Production Of Phytochemicals Research Articles

Mechanical and phytochemical protection mechanisms of Calligonum comosum in arid deserts.

Unlike animals, plants are sessile organisms, lacking circulating antibodies and specialized immune cells and are exposed to various harsh environmental conditions that make them at risk of being attacked by different pathogens and herbivores. Plants produce chemo-signals to respond to the surroundings and be able to distinguish between harmless and harmful signals. In this study, the production of phytochemicals as plant signaling mechanisms and their defensive roles in disease resistance and repelling herbivores are examined in Calligonum comosum. C. comosum is a leafless standalone perennial shrub widespread in sand dunes. The plant has the ability to survive the drastic environmental conditions of the arid/ hyperarid deserts of the Arabia. Structural anatomy and phytochemicals analyses were used to identify both mechanical and chemical defensive mechanisms in C. comosum. Microscopy-based investigations indicated that stems of this species developed hard structures in its outer layers including sclerenchyma and cluster crystals of calcium oxalate (CaOx). Sclerenchyma and CaOx are difficult to be eaten by herbivores and insects and can harm their mouthparts. On the other hand, the plant developed both short-distance (local) and long-distance (systematic over limited sphere) phytochemicals-producing cells located at its outer regions that is surrounding the inner nutrient-rich vascular system (VS). Local chemical was represented by phenolic idioblasts that were released in response to plant cutting. Systematic chemical was represented by toxic volatile oil containing ~50% benzaldehyde derivative (cuminaldehyde). The oil caused strong killing effect on both mammalian cells and microbial pathogens via either direct addition or indirect exposure to its vapor. The plants lost the oil content and allowed fungal growth once cut and dried. The localization of both defensive mechanisms to the outer region of the plant seemed to protect the inner nutrient-rich VS and hence maintained the plant survival. Surprisingly, in relation to traditional folklore use as medicine, local people use only green parts of the plant and only during the winter, where the plant found devoid of volatile oil and phenolic idioblasts. Moreover, it turns into recommendations for local people to avoid any health problems caused by the plant supply.

Cultured cambial meristematic cells - a sustainable route towards in vitro production of phytochemicals.

Cultured cambial meristematic cells - a sustainable route towards in vitro production of phytochemicals.

Phytochemical Screening of the Hydro Ethanolic Extract of Brassica oleracea Var. Italica Plant Extract

Objective: This study was designed elucidate the Phytochemicals of the widely-used plant Brassica oleracea var. Italica. Method: Hydroethanolic extracts of Brassica oleracea var. Italica plant extract was investigated. A small portion of the hydroethanolic extracts of Brassica oleracea var. Italica was subjected to the phytochemical test using Trease and Evans and Harbourne methods to test for the presence of alkaloids, tannins, reducing sugars, saponins, terpenoids, phenols, flavonoids and Anthraquionones. Result: The phytochemical analysis revealed the presence of alkaloids, saponins, tannins, flavonoids, terpenoids, coumarins, quinines, cardiac glycosides, Xanthoproteins, glycosides, steroids, phenols, resins, carboxylic acid group in varying concentrations. The present study provides evidence that Hydro ethanolic extracts of Brassica oleracea var. Italica contains medicinally important bioactive compounds and this justifies the use of plant species as traditional medicine for treatment of various diseases. Conclusion: Thus, from the present study the plant leaf extracts of Brassica oleracea var. Italica showed an abundant production of Phytochemicals as secondary metabolites and they can be used in the pharmaceutical industries for producing a potent drug. The studies result of the above two plants gives a basis of its use in traditional medicine to manage ailments and disorders.

The Role of Shade and Nitrogen on Physiological Traits and Secondary Metabolites of Piper betle L

Piper betle, better known as purple betel has received a universal attention due to its increasing medicinal value. There is enormous potential to discover various new medicinal compounds in this species and an instantaneous need for the techniques to facilitate the production of high quality. In that connection, shade and nitrogen could play a significant role in the development of a phytochemical production. Therefore, the current study utilized the different levels of shade and rates of nitrogen. After harvesting of three months Piper betle seedlings, different physio-chemical traits were observed including photosynthetic rate, stomata conductor, transpiration rates, chlorophyll content (a, b, ab) and secondary metabolites ( total phenolics contents (TPC) concentration, total flavonoids contents (TFC) concentration and antioxidant activate (DPPH and FRAP). The chlorophyll content was more under 30% and 50% shades with 100 kg/ ha of N, whereas full sunlight with 0 kg/ha of N was unsuitable for the Piper betle crop as the high light intensity caused scorching on leaves and stunted in growth, accumulation secondary metabolites and Antioxidant activates. The results of the present study revealed the establishment of shade techniques which influenced the production parameters of the piper species. These findings are an overview of the recent advances and could be further helpful in in vivo studies of Piper betel in regard to their physiological parameters.

Fitoquímicos, nutrimentos y factores ambientales asociados a la rugosidad de la piel del aguacate ‘Hass’ de tres regiones de México

El ambiente en el que se desarrolla el fruto de aguacate (Persea americana Mill.) cv. Hass determina las características de su piel la cual representa la primera barrera natural y el tejido que interactúa directamente con el ambiente. El fruto está expuesto a muchos tipos de estrés y para protegerse produce una amplia variedad de fitoquímicos, llamados metabolitos secundarios. Se desconoce si aspectos como la composición nutrimental y morfología de la piel también podrían ser afectados. El estudio se realizó en huertos comerciales de aguacate ‘Hass’ con el manejo estándar de cada productor con el objetivo de determinar las concentraciones de fitoquímicos (compuestos fenólicos totales, clorofilas totales, carotenoides totales y lignina) y nutrimentos (N, P, K, Ca, Mg, S, Fe, Cu, Mn, Zn y B), y establecer su relación con la rugosidad de la piel de los frutos durante su desarrollo en tres regiones con distinto tipo de clima. El tipo de clima (cálido, semicálido o templado) influyó sobre la producción de fitoquímicos y en la rugosidad de la piel de ‘Hass’. El análisis global mostró que los frutos del clima cálido presentaron la mayor concentración de carotenoides totales y mayor rugosidad, mientras que en el clima templado, la rugosidad fue menor. El grosor de las protuberancias de la piel y el corcho se correlacionó significativamente con la concentración de lignina y macronutrimentos.

Recent Advances in the Recombinant Biosynthesis of Polyphenols.

Plants are the source of various natural compounds with pharmaceutical and nutraceutical importance which have shown numerous health benefits with relatively fewer side effects. However, extraction of these compounds from native producers cannot meet the ever-increasing demands of the growing population due to, among other things, the limited production of the active compound(s). Their production depends upon the metabolic demands of the plant and is also subjected to environmental conditions, abundance of crop species and seasonal variations. Moreover, their extraction from plants requires complex downstream processing and can also lead to the extinction of many useful plant varieties. Microbial engineering is one of the alternative approaches which can meet the global demand for natural products in an eco-friendly manner. Metabolic engineering of microbes or pathway reconstruction using synthetic biology tools and novel enzymes lead to the generation of a diversity of compounds (like flavonoids, stilbenes, anthocyanins etc.) and their natural and non-natural derivatives. Strain and pathway optimization, pathway regulation and tolerance engineering have produced microbial cell factories into which the metabolic pathway of plants can be introduced for the production of compounds of interest on an industrial scale in an economical and eco-friendly way. While microbial production of phytochemicals needs to further increase product titer if it is ever to become a commercial success. The present review covers the advancements made for the improvement of microbial cell factories in order to increase the product titer of recombinant polyphenolic compounds.

Triterpene Structural Diversification by Plant Cytochrome P450 Enzymes.

