Plant Cell Cultures Research Articles

Microfluidic capillary zone electrophoresis mass spectrometry analysis of alkaloids in Lobelia cardinalis transgenic and mutant plant cell cultures.

Application of a microfluidic CE* device for CZE-MS allows for fast, rapid, and in-depth analysis of large sample sets. This microfluidic CZE-MS device, the 908 Devices ZipChip, involves minimal sample preparation and is ideal for small cation analytes, such as alkaloids. Here, we evaluated the microfluidic device for the analysis of alkaloids from Lobelia cardinalis hairy root cultures. Extracts from wild-type, transgenic, and selected mutant plant cultures were analyzed and data batch processed using the mass spectral processing software MZmine2 and the statistical software Prism 8. In total 139 features were detected as baseline resolved peaks via the MZmine2 software optimized for the electrophoretic separations. Statistically significant differences in the relative abundance of the primary alkaloid lobinaline (C27 H34 N2 ), along with several putative "lobinaline-like" molecules were observed utilizing this approach. Additionally, a method for performing both targeted and untargeted MS/MS experiments using the microfluidic device was developed and evaluated. Coupling data-processing software with CZE-MS data acquisition has enabled comprehensive metabolomic profiles from plant cell cultures to be constructed within a single working day.

Exploiting plant cell culture for natural product formation

The initiation of callus cultures and in vitro cultivation of plant cells, even in large bioreactors, has become a routine task. Despite the fact, that permanent cell and organ cultures can produce a whole range of small natural compounds (SNAPs) used in medicine, only a few could be produced at commercial scale. However, plant cell cultures provide very useful systems to study the biosynthetic pathways leading to SNAPs at the enzyme and gene level. They turned out to be ‘a pot of gold’ for those chasing the enzymes and genes involved in natural product formation. The use of genetically modified yeast and bacteria for the production of SNAPs is an emerging technique that will take research into the coming decades. This review contemplates and revisits developments in the field of using plant cell and tissue culture as tools to elucidate SNAP formation and means to produce bioactive plant natural products over the past 40 years alongside my own work.

Nanoharvesting of bioactive materials from living plant cultures using engineered silica nanoparticles.

Plant secondary metabolites are valuable therapeutics not readily synthesized by traditional chemistry techniques. Although their enrichment in plant cell cultures is possible following advances in biotechnology, conventional methods of recovery are destructive to the tissues. Nanoharvesting, in which nanoparticles are designed to bind and carry biomolecules out of living cells, offers continuous production of metabolites from plant cultures. Here, nanoharvesting of polyphenolic flavonoids, model plant-derived therapeutics, enriched in Solidago nemoralis hairy root cultures, is performed using engineered mesoporous silica nanoparticles (MSNPs, 165 nm diameter and 950 m2/g surface area) functionalized with both titanium dioxide (TiO2, 425 mg/g particles) for coordination binding sites, and amines (NH2, 145 mg/g particles) to promote cellular internalization. Intracellular uptake and localization of the nanoparticles (in Murashige and Skoog media) in hairy roots were confirmed by tagging the particles with rhodamine B isothiocyanate, incubating the particles with hairy roots, and quenching bulk fluorescence using trypan blue. Nanoharvesting of biologically active flavonoids was demonstrated by observing increased antiradical activity (using 2,2-diphenyl-1-picrylhydrazyl radical scavenging assay) by nanoparticles after exposure to hairy roots (indicating general antioxidant activity), and by the displacement of the radio-ligand [3H]-methyllycaconitine from rat hippocampal nicotinic receptors by solutes recovered from nanoharvested particles (indicating pharmacological activity specific to S. nemoralis flavonoids). Post-nanoharvesting growth suggests that the roots are viable after nanoharvesting, and capable of continued flavonoid synthesis. These observations demonstrate the potential for using engineered nanostructured particles to facilitate continuous isolation of a broad range of biomolecules from living and functioning plant cultures.

Production of Tetrahydrocurcumin From Curcumin by Cultured Plant Cells and Inhibitory Effects of Curcuminoids on NF-κB Activity

Biotransformation of curcumin was investigated using cultured plant cells of Morus sp. Cultured Morus cells converted curcumin into dihydrocurcumin and tetrahydrocurcumin. The activity of nuclear factor-κB was inhibited by curcumin and tetrahydrocurcumin.

