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The genus Narcissus has several endemic, rare and/or threatened species in the Iberian Peninsula and North Africa. In vitro propagation is a useful tool for threatened plants conservation used in ex situ strategies. Thus, the aim of this work was to study the propagation in vitro of bulb scale explants of five endemic, rare and/or endangered Narcissus species from the Iberian Peninsula, treated with different PGR combinations. Initiation was achieved in half-strength Murashige and Skoog (MS) basal salts and vitamins, 10 g/L sucrose, 500 mg/L casein hydrolysate, 2 mg/L adenine, 10 mg/L glutathione and 5.5 g/L plant agar. In the multiplication phase, the highest bulblet proliferation was obtained in MS medium supplemented with 30 g/L sucrose and the combination of 10 μM 6-Benzylaminopurine (BAP) + 5 μM α-Naphthaleneacetic acid (NAA) in N. alcaracensis, N. eugeniae and N. hedraeanthus; 20 μM BAP + 5 μM NAA in N. jonquilla and N. yepesii. The highest rooting was obtained with 5 μM NAA + 1 μM Indole-3-butyric acid (IBA) for all species (>75%) and more than 80% of the produced bulblets were successfully acclimatized. [ABSTRACT FROM AUTHOR]

Lapiedra martinezii Lag. is a potential medicinal and ornamental plant facing conservation challenges. Thus, this study was focused on determining the conditions for culture initiation and propagation using in vitro techniques. The optimal sterilization procedure combined thermotherapy at 54°C for 60 min and immersion in 7% (w/v) Ca(ClO)2 solution for 20 min. The most suitable medium to initiate bulb scales cultures was Gamborg B5 medium containing 500 mg L−1 casein, 2 mg L−1 adenine, 10 mg L−1 glutathione and 10 g L−1 sucrose. The most productive multiplication medium tested was Murashige and Skoog medium containing 30 g L−1 sucrose, 4.0 mg L−1 6-benzylaminopurine, and 0.12 mg L−1 1-naphtaleneacetic acid. Most plants developed in vitro rooted spontaneously in the multiplication phase. The vast majority of the plants (89%) were successfully transferred to ex vitro conditions, and 100% survived over 1 yr of cultivation outdoors. Sucrose at a concentration of 60 g L−1 was the most effective treatment to increase the biomass of bulblets. High auxin/cytokinin ratios produced the highest callus induction efficiency. The vast majority of callus developed in dark conditions, but none regenerated in the combinations of growth regulators previously tested. The plants obtained by micropropagation did not show significant differences in morphometric traits compared with the wild specimens, which supported the stability of the materials produced in vitro. This is the first report on cell cultures and micropropagation of L. martinezii, and the results can be applied to other Amaryllidaceae for industrial or conservation purposes. [ABSTRACT FROM AUTHOR]

Chitin is one of the most abundant biopolymers and is present in many organisms in different forms. Its resistance to degradation has caused many problems in industry (waste decomposition) and agriculture (as protective structures in pests); this has led to increased interest in chitin-hydrolyzing enzymes: chitinases. Chitinases are enzymes that break down the 1→4 β-glycoside bond of N-acetyl d-glucosamine in chitin to produce mono- and oligomers. The inducible nature of chitinases, low activity of synthesized enzymes, and inertia of the substrate are only a few of the problems that can be solved by biotechnology to meet industry demands for green, energy-efficient, pollution-free, and economically profitable chitin use. This review aims to present the pitfalls and successes in research and production of chitinolytic enzymes, as well as to promote the use of chitinases in everyday practice. The focus is on the biosynthesis of chitinases: inducers, type of fermentation, and media composition. Methods for purification and future perspectives are also discussed. [ABSTRACT FROM AUTHOR]

Gardenia jasminoides Ellis is an aromatic and medicinal plant of high economic value. Much research has focused on the phytochemistry and biological activities of Gardenia fruit extracts; however, the potential of the Gardenia plant in vitro cultures used as mass production systems of valuable secondary metabolites has been understudied. This paper presents data on metabolite profiling (GC/MS and HPLC), antioxidant activities (DPPH, TEAC, FRAP, and CUPRAC), and SSR profiles of G. jasminoides plant leaves and in vitro cultures with different levels of differentiation (shoots, callus, and cell suspension). The data show strong correlations (r = 0.9777 to r = 0.9908) between antioxidant activity and the concentrations of chlorogenic acid, salicylic acid, rutin, and hesperidin. Eleven co-dominant microsatellite simple sequence repeats (SSRs) markers were used to evaluate genetic variations (average PIC = 0.738 ± 0.153). All of the investigated Gardenia in vitro cultures showed high genetic variabilities (average Na = 5.636 ± 2.157, average Ne = 3.0 ± 1.095). This is the first report on a study on metabolite profiles, antioxidant activities, and genetic variations of G. jasminoides in vitro cultures with different levels of differentiation. [ABSTRACT FROM AUTHOR]

Abstract: Calli, shoot-clumps and regenerated plants were initiated from young fruits of Pancratium maritimum L. Their genetic stability was monitored by flow cytometry before chemical studies. Apolar metabolites (alkaloids extracted at pH > 7, free fatty acids and fatty alcohols, sterols etc.) were qualitatively and quantitatively analyzed by GC–MS. The results clearly demonstrated that alkaloid synthesis in P. maritimum is closely related with tissue differentiation. The highest amounts of alkaloids and presence of homolycorine and tazettine type compounds (end products of the biosynthetic pathway of the Amaryllidaceae alkaloids) were found in highly differentiated tissues. Galanthamine accumulated in the leaves of plantlets. The amount of hordenine, a protoalkaloid, is related with the ability of tissues to synthesize alkaloids. Saturated fatty acids were found in considerably higher levels in undifferentiated callus cultures and partially differentiated shoot-clumps than in regenerated plants. Mono- and dienoic fatty acids were found at higher levels in non-photosynthesizing tissues – calli, and in vitro and intact bulbs, while α-linolenic acid (trienoic acid) was found in higher amounts in the photosynthesizing leaves of shoot-clumps and regenerated plants than in bulbs and calli. Fatty alcohols were found mainly in leaves, while sterols tended to accumulate in photosynthesizing and undifferentiated tissues. [ABSTRACT FROM AUTHOR]

“Hairy root” systems, obtained by transforming plant tissues with the “natural genetic engineer” Agrobacterium rhizogenes, have been known for more than three decades. To date, hairy root cultures have been obtained from more than 100 plant species, including several endangered medicinal plants, affording opportunities to produce important phytochemicals and proteins in eco-friendly conditions. Diverse strategies can be applied to improve the yields of desired metabolites and to produce recombinant proteins. Furthermore, recent advances in bioreactor design and construction allow hairy root-based technologies to be scaled up while maintaining their biosynthetic potential. This review highlights recent progress in the field and outlines future prospects for exploiting the potential utility of hairy root cultures as “chemical factories” for producing bioactive substances. [ABSTRACT FROM AUTHOR]

Abstract: The callus induction from young fruits of Leucojum aestivum was performed on Murashige–Skoog nutrient medium supplemented with 4mg/L 2,4 dichlorphenoxyacetic and 2mg/L 6-benzylamynopurine. Further, by planting the obtained calluses on the same nutrient medium supplemented with 1.15mg/L α-naphtylacetic acid and 2.0mg/L 6-benzylamynopurine shoot cultures were established. The growth and galanthamine accumulation of obtained L. aestivum in vitro systems were studied. It was established that the amount of accumulated galanthamine strongly depended on the level of the differentiation. The maximum yield of biomass (17.8g/L) and the maximum amount of accumulated galanthamine (2.5mg/L) were achieved after day 35 of submerged cultivation of L. aestivum 80 shoot culture, performed under illumination. Data concerning the time courses of the utilization of the main nutrient components of the medium during cultivation of L. aestivum shoot culture are presented as well. [Copyright &y& Elsevier]

Abstract: The process of betalains biosynthesis by Beta vulgaris L. hairy root culture in a temporary immersion system was studied. It was established that temporary immersion technique is suitable for the cultivation of hairy roots. The immersion frequency had significant effect on the biomass accumulation and the yields of betalains. The hairy root culture biosynthesized 18.8mgg−1 dry biomass (DB) betalains (9.6mgg−1 DB betacyanins and 9.2mgg−1 DB betaxanthins) at immersion frequency with 15min flooding and 60min stand-by periods. The maximal growth index (16.6) was achieved at 15min flooding and 75min stand-by periods. Data on the relationships in the biological system “Nutrient medium–B. vulgaris L. hairy root culture–Betalains” are presented. [Copyright &y& Elsevier]

While, the application of microalgae in various areas like environmental clean-up and improvement of bioreactor technologies for micropropagation of economically valuable plant species are also currently receiving considerable attention. The authors have paid special attention to the simultaneous application of more than one elicitor as part of the integrated approach for optimization of secondary metabolites production. Given the fact that wastewater clean-up is a global problem, the authors have summarized extremophilic microalgae-based wastewater treatment technologies for industrial wastewater sources that have already been put into practice. [Extracted from the article]

Abstract: Studies were conducted on low molecular metabolites (volatiles and polar compounds) produced by Rosa damascena Mill 1803 cell suspension culture, cultivated under different regimes: as a free suspension (in flasks and in bioreactor) and in a two-phase system (in the presence of Amberlite XAD-4 as a second phase). It was established that the main groups of volatiles were hydrocarbons and free acids and their esters and only traces of terpenoids were found. The main components of polar fraction were free acids, especially amino acids and oxidized acids. Depending on the culture conditions, significant differences were established in the amounts of all compounds under study in biomasses, culture media and adsorbed on the second phase (Amberlite XAD-4). [Copyright &y& Elsevier]

The time course of growth and betalains biosynthesis by Beta vulgaris L. (cv. Detroit Dark Red) hairy root culture were investigated. It was established that hairy root culture produced 42.2 mg/flasks betalains (26.2 mg/flasks betaxanthins and 16.0 betacyanins) between 6th and 15th days of cultivation. The uptake of sucrose, nitrate and phosphate ions were followed during the cultivation and data on the physiology of the B. vulgaris hairy root culture are presented. [Copyright &y& Elsevier]

Carotenes have been identified as the most essential carotenoids in the human body, together with lycopene, lutein, and β-cryptoxanthin due to their beneficial antioxidant activities. Because of these properties, we have designed and conducted in 2018--2021 an experiment where various tomato accessions were metabolically profiled for their antioxidant content and other fruit parameters for further usage in crop improvement programs. Same genotypes were phenotyped and profiled with DNA markers to characterize their relative genetic distance. Aggregated data from metabolic and genetic profiling were complemented by the phenotypic data to select for the most promising cross (Pl. karotina x L1116) that was performed and developed into F1 and F2 generations. After phenotyping and genetic profiling of the 104 F2 individuals obtained from that cross, 24 individuals were selected for metabolic profiling for antioxidant contents. In relation to carotenoids accumulated in fully matured fruits the highest lycopene concentration detected in our study was 587.03 µg/g DW, while β-carotene reached 440.29 µg/g DW. Our results support the use of such integrative approach to accelerate tomato breeding when improved metabolic content of the fruits is aimed. [ABSTRACT FROM AUTHOR]

Plants have been used as the main source of phytochemicals with nutritional, medicinal, cultural and cosmetic applications since times immemorial. Nowadays, achieving sustainable development, global climate change, restricted access to fresh water, limited food supply and growing energy demands are among the critical global challenges faced by humanity. Plant cell culture technology has the potential to address some of these challenges by providing effective tools for sustainable supply of phyto‐ingredients with reduced energy, carbon and water footprints. The main aim of this review is to discuss the recent trends in the development of plant cell culture technologies for production of plant‐derived substances with application in food products and cosmetic formulations. The specific technological steps and requirements for the final products are discussed in the light of the advances in cultivation technologies used for growing differentiated and undifferentiated plant in vitro systems. Future prospects and existing challenges of the commercialization of plant cell culture‐derived products have been outlined through the prism of the authors' point of view. We expect this review will encourage scientists, policymakers and business enterprises to join efforts for speeding‐up the mass commercialization and popularization of plant cell culture technology as an eco‐friendly alternative method for sustainable production of plant‐derived additives with application in food and cosmetic products. [ABSTRACT FROM AUTHOR]

Two‐phase bioreactor cultivation system was developed and applied for in sito recovery of extracellular galanthamine during the cultivation of Leucojum aestivum L. shoot culture in a modified column bioreactor system. The inclusion of an external circulation column with adsorbent resin Amberlite XAD‐4 as a second phase, on the 21st day of the beginning of cultivation resulted in 1.25 folds increase in biomass accumulation and maximal amounts of accumulated galanthamine of 6 mg/L (3.1 mg/L intracellular and 2.9 mg/L extracellular). It was demonstrated that the inclusion of a second phase at the cultivation of the L. aestivum shoot culture in a bubble column bioreactor with internal sections redirected the alkaloid metabolism to galanthamine synthesis and inhibits the synthesis of hemanthamine and lycorine type alkaloids. Our research demonstrated that the application of the two‐phase cultivation systems could be an important tool to increase the yields of valuable secondary metabolites in plant tissue culture‐based bioprocess. [ABSTRACT FROM AUTHOR]

The effects of growth regulators on shoot development and rooting have been studied in Fabiana imbricata Ruiz et Pav. micropropagation. Plants were cultured in vitro on media with 0.1, 0.25 and 0.5 mg l-1 6-benzylaminopurine (BAP) and 0.01 mg l-1 indole-3-butyric acid (IBA). In all the treatments similar values for the mean height of the plants and 100% rooting were established. Data analysis showed that within 28 days the proliferation value in the hormone-free variant was 5.04±0.68 compared to the treatment with 0.5 mg l-1 BAP and 0.01 mg l-1 IBA (5.64±0.70), where symptoms of hyperhydricity occurred. Application of 0.3% activated charcoal (AC) influenced the growth and development of the plants positively, resulting in almost two-fold higher proliferation (9.04±0.54), overcoming this physiological disorder. Moreover, the optimized propagation system allowed the organogenesis of both, shoots and roots simultaneously, as approach to accelerate the micropropagation process and reduce production costs. Although the percentage of surviving plants was dependent on root system morphology, the lack of growth regulators in the last subculture before adaptation could be a useful prerequisite for the hardening of process, resulting in successful plant survival. [ABSTRACT FROM AUTHOR]

The current scientific research is focused on the grape seeds - a waste product retrieved after alcoholic fermentation of wine, in the form of grape seeds flakes. Recently, the grape residue has been considerated as a functional food product, a source of bioactive compounds (BAC) including antioxidants. The present study presents data about the antioxidant activity of grape seeds flakes of different grape varieties of Bulgarian origin, analysed via four analytical methods differ in mechanism and reaction conditions, namely DPPH, ABTS, FRAP and CUPRAC. The obtained results were expressed as mM TE/g extract (842.50 ± 34.75; 959.63 ± 65.05; 94.73 ± 10.42 and 200.24 ± 48.60, respectively) and mM TE/g flakes (as follows 141.40 ± 5.83; 161.06 ± 10.92; 94.89 ± 10.44 and 200.56 ± 48.68). The grape seeds flakes were stored at room temperature - 25°? and relative air humidity of 75%. The storage regime was optimized to one month by means of packaging the product in a co-extruded barrier film with copolymer covering for heat sealing. During the storage of grape seeds flakes, analysis of their microbial load, granulometric composition and moisture content were performed. [ABSTRACT FROM AUTHOR]

The Inter-Simple Sequence Repeat marker technique was applied to a set of tomato genotypes that were found to diverge in antioxidant compounds in fruits. As marker-assisted selection is becoming a tool that brings major advantages for breeding programs need is rising to use it to quickly respond to consumer demand. Being part of a system for effi cient plant breeding in many crops the aim of the present study was to test Inter-Simple Sequence Repeats as candidate tool for incorporating in a breeding program aiming to improve antioxidant properties of tomato fruits. Application of Inter-Simple Sequence Repeat markers resulted in obtaining adequate representation of every accession from the selected group. This opens the opportunity for using the system in confirming the hybrid nature of the F1 progenies and further accelerating the breeding of genotypes for use as healthy food [ABSTRACT FROM AUTHOR]

Plant tissue and organ cultures in vitro usually face technological challenges. When submerged cultivation of plant cells in a controlled environment is desired, the characteristic growth morphology and physiology of differentiated organ cultures present a problem in process scale-up. Temporary immersion systems (TIS) were developed several decades ago. These systems are providing the most natural environment for in vitro culture of plant shoots and seedlings. Over the past few years, TIS have been recognized as a perspective technology for plant micropropagation, production of plant-derived secondary metabolites, expression of foreign proteins, and potential solutions in phytoremediation. Nowadays, several TIS, operating on similar or divergent technological principles, have been developed and successfully applied in the cultivation of various plant in vitro systems, including somatic embryos and transformed root cultures. In this article, the operational principle and technological design of the most popular TIS are reviewed. In addition, recent examples of the application of temporary immersion technology for in vitro cultivation of plant tissue and organ cultures at laboratory and pilot scales are discussed. Finally, future prospects and challenges to the industrial realization of that fast-developing technique are outlined. [ABSTRACT FROM AUTHOR]

The Inter-Simple Sequence Repeat marker technique was applied to parental tomato genotypes and their F1 hybrids differing in the antioxidant compounds in the fruits. As the marker-assisted selection is becoming a tool that brings major advantages for breeding programs, the need to use it to quickly respond to the consumer demand has greatly increased. Modern genetic technology opens the possibility for effective application of a system for efficient plant breeding in many crops, including tomatoes. The aim of the present study was to test the Inter-Simple Sequence Repeats as a candidate tool to use in Marker-Assisted Selection in tomatoes. The application of the Inter-Simple Sequence Repeat markers allowed for efficient verification of the hybrid origin of the progeny from parental genotypes that were crossed for improving the antioxidant content in tomato fruits. [ABSTRACT FROM AUTHOR]

Abstract: The effects of methyl jasmonate and jasmonic acid on galanthamine production, phenolic acid content and growth of Leucojum aestivum L. shoot culture, cultivated in submerged conditions were investigated. The best time-point for addition of elicitors was during the exponential phase of the culture growth. The maximal contents of galanthamine and lycorine (226.9μg/flask and 491.4μg/flask, 1.36 and 1.67-fold higher compared to the control, respectively) were achieved after elicitation with jasmonic acid, whereas the elicitation with methyl jasmonte resulted in maximal accumulation of phenolic acids. It was demonstrated that the boosting effect of jasmonic acid on Amaryllidacea alkaloid biosynthesis was due to induction of the activity of tyrosine decarboxylase, whereas methyl jasmonate stimulates the biosynthesis of phenolic acids by inducing mainly the activity of phenylalanine ammonia-lyase. [Copyright &y& Elsevier]