Cytochrome P450 monooxygenases (P450s) represent the largest enzyme family of the plant metabolism. Plants typically devote about 1% of the protein-coding genes for the P450s to execute primary metabolism and also to perform species-specific specialized functions including metabolism of the triterpenes, isoprene-derived 30-carbon compounds. Triterpenes constitute a large and structurally diverse class of natural products with various industrial and pharmaceutical applications. P450-catalyzed structural modification is crucial for the diversification and functionalization of the triterpene scaffolds. In recent times, a remarkable progress has been made in understanding the function of the P450s in plant triterpene metabolism. So far, ∼80 P450s are assigned biochemical functions related to the plant triterpene metabolism. The members of the subfamilies CYP51G, CYP85A, CYP90B-D, CYP710A, CYP724B, and CYP734A are generally conserved across the plant kingdom to take part in plant primary metabolism related to the biosynthesis of essential sterols and steroid hormones. However, the members of the subfamilies CYP51H, CYP71A,D, CYP72A, CYP81Q, CYP87D, CYP88D,L, CYP93E, CYP705A, CYP708A, and CYP716A,C,E,S,U,Y are required for the metabolism of the specialized triterpenes that might perform species-specific functions including chemical defense toward specialized pathogens. Moreover, a recent advancement in high-throughput sequencing of the transcriptomes and genomes has resulted in identification of a large number of candidate P450s from diverse plant species. Assigning biochemical functions to these P450s will be of interest to extend our knowledge on triterpene metabolism in diverse plant species and also for the sustainable production of valuable phytochemicals.

Ecophysiological and phytochemical characterization of wild populations of Inula montana L. (Asteraceae) in Southeastern France

Inula montana is a member of the family Asteraceae and is present in substantial numbers in Garrigue country (calcareous Mediterranean ecoregion). This species has traditionally been used for its anti-inflammatory properties as well as Arnica montana. In this study, three habitats within Luberon Park (southern France) were compared regarding their pedoclimatic parameters and the resulting morpho-physiological response of the plants. The data showed that I. montana grows in south-facing poor soils and tolerates large altitudinal and temperature gradients. The habitat conditions at high elevation appear to affect mostly the morphology of the plant (organ shortening). Although the leaf contents of total polyphenols and flavonoids subclass essentially followed a seasonal pattern, many sesquiterpene lactones were shown to accumulate first at the low-elevation growing sites that suffered drought stress (draining topsoil, higher temperatures and presence of a drought period during the summer). This work highlights the biological variability of I. montana related to the variation of its natural habitats which is promising for the future domestication of this plant. The manipulation of environmental factors during cultivation is of great interest due to its innovative perspective for modulating and exploiting the phytochemical production of I. montana.

Anthocyanin Contents Enhancement with Gamma Irradiated Mutagenesis in Blackberry (Rubus fructicosus)

Effects of radiation processing on the endogenous phytochemical production in blackberry ( Rubus fructicosus L.) were investigated through a metabolites analysis, using high-performance liquid chromatography with a diode array detector and a triple quadrupole mass spectrometer (HPLC-DAD-ESI-MS/MS). Four mutant cultivar lines (B, C, D, and E) were developed through the treatment of various doses of gamma irradiation (80, 60, 40, and 20 Gy) on the control blackberry cultivar (V3). These mutant cultivar lines were selected according to their different morphological variations in leaf shape and branch color. In the present study, we performed quantitative analyses of the four standard compounds in blackberry mutant cultivars: cyanidin-3- O-glucoside, cyanidin-3- O-xyloside, cyanidin-3- O-malonylglucoside, and cyanidin-3- O-dioxalylglucoside. The HPLC fingerprints displayed different profiles for six blackberry cultivar lines, demonstrating that the accumulation of total anthocyanins was affected by the gamma irradiation. The contents of cyanidin-3- O-glucoside and cyanidin-3- O-xyloside were high in the gamma-irradiated mutant cultivars of blackberries, compared to those of the control cultivar (V3) and the cross breeding cultivar ( R. fructicosus × R. parvifolius; CB). Cyanidin-3- O-malonylglucoside and cyanidin-3- O-dioxalylglucoside were also analyzed for their quantification in six cultivar lines, as metabolites contributing to the discrimination of six cultivar lines. Among gamma irradiated mutant blackberry cultivars, the highest dose level of gamma irradiation (80 Gy) resulted in the highest cyanidin-3- O-glucoside content enhancement. Therefore, these results can be useful to determine the optimized gamma irradiated value at which anthocyanins reaches an abundant level in Rubus species.

