Plant Cell Culture Research Papers (original) (raw)

2025

Artemisinin is a sesquiterpene lactone extracted from the medicinal plant Artemisia annua L. (sweet wormwood). It has traditionally been utilized in artemisinin-based combination therapies (ACTs) for the malarial parasite, including drug-resistant strains. Natural artemisinin extraction is costly with low yields. Due to its effectiveness, there is a significant rise in the demand for artemisinin production. In vitro cell suspension culture offers a cost-effective and viable technique for artemisinin production. Therefore, this study aimed to optimize a protocol for cell suspension culture of A. annua L. to enhance biomass and artemisinin production. A successful cell suspension culture was initiated from induced callus. The highest cell biomass was obtained in suspension cultures grown with an initial inoculum size of 0.1 g of mixed type cell aggregates, in media with a pH of 6.2 and a rotation speed of 90 rpm. Macronutrient concentrations influenced both biomass and artemisinin content, with optimal biomass achieved at 19 mM KNO 3 and 1.56 mM KH 2 PO 4 . The absence of these nutrients resulted in the highest artemisinin levels. Different LED wavelengths also significantly influenced biomass and artemisinin production. Red + blue LED increased cell biomass, while the highest artemisinin content was observed under red LED. The upscaling of the culture indicated a variation in biomass yield pattern, but the highest growth index was achieved in the 500 mL Erlenmeyer flask. This study successfully established a cell suspension culture for A. annua L., demonstrating the influence of macronutrients and red LED on biomass and artemisinin production, providing insights for potential large-scale production.

2025, Journal of Pharmacognosy and Phytochemistry

L-dopa (3, 4-dihydroxy L-phenylalanine) is a neurotransmitter precursor being used for symptomatic relief of Parkinson’s disease. It is one of the most highly active allele chemicals present in Mucuna species. L-Dopa is produced via the oxidation of tyrosine by the copper containing enzyme tyrosine hydroxylase. The effect of elicitors and precursor molecules on the activity of tyrosine hydroxylase and their effect on the production of L-Dopa in cell suspension cultures of Mucuna prurita Hook were studied. Callus cultures were initiated on MS medium supplemented with BAP and IAA. The callus was transferred to MS liquid medium supplemented with BAP, IAA and elicitors methyl jasmonate, chitin, pectin, yeast extract and also a precursor L-tyrosine at different concentrations. Compared to the control, several fold increase in the activity of tyrosine hydroxylase was observed. With the progressive increase in the activity of tyrosine hydroxylase also, several fold increase in L-Dopa produ...

2025, Effect of gamma irradiation on seed germination, survivability and growth performances of Calotropis gigantea (Vara)

Physical mutagenesis is an effective mutational breeding method for improving various morphological characteristics of agricultural and medicinal crops. The objective of the conducted experiment was determining the effective dose of gamma radiation to induce mutations in Calotropis gigantea. Well matured seeds of C. gigantea were exposed to Gamma irradiation using "Gamma chamber 1200 Cobalt-60" research irradiator and these treatments were carried out at the Horticultural Crop Research and Developmental Institute at Gannoruwa, Sri Lanka. Mature seeds were subjected to seven treatments as 150Gy, 300Gy, 450Gy, 600Gy, 750Gy, 900Gy and non-irradiation (0 Gy) as a control to study their effect on germination percentage, survival rate and growth performances of C. gigantea. Treated seeds were arranged under shade house condition in Completely Randomized Design with four replications and each replication contained twenty-four seeds. The mutagenic treatments were tested for lethal dose of 50% and the dose at which 50% of the survival at four months was considered as LD50 value. Data were analyzed by ANOVA using Minitab 17 software and compared the treatment means using Dunnett's test at 0.05 significant level. The study revealed that the higher dosages of gamma irradiation was significantly decreased the germination percentage, survivability and growth performances (P<0.05). Plant survivability, number of leaves and plant height were significantly (P<0.05) reduced with the higher dosages of 450 Gy to 900 Gy. Lethality level of gamma irradiation for Calotropis gigantea was found as 395 Gy at maturity stage and it could be concluded that the radiation below 395 Gy should be imposed to induce mutations in Calotropis gigantea.

2025, Biotechnology and Bioprocess Engineering

Plant cell culture provides a viable alternative over whole plant cultivation for the production of secondary metabolites. In order to successfully cultivate the plant cells at large scale, several engineering parameters such as, cell aggregation, mixing, aeration, and shear sensitivity are taken into account for selection of a suitable bioreactor. The media ingredients, their concentrations and the environmental factors are optimized for maximal synthesis of a desired metabolite. Increased productivity in a bioreactor can be achieved by selection of a proper cultivation strategy (batch, fed-batch, two-stage etc.), feeding of metabolic precursors and extraction of intracellular metabolites. Proper understanding and rigorous analysis of these parameters would pave the way towards the successful commercialization of plant cell bioprocesses.