Editorial - 100 years Journal of Applied Botany

This year we want to celebrate the 100th birthday of our journal, which was first published as (= Applied Botany) in 1919. It started as journal for members of the “Vereinigung fur (German Society for Applied Botany)” only. On a monthly basis, it delivered the newest research results in the field of applied botany as well as short communications, literature reviews and reports from scientific meetings to the German speaking research community. After a short publication hiatus in the 1940ies, society members continued to report their findings, quickly exhausting the available print pages. Throughout the years, English titles and summaries where added, making the journal accessible for a wider, non-German speaking audience. final step into the international research community was the opening of the journal for authors, who were not society members in the 1980ies. In 2004, the “Deutsche Gesellschaft fur Qualitatsforschung - Pflanzliche Lebensmittel (German Society for Quality Research in Plant Foods)” joined the journal due to partial overlap in their research focus, thus, making it the Journal of Applied Botany and Food Quality as we know it today. latest change happened in 2013, with the switch from a subscription based print journal to an online-only gold open access journal. This once again broadened the readership and gained the attention of even more international researchers, which nowadays make up the majority of our journal authors. The first article in Angewandte Botanik published in 1919 was submitted by Appel, a phytopathologist well known for his discoveries on bacterial and fungal diseases of crops such as potato and cereals. His work ranged from fundamental research to applied research and reflected in his long-term membership in the board of the Association for Applied Botany. In the first contribution of this special jubilee section Hartwig Schulz and Heike Riegler report about Otto Appel and his contributions to food quality and safety at the beginning of the 20th and give insights into the major challenges in phytopathology at the early years of the 20th century and discoveries by Appel in this Maik Kleinwachter and Dirk Selmar prepared a contribution about Modern Applied Botany - Changes in the perception of applied botanists to themselves and others during the last century. Three recent examples of the scientific potential of this field. They give a short outline on the history of applied research in plant biology in and illustrate the relevance of modern Applied Botany in three relevant post-harvest processes. They state that interdisciplinary work and intensive cooperation with growers and producers is an integral part of developing feasible and economically acceptable solutions that can be successfully transferred into practice. major challenge in Applied Botany today is the implementation of new concepts and ideas into product-related research. In the third article in the special section Claudia Bahmann, Thomas Rinas, Nicolas Niemenak, Elsa Hegmann, Bernward Bisping, David Seigler and Reinhard Lieberei describe how Cacao seed tests controls transport processes during germination and fermentation-like incubation. contribution of Muna Ali Abdalla and Karl H. Muhling leads us to Plant-derived sulfur containing natural products produced as a response to biotic and abiotic stresses. A review of their structural diversity and medicinal importance and reveals the diversity in structure and function of secondary compounds in plants. In the article written by Wolfgang Kreis Exploiting plant cell culture for natural product formation we get insights into the high expectations researchers and industry had on the exploitation of plant cell cultures from a historical and personal point of view. Imke Hutter and Carolin Schneider report on the current state of Commercial micropropagation in Germany from the applied site. In an interdisciplinary project, Jan Philipp Schuchardt, Andreas Hahn, Theresa Greupner, Paulina Wasserfurth, Maria Rosales-Lopez, Johann Hornbacher and Jutta Papenbrock work on different aspects concerning Watercress – cultivation methods and health effects. Georg Langenkamper and Christian Zorb submit an article about Modern aspects of wheat grain storage proteins. Finally, on the interaction of plants with microorganisms, in this case fungi, is shed light on by Oluwatosin Abdulsalam, Erika Kothe and Katrin Krause in The parasitic-neutral-mutual continuum of plant-fungal interactions.

Influence of biotic elicitor Aspergillus niger on salicylic acid products in callus cultures of Calendula officinalis L. plant

The study was carried out in the college of education for pure sciences / department of biology / plant tissue and cell culture laboratory to study the effect of biotic elictors Aspergillus niger in the production of salicylic acid in callus cultures of Calendula officinalis L. plant. the results showed the best induction of the explants, which included the leaves and cotyledon obtained from culturing on the solidified MS medium supplemented with 2.0 mg.L1- of 2,4- Dichlorophenoxy acetic acid (2,4-D) interacted with 0.5 mg.L−1 Kinetin, which reached the highest fresh weight of callus was 2.2350 and 2.7125 g, respectively, after four weeks culture. The callus induction from the leaves its compact, in contrast to the callus induced by the cotyledon, which is characterized its friable.

329 Effective inhibition of Th17 pathway in 2D and 3D models of psoriasis by Celastrol enriched plant cell culture extract (CEE)

329 Effective inhibition of Th17 pathway in 2D and 3D models of psoriasis by Celastrol enriched plant cell culture extract (CEE)

051 Effective inhibition of Th17/Th22 pathway in in vitro and ex vivo models of psoriasis by Celastrol enriched plant cell culture extract (CEE)

051 Effective inhibition of Th17/Th22 pathway in in vitro and ex vivo models of psoriasis by Celastrol enriched plant cell culture extract (CEE)

Fabrication of innocuous gold nanoparticles using plant cells in culture

Plant extracts and their different growth phases have been manipulated for the fabrication of nanomaterials, which can be an eco-friendly alternative to the chemical methods that produce hazardous by-products. However, practical difficulties in isolation of the nanoparticles obtained through biological methods and the scanty control that these methods allow over their shapes and sizes impose limitations in their utility. For the first time, we report here a versatile system using cell suspension culture of Medicago sativa, which ensures control over the reaction to regulate size of the particles as well as their easier recovery afterwards. Isolated nanoparticles were characterized for their shape, size and functions. The particles varied in shapes from isodiametric spheres to exotic tetrahedrons, pentagons and pentagonal prisms. They clearly demonstrated catalytic activity in the reduction reaction of methylene blue by stannous chloride. Interestingly, the cell culture-derived particles were found less cytotoxic to healthy human cell line HEp-2 while more cytotoxic to the cancer cell line 4T-1 in comparison to those synthesized through citrate method. However, when administered in mice, these nanoparticles elicited similar inflammatory responses as those produced by chemically synthesized counterparts. These results envisage the utility of these particles for various biological applications.

Cell selection with heavy metal ions for obtaining salt tolerant plant cell cultures

Cell selection with heavy metal ions for obtaining salt tolerant plant cell cultures

Improvement of bioactive compound accumulation in cell cultures of Orostachys cartilaginous A. Bor. through elicitation with salicylic acid and effect of cell extract on bioactive activity

Orostachys cartilaginous A. Bor. has various pharmacological effects, which is mainly distributed in Changbai Mountain area of China, however, the resource of O. cartilaginous is difficult to satisfy the market demand. Plant cell culture can be an alternative approach for alleviating the resource shortage but a problem that relative low yield of bioactive compounds in cell cultures should be solved. Therefore, this study used salicylic acid (SA) as an elicitor to treat 25-day-old bioreactor cultured O. cartilaginous cells and the effects of concentrations and durations of SA treatment on bioactive compound accumulation were investigated to select a suitable strategy of SA elicitation. Results showed that SA concentrations from 25 μM to 150 μM obviously enhanced the bioactive compound accumulation without a decrease of cell biomass. At 100 μM SA, the total polysaccharide (218.5 mg/g DW), phenolic (27.2 mg/g DW), and flavonoid (35.4 mg/g DW) contents reached their maximum levels. Bioactive compound accumulation increased with prolonged SA treatment duration from 0 h to 48, reached the maximum at 48 h, and decreased afterwards. During the SA elicitation, the bursts of hydrogen peroxide (H2O2) and nitric oxide (NO) in cells were detected at the early stage, and peaks of H2O2 and NO contents occurred at 4 h and 6 h, respectively. Furthermore, the maximum contents of flavonoid monomers, including quercetin, kaempferide, epicatechin gallate, quercetin-3-O-glucose, and kaempferol-3-rutinoside were determined at 48 h, thereby speculating that H2O2 and NO might the upstream signaling molecules involved in SA-induced flavonoid synthesis in O. cartilaginous cells. In addition, the O. cartilaginous cell extract (OCE) inhibited growth of Bacillus subtilis and Pseudomonas aeruginosa, and the anti-bacterial activity against B. subtilis was significantly higher than that against P. aeruginosa. Meanwhile, tyrosinase activity was inhibited by the OCE, reaching 61.7% of the inhibitory rate at 1.4 mg/mL OCE, which demonstrated that OCE possessed an anti-melanin ability. This study concluded that 25-day-old bioreactor cultured O. cartilaginous cells can be treated with 100 μM SA for 48 h for the maximum production of polysaccharides, phenolics, and flavonoids; the O. cartilaginous cell cultures could be used as a potential material in the production of anti-bacterial and anti-melanin related products.

Sekonder Bitki Metabolitlerinin In Vitro Koşullarda Üretimi

Sekonder metabolitler, bitkiler tarafından biyotik ve abiyotik faktörlere karşı bir tepki olarak ve savunma mekanizması oluşturmak amacıyla üretilen farklı bir organik bileşik grubudur. Sekonder metabolitler biyosentetik orijinlerine ve kimyasal yapılarına göre farklı sınıflara ayrılırlar. Sekonder metabolitler, günümüzde ilaç hammaddesi, tarım ilacı, tat verici, koku maddesi, renk maddesi ve gıda katkı maddesi gibi pek çok alanda yaygın bir şekilde kullanılmaktadır. Sekonder metabolit üretimi, geleneksel olarak ya doğadan bitki toplamaya veya söz konusu bitkilerin tarla koşullarında kültür edilmesine dayanmaktadır. Fakat bu şekilde geleneksel sekonder metabolit üretimi düşük verim, coğrafik, mevsimsel ve çevresel varyasyonlar nedeniyle bitkinin sekonder metabolit içeriğinin çok değişken olması, sekonder metabolit üreten bitkilerin yetiştiriciliği için arazi ve yoğun işgücüne gereksinim duyulması gibi dezavantajlara sahiptir. Bu nedenle, bitki hücre ve organ kültürleri sekonder metabolitlerin üretiminde tüm bitkiden elde etmeye alternatif bir yöntem olarak ortaya çıkmıştır. Bu literatür derlemesinde, bitki hücre, doku ve organ kültürleri ile sekonder metabolit üretiminin mevcut durumu, sorunları ve sorunların çözümüne yönelik olarak yapılan çalışmalar irdelenmiştir.

Chemical characterization of leaves and calli extracts of Rosmarinus officinalis by UHPLC-MS.

Rosmarinus officinalis L. (Lamiaceae) is an aromatic plant widely popular mainly due to its uses in traditional medicine as an anti-inflammatory, diuretic and antimicrobial, as well as in the prevention and treatment of diseases. These biological activities are mainly related to the presence of phenolic and terpenic compounds. This work reports a chemical profile analysis of extracts from leaves and calli of rosemary obtained by both pressurized liquid extraction and maceration. Chemical profiles were determined on calli extracts of 3, 6, 9, and 15days of culture; chemical characterization and quantification of compounds was carried out using ultrahigh performance liquid chromatography-mass spectrometry. A total of 53 metabolites were identified in callus and 47 compounds in leaf extracts, of which 25 correspond to phenolic compounds, mainly flavonoids and flavones, 13 terpenes that include phenolic terpenes and one diterpenolactone, two glycosides which correspond to 6-O-caffeoyl-β-D-fructofuranosil-(2→1)-α-D-glucopyranoside and primulaverin, an aromatic compound identified as fenantrenone and a growth regulator 12-hydroxy jasmonic acid. These results showed that undifferentiated rosemary cells accumulate the same compounds identified mainly in highly specialized tissues such as leaves. The plant cell culture supply the possibility of developing biotechnological processes to obtain compounds of commercial interest.

Elicitation effect on the production of asiaticoside and asiatic acid in shoot, callus, and cell suspension culture of Centella asiatica

Centella asiatica is an important medicinal plant which contains various phytocompounds. Asiatic acid and asiaticoside are two major compounds which are responsible for its various pharmaceutical activities. The present study analyzes the effect of elicitor i.e. methyl jasmonate on the synthesis of asiaticoside and asiatic acid in shoot, callus and cell suspension cultures of Centella asiatica. A high performance liquid chromatography analysis showed that elicitation with 100 µM concentration of methyl jasmonate enhanced asiaticoside content by 69 fold in callus culture and 39 fold in shoot cultures whereas asiatic acid was enhanced by 1.9 folds in callus culture. Therefore, elicitation with methyl jasmonate is an effective method to increase the biosynthesis of asiaticoside and asiatic acid in plant cell cultures of Centella asiatica.