Identification of genic SSRs in jute (Corchorus capsularis, Malvaceae) and development of markers for phenylpropanoid biosynthesis genes and regulatory genes

In jute (Corchorus spp.), phenylpropanoid biosynthesis pathway is associated with lignocellulosic bast fibre development and production of various phytochemicals. Development of phenylpropanoid-related genic markers is thus a key component of gene-specific precision molecular breeding in jute. We identified a total of 12,772 SSRs in 10,041 unigenes and designed genic SSR markers associated with phenylpropanoid biosynthesis from bast transcriptome of Corchorus capsularis cv. ‘JRC-212’. A total of 39 and 14 SSR markers were developed from key genes of phenylpropanoid biosynthesis and bast fibre formation, respectively. Further, we developed 457 SSR markers associated with different regulatory gene sequences. Of these, 137 markers were for transcription factors such as WRKY, MYB, MYB-related, bHLH and zinc finger, which are key regulators and/or modulators of the biosynthesis of phenylpropanoids. Additionally, 600 SSR primer pairs were also designed for other unigenes containing SSR. Diversity analysis based on a set of 41 genic SSR markers in 35 genotypes revealed high polymorphism. The genic SSRs described here are expected to be useful for molecular breeding targeting fibre quality, phytochemical-related and stress tolerance traits in jute.

Transcriptomic Response of Purple Willow (Salix purpurea) to Arsenic Stress.

Arsenic (As) is a toxic element for plants and one of the most common anthropogenic pollutants found at contaminated sites. Despite its severe effects on plant metabolism, several species can accumulate substantial amounts of arsenic and endure the associated stress. However, the genetic mechanisms involved in arsenic tolerance remains obscure in many model plant species used for land decontamination (phytoremediation), including willows. The present study assesses the potential of Salix purpurea cv. ‘Fish Creek’ for arsenic phytoextraction and reveals the genetic responses behind arsenic tolerance, phytoextraction and metabolism. Four weeks of hydroponic exposure to 0, 5, 30 and 100 mg/L revealed that plants were able to tolerate up to 5 mg/L arsenic. Concentrations of 0 and 5 mg/L of arsenic treatment were then used to compare alterations in gene expression of roots, stems and leaves using RNA sequencing. Differential gene expression revealed transcripts encoding proteins putatively involved in entry of arsenic into the roots, storage in vacuoles and potential transport through the plant as well as primary and secondary (indirect) toxicity tolerance mechanisms. A major role for tannin as a compound used to relieve cellular toxicity is implicated as well as unexpected expression of the cadmium transporter CAX2, providing a potential means for internal arsenic mobility. These insights into the underpinning genetics of a successful phytoremediating species present novel opportunities for selection of dedicated arsenic tolerant crops as well as the potential to integrate such tolerances into a wider Salix ideotype alongside traits including biomass yield, biomass quality, low agricultural inputs and phytochemical production.

Antimycobacterial Potential of Essential Oils Used Traditionally for Cough Remedy

Tuberculosis (TB) is currently a major health hazard due to multidrug-resistant forms of bacilli. Global efforts are underway to eradicate TB using new drugs with new modes of action, higher activity, and fewer side effects. Essential oils from some dietary herbal species have been used as sources of medicine and food preservatives for over 4000 years. The objective of this research is to investigate the antimycobacterial activity of the essential oils of Boswellia papyrifera (Del) Hochst olibanum, Nigella sativa seeds and Eucalyptus camaldulensis Dehnh leaves, which are used traditionally in treatment of cough and cold. The essential oils from herbal materials were prepared using hydrodistillation method. The essential oils were analyzed for identification of their chemical composition using GC-MS technique. The essential oils were tested in vitro for their activity against nine clinical isolates and a reference susceptible strain (H37Rv) M. tuberculosis in Lowenstein-Jensen (LJ) medium, at concentrations of 75, 50, 30 and 15 μl/mL. The main constituents identified in the investigated essential oils were p-cymene (45.26%) and thymoquinone (35.35 %) in N. sativa oil; octyl acetate (37.3%) and octyl formate (13.3%) in the B. papyrifera oil; 1,8-cineole (75%) and p-cymene (16.7%) in E. camaldulensis oil. The investigated essential oils exhibited anti-tubercular activity, in LJ medium, up to concentration of 15 μl/ml for each oil, towards all the tested strains, three of which were multi-drug resistant strains (MDR-TB), as they resist rifampicin, a marker drug for MDR-TB. This research findings may be useful in developing future powerful drugs for MDR-TB from essential oils, the bioactive phytochemical natural products.

The influence of light quality on phenolic acid and biflavonoid production in Anacardium othonianum Rizz. seedlings grown in vitro

Anacardium othonianum Rizz., popularly known as the Cerrado cashew tree, is widely used in medicine due to its therapeutic properties, including its antioxidant, anti-inflammatory, and antidiarrheal activities. Light is an important regulator of the growth, development of plants differentiation, and metabolism. Considering the influence of light on the production of phytochemicals, the aim of the present study was to evaluate production of gallic and 3,4-dihydroxybenzoic acids and amentoflavone in A. othonianum seedlings grown in vitro exposure to white, blue, green, red, and yellow light. The seedlings were evaluated with respect to the length of the aerial parts and the number of leaves, at 0, 20, and 40 days of treatment. The fresh and dry weights of the aerial parts and the levels of phenolic acid and biflavonoid were evaluated at 40 days via high-performance liquid chromatography with a diode array detector (HPLC-DAD). Regardless of the light wavelength used for the growth, the largest length of the aerial parts (5.54 cm) and an increase in the number of leaves (2.86-fold) were observed at 40 days. The largest accumulations of fresh weight (325.5 mg and 335.1 mg per seedling) and dry weight (83.3 mg and 84.5 mg per seedling) were observed in seedlings grown under red and yellow light, respectively. Relatively higher levels of gallic acid (0.66 mg.g-1dry weight) and amentoflavone (1.41 mg.g-1dry weight) were found in seedlings grown under blue light. In contrast, the 3,4-dihydroxybenzoic acid levels of the seedlings under red light increased (8.22 mg.g-1 dry weight).

Impact of pulsed light on colour, carotenoid, polyacetylene and sugar content of carrot slices

Abstract Carrot slices were subjected to pulsed light (PL) treatments on both sides (total PL fluences of 2.26, 4.52, 5.41, 9.38 and 13.15 J/cm 2 ) with the objective to evaluate their effect on the colour, β-carotene, polyacetylene and soluble sugar content. Higher concentrations of β-carotene were found after processing of carrot slices to the fluences of 2.26 and 4.52 J/cm 2 as compared with untreated carrots. This behaviour was partly related to the strongly colour observed in the cortex tissue of samples when similar PL treatment conditions were applied. Moreover, carrot slices treated with PL doses of 2.26 J/cm 2 also showed the highest falcarindiol (127.1%), falcarinol (94.8%) and falcarindiol-3-acetate (84.7%) retention levels. The same PL dose also led to an increment in the β-glucose in carrot slices after the treatment. Industrial relevance The pre-treatment of carrot slices with pulsed light would enhance the level of polyacetylene and carotenoid content as a stress response without affecting negatively the colour characteristics of the fresh material. Pulsed light technology could be useful for the food industry in production of high value phytochemical vegetable products.

Growth and Bioactive Compound Synthesis in Cultivated Lettuce Subject to Light-quality Changes

This study aimed to determine the effect of changes in light quality on the improvement of growth and bioactive compound synthesis in red-leaf lettuce (Lactuca sativa L. ‘Sunmang’) grown in a plant factory with electrical lighting. Lettuce seedlings were subjected to 12 light treatments combining five lighting sources: red (R; 655 nm), blue (B; 456 nm), and different ratios of red and blue light combined with three light-emitting diodes [LEDs (R9B1, R8B2, and R6B4)]. Treatments were divided into control (continuous irradiation of each light source for 4 weeks), monochromatic (changing from R to B at 1, 2, or 3 weeks after the onset of the experiments), and combined (changing from R9B1 to R8B2 or R6B4 at 2 or 3 weeks after the onset of the experiments). Growth and photosynthetic rates of lettuce increased with increasing ratios of red light, whereas chlorophyll and antioxidant phenolic content decreased with increasing ratios of red light. Individual phenolic compounds, including chlorogenic, caffeic, chicoric, and ferulic acids, and kaempferol, showed a similar trend to that of total phenolics. Moreover, transcript levels of phenylalanine ammonia-lyase (PAL) and chalcone synthase (CHS) genes were rapidly upregulated by changing light quality from red to blue. Although the concentration of bioactive compounds in lettuce leaves enhanced with blue light, their contents per lettuce plant were more directly affected by red light, suggesting that biomass as well as bioactive compounds’ accumulation should be considered to enhance phytochemical production. In addition, results suggested that growth and antioxidant phenolic compound synthesis were more sensitive to monochromatic light than to combined light variations. In conclusion, the adjustment of light quality at a specific growth stage should be considered as a strategic tool for improving crop yield, nutritional quality, or both in a plant factory with electrical lighting.

Antibacterial and Phytochemical Study of Iraqi Salvia officinalis Leave Extracts

Sage (Salvia officinalis), belong to Labiatae family is indigenous to Iraq and other Mediterranean areas but now cultivated world- wide, principally for its use as culinary herb. In the present study preliminary screening for the important phytochemical natural product groups indicated the presence of flavonoid, saponin, hyrolysable and condensed tannin groups. The antibacterial activity of two concentrations 10 mg/ml and 100 mg/ml of chloroform and hydroalcoholic extracts from Salvia officinalis leaves was evaluated against four strains of gram negative bacteria (Escherichia coli, Pseudomonas arigenossa, Klebsiella pneumonia, and Proteus spp) and two strains of gram positive bacteria (Staphylococcus aureus, Bacillus cerus) using agar well diffusion method. Chloroform extract 100mg/ml was found active against two types of bacteria, Staphylococcus aureus with MIC of 90 mg/ml and Proteus spp with MIC of 80 mg/ml. Bioassay guided separation using TLC led to the separation of 6 constituents from the active extract, one of them was identified as thujone.
 Key words: Salvia officinalis, Antibacterial, TLC, Thujone.

Induction of milk thistle (Silybum marianum L. Gaertn) growth and phytochemicals production by natural stimulants

Two field experiments were carried out through 2013/2014 and 2014/2015 seasons in Faculty of Agriculture experimental farm, Tanta University, to investigate the influence of chicken manure (CM) and yeast extract (YE) on Silybum marianum (milk thistle) growth, yield and phytochemical constituents. CM was applied to soil with rates of 0.0, 23.81, 35.71 and 47.62m3/hectare (ha) and YE was applied after 15 and 75days of seed sowing, with rates of 0, 25, 50 and 100g/L as foliar spray. The design of the experiment was as split plot with CM as the main plot. Results indicated that the applied bio-stimulants improved the vegetative traits and seed yield, but without obvious effect on chlorophyll content, though both treatments had resulted in a significant enhancement in PSII photochemical efficiency. Also, both bio-stimulants possessed positive impact on the antioxidant status of seed extracts and silymarin production, as well as the phytochemical constituents. However, these treatments caused a pronounced fluctuation in amino acids profile.