DOI: 10.1002/elsc.202000078 The cover picture shows a process of submerged cultivation of hairy root culture of red beet in a stirred tank bioreactor, different types I Beta vulgaris i L. in vitro cultures and their ploidity levels. Among the investigated red beet in vitro cultures, hairy roots showed the best potential for production of betalains. [Extracted from the article]

Plants belonging to the monocotyledonous Amaryllidaceae family include about 1100 species divided among 75 genera. They are well known as medicinal and ornamental plants, producing pharmaceutically important alkaloids, the most intensively investigated of which are galanthamine and lycorine. Amaryllidaceae alkaloids possess various biological activities, the most important one being their anti-acetylcholinesterase activity, used for the treatment of Alzheimer's disease. Due to increased demand for Amaryllidaceae alkaloids (mainly galanthamine) and the limited availability of plant sources, in vitro culture technology has attracted the attention of researchers as a prospective alternative for their sustainable production. Plant in vitro systems have been extensively used for continuous, sustainable, and economically viable production of bioactive plant secondary metabolites. Over the past two decades, a significant success has been demonstrated in the development of in vitro systems synthesizing Amaryllidaceae alkaloids. The present review discusses the state of the art of in vitro Amaryllidaceae alkaloids production, summarizing recently documented plant in vitro systems producing them, as well as the authors' point of view on the development of biotechnological production processes with a focus on the future prospects of in vitro culture technology for the commercial production of these valuable alkaloids. [ABSTRACT FROM AUTHOR]

During the last few years there is an increasing demand to the natural biologically active compounds. According to the World Health Organization (WHO) about 11% of the conventional medicines are of plant origin. Nowadays, plant biotechnologies are modern and reliable tool for producing valuable bioactive compounds. Recently, the potential of plant cells as foods also was confirmed. The advantages of plant in vitro systems over the intact plants are well known: growing under controlled and optimized laboratory conditions; independence of climatic and soil differences; preservation of rare and endangered plant species; cultivation in diverse bioreactor systems for increasing production yields of target metabolites. There have been developed many in vitro systems for production of various plant bioactive compounds with potential application in food industries. But potential for industrial implementation of this technology depends on solving problems with the scale‐up of bioreactor cultivation, development of additional approaches for improving/modification of bioactivities of the target plant secondary metabolites, and to find way to exclude or replace in the culture media the carcinogenic plant growth regulator 2,4‐dichlorophenoxyacetic acid (2,4‐D) with its safety analogs, such as α‐naphtaleneacetic acid (NAA) and/or indole‐3‐butyric acid (IBA). The aim of the current mini review is to summarize information about different in vitro systems of edible plants from the Balkan Peninsula with potential for producing food additives and biologically active substances and to describe prospects for successful industrial implementation of this technology. [ABSTRACT FROM AUTHOR]

Plants have always been the main source for active cosmetic ingredients, having proven health beneficial effects on human, such as anti‐aging, antioxidant, anti‐inflammatory, UV‐protective, anti‐cancer, anti‐wrinkle, skin soothing, whitening, moisturizing, etc. Extracts from herbal, aromatic and/or medicinal plants have been widely used as effective active ingredients in cosmeceuticals or nutricosmetics, especially in products for topical application and skin‐care formulations. However, over the past decade, there has been an increasing interest to plant cell culture – derived active cosmetic ingredients. These are "new generation" of high quality natural products, produced by the modern plan biotechnology methods, which usually showed stronger activities than the plant extracts obtained by the classical methods. In this review, the advantages and the current progress in plant cell culture technology for the production of active cosmetic ingredients have been summarized, and discussed in details within a presented case study for calendula stem cell product development. [ABSTRACT FROM AUTHOR]

