Structure Elucidation and Toxicity Analysis of the Byproducts Formed after Biodegradation of Aflatoxins B1 and B2 Using Extracts of Mentha arvensis.

The aqueous extracts of leaves and shoots of Mentha arvensis were checked for their potential to biodegrade aflatoxin B1 and B2 (AFB1; 100 µg/L and AFB2; 50 µg/L) through in vitro assays. Overall, the results showed that leaf extract degrades aflatoxins more efficiently than the shoot extract. First, the pH, temperature and incubation time were optimized for maximum degradation by observing this activity at different temperatures between 25 and 60 °C, pH between 2 and 10 and incubation time from 3 to 72 h. In general, an increase in all these parameters significantly increased the percentage of biodegradation. In vitro trials on mature maize stock were performed under optimized conditions, i.e., pH 8, temperature 30 °C and an incubation period of 72 h. The leaf extract resulted in 75% and 80% biodegradation of AFB1 and AFB2, respectively. Whereas the shoot extract degraded both toxins up to 40-48%. The structural elucidation of degraded toxin products by LCMS/MS analysis showed seven degraded products of AFB1 and three of AFB2. MS/MS spectra showed that most of the products were formed by the loss of the methoxy group from the side chain of the benzene ring, the removal of the double bond in the terminal furan ring and the modification of the lactone group, indicating less toxicity compared to the parent compounds. The degraded products showed low toxicity against brine shrimps, confirming that M. arvensis leaf extract has significant potential to biodegrade aflatoxins.

Classification of vine-shoots for use as enological additives.

BACKGROUND: Toasted vine shoots have recently been proposed as enological additives with the aim of improving the sensorial profile of wines. However, so far, there is no simple method for classifying vine shoots for this innovative enological practice. In this study, therefore, an enological aptitude classification for toasted vine shoots has been proposed for the first time. Moreover, given the need for quick techniques to be used in wineries to determine the main phenolic compounds of vine shoots, near-infrared (NIR) spectroscopy has been calibrated and validated. RESULTS: By means of a detailed statistical analysis, an enological classification of toasted vine shoots has been proposed based on their total polyphenol index and (+)-catechin, (-)-epicatechin, ellagic acid, and trans-resveratrol. Moreover, the NIR methodology that was developed showed good validation statistics and acceptable accuracy. CONCLUSIONS: This work proposes the first enological toasted vine-shoot classification and it provides a tool for rapid screening, mainly of phenolic compounds, in toasted vine shoots. © 2021 The Authors. Journal of The Science of Food and Agriculture published by John Wiley & Sons Ltd on behalf of Society of Chemical Industry.

In-vitro propagation and phytochemical profiling of a highly medicinal and endemic plant species of the Himalayan region (Saussurea costus).

Efficient protocols for callus induction and micro propagation of Saussurea costus (Falc.) Lipsch were developed and phytochemical diversity of wild and in-vitro propagated material was investigated. Brown and red compact callus was formed with frequency of 80-95%, 78-90%, 70-95% and 65-80% from seeds, leaf, petiole and root explants, respectively. MS media supplemented with BAP (2.0 mgL-1), NAA (1.0 mgL-1) and GA3 (0.25 mgL-1) best suited for multiple shoot buds initiation (82%), while maximum shoot length was formed on media with BAP (1.5 mgL-1), NAA (0.25 mgL-1) and Kinetin (0.5 mgL-1). Full strength media with IAA (0.5 mgL-1) along with IBA (0.5 mgL-1) resulted in early roots initiation. Similarly, maximum rooting (87.57%) and lateral roots formation (up to 6.76) was recorded on full strength media supplemented with BAP (0.5 mgL-1), IAA (0.5 mgL-1) and IBA (0.5 mgL-1). Survival rate of acclimatized plantlets in autoclaved garden soil, farmyard soil, and sand (2:1:1) was 87%. Phytochemical analysis revealed variations in biochemical contents i.e. maximum sugar (808.32 µM/ml), proline (48.14 mg/g), ascorbic acid (373.801 mM/g) and phenolic compounds (642.72 mgL-1) were recorded from callus cultured on different stress media. Nonetheless, highest flavenoids (59.892 mg/g) and anthocyanin contents (32.39 mg/kg) were observed in in-vitro propagated plants. GC-MS analysis of the callus ethyl acetate extracts revealed 24 different phytochemicals. The variability in secondary metabolites of both wild and propagated plants/callus is reported for the first time for this species. This study may provide a baseline for the conservation and sustainable utilization of S. costus with implications for isolation of unique and pharmacologically active compounds from callus or regenerated plantlets.

A metabolomics approach to evaluate the effect of lyophilization versus oven drying on the chemical composition of plant extracts.

Lyophilization is the "gold standard" for drying plant extracts, which is important in preserving their quality and extending their shelf-life. Compared to other methods of drying plant extracts, lyophilization is costlier due to equipment, material and operational expenses. An alternative method is post-extraction oven-drying, but the effects of this process on extract quality are unknown. In this study, crude extracts from Arthrocnemum macrostachyum shoots were compared using three post-extraction drying methods (lyophilization and oven drying at 40 and 60 °C) and two extraction solvents (water and aqueous 50% ethanol). Untargeted metabolomics coupled with chemometrics analysis revealed that post extraction oven-drying resulted in the loss of up to 27% of molecular features when compared to lyophilization in water extracts only. In contrast, only 3% of molecular features were lost in aqueous 50% ethanol extracts when subjected to oven drying. That is to say, ethanol used as a solvent has a stabilizing effect on metabolites and enhances their resistance to thermal transformation in the oven. Collectively, oven-drying of extracts was as effective as lyophilization in preserving metabolites in extracts only when 50% ethanol was used as a solvent. The results presented in this paper demonstrate the value of selecting solvent-appropriate post-extraction drying methods.

Micropropagation of Rare Scutellaria havanensis Jacq. and Preliminary Studies on Antioxidant Capacity and Anti-Cancer Potential.

We report the development of in vitro propagation protocols through an adventitious shoot induction pathway for a rare and medicinal Scutellaria havanensis. In vitro propagation studies using nodal explants showed MS medium supplemented with 10 µM 6-Benzylaminopurine induced the highest number of adventitious shoots in a time-dependent manner. A ten-day incubation was optimum for shoot bud induction as longer exposures resulted in hyperhydricity of the explants and shoots induced. We also report preliminary evidence of Agrobacterium tumefaciens EHA105-mediated gene transfer transiently expressing the green fluorescent protein in this species. Transformation studies exhibited amenability of various explant tissues, internode being the most receptive. As the plant has medicinal value, research was carried out to evaluate its potential antioxidant capacity and the efficacy of methanolic leaf extracts in curbing the viability of human colorectal cancer cell line HCT116. Comparative total polyphenol and flavonoid content measurement of fresh and air-dried leaf extract revealed that the fresh leaf extracts contain higher total polyphenol and flavonoid content. The HCT 116 cell viability was assessed by colorimetric assay using a 3-(4,5-dimethyl-thiazol-2-yl)-2,5-diphenyltetrazolium bromide, showed a steady growth inhibition after 24 h of incubation. Scanning electron microscopy of leaf surface revealed a high density of glandular and non-glandular trichomes. This research provides a basis for the conservation of this rare plant and future phytochemical screening and clinical research.

Oral and Topical Anti-Inflammatory and Antipyretic Potentialities of Araucaria bidiwillii Shoot Essential Oil and Its Nanoemulsion in Relation to Chemical Composition.

Different parts of Araucaria bidiwillii (bunya pin) trees, such as nuts, seeds, bark, and shoots, are widely used in cooking, tea, and traditional medicines around the world. The shoots essential oil (EO) has not yet been studied. Herein, the chemical profile of A. bidiwillii shoots EO (ABSEO) was created by GC-MS analysis. Additionally, the in vivo oral and topical anti-inflammatory effect against carrageenan-induced models, as well as antipyretic potentiality of ABSEO and its nanoemulsion were evaluated. Forty-three terpenoid components were identified and categorized as mono- (42.94%), sesqui- (31.66%), and diterpenes (23.74%). The main compounds of the ABSEO were beyerene (20.81%), α-pinene (16.21%), D-limonene (14.22%), germacrene D (6.69%), ß-humulene (4.14%), and sabinene (4.12%). The ABSEO and its nanoemulsion exhibited significant inflammation suppression in carrageenan-induced rat paw edema model, in both oral (50 and 100 mg/kg) and topical (5% in soyabean oil) routes, compared to the control and reference drugs groups. All the results demonstrated the significant inflammation reduction via the inflammatory cytokines (IL-1ß and IL8), nitrosative (NO), and prostaglandin E2 (PGE2) supported by the histopathological studies and immunohistochemical assessment of MMP-9 and NF-κß levels in paw tissues. Moreover, the oral administration of ABSEO and its nanoemulsion (50 and 100 mg/kg) exhibited antipyretic activity in rats, demonstrated by the inhibition of hyperthermia induced by intramuscular injection of brewer's yeast. These findings advised that the use of ABSEO and its nanoemulsion against numerous inflammatory and hyperthermia ailments that could be attributed to its active constituents.

Precursor-Boosted Production of Metabolites in Nasturtium officinale Microshoots Grown in Plantform Bioreactors, and Antioxidant and Antimicrobial Activities of Biomass Extracts.

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

Pseudotsuga menziesii (Pinaceae): Volatile Profiles, Antimicrobial Activity and Toxicological Evaluation of Its Essential Oil.

The present article investigates the chemical composition of volatiles of essential oil (EO) and headspace (HS) fraction, as well as biological activities of EO obtained from needles with twigs of Pseudotsuga menziesii var. menziesii cultivated in Serbia. The major class of compounds was monoterpene hydrocarbons with α-terpinolene, sabinene and ß-pinene (EO), and sabinene, α-terpinolene and ß-pinene (HS) as the dominant volatiles. Tested EO exhibited mostly low antimicrobial potential against investigated strains (ATCC and respiratory isolates), where MICs ranged 1.25-20.00 mg/mL. Nevertheless, based on presented results, where antimicrobial testing was done for the first time on human respiratory system isolates, there is a potential of this EO to be used as an adjuvant in the treatment of human respiratory infections, especially those caused by Pseudomonas aeruginosa or Candida albicans strains. Regarding toxicological evaluation, EO showed moderate toxicity in Artemia salina toxicity bioassay (LC50 =347.41, after 24 h) as well as week toxicity against Drosophila melanogaster with the ability only to moderately delay larval and pupal development.

Evaluation of Hepatoprotective Activity and Oxidative Stress Reduction of Rosmarinus officinalis L. Shoots Tincture in Rats with Experimentally Induced Hepatotoxicity.

Rosmarinus officinalis L. is a widely known species for its medicinal uses, that is also used as raw material for the food and cosmetic industry. The aim of the present study was to offer a novel perspective on the medicinal product originating from this species and to test its hepatoprotective activity. The tested sample consisted in a tincture obtained from the fresh young shoots. Compounds that are evaluated for this activity are polyphenols and terpenoids, that are identified and quantified by HPLC-UV-MS and GC-MS. Antioxidant activity was assessed in vitro, using the DPPH, FRAP and SO assays. Hepatoprotective activity was tested in rats with experimentally-induced hepatotoxicity. In the chemical composition of the tincture, phenolic diterpenes (carnosic acid, carnosol, rosmanol, rosmadial) and rosmarinic acid were found to be the majority compounds, alongside with 1,8-cineole, camphene, linalool, borneol and terpineol among monoterpenes. In vitro, the tested tincture proved significant antioxidant capacity. Results of the in vivo experiment showed that hepatoprotective activity is based on an antioxidant mechanism. In this way, the present study offers a novel perspective on the medicinal uses of the species, proving significant amounts of polyphenols and terpenes in the composition of the fresh young shoots tincture, that has proved hepatoprotective activity through an antioxidant mechanism.

Mass propagation of Juniperus procera Hoechst. Ex Endl. From seedling and screening of bioactive compounds in shoot and callus extract.

BACKGROUND: Juniperus procera Hoechst. ex Endl. is a medicinal tree in Saudi Arabia, primarily in the Enemas region, but it is locally threatened due to die-back disease and difficulties regarding seed reproduction (seed dormancy and underdeveloped embryonic anatomy, and germination rate < 40%). Hence, the alternative methods for reproduction of Juniperus procera are really needed for conservation and getting mass propagation for pharmaceutical uses. RESULTS: In this manuscript, we articulated the successful in vitro shoot multiplication and callus induction of J. procera by using young seedling as explants and detected an important antibacterial and antitumor product. Explants were grown on different types of media with the supplement of different combinations of Plant Growth Regulators (PGRs) at different concentrations. The best media for shoot multiplication was Woody Plant Media (WPM) supplemented with PGRs (0.5 µM of IAA and 0.5 µM BAP or 0.5 µM IBA and 0.5 µM BAP). Whereas for callus induction and formation Woody Plant Media (WPM) with the addition of PGRs (0.5 µM 2,4-D and 0.5 µM BAP) was better than the Chu Basal Salt Mixture (N6), Gamborg's B-5 Basal Medium (B5), and Murashige and Skoog media. The possibility of multiplication of J. procera in vitro creates significant advantages to overcome the difficulties of seeds dormancy for the reproduction of plants, conservation of trees, and getting mass propagation material for pharmaceutical studies. The shoot and callus extract of J. procera was detected using gas chromatography-mass spectrometry analysis and revealed more than 20 compounds related to secondary metabolites, which contained antibacterial and antitumor agents, such as ferruginol, Retinol, and Quinolone as well as confirmed by Direct Analysis in Real Time, Time of Flight Mass Spectrometry (DART-ToF-MS). Podophyllotoxin (PTOX) was detected in callus material by HPLC with sigma standard and confirmed by DART-ToF-MS and UV spectra. CONCLUSION: We successfully conducted in vitro shoot multiplication and callus induction from J. procera seedlings using WPM and a different combination of PGRs and, detected an important antibacterial and antitumor product such as ferruginol and podophyllotoxin. According to our findings, J. procera has become a new natural source of novel bioactive compounds.

Sarracenia alata (Alph.Wood) Alph.Wood Microcuttings as a Source of Volatiles Potentially Responsible for Insects' Respond.

Rare carnivorous plants representing the genus Sarracenia are perceived as very interesting to scientists involved in various fields of botany, ethnobotany, entomology, phytochemistry and others. Such high interest is caused mainly by the unique capacity of Sarracenia spp. to attract insects. Therefore, an attempt to develop a protocol for micropropagation of the Sarracenia alata (Alph.Wood) Alph.Wood, commonly named yellow trumpets, and to identify the specific chemical composition of volