as extracts from leaves of the parent plant, focusing on anti-inflammatory, antioxidant, anticancer, and antimicrobial activities. The phytochemical analysis included the isolation and quantification of 17 compounds from dibenzocyclooctadiene, aryltetralin lignans, and neolignans using centrifugal partition chromatography (CPC), HPLC-DAD, and UHPLC-MS/MS tandem mass spectrometry with triple quadrupole mass filter methods. Higher contents of compounds were found in microshoots extracts (max. 543.99 mg/100 g DW). The major compound was schisantherin B both in the extracts from microshoots and the leaves (390.16 and 361.24 mg/100 g DW, respectively). The results of the anti-inflammatory activity in terms of the inhibition of COX-1, COX-2, sPLA2, and LOX-15 enzymes indicated that PlantForm microshoot extracts showed strong activity against COX-1 and COX-2 (for 177 mg/mL the inhibition percentage was 76% and 66%, respectively). The antioxidant potential assessed using FRAP, CUPRAC, and DPPH assays showed that extracts from microshoot cultures had 5.6, 3.8, and 3.3 times higher power compared to extracts from the leaves of the parent plant, respectively. The total polyphenol content (TPC) was 4.1 times higher in extracts from the in vitro culture compared to the leaves. The antiproliferative activity against T-cell lymphoblast line Jurkat, breast adenocarcinoma cultures (MCF-7), colon adenocarcinoma (HT-29), and cervical adenocarcinoma (HeLa), showed that both extracts have considerable effects on the tested cell lines. The antimicrobial activity tested against strains of Gram-positive and Gram-negative bacteria and fungi showed the highest activity towards H. pylori (MIC and MBC 0.625 mg/mL). Full article

The genus Sorbus has maintained an extremely relevant role over time from a landscape and environmental perspective in many countries in the Mediterranean and Central Europe. Based on the requirements coming from the environmental policies provided in the European strategy Next Generation EU, Sorbus has been considered a valuable species to be introduced in urban and peri-urban areas. The purpose of this study was to propagate four Sorbus accessions selected in the Sicilian territory, Southern Italy, using the liquid substrate in temporary immersion bioreactors Plantform™. The results obtained showed that the presence of 1 mg L⁻¹ mT in the substrate in combination with IBA 0.05 mg L⁻¹ produced a significant number of shoots (4.7) and a greater length (2.2 cm). Among the accessions, there were statistically significant differences; the accession SN2 and SN1 produced more shoots (respectively, 4.0 and 3.6), and a greater length of the shoots was observed in the selections SN4 and SN3 (respectively, 2.4 cm and 2.3 cm). The relative growth rate (RGR) was significantly influenced by the presence of the culture substrate of the combination of cytokines and auxin; SN4 selection showed the best RGR results of 8.3 mg⁻¹ d⁻¹. The use of the bioreactor Plantform™ in Sorbus domestica L. has favored a better development of plants obtained in vitro, demonstrating that this system is a valid alternative for the micropropagation of Sorbus. Full article

In this study, the solid culture method, and Plantform™ and SETIS™ temporary immersion bioreactor systems were used comparatively to propagate, root, and acclimatize ‘Grande Naine’ and ‘Azman’ banana varieties for rapid, cheap, and mass production in in vitro conditions. Micropropagation rate, plant height, number of leaves, and fresh and dry weight parameters were investigated in the micropropagation stage across eight subcultures. Rooting rate, plant height, number of leaves, number of roots/plant, root length, fresh and dry weight parameters were investigated in the rooting stage. Photosynthetic pigment analyses and stoma examinations were performed throughout all stages. In the micropropagation stage, a 20% increase in the Plantform™ system, a 12% increase in the SETIS™ system in ‘Grande Naine’, an 82% increase in the Plantform™ system, and a 98% increase in SETIS™ system in ‘Azman’ were determined compared to the solid culture. At the rooting stage, higher data were obtained from bioreactor systems than solid culture. Plants from bioreactor systems acclimatized faster and developed healthier in the greenhouse stage. It was determined that stomata were more active, and pigment accumulation was higher in bioreactor systems. Genetic variations across subcultures are among the most critical issues in banana clonal propagation. Leaf samples were taken from each system, and plant variation was investigated using SSR (Simple Sequence Repeat) markers. No variation was observed from the initial stage to the greenhouse stage. As a result, it has been determined that bioreactor systems are an essential alternative for the mass production of bananas. Full article