Effects of methyl jasmonate and salicylic acid on the production of camphor and phenolic compounds in cell suspension culture of endemic Turkish yarrow(Achilleagypsicola) species

Elicitors have been widely used as biotic and abiotic stimulants in triggering the production of secondary metabolites in plant cell culture. The present study aimed to enhance the production of camphor and phenolic compounds and cell growth using methyl jasmonate (MeJA) and salicylic acid (SA) in the cell suspension culture of Turkish endemic species Achillea gypsicola. Various concentrations (0, 10, 50, and 100 μM) of MeJA and SA were applied to 8-day-old cell cultures. The camphor and phenolic compound contents were determined using a headspace gas chromatographic-mass spectrometer device and spectrophotometer. Increasing doses of MeJA and SA significantly enhanced the accumulation of the camphor and phenolic compounds in general. The highest amount of camphor accumulation occurred in cells treated with 100 μM MeJA (0.3449 μg/g) and 50 μM SA (0.3816 μg/g). Increasing concentrations of MeJA resulted in a significant decrease in the total anthocyanin when compared to the initial culture. The present study showed that MeJA and SA could effectively be used as potent elicitors to enhance the production of camphor and phenolic compounds, along with cell growth, in cell suspension cultures of the endemic Turkish yarrow species Achillea gypsicola.

Rational metabolic engineering for enhanced alpha-tocopherol production in Helianthus annuus cell culture

Alpha-tocopherol, an essential dietary supplement, synthesized by photosynthetic organisms is the most biologically active antioxidant component of vitamin E in humans. Attempts to improve the yield of alpha-tocopherol using plant cell cultures has gained significance in recent years. Here, we developed a high alpha-tocopherol yielding cell line of Helianthus annuus using a model based metabolic engineering approach. To this end, we adapted an available genome-scale model of Arabidopsis for simulating H. annuus metabolism using constraint-based analysis to identify and rank suitable enzyme targets for overexpression. Of the various model-predicted enzyme targets, majority belonged to the vitamin E pathway and the MEP pathway while the others included reactions from the nucleotide biosynthesis and amino acid metabolism. Experimental validation of the top strategy (overexpression of p-hydroxyphenylpyruvate dioxygenase,) resulted in a high alpha-tocopherol yielding transformed cell line (up to 240 μg g−1), which was ≈10-fold more than in the untransformed cell line. A cell suspension was developed from the selected transformed cell line for in vitro production of alpha-tocopherol, which resulted in a maximum alpha-tocopherol yield of 412.2 μg g−1 and titre of 6.4 mg L−1.We thus demonstrate the utility of model-based metabolic engineering for multi-fold yield enhancement of commercially important plant secondary metabolites.

Advances and challenges on the in vitro production of secondary metabolites from medicinal plants

ABSTRACT The production of secondary metabolites from medicinal plants, also called Plant-Derived Medicinal Compounds (PDMC), is gaining ground in the last decade. Concomitant to the increase in the knowledge about pharmacological properties of these compounds, horticultural plants are becoming the most important, sustainable and low-cost biomass source to obtain high-complex PDMCs to be used as medicaments. Biotechnological tools, including plant cell and tissue culture and plant genetic transformation, are increasingly being employed to produce high quality and rare PDMC under in vitro conditions. The proper use of these technologies requires studies in organogenesis to allow for better control of in vitro plant development and, thus, to the production of specific tissues and activation of biochemical routes that result in the biosynthesis of the target PDMCs. Either biotic or abiotic factors, called elicitors, are responsible for triggering the PDMC synthesis. In vitro techniques, when compared to the conventional cultivation of medicinal plants in greenhouse or in the field, have the advantages of (1) producing PDMCs in sterile and controlled environmental conditions, allowing better control of the developmental processes, such as organogenesis, and (2) producing tissues with high PDMC contents, due to the efficient use of different biotic and abiotic elicitors. Nevertheless, the process has many challenges, e.g., the establishment of step-by-step protocols for in vitro biomass and PDMC production, both involving and being affected by many factors. Other limitations are the high costs in opposition to the relatively cheaper alternative of growing medicinal plants conventionally. This paper aims to quickly review the general origin of plant secondary metabolites, the leading techniques and recent advances for PDMC in vitro production, and the challenges around the use of this promising technology.