PROSPECTS FOR USE OF CONDENSED GASES AND SUPERCRITICAL FLUIDS IN PHYTOCHEMICAL PRODUCTION

In the given review article analysis of the literature and patent sources concerning main methods for intensification of extraction processes of medicinalvegetative raw materials – use of condensed gases and supercritical fluids (SCF) on more acceptable extractants has been carried out for last20 years. Urgency of the specified technologies consists in need for replacement of traditional extraction methods on power- and time-saving ones, and also in use of nontoxic, fire-proof and low-boiling solvents because the most of routine organic solvents (ethanol, methanol, acetone, chloroform, ethylacetate, etc.) are toxic and/or flammable or expansive and rather hard to evaporate out from extracts obtained. The abovementioned trends are the most universal for intensification of extraction processes and sometimes purification of final or intermediate products acceptable for commercial scale of manufacture. The main advantages and disadvantages of the givenmethodsare compared for different plant species and groups of biologically active substances (BAS). It has been shown that in most cases supercritical СО2(SC-СО2)are inferiorin its dissolving ability to number of condensed gases and, besides, such technology is much more expensive. The range of BAS taken with SC-СО2 is limited to mainly lipophilic compounds because of zero electrical dipole moment of SC-СО2 and its low polarity. As extractants alternative to SC-СО2 with higher dissolving ability SC - ethane, nitrogen monoxide, freons - R134а, R23, R32, R408 and number of others can be used. Also to enlarge range of extractable BAS it is possible to add different cosolvents, mainly ethanol or methanol in quantity up to 20%. At the same time in phytochemical production prospective alternatives to liquid or supercritical СО2 are certain condensed gases with wider range of physico-chemical properties: fluorinated derivatives of hydrocarbons (freons), liquid ammonia, dimethyl ether (DME), sulfur hexafluoride (insulating gas) or their mixtures, etc. Their major characteristics include lower vapor pressure if compared with liquid СО2, antimicrobic activity allowing to solve one the main problem in phytochemical production – microbial contamination of extracts (and other herbal drug preparations), possibility to extract not only lipophilic, but also more polar substances depending on choice of solvents or their mixes and their higher extraction rate.It has been found that some kinds of freons (for example, R22) due to their higher polarity are able to take wider spectrum of BAS than liquid СО2: essential and fat oils, fat-soluble vitamins, coumarins, carotenoids, phenolic alcohols, valrates, iridoids, some alkaloids and flavonoids. Besides, certain freons (for example, С318) have very high selectivity allowing to extract essential oils without accompanying fats. Some condensed gases, such as liquid ammonia, dimethyl ether and difluoromethane (freon R32) can be used as well for obtaining of hydrophilic BAS (saponins, alkaloids, flavonoids). Thus such solvents should be polar enough or can be mixed with polar modifiers. Supercriticalfluids and some subcritical condensed gases are suitable for fractionatingof primary (crude) extracts because their selectivity considerably depends on temperature, pressureand composition (in case of mixtures with each other or with cosolvents). Also high selectivity of condensed gas and SCFs is shown in near-critical areas. Very important property of most of condensed gases and SCFs is their ability to considerably reduce microbial contamination of extracts in comparison with initial plant raw materials.Conclusions. Among existing methods for intensification of stage of plant extraction the most applicable for commercial scale is use of condensed gases and supercritical fluids as extractants. It is found that for today in the world the most widespread SCF is carbon dioxide. The main lacks of СО2 as an extractant are high working pressure and narrow spectrum of extractable BAS which is limited only to lipophilic substances. This induces the search for alternative condensed gases and SCF among which fluorohydrocarbons (freons) are of the greatest interest. Also perspective methods of extraction intensification are applications of ultrasound, microwave field and some other methods of raw material processing, but meanwhile they have not got industrial value because of insufficient scientific base.

Jasmonic and Salicylic Acids Enhanced Phytochemical Production and Biological Activities in Cell Suspension Cultures of Spine Gourd (Momordica dioica Roxb)

In vitro cell suspension culture was established for the production of commercially valuable phytochemicals in Momordica dioica. The influence of elicitors in jasmonic acid (JA) and salicylic acid (SA) increased their effect on phytochemical production and biomass accumulation in M. dioica. The results indicate that compared with non-elicited cultures, JA- and SA-elicited cell suspension cultures had significantly enhanced phenolic, flavonoid, and carotenoid production, as well as antioxidant, antimicrobial, and antiproliferative activities. Furthermore, elicited cultures produced 22 phenolic compounds, such as flavonols, hydroxycinnamic acids, and hydroxybenzoic acids. Greater biomass production, phytochemical accumulation, and biological activity occurred in JA- than in SA-elicited cell cultures. This study is the first to successfully establish M. dioica cell suspension cultures for the production of phenolic compounds and carotenoids, as well as for biomass accumulation.

Establishment of Axenic Explants and Callus Induction of Vernonia amygdalina Del.

Vernonia amygdalina is a plant with great medicinal potentials due to the phytochemicals it possesses. The establishment of axenic culture and callus induction are important as the primary steps to acquire callus and cell suspension cultures which are prospective to be used as alternatives in the production of phytochemicals. The effect of different regimes of sterilization unveiled that explants treated with mercuric chloride showed significant difference in the mean number of axenic explants, but observations of serious necrosis in all explants that later lead to death were recorded. Whilst, the explants treated with Clorox® were not significantly difference in the mean number of axenic explants but the explants obtained were healthy and the growth was sustained. For callus induction, there was no significant difference in the callus induction frequency (CIF) among different concentrations of 2,4-D or picloram. In conclusions, the explants were recommended to be surface sterilized with 15% of Clorox® for 15 minutes as it was more economical and less time consuming. Whereas, MS media supplemented with 0.5 mg/L 2,4-D or 0.5 mg/L picloram would be a better choice for callus induction due to observation of root formation at lower concentrations of 2,4-D.