2025, Annals of Agricultural & Crop Sciences

Location of azulenes in plant cell has been studied on three clover species Trifolium repens L., T. pratense L., and T. medium L. (fam. Fabaceae). By using of microspectrophotometer/ microspectrofluorimeter the absorbance spectra were recorded from the surface of intact leaves and intact isolated chloroplasts. Here the maxima in the range 580-620 nm, peculiar to blue pigments azulenes, were observed in all cases. The pigments’ picks 590-610 nm also have been found in the absorbance spectra of ethanolic extracts from the leaf surface and from the chloroplast suspension. The protector function of the azulenes for the plant cell has been discussed. Clovers studied are recommended for agriculture and pharmacy as useful resource of azulenes.

2024, Process Biochemistry

A cell suspension culture was developed from calli of grape rootstock 41B in order to study the bioproduction of resveratrol. While 41B grape cultures produced no resveratrol, methyljasmonate (MeJA) elicitor treatment activated its production in a dose dependent manner. The concentration of 0.2 mM MeJA was optimal for efficient production and high accumulation of resveratrol (150 mg/L) in flask experiments. Microscopic analysis of cells monitored for viability showed that MeJA elicitor triggered expression of resveratrol fluorescence within the cells. These results led to scale-up of the culture in a 2 L stirred bioreactor where a resveratrol production of 209 mg/L being secreted into the liquid medium, corresponding to 90% of the total production. Liquid/liquid extraction of the culture medium and a solid/liquid extraction of the cells showed that other stilbenes were also produced. For the first time, trans-viniferin, trans-␦-viniferin, and a trans-3-methylviniferin as well as trans-piceatannol were identified in a 2 L bioreactor cell cultures of grapevine. Furthermore, a one step FPLC method was developed for the purification of resveratrol and-viniferin.

2024, Process Biochemistry

2024

In-situ cell density monitoring and apoptosis detection in adherent Vero cell bioreactor cultures

2024, Journals International Journal of Pharmacy and Pharmaceutical Sciences

Objective: The production of L-Dopa in callus cultures of Mucuna pruriens is influenced by substances that regulate calcium channels. Methods: M. pruriens seeds were gathered from the Western Ghats area in Tamil Nadu. L-Dopa was obtained by the Brain (1976) method, A. niger cultures were used for fungal elicitor preparation, and the assay of Ca 2+ ATPase was studied. Results: The use of fungal triggers resulted in increased L-Dopa levels on the 9th day but reduced levels on the 24th day of culture. When calcium channel modulators like verapamil and chlorpromazine were added, it decreased the growth of calli and L-Dopa production, suggesting the role played by calcium ion channels in L-Dopa synthesis. The calcium ionophore A23187 enhanced calli growth and L-Dopa production, increasing Ca 2+ ATPase activity. Particularly on the 12 th d, Ca 2+ ATPase was notably active, while the presence of calcium ionophore boosted L-Dopa biosynthesis. Conclusion: The use of fungal elicitors in in vitro cultures of Mucuna pruriens is recommended as a potential method to increase the production of secondary metabolites.

2024, Research Square (Research Square)

The berries of Embelia ribes Burm f. are a rich source of embelin, a compound known for its anthelmintic, antidiabetic, and anticancer activity. Due to over-exploitation, the natural habitat of Embelia ribes is now considered vulnerable. Traditional propagation methods are insu cient to meet current demands, necessitating alternative production methods. This study was designed to explore in vitro culture as a viable alternative for the production of embelin. While numerous studies have focused on extracting embelin from callus cultures, there is a lack of research on enhancing the embelin content in both organogenic and embryogenic callus cultures. The objective of this study was to enhance embelin production in callus cultures by using both biotic and abiotic elicitors. Additionally, efforts were made to optimize rapid in vitro shoot induction. We achieved the best response rate for the induction of organogenic and embryogenic calli using MS basal medium supplemented with TDZ. Among the various elicitors tested, chitosan at 200 mg/L was the most effective, yielding the highest embelin content, at 6.44% in embryogenic calli and 5.72% in organogenic calli. Subsequent subculturing enabled successful differentiation of callus cultures into shoot buds on MS medium supplemented with a combination of BAP (6-benzylaminopurine) and IAA (indole-3-acetic acid) at concentrations of 2.0 mg/l and 0.1 mg/l, respectively. An effective protocol has been developed for obtaining the highest embelin content from embryogenic and organogenic callus cultures, coupled with a high frequency of shoot multiplication. The protocol can be instrumental for large-scale embelin production, ex-situ conservation, sustainable utilization, and industrial applications. Key message This study demonstrates a successful protocol for enhancing embelin production in Embelia ribes Burm f. through in vitro callus cultures using biotic and abiotic elicitors. Chitosan was found to be the most effective elicitor, signi cantly increasing embelin content in both organogenic and embryogenic calli. The developed protocol also ensures e cient shoot induction and multiplication, providing a viable solution for large-scale embelin production, ex-situ conservation, and sustainable utilization of this vulnerable species.

2024, Lat. Am. J. …

KEY WORDS: In vitro culture, Galipea longiflora, Quinoline alkaloids. PALABRAS CLAVE: Alcaloides quinolínicos, Cultivo in vitro, Galipea longiflora. * Autor a quien dirigir la correspondencia. E-mail: Magali-PazGarcia@biociencias.org ...... more

2024

Kiralni alkoholi su važne građevne jedinice za sintezu farmaceutskih pripravaka, pesticida, feromona, aroma, mirisa i materijala poput tekucih kris

2024

Recently, through modern biotechnology, it is now recognized that plants are potentially a new source of pharmaceutical proteins including vaccines, antibodies, blood substitutes and other therapeutic entities. Unlike mammalian-derived rDNA drugs, plant-derived antibodies, vaccines and other proteins are particularly advantageous since they are free of mammalian viral vectors and human pathogens. Advantages offered by plants include also low cost of cultivation and high biomass production, relatively fast “gene to protein” time, low capital and operating costs, excellent scalability, eukaryotic posttranslational modifications and a relatively high protein yield. Crop plants can synthesize a wide variety of proteins that are free of mammalian toxins and pathogens. Crop plants produce large amounts of biomass at low cost and require limited facilities. Since plants have long been used as a source of medicinal compounds, molecular farming represents a novel source of molecular medicine...

2024, Australian Journal of Crop Science

In this study, the effect of chitosan and pectin on the accumulation of anthraquinones (AQ), phenolics and flavonoids was investigated by adventitious root suspension cultures of Morinda citrifolia in shake flasks. Adventitious root cultures elicitor treated with various combinations of chitosan and pectin or chitosan alone resulted in enhanced biosynthesis of secondary metabolite but inhibited root growth. The strong inhibition of root growth might be due to the lethal effect of elicitor as evidenced by 36-79% cell death was measured. The optimum concentration of elicitor for enhancing metabolite biosynthesis was found at the concentration of 0.2 mg ml -1 chitosan, in which 103.16, 48.57 and 75.32 mg g -1 dry weight (DW) of AQ, phenolics and flavonoids, respectively were achieved. To elucidate the optimum contact period and reduction of inhibitory effect of chitosan on root growth, a two-stage culture system was adopted. The optimum contact period of root to elicitor was observed w...

2024, In Vitro Cellular & Developmental Biology - Plant

Introduction: Hyoscyamine, because of its medicinal properties, is an important tropane alkaloid. In order to implement a biotechnological process for its production, hairy roots of Datura species resulting from genetic transformation by Agrobacterium rhizogenes A4 strain have been the subject of this work. In the in vitro alkaloids production programs, optimization of the transformation is a crucial step for obtaining a large number of root lines allowing the selection of efficient lines. Methods: The conditions for hairy roots induction were explored by studying the effect of Datura sp. in vitro seedling etiolation on the genetic transformation. The first step was the establishment of Datura in vitro plantlet cultures followed by the hairy roots induction with A4 strain of A. rhizogenes. The confirmation of the genetic transformation was performed by polymerase chain reaction (PCR) analysis of rolB gene in the roots. After hyoscyamine extraction, it was analyzed (quantitatively and qualitatively) by gas chromatography. Results: 343 root lines were obtained in total, 96 root lines from non-etiolated Datura in vitro seedlings and 247 root lines from etiolated in vitro plantlets. After the selection of six transgenic root lines, tow root lines from each species (D. tramonium, D. tatula and D. innoxia), one from non-etiolated in vitro plantlet and one from etiolated in vitro plantlet, the most hyoscyamine content was 8.43 mg/g D.W. obtained with D. tatula etiolated in vitro seedling. Conclusion: The etiolated in vitro plantlets seem more favorable to hairy roots induction.

2024, Journal of Chromatography A

An HPLC method was developed for quick scanning of taxanes from large numbers of plant cell suspension samples. The method was optimized for analysis of a range of taxanes of differing polarity. Identification of a standard mixture of paclitaxel and 12 related taxanes was achieved in less than 15 min using a gradient mode and a Microsorb-MV C column. 8 The method was used to investigate the influence of several factors on stability and recovery of paclitaxel from suspension media and cultures of Taxus cuspidata cv Densiformis. Incubation time had the most significant influence on stability of paclitaxel, contributing 88% to the total variation. Shaking contributed 6% to the total variation. Light contributed only 0.25% to the total variation. Analysis of test samples of suspension cultures of T. cuspidata cv Densiformis over a 4 week period show paclitaxel, 10-deacetylbaccatin III,

2024, Revista …

CAPATAZ TAFUR, Jacqueline; OROZCO SANCHEZ, Fernando; VERGARA RUIZ, Rodrigo and HOYOS SANCHEZ, Rodrigo. ANTIFEEDANT EFFECT OF CELL SUSPENSION EXTRACTS OF Azadirachta indica ON Spodoptera frugiperda JE Smith UNDER ...

2024, Plants

This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY

2024, Pharmacognosy magazine

The biosynthesis of artemisinin derivatives is one of the interesting subjects. Artemisinic acid (AA) has been widely studied as a supposed intermediate in the biosynthetic pathway leading to artemisinin in Artemisia annua. To investigate the bioconversion of AA by transgenic crown galls of Panax quinquefolium. AA was administered into crown galls of P. quinquefolium and co-cultured for 2 days. The methanol extract was separated by column chromatography, and the structures of two biosynthesis products were elucidated by physicochemical and spectroscopic methods. Co-culture time curves on conversion were also established. In addition, the effects of AA on the growth and ginsenosides production of crown galls of P. quinquefolium were investigated. Furthermore, the in vitro antitumor activities of AA and two glycosides against HepG2 cell line were evaluated by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl-tetrazolium bromide (MTT) assay. Glycosylation of AA by crown galls of P. quinquefoli...

2024, Plant Cell, Tissue and Organ Culture (PCTOC)

The utility of hairy root cultures to produce valuable phytochemicals could be improved by repartitioning more of the desired phytochemical into the spent culture media, thereby simplifying the bioprocess engineering associated with the purification of the desired phytochemical. The majority of nicotine produced by tobacco hairy root cultures is retained within roots, with lesser amounts exuded into the spent culture media. Reduced expression of the tobacco nicotine uptake permease (NUP1) results in significantly more nicotine accumulating in the media. Thus, NUP1-reduced expression lines provide a genetic means to repartition more nicotine into the culture media. The present study examined a wild type and a NUP1-reduced expression hairy root line during a variety of treatments to identify culture conditions that increased nicotine accumulation in the media. The NUP1reduced expression line grew faster, used less oxygen, and exuded more nicotine into the media. Basification of the culture media associated with root growth resulted in a dramatic reduction in nicotine accumulation levels in the media, which was reversed by decreasing the pH of the media. Kinetic analysis of hairy root growth and nicotine accumulation in the media revealed a potential improvement in nicotine yields in the media by stimulating the branching of tobacco hairy roots. Keywords Nicotiana tabacum Á Nicotine uptake permease (NUP) Á Aeration Á Media basification Á Ion trapping Á Kinetics Electronic supplementary material The online version of this article (

2024, IFAC Proceedings Volumes

2024, Journal of Natural Products

Plant cell culture systems represent a potential renewable source of valuable medicinal compounds, flavours, fragrances, and colorants, which cannot be produced by microbial cells or chemical synthesis. The evolving commercial importance of the secondary metabolites has in recent years resulted in a great interest, in secondary metabolism, and particularly in the possibility to alter the production of bioactive metabolites by means of cell culture technology. The principle advantage of this technology is that it may provide continuous, reliable source of plant pharmaceuticals and could be used for the large scale culture of plant cells from which these metabolite can be extracted. Plant cell and tissue cultures hold great promise for controlled production of myriad of useful secondary metabolites on demand. The current yield and productivity cannot fulfill the commercial goal of plant cell-based bioprocess for the production of most secondary metabolites. In order to stretch the boundary, recent advances, new directions and opportunities in plant cell based processes are being critically examined.

2024, FEBS Letters

A novel technique is described for the immobilisation of plant cells. The method is simple and does not require the use of potentially toxic gels for the entrapment process. Liquid-suspended cells of two species, Capsicum frutescens Mill. and Daucus carota L. were found to invade, and were strongly retained in, polyurethane foam particles over a 21 day culture period. The viability of the immobilised cells was high (70-80% were alive after 21 days). On agitation of the loaded foam particles in fresh liquid medium, at least 95% of the cells remained immobilised after 3 or 4 days. Cell immobilisation Secondary metabolite production Cell differentiation Polyurethane foam Capsicum frutescens Mill. Daucus carota L.

2024, Current Science

Methoxybenzaldehydes in plants are one of the important groups of benzoate derivatives. Some of them, exhibiting refreshing fragrance can be used as flavouring ingredients in food and cosmetics. Therefore, they have important roles in food and cosmetic industries. Methoxybenzaldehydes also exhibit significant medicinal properties and thus have certain prospects in pharmaceutical industry. Biosynthesis of benzoic acid in plants has been explored in the last decade. There has been focus on benzaldehyde and methoxybenzaldehyde biosynthesis as well. There have been several studies regarding the biosynthetic route and mechanism of formation of benzaldehyde via benzoic acid from cinnamate and further addition of 'methoxy' group to it in the last few years. Still there are many ambiguities regarding the medicinal properties and biosynthesis of methoxybenzaldehydes. This review highlights the latest advances in fragrant methoxybenzaldehyde research and the knowledge gaps till date. The review also discusses the occurrence of methoxybenzaldehydes in plants, their separation methods, medicinal properties and biosynthesis.

2024

Podophyllin, an ethanolic extract of Podophyllum peltatum L. or P. emodi Wall (syn. P. hexandnum Royle), is a good source of the aryltetralin-type lignan, podophyllotoxin. The latter compound, as well as its congeners and derivatives exhibit pronounced biological activity mainly as strong antiviral agents and as antineoplastic drugs. The podophyllotoxin derivatives etoposide, etopophos (etoposide phosphate), and teniposide are thus successfully utilized in the treatment of a variety of malignant conditions. Continued research on the Podophyllum lignans is currently focused on structure optimization to generate derivatives with superior pharmacological pro®les and broader therapeutic scope, and the development of alternative and renewable sources of podophyllotoxin.

2024, Brazilian Journal of Plant Physiology

Phytoalexin accumulation is one of a myriad of plant defense responses; these responses can be elicited by pathogens or molecules such as oligogalacturonides (OGAs). Phytoalexin production has been considered a vital component of the resistance mechanisms that determine the outcome of many plant-microbe interactions. Besides inducing defense responses, OGAs have been shown to affect plant development, which normally is controlled by plant hormones, particularly auxin. In this work we measured phytoalexin accumulation in roots of bean (Phaseolus vulgaris L.) seedlings grown in the presence or absence of the auxin 3-naphtalenacetic acid (NAA) and treated with a decagalacturonide (OGA10). We found that OGA10 (0.01 mM) caused phytoalexin production and also inhibited main root elongation and the formation of secondary roots by ca. 33%. Expression of Cycb 2-2 was also inhibited, while pal and chs were highly expressed. The root growth inhibition was not overcome by the addition of a stim...

2024, Engineering in Life Sciences

Plant cells contain a wide range of interesting secondary metabolites, which are used as natural pigments and flavoring agents in foods and cosmetics as well as phyto‐pharmaceutical products. However, conventional industrial extraction from whole plants or parts of them is limited due to environmental and geographical issues. The production of secondary metabolites from in vitro cultures can be considered as alternative to classical technologies and allows a year‐round cultivation in the bioreactor under optimal conditions with constant high‐level quality and quantity. Compared to plant cell suspensions, differentiated plant in vitro systems offer the advantage that they are genetically stable. Moreover, the separation of the biomass from culture medium after fermentation is much easier. Nevertheless, several investigations in the literature described that differentiated plant in vitro systems are instable concerning the yield of the target metabolites, especially in submerged culti...

2024, Plants

Plant cell suspension cultures are widely used as a tool for analyzing cellular and molecular processes, metabolite synthesis, and differentiation, bypassing the structural complexity of plants. Within the range of approaches used to increase the production of metabolites by plant cells, one of the most recurrent is applying elicitors capable of stimulating metabolic pathways related to defense mechanisms. Previous proteomics analysis of tamarillo cell lines and cell suspension cultures have been used to further characterize and optimize the growth and stress-related metabolite production under in vitro controlled conditions. The main objective of this work was to develop a novel plant-based bioreactor system to produce hydrolytic enzymes using an elicitation approach. Based on effective protocols for tamarillo micropropagation and plant cell suspension culture establishment from induced callus lines, cell growth has been optimized, and enzymatic activity profiles under in vitro con...

2024, Plant Cell Tissue and Organ Culture

The production of an antifungal spirostanol saponin designated SC-1 has been detected in cell suspension cultures of the Mexican species Solanum chrysotrichum. Batch cultures of a cell suspension obtained from hypocotyl derived calluses of this species were grown for 25 days in shake flasks containing Murashige & Skoog (MS) medium. Throughout the growth cycle, fresh and dry weight, SC-1 yield,

2024, Functional Plant Science and Biotechnology

Biotechnology can offer useful tools to complement conventional breeding programs. The techniques of in vitro selection, that exploit phenomenon, called somaclonal variation, by screening cell cultures for resistance to herbicides, different types of stress or diseases, already helped to obtain tolerant lines of flax (Linum usitatissimum), which were used in further commercial breeding programs. The other important techniques in flax biotechnology are anther and immature embryo cultures. Development of haploid or dihaploid lines based on a regeneration capacity of microspores in immature anthers can speed up conventional breeding. Embryo rescue helps to overcome the post-zygotic incompatibility mechanisms after wide crosses for the transfer of desired traits. One of the main objectives of tissue culture studies is to obtain high-frequency shoot regeneration or somatic embryo formation. Age and viability of the explant, the tissue source and genotype of donor plant from which the explant was excised, composition of the culture medium and plant growth regulator supplementation, are very important for the effectiveness and the direction of morphogenic responses. This review highlights main studies devoted to flax biotechnology and the main factors controlling morphogenesis of L. usitatissimum.

2024, Medicinal and Aromatic Plant Science and Biotechnology

Hyoscyamus niger is an important medicinal plant belonging to the Solanaceae family, commonly known as henbane and locally as Bazir bangh. A study was undertaken to investigate the effects of different plant growth regulators (PGRs) at various concentrations on shoot proliferation from shoot tip explants. Full-strength Murashige and Skoog (MS) medium supplemented with different concentrations of 6benzyladenine (BA), kinetin (Kn) and thidiazuron (TDZ) were tested. Amongst the PGRs tested, BA at 12.5 μM showed the best shoot proliferation and highest number of indirect multiple shoots while TDZ at 10 μM formed intense disorganized friable light-greenish callus only; there was no response with Kn. In vitro shoots were separated and rooted under varying concentrations of different auxins viz. indole-3-butyric acid (IBA), indole-3-acetic acid (IAA) and Į-naphthaleneacetic acid (NAA). Best rooting response was recorded with 4.0 μM IBA and 2.5 μM NAA. However, no response was noted with any concentration of IAA. Plantlets transferred to field conditions showed 50% survival.

2024, Medicinal and Aromatic Plant Science and Biotechnology

Conventionally, pharmaceutically important secondary metabolites such as flavonoids were extracted directly from whole plants collected from the wild. This conventional method is not cost effective and could even lead to extinction of some endangered plant species. Furthermore, the production of uniform quantity and quality secondary metabolites will be affected as their production is greatly influenced by geographical, seasonal and environmental variations. Biotechnological approaches, specifically, plant tissue culture techniques, have been considered as an attractive solution to the problems of extracting secondary metabolites for industrial applications. Nevertheless, the commercial implementation of pilot scale plant cell suspensions for chemical production is still in the development stage with a few exceptional cases. The major setback is the failure of cell cultures to accumulate significant amounts of secondary metabolites compared to whole plants or organ cultures. Even so, several strategies can be applied in order to substantially increase the yields of secondary metabolites in plant cell cultures. This paper discussed the strategies of nutrient manipulation, precursor feeding and elicitation in enhancing the production of secondary metabolites.

2024, Cross Ref

overall productivity in plant tissue culture. Two auxin and one
cytokine used to manipulate the media and the effect was studies in
terms of cell growth. Secondary metabolite production was analyzed by
HPLC.
The calcium alginate concentration was optimized on the basis of cell
viability and cell number increase was observed in immobilized
condition.

2024, Food research international (Ottawa, Ont.)

Plant cell cultures from cloudberry, lingonberry and stoneberry were studied in terms of their nutritional properties as food. Carbohydrate, lipid and protein composition, in vitro protein digestibility and sensory properties were investigated. Dietary fibre content varied between 21.2 and 36.7%, starch content between 0.3 and 1.3% and free sugar content between 17.6 and 33.6%. Glucose and fructose were the most abundant sugars. High protein contents between 13.7 and 18.9% were recorded and all samples had a balanced amino acid profile. In vitro protein digestion assay showed hydrolysis by digestive enzymes in fresh cells but only limited hydrolysis in freeze-dried samples. The lipid analysis indicated that the berry cells were rich sources of essential, polyunsaturated fatty acids. In sensory evaluation, all fresh berry cells showed fresh odour and flavour. Fresh cell cultures displayed a rather sandy, coarse mouthfeel, whereas freeze-dried cells melted quickly in the mouth. All in...

2024, Current Biology

Recent significant contributions to the design of large-scale plant cell cultures for secondary metabolite production include: the development of a strategy to control the concentration of dissolved gases at constant shear; a surface immobilization technique to retain cell mass at high mixing rates; and a modified stirred-tank reactor with a mesh cage to prevent damage of hairy root cultures.

2024, Plant Cell Reports

A plant cell suspension culture of Alfalfa (Medicago sativa L.) was grown in a bioreactor using a batch procedure. The cytoplasmic esterase activity (EC 3.1) was extracted from the cells and measured during cultivation using fluorescein diacetate as the fluorogenic substrate. This enzymatic activity was conclusively found to be correlated to cell viability assessed with the membrane integrity test using the trypan blue dye. This new viability determination method is convenient, simple and can be reproduced because: (1) the difficult step of counting the cells when using the trypan blue exclusion method is avoided and (2) the esterase activity level per viable cell constituted of numerous enzymes depends on cell viability but is independent of cellular metabolism.

2024, Current Opinion in Plant Biology

High stability of the production of secondary metabolites is an interesting characteristic of hairy root cultures. For 25 years, hairy roots have been investigated as a biological system for the production of valuable compounds from medicinal plants. A better understanding of the molecular mechanism of hairy root development, which is based on the transfer of Agrobacterium rhizogenes T-DNA into the plant genome, has facilitated its increasing use in metabolic engineering. Hairy roots can also produce recombinant proteins from transgenic roots, and thereby hold immense potential for the pharmaceutical industry. In addition, hairy roots offer promise for phytoremediation because of their abundant neoplastic root proliferation. Recent progress in the scaling-up of hairy root cultures is making this system an attractive tool for industrial processes. Addresses 1 UPRES EA 2106 'Biomolé cules et Biotechnologies Vé gé tales',