Inhibition of Autolysosome Formation Improves rrhGAA Production Driven by RAmy3D Promoter in Transgenic Rice Cell Culture

Although plant cell cultures produce low yields of recombinant proteins compared to other production systems, a dramatic increase of the heterologous protein production in transgenic rice cells was achieved with the alpha-amylase isozyme 3D (RAmy3D) promoter system. However, this expression system has inherent limitations in that gene expression is initiated by sucrose/glucose deprivation, concurrently triggering starvation-derived autophagy and rapid cell death. Decreased viability and culture longevity subsequently prevent further increment of production. In this study, we introduced autophagy inducers and inhibitors in the rrhGAA-producing transgenic rice cell cultures in order to explore their effects on production controlled by the RAmy3D promoter. The autophagy inducers rapamycin and CCI-779 increased autophagosome and autolysosome while concanamycin A1 and bafilomycin A successfully decreased autolysosome. Interestingly, autophagy inhibitors improved viability, DCW loss, and rrhGAA production, while autophagy inducers deteriorated these profiles compared to the control. As the production conditions under the death phase may facilitate protein degradation, and subsequently exacerbate functional activity, the size variant distribution and enzyme activity of the purified rrhGAAs were evaluated. However, no significant difference in rrhGAA degradation as well as GAA activity was observed compared to the control condition, thus indicating that the autophagy regulation is an efficient approach to increase protein yield in rice cell culture system for rrhGAA production.

Biotechnological production of ruscogenins in plant cell and organ cultures of Ruscus aculeatus

Ruscus aculeatus is a threatened medicinal plant whose main bioactive components, the ruscogenins, have long been used in the treatment of hemorrhoids and varicose veins, but recently demonstrated activity against some types of cancer. Plant cell biofactories could constitute an alternative to the whole plant as a source of ruscogenins. In this pipeline, despite the in vitro recalcitrance of R. aculeatus, after many attempts we developed friable calli and derived plant cell suspensions, and their ruscogenin production was compared with that of organized in vitro plantlet and root-rhizome cultures. Root-rhizomes showed a higher capacity for biomass and ruscogenin production than the cell suspensions and the yields were greatly improved by elicitation with coronatine. Although ruscogenins accumulate in plants mainly in the root-rhizome, it was demonstrated that the aerial part could play an important role in their biosynthesis, as production was higher in the whole plant than in the root-rhizome cultures.

Biogenic zinc oxide nanoparticles-enhanced biosynthesis of lignans and neolignans in cell suspension cultures of Linum usitatissimum L

Zinc oxide nanoparticles (NPs) have emerged as a novel elicitor for enhanced biosynthesis of secondary metabolites in in vitro plant cell cultures. The current study was aimed to explore elicitation abilities of ZnO-NPs for enhanced accumulation of lignans and neolignans in cell cultures of Linum usitatissimum. We optimized concentration of zinc oxide NPs before carrying out a full-fledged experiment. Subsequently, an optimum dose of 100 mg/l was introduced into the culture medium on day 0, days 0 and 15, and finally days 0 and 25. We observed that repeated elicitation stimulated various parameters and physiological responses in Linum usitatissimum cell cultures than one-time elicitation. Repeated elicitation of cell cultures on day 0 and 15 resulted in highest fresh weight (412.16 g/l) and lignans production (secoisolariciresinol diglucoside 284.12 mg/l: lariciresinol diglucoside 86.97 mg/l). Contrarily, repeated elicitation on day 0 and 25 resulted in highest DW (13.53 g/l), total phenolic production (537.44 mg/l), total flavonoid production (123.83 mg/l) and neolignans production (dehydrodiconiferyl alcohol glucoside 493.28 mg/l: guaiacylglycerol-β-coniferyl alcohol ether glucoside 307.69 mg/l). Enhancement in plant growth and secondary metabolites accumulation was several fold higher than controls. Furthermore, a linear relationship existed between total phenolic and flavonoid contents which in turn was correlated with higher antioxidant activities.