This study developed an efficient protocol for the in vitro propagation of giant reed (Arundo donax L.) biomass, defining a complete cycle of the induction of somatic embryogenesis from immature inflorescences, followed by the maturation of somatic embryos and the subsequent multiplication of the derived shoots in liquid culture in a temporary immersion system (TIS). The best explants were found to be 30 cm long immature inflorescences, preferably collected in spring. Such an explant type was easy to decontaminate, and the spikelets isolated from it provided over 100 embryogenic callus lines. Among the callus induction media tested, gelled MS medium supplemented with 1.1 mg/L 2,4-D provided the highest percentage of responsive spikelets and the highest density of embryogenic callus. Maturation of the embryogenic callus was easily triggered on gelled MS medium devoid of plant growth regulators. The obtained shoots could be further multiplied on previously optimized gelled DKW medium supplemented with 30 g/L sucrose, 5 mg/L BA, 0.1 mg/L IBA, and 6.8 g/L plant agar. Subsequent high multiplication of the developed shoots was achieved in liquid culture in TIS using a Plantform™ bioreactor, with an immersion cycle of 12 min every 8 h. Full article

The present work focuses on in vitro cultures of Ruta montana L. in temporary immersion Plantform^TM bioreactors. The main aim of the study was to evaluate the effects of cultivation time (5 and 6 weeks) and different concentrations (0.1–1.0 mg/L) of plant growth and development regulators (NAA and BAP) on the increase in biomass and the accumulation of secondary metabolites. Consequently, the antioxidant, antibacterial, and antibiofilm potentials of methanol extracts obtained from the in vitro-cultured biomass of R. montana were evaluated. High-performance liquid chromatography analysis was performed to characterize furanocoumarins, furoquinoline alkaloids, phenolic acids, and catechins. The major secondary metabolites in R. montana cultures were coumarins (maximum total content of 1824.3 mg/100 g DM), and the dominant compounds among them were xanthotoxin and bergapten. The maximum content of alkaloids was 561.7 mg/100 g DM. Concerning the antioxidant activity, the extract obtained from the biomass grown on the 0.1/0.1 LS medium variant, with an IC₅₀ 0.90 ± 0.03 mg/mL, showed the best chelating ability among the extracts, while the 0.1/0.1 and 0.5/1.0 LS media variants showed the best antibacterial (MIC range 125–500 µg/mL) and antibiofilm activity against resistant Staphylococcus aureus strains. Full article

Microshoot agitated and bioreactor cultures (PlantForm bioreactors) of three Hypericum perforatum cultivars (Elixir, Helos, Topas) were maintained in four variants of Murashige and Skoog medium (MS) supplemented with 6-benzylaminopurine (BAP) and 1-naphthaleneacetic acid (NAA) (in the range of 0.1–3.0 mg/L). In both types of in vitro cultures, the accumulation dynamics of phenolic acids, flavonoids, and catechins were investigated during 5- and 4-week growth cycles, respectively. The contents of metabolites in methanolic extracts from biomasses collected in 1-week intervals were estimated by HPLC. The highest total contents of phenolic acids, flavonoids, and catechins were 505, 2386, and 712 mg/100 g DW, respectively (agitated cultures of cv. Helos). The extracts from biomass grown under the best in vitro culture conditions were examined for antioxidant and antimicrobial activities. The extracts showed high or moderate antioxidant activity (DPPH, reducing power, and chelating activity assays), high activity against Gram-positive bacteria, and strong antifungal activity. Additionally, experiments with phenylalanine feeding (1 g/L) in agitated cultures were performed reaching the highest enhancement of the total contents of flavonoids, phenolic acids, and catechins on day 7 after the addition of the biogenetic precursor (2.33-, 1.73- and 1.33-fold, respectively). After feeding, the highest accumulation of polyphenols was detected in the agitated culture of cv. Elixir (4.48 g/100 g DW). The high contents of metabolites and the promising biological properties of the biomass extracts are interesting from a practical point of view. Full article