After eight years of cultivation of Sea daffodil (Pancratium maritimum L.) shoot cultures at in vitro conditions, bacterial contamination on the plant tissues and medium surface appeared. The contamination was due to endophytic bacterium later isolated as a bacterial strain PM_YT. The colonial characteristics and cell morphology of the isolate were determined after cultivation on YEB-agar medium and coloring of microscopic preparations. The strain identification was implemented by biochemical methods (oxidase and catalase standard strip tests) and nucleotide sequencing of the 16S rRNA gene. The results from the morphological and cultural identification showed that the isolate was Gram-positive bacterium presented from small, short rods, forming small (size of 1-3 mm), round-shaped, yellow- to slightly orange-colored colonies. The results from biochemical identification showed that the strain was oxidase-negative and catalasepositive. The comparative 16S rRNA gene sequence-based phylogenetic analysis revealed 99% pairwise similarity of the strain PM_YT to the bacterial species Curtobacterium flaccumfaciens. Antibiotic susceptibility was implemented by Bauer-Kirby disc diffusion method and the results demonstrated that the isolate was most sensitive to the antibiotics belonging to inhibitors of protein synthesis group, especially to tetracycline, doxycycline, amikacin, rifampin, lincomycin, chloramphenicol, erythromycin, streptomycin and ciprofloxacin. [ABSTRACT FROM AUTHOR]

The article profiles editors Thomas Bley, Atanas Pavlov and Kenneth F. Reardon, who contributed to the editing of the journal.

The article profiles editors of the periodical "Engineering in Life Sciences," including Thomas Bley, An-Ping Zeng, and Atanas Pavlov.

The influence of Salvia officinalis L. hairy roots derived phenolic acids on the growth of two typical cellulose degrading moulds, Chaetomium globosum and Trichoderma viride, was examined. For this, 26 hairy root lines of S. officinalis were induced and investigated with regard to the production of phenolic acids. Pure, commercial phenolic acids, analogues to that biosynthesised in the hairy root cultures, were solved in 96% ethanol and applied to malt extract agar plates. The fungal cultures were transferred to the agar plates, and fungal growth was observed 5 and 12 days after inoculation. The hairy root line A produced the highest amount of total phenolic acids (1582.0µg/g), while line B produced the highest amount of sinapic acid (262.8µg/g). Trans-cinnamic and salicylic acid showed antifungal activity to the growth of C. globosum. The growth of this strain was inhibited by vanillic, pcoumaric and sinapic acid. Moreover, it was observed that sinapic acid prevented the spore formation of C. globosum. Protocatechuic, sinapic, and salicylic acid inhibited the growth of T. viride minimal. Furthermore, only sinapic acid inhibited the spore formation of this mould. The investigations have shown antifungal activity of six S. officinalis hairy root derived phenolic acids on malt extract agar plates. [ABSTRACT FROM AUTHOR]

GS-MS based metabolite profile of fenugreek (Trigonella foenum-graecum L.) of three different trademarks was investigated. In polar fractions totally 40 primary metabolites (carbohydrates, organic and amino acids) and 9 phenolic acids (with caffeic and 4-hydroxybenzoic acids as major) were identified. The dominant metabolites in non-polar fraction were palmitic, linoleic, oleic and stearic acids. Two untypical triterpenic acids (oleanolic and ursolic acid) for family Fabaceae were identified in one of the analyzed sample. The presence of these triterpenoids in the sample of fenugreek indicates for probably adulteration, during the production process, with spices from other families, such as Lamiaceae or Rosaceae. The data of provided GC-MS analyses and subsequent principal component analysis (PCA) and hierarchical cluster analysis (HCA) revealed that investigated samples of the same species from three different trademarks had significant differences in their phytochemical content. Thus could be base for developing a protocol for quality control based on metabolomics approach. [ABSTRACT FROM AUTHOR]

Chemical composition and bioactivity of essential oil from Salvia tomentosa Mill. natively grown in Bulgaria were investigated. GC-MS analysis identified 60 compounds which represented 98% of the oil constituents. The prevalent constituents were monoterpenes with eight dominant compounds being identified: borneol (10.3%), β-pinene (9%), camphor (7.9%), α-pinene (6%), camphene (4%), 1.8-cineole (3.8%), α-limonene (3.5%) and β-caryophyllene (3%). The essential oil showed considerable acetylcholinesterase inhibitory activity (IC50=0.28±0.06 μg/mL), comparable with that of galanthamine. Study of antioxidant activity strongly suggested that the hydrogen atom transfer reaction was preferable over the electron transfer (ORAC=175.0±0.40 μM Trolox equivalents/g oil and FRAP=1.45±0.21 mM Trolox equivalents/g oil). The essential oil showed moderate antifungal and antibacterial activities against Candida albicans and Gram-positive bacteria, whereas it was almost inactive against the investigated Gram-negative strains. The results suggested that the essential oil of Bulgarian S. tomentosa could be considered as a prospective active ingredient for prevention of oxidative stress-related and neurodegenerative disorders in aromatherapy. Because of the high antioxidant capacity, the oil could be considered as natural supplement or antioxidant in cosmetics and food products. [ABSTRACT FROM AUTHOR]

The purpose of the present study was to investigate the antioxidant activity and fructans content in ethanol and water extracts of roots of elecampane (Inula helenium L.), a Bulgarian medicinal plant. The extraction procedure included 95% (v/v) ethanol extraction and subsequent water treatment. The antioxidant activity was evaluated by several reliable methods such as DPPH-, ABTS-, FRAP- and CUPRAC-assays, as well as the total phenolic content. In addition, the total fructans and sugar content were determinated by spectrophotomeric, TLC and HPLC-RID methods. The level of fructans in ethanol extracts was 14.1 g/100g dry weight, as nystose and 1-kestose were only 0.3g/100g dry weight, and 0.5g/100g dry weight, respectively. The absence of fructooligosacharides and sugars in water extracts after the ethanol pretreatment was established. Inulin content was evaluated to be 32g/100g dry weight. The metabolites profile of roots revealed their potential application as radical scavengers due to the presence of polyphenols. Therefore, the root extracts of elecampane could be assumed as a rich source of biologically active substance, in particular dietary fiber with potential prebiotic effect, due to the presence of polysaccharide inulin and fructooligosacharides. [ABSTRACT FROM AUTHOR]

The Amaryllidaceae alkaloid galanthamine (Gal) is a long-acting, selective, reversible, and competitive acetylcholinesterase inhibitor used for the treatment of early- to mid-stage Alzheimer's disease, poliomyelitis, and other neurological diseases. Currently, Gal is produced by extraction from plants such as daffodils ( Narcissus cultivar Carlthon), snowflake ( Leucojum aestivum), 'red-tubed lily' ( Lycoris radiata), and Ungernia victoria, and alternatively by chemical synthesis. Due to the increased demand by the generic pharmaceutical companies and the limited availability of plant sources, the biosynthesis of Gal by plant in vitro systems has been considered as an alternative approach for its sustainable production. The present article reviews the state of the art of in vitro Gal biosynthesis including growth regulators, medium components, culture conditions, elicitation, and bioreactor systems. It may be used as a starting point for further studies in this area leading to a progress in biotechnological production of this valuable alkaloid. [ABSTRACT FROM AUTHOR]

The aim of this study was profiling of primary metabolites of five Bulgarian Fumaria species (F. officinalis L., F. thuretii Boiss., F. kralikii Jord., F. rostellata Knaf. and F. schrammii Velen.) by GC-MS analyses. In polar fractions ten carbohydrates, one polyol, ten amino acids and six organic acids were identified. Apolar (lipid) fractions showed the presence of four free fatty acids, two esters of fatty acids with glycerol and two fatty alcohols. Sucrose and fructose were in the highest relative concentrations of identified carbohydrates. Citric acid was the dominant organic acid in polar fractions of five Fumaria species. Predominant compounds in lipid fractions were palmitic acid and 1- stearoyl-glycerol. Principal component analysis (PCA) of GC-MS data of polar and apolar fractions of five Bulgarian Fumaria species differentiates them in two groups (F. officinalis and F. thuretii; F. rostellata and F. schrammii, respectively), while F. kralikii had phythochemical similarity with plants of both distinguished groups. The obtained results of PCA of primary metabolites could be proposed as chemotaxonomic markers for plants of the genus Fumaria. [ABSTRACT FROM AUTHOR]

The constituents of essential oils, obtained from different aerial parts (leaves and flowers) of Salvia ringens Sibth. et Sm. growing wild in Bulgaria, have been analyzed by GC-MS. The yields of leaves and flowers oils were similar (0.03% v/w and 0.04% v/w). 60 compounds were identified in both oils, which correspond to 97.7% and 97.3% of the total oils constituents for leaves and flower oils, respectively. Both leaves and flowers oils showed similar chemical compositions, with predominance of camphor (17.2% and 18.8%) and borneol (7.2% and 8.7%). Further analyses of minor component distributions proves that the both leaves and flowers could be equally used as a raw material for obtaining of camphor reach essential oil from Salvia ringens Sibth. et Sm. [ABSTRACT FROM AUTHOR]

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 cultivations. Other major problems are associated with the challenges of cultivation conditions and bioreactor design as well as upscaling of the process. This article reviews bioreactor designs for cultivation of differentiated plant in vitro systems, secondary metabolite production in different bioreactor systems as well as aspects of process control, management, and modeling and gives perspectives for future cultivation methods. [ABSTRACT FROM AUTHOR]

A rapid and accurate TLC-densitometry method for qualitative and quantitative determination of protopine has been developed. The best separation was achieved using a mobile phase chloroform - ethyl acetate - methanol - ammonium hydroxide (80:80:40:0.05, v/v/v/v). The results obtained by this method (CV% 3.4) corresponded well with those obtained by using a HPLC method. The reliability of the proposed method was proved through reproducibility test with alkaloid extracts from Fumaria spp [ABSTRACT FROM AUTHOR]

Biomass accumulation and tropane alkaloids production by diploid and tetraploid hairy root cultures of Datura stramonium L. cultivated in stirred tank bioreactor at different aeration rates were investigated. The maximal growth for both hairy root cultures (ADB = 8.3 g/L and 6.8 g/L for diploid and tetraploid line, respectively) was achieved at aeration rate of 15.0 L/(L.h). The corresponding growth indexes were remarkably high (GIDW = 9.0 and 7.8 for diploid and tetraploid line, respectively) compared to the values, usually reported for other hairy root cultures. The optimal aeration rate for biomass accumulation was also optimal for alkaloids biosynthesis. According to our survey, the achieved maximal amounts of accumulated hyoscyamine (35.0 mg/L and 27.0 mg/L for diploid and tetraploid line) were the highest reported in the scientific literature for D. stramonium L. hairy roots. During the cultivation in stirred tank bioreactor, the hairy roots biosynthesized pharmaceutically important alkaloid scopolamine in minor concentrations. This is an important observation since scopolamine was not detected during submerged cultivation of these hairy root lines in other bioreactor types. However, the ploidy level was found to be the most important factor concerning scopolamine production by D. stramonium L. hairy roots cultures. The present work demonstrated the effect of ploidity levels on biomass accumulation and tropane alkaloids production by D. stramonium L. hairy roots cultivated in stirred tank bioreactor. This investigation show that the stirred tank bioreactor could be successfully applied for both maximal biomass accumulations, as well as for manipulation of tropane alkaloids production by diploid and tetraploid D. stramonium L. hairy root cultures. [ABSTRACT FROM AUTHOR]

Shoot culture of summer snowflake ( Leucojum aestivum L.) was successfully cultivated in an advanced modified glass-column bioreactor with internal sections for production of Amaryllidaceae alkaloids. The highest amounts of dry biomass (20.8 g/L) and galanthamine (1.7 mg/L) were achieved when shoots were cultured at 22°C and 18 L/(L·h) flow rate of inlet air. At these conditions, the L. aestivum shoot culture possessed mixotrophic-type nutrition, synthesizing the highest amounts of chlorophyll (0.24 mg/g DW (dry weight) chlorophyll A and 0.13 mg/g DW chlorophyll B). The alkaloids extract of shoot biomass showed high acetylcholinesterase inhibitory activity ( IC50 = 4.6 mg). The gas chromatography-mass spectrometry (GC/MS) profiling of biosynthesized alkaloids revealed that galanthamine and related compounds were presented in higher extracellular proportions while lycorine and hemanthamine-type compounds had higher intracellular proportions. The developed modified bubble-column bioreactor with internal sections provided conditions ensuring the growth and galanthamine production by L. aestivum shoot culture. [ABSTRACT FROM AUTHOR]

Abstract: The alkaloid patterns in Leucojum aestivum L. shoot culture cultivated at temporary immersion conditions were investigated using gas chromatography–mass spectrometry. 18 alkaloids were identified, and galanthamine, hamayne and lycorine were dominant. The L. aestivum 80 shoot culture, cultivated at temporary immersion conditions, is a prospective biological matrix for obtaining wide range Amaryllidaceae alkaloids, showing valuable biological and pharmacological activities. The temperature of cultivation influenced enzyme activities, catalyzing phenol oxidative coupling of 4′-O-methylnorbelladine and formation of the different groups Amaryllidaceae alkaloids. Decreasing the temperature of cultivation of L. aestivum 80 shoot culture led to activation of para-ortho’ phenol oxidative coupling (formation of galanthamine type alkaloids) and inhibited ortho-para’ and para-para’ phenol oxidative coupling (formation of lycorine and haemanthamine types alkaloids). [Copyright &y& Elsevier]

The growth and hyosciamine biosynthesis by diploid and tetraploid Datura stramonium L. hairy rootlines in temporary immersion RITA" system were investigated. The immersion frequency had significant effect on hyosciamine production and accumulation of biomass for both hairy root cultures. The maximum amount of hyosciamine (6.53 mg.g-1 DW) was achieved during cultivation of diploid Datura stramonium L. hairy root culture at immersion frequency with 15 rain flooding and 105 min stand-by periods. Data for utilization of the main inorganic compounds during cultivation process in temporary immersion RITA" system are presented as well. [ABSTRACT FROM AUTHOR]

Abstract: Three often cited systems for the extraction of plant nuclei for flow cytometric measurement (CyStain PI, Partec GmbH, Münster, Germany, the method of Arumuganathan and Earle, and LB01 buffer) failed, when applied to the hairy roots of red beet (Beta vulgaris). By combining these systems and introducing a centrifugation step, the extraction, staining, and analysis of nuclei from this tissue were performed successfully. [Copyright &y& Elsevier]