2024, International Journal of Pharmacology

2024

ii supplemented with 1.0 mg/L BAP after 3 weeks. Besides, optimum rooting (3.20 roots per shoot) was achieved in half-strength MS medium containing 0.1 mg/L IBA. Platelets were successfully acclimatized to ex vitro conditions with 85% of survival percentage. Simple Sequence Repeat (SSR) and Inter Simple Sequence Repeat (ISSR) markers were tested to assess the genetic fidelity of the in vitro raised clones of E. longifolia. Out of the 12 SSR primers screened, nine primers produced 15 amplicons (1.7 bands in average) ranging from 100 to 800 bp, whereas eight ISSR primers generated 27 bands with an average of 3.4 bands ranging between 300 to 1000 bp. The monomorphic banding pattern confirmed the clonal homogeneity of the tissue culture-raised E. longifolia plantlets and reliability of the multiplication system used. Furthermore, different solvents were used for root extraction of in vitro and in vivo plants to determine the total antioxidant activity, total phenolic and flavonoid contents. Results of 1,1-diphenyl-2picrylhydrazyl (DPPH) free radical scavenging method showed that in vitro root extract at 25°C exhibited maximum antioxidant activity with 0.114 mg TE/g DW. Meanwhile, Ferric Reducing Antioxidant Potential (FRAP) showed the highest antioxidant content with 0.075 mg TE/g DW in in vivo root extracts at 25ºC. On the other hand, in vivo root extracts with 0.09 mg GAE/g DW had maximum phenolic content at 4°C. Whereas, in vitro root extract at 25°C showed the highest total (0.31 mg GAE/g DW and 0.10 mg RE/g DW) of both polyphenol and flavonoid content respectively. From this study, the successful in vitro propagation of E. longifolia could provide a potential of large-scale production of planting materials in meeting the industrial and domestic demands.

2024, Frontiers in Plant Science

Microalgae are currently used for the production of food compounds. Recently, few microalgae species have been investigated as potential biofactories for the production of biopharmaceuticals. Indeed in this context, microalgae are cheap, classified as Generally Recognized As Safe (GRAS) organisms and can be grown easily. However, problems remain to be solved before any industrial production of microalgae-made biopharmaceuticals. Among them, post-translational modifications of the proteins need to be considered. Especially, N-glycosylation acquired by the secreted recombinant proteins is of major concern since most of the biopharmaceuticals are N-glycosylated and it is well recognized that glycosylation represent one of their critical quality attribute. Therefore, the evaluation of microalgae as alternative cell factory for biopharmaceutical productions thus requires to investigate their N-glycosylation capability in order to determine to what extend it differs from their human counterpart and to determine appropriate strategies for remodeling the microalgae glycosylation into human-compatible oligosaccharides. Here, we review the secreted recombinant proteins which have been successfully produced in microalgae. We also report on recent bioinformatics and biochemical data concerning the structure of glycans N-linked to proteins from various microalgae phyla and comment the consequences on the glycan engineering strategies that may be necessary to render those microalgae-made biopharmaceuticals compatible with human therapy.

2024, Frontiers in Plant Science

2024, Microbial Physiology

Recently, biotechnological opportunities have been found in non-Saccharomyces yeasts because they possess metabolic characteristics that lead to the production of compounds of interest. It has been observed that Kluyveromyces marxianus has a great potential in the production of esters, which are aromatic compounds of industrial importance. The genetic bases that govern the synthesis of esters include a large group of enzymes, among which the most important are alcohol acetyl transferases (AATases) and esterases (AEATases), and it is known that some are present in K. marxianus, because it has genetic characteristics like S. cerevisiae. It also has a physiology suitable for biotechnological use since it is the eukaryotic microorganism with the fastest growth rate and has a wide range of thermotolerance with respect to other yeasts. In this work, the enzymatic background of K. marxianus involved in the synthesis of esters is analyzed, based on the sequences reported in the NCBI database.

2024, Plant Science

Suspension cells of Dioscorea deltoidea Wall (strain D-1) were maintained in a semicontinuous culture (SCC) in shake flasks at a high growth rate. It was shown that continuous propagation growth of this culture is unstable on Murashige's and Skoog's (MS) medium due to P starvation. On a P-enriched MS-medium the SCC was stable even at mean specific growth rates \ 0.3 day − 1. Highest volumetric concentrations of furostanol glycosides were obtained, when a P-enriched SCC was not further subcultivated but fed with sucrose. The investigated culture is able to control phosphate uptake and to prevent toxicity on media with excess P. High concentrations of cellular P i did not effect the ratio of furostanol to starch.

2024, Records of Natural Products

A novel 2,3-dioxygenated flavanone, dysosmaflavanone (2), along with five known phenolic compounds including podophyllotoxin (1), podoverin A (3), kaempferol (4), 8,2′-diprenyl quercetin 3-methyl ether (5), and ethyl β-D-glucoside (6) were isolated from the roots of the plant Dysosma difformis. Their structures were elucidated via spectroscopic analysis. Besides podophyllotoxin and kaempferol, the rest of the compounds were isolated from the genus Dysosma for the first time. Dysosmaflavanone, which possesses a rare 2,3-dioxygenated skeleton, could be regarded as an important chemotaxonomic marker. The antioxidant and antidiabetic activities of the isolated compounds were evaluated.