The study investigated the effect of elicitation with: chitosan (CH) (200 mg/L), yeast extract (YeE) (3000 mg/L), ethephon (ETH) (25 µM/L), and methyl jasmonate (MeJA) (50 µM/L), on lignan accumulation in agitated and bioreactor (Plantform temporary immersion systems) microshoot cultures of female (F) and male (M) Schisandra rubriflora Rehd. et Wils. (Schisandraceae) lines. The elicitors were supplemented on the 10th day of culture. Biomasses were collected at 24 h and 48 h, and 4, 6, and 8 days after the addition of each elicitor. The 24 compounds from the dibenzocyclooctadiene, aryltetralin, dibenzylbutane, and tetrahydrofuran lignans and neolignans were determined qualitatively and quantitatively in biomass extracts using the UHPLC–MS/MS method. The highest total contents [mg/100 g DW] of lignans were: for CH-95.00 (F, day 6) and 323.30 (M, 48 h); for YeE 104.30 (F, day 8) and 353.17 (M, day 4); for ETH 124.50 (F, 48 h) and 334.90 (M, day 4); and for MeJA 89.70 (F, 48 h) and 368.50 (M, 24 h). In the biomass extracts of M cultures grown in bioreactors, the highest total lignan content was obtained after MeJA elicitation (153.20 mg/100 g DW). The maximum total lignan contents in the biomass extracts from agitated and bioreactor cultures were 3.29 and 1.13 times higher, respectively, than in the extracts from the non-elicited cultures. The poor understanding of the chemical composition and the lack of studies in the field of plant biotechnology of S. rubriflora emphasize the innovativeness of the research. Full article

In vitro culture has become a dependable approach for the mass production of plant material as the market for innovative plant-derived medicinal approaches has grown significantly. Furthermore, because it permits manipulation of biosynthetic routes to boost the production and accumulation of certain compounds, this technology has enormous potential for the manufacture of natural bioactive chemicals. As a result, the goal of this study was to develop an efficient micropropagation system for biomass production and to investigate the accumulation of bioactive compounds from Vaccinium corymbosum L., Duke and Hortblue Petite cultivars. Two in vitro plant tissue culture systems were used for shoots production: a solid medium (5 g/L Plant agar) and liquid medium (Plantform bioreactor). The culture medium used was Woddy Plant Medium (WPM) supplemented with two growth regulators: 0.5 mg/L and 1 mg/L zeatina (Z) and 5 mg/L N6-(2-Isopentenyl) adenine (2iP). The content of phenolic compounds, carotenoids, and chlorophylls of the in vitro shoot extracts were examined via the HPLC-DAD-MS/MS technique. The results showed that cv. Hortblue Petite produced a higher amount of biomass compared with cv. Duke, on all variants of culture media in both systems (solid and liquid), while the shoots extract of the Duke variety in the liquid culture system (under all concentrations of growth regulators) had the highest content of total phenolic compounds (16,665.61 ± 424.93 μg/g). In the case of the lipophilic compounds analysed (chlorophylls and carotenoids), the solid medium reported the highest values, whereas media supplemented with 0.5 mg/L Z was proved to have the richest total content for both cultivars. Full article

The study demonstrated the effects of precursor feeding on the production of glucosinolates (GSLs), flavonoids, polyphenols, saccharides, and photosynthetic pigments in Nasturtium officinale microshoot cultures grown in Plantform bioreactors. It also evaluated the antioxidant and antimicrobial activities of extracts. L-phenylalanine (Phe) and L-tryptophan (Trp) as precursors were tested at 0.05, 0.1, 0.5, 1.0, and 3.0 mM. They were added at the beginning (day 0) or on day 10 of the culture. Microshoots were harvested after 20 days. Microshoots treated with 3.0 mM Phe (day 0) had the highest total GSL content (269.20 mg/100 g DW). The qualitative and quantitative profiles of the GSLs (UHPLC-DAD-MS/MS) were influenced by precursor feeding. Phe at 3.0 mM stimulated the best production of 4-methoxyglucobrassicin (149.99 mg/100 g DW) and gluconasturtiin (36.17 mg/100 g DW). Total flavonoids increased to a maximum of 1364.38 mg/100 g DW with 3.0 mM Phe (day 0), and polyphenols to a maximum of 1062.76 mg/100 g DW with 3.0 mM Trp (day 0). The precursors also increased the amounts of p-coumaric and ferulic acids, and rutoside, and generally increased the production of active photosynthetic pigments. Antioxidant potential increased the most with 0.1 mM Phe (day 0) (CUPRAC, FRAP), and with 0.5 mM Trp (day 10) (DPPH). The extracts of microshoots treated with 3.0 mM Phe (day 0) showed the most promising bacteriostatic activity against microaerobic Gram-positive acne strains (MIC 250–500 µg/mL, 20–21 mm inhibition zones). No extract was cytotoxic to normal human fibroblasts over the tested concentration range (up to 250 μg/mL). Full article

Mertensia maritima is a commercially interesting herb with edible leaves and flowers, characterized by oyster flavor and taste. Plant propagation and traditional cultivation are challenging for this species. Therefore, the main purpose of the present study was to establish successful protocols aimed at ensuring oyster plant shoot propagation, rooting and in vivo acclimatization. Both micropropagation and rooting were tested, comparing the traditional in vitro solid substrate in jar vs. the liquid culture in a temporary immersion system (TIS) bioreactor (Plantform™). A Murashige and Skoog (MS) medium added with 4-μM thidiazuron (TDZ) and 1-μM α-naphthaleneacetic acid (NAA) was employed for micropropagation, while a half-strength MS medium supplemented with 4-μM indole−3-butyric acid (IBA) was used for rooting. Different acclimatization conditions in the greenhouse or in growth chamber were tested. Morphometric and microscopical analyses were performed on the oyster plant leaves at the propagation, rooting and acclimatization stages both in a jar and in a TIS. Micropropagation in a TIS allowed to obtain large shoots, while a great number of shoots was observed in the jar. M. maritima shoots rooted in TIS produced more developed roots, leaves with more developed waxy glands and well-formed stomata; moreover, the plants coming from the TIS showed the best acclimatization performances. Full article

Transformed shoots of the Tibetan medicinal plant Dracocephalum forrestii were cultured in temporary immersion bioreactors (RITA and Plantform) and in nutrient sprinkle bioreactor (NSB) for 3 weeks in MS (Murashige and Skoog) liquid medium with 0.5 mg/L BPA (N-benzyl-9-(2-tetrahydropyranyl)-adenine) and 0.2 mg/L IAA (indole-3-acetic acid). The greatest biomass growth index (GI = 52.06 fresh weight (FW) and 55.67 dry weight (DW)) was observed for shoots in the RITA bioreactor, while the highest multiplication rate was found in the NSB (838 shoots per bioreactor). The levels of three phenolic acids and five flavonoid derivatives in the shoot hydromethanolic extract were evaluated using UHPLC (ultra-high performance liquid chromatography). The predominant metabolite was rosmarinic acid (RA)—the highest RA level (18.35 mg/g DW) and total evaluated phenol content (24.15 mg/g DW) were observed in shoots grown in NSB. The NSB culture, i.e., the most productive one, was evaluated for its antioxidant activity on the basis of reduction of ferric ions (ferric reducing antioxidant power, FRAP) and two scavenging radical (O₂^•– and DPPH, 1,1-diphenyl-2-picrylhydrazyl radical) assays; its antibacterial, antifungal, and antiproliative potential against L929 cells was also tested (3-[4,5-dimethylthiazole-2-yl]-2,5-diphenyltetrazolium bromide (MTT) test). The plant material revealed moderate antioxidant and antimicrobial activities and demonstrated high safety in the MTT test—no cytotoxicity at concentrations up to 50 mg/mL was found, and less than a 20% decrease in L929 cell viability was observed at this concentration. Full article

Goji (Lycium barbarum L.) has recognized nutritive and antioxidant properties and many products are commercialized for health in food market. Besides its food use, goji has been the subject of more than 2000 years of traditional Chinese medicine, using berries, root bark, and leaves. Here, the potential of the liquid culture in temporary immersion system (TIS) by using the bioreactor Plantform^TM was tested for the large-scale production of high-quality goji shoots and the subsequent production of total phenols and flavonoids. The three tested immersion cycles differently influenced the shoot quality in terms of proliferation and hyperhydricity. The best immersion cycle (time and frequency) was proven to have the shortest daily immersion time (6 min every 24 h) which ensured good levels of relative growth and multiplication rate, very limited onset of hyperydricity, and the longest shoots, promoting direct rooting after only 30 days of culture. In comparison with the semisolid culture, the TIS culture resulted in an increase of the total phenolic content (TPC) and in a lower value of the total flavonoid content (TFC). However, considering the higher quantity of biomass produced in the Plantform^TM bioreactor, the difference in terms of TFC productivity between semisolid medium and TIS liquid culture was proven to be statistically equivalent. Full article