2024, Journal of Forest Science

For improving the seed germination of Prosopis juliflora and Dalbergia sissoo different treatments were tested, including side cutting, abrasion, overnight soaking in boiling water, scalding in actively boiling water for 1 minute and immersion in 30%, 60% and 95% H2SO4 solution. Results showed that abrasion with sandpaper and side cutting were the most effective methods to break seed dormancy in both species, while scalding in actively boiling water for 1 minute, overnight soaking and different concentrations of H2SO4 gave low to zero seed germination. Based on the positive effects of scarification it was concluded that seed dormancy in both species was due to water impermeability of the seed coat. Mutation breeding involves the treatment of large quantities of seeds, therefore abrasion with sandpaper was the most efficient and less labour-intensive method; this method was subsequently used for determination of LD50 as it is a prerequisite in a mutation breeding program. Abrasion be...

2024, IEEE Transactions on Automatic Control

A homotopy approach for solving constmined parameter optimization problems is examined. The first-ordcr neasslvg conditions, with the complementarity conditions reprerunted using a technique due to M q p s w b , arc solved. The equations arc augmented to avoid singnlaritie-s which occur when the d v e constrrint changes. The Chow-Yorke algorithm is used to thck the homotopy path lepdiag to the solution to the desind problem at the terminal point. A elmple example which illustrates the technique, and an application to a fuel optirml orbital transfer problem a n presented.

2024, Iranian Journal of Biotechnology

Background When using a complex system, several experimental factors have to be optimized so it is essential to evaluate alternative analytical procedures according to multiple criteria (1, 2). The determination of the optimum conditions for the input variables requires the simultaneous consideration of all responses. This is called a multiresponse problem (3, 4) based on Multicriteria Decision Making. To obtain a satisfactory compromise, the desirability approach is used as a powerful tool in multi-response systems (4). The desirability function, defined by Harrington (1965) (5) and by Derringer and Suich (1980) (3), is one of the approaches used for factor optimization in such systems (6-8). It is based on the transformation of all the obtained responses from different scales into a scale-free value. The values of desirability functions lie between 0 and 1. The value 0 is attributed when the factors give an undesirable response, while the value 1 corresponds to the optimal performance for the studied factors (3, 4).

2024, Journal of Applied Bacteriology

2. Properties of plant cell cultures, 105s 2.1 Cell suspension cultures, 105s 2.2 Immobilized plant cell cultures, 109s 3. Plant organ cultures and secondary product formation, 110s 4. Conclusions, 113s 5. Acknowledgements, 11 3s 6.... more

2024, Planta Medica

In this study, three semisynthetic betulonic acid-based compounds, 20(29)-dihydrolup-2-en[2,3-d]isoxazol-28-oic acid, 1-betulonoylpyrrolidine, and lupa-2,20(29)-dieno[2,3-b]pyrazin-28-oic acid, were studied in biotransformation experiments using Nicotiana tabacum and Catharanthus roseus cell suspension cultures. Biotransformation was performed using cyclodextrin to aid dissolving poorly water-soluble substrates. Several new derivatives were found, consisting of oxidized and glycosylated (pentose- and hexose-conjugated) products.

2024, Shokubutsu soshiki baiyō

Root and cell suspension cultures of Rheum palmatum were established and analyzed for the production of (+)-catechin. The effects of media and growth regulators on the growth and (+)catechin formation in the root and cell cultures were also investigated. The content of (+)catechin in the cells, cultured in MS liquid medium containing 1 mg/l 2, 4-D, reached the level of 0.38% (dry weight, dw) at week 7 of the culture. In contrast, the root cultured in MS liquid medium containing 2 mg/l NAA showed the highest level of (+)-catechin (0.24%, dw) at week 1 of the culture. Rhubarb (Daio' in Japanese), the rhizome and root of Rheum spp. (Polygonaceae), is one of the most important traditional Chinese medicines used in combination with other crude drugs for bloodstasis syndrome, hypertension, renal disorder, diarrhea, and so on. Recently, some new biological activities such as psychotropic, l) improvement of nitrogen metabolism, 2-5) and inhibition of angiotensinconverting enzyme6) were discovered and proved to be originated from its tannin constituents (rhatannins and RG-tannin). In spite of many phytochemical, biochemical, and pharmacological examinations, there have been few reports on tissue culture products (anthraquinones7) and sennosides8)) of rhubarb plants. We have succeeded in the establishment of root and cell suspension cultures of R. palmatum and in vitro formation of (+)-catechin (1), which is the structural element of rhatannins and RG-tannin. We also determined the effects of media and growth regulators on the growth and production of 1 in the root and cell cultures by HPLC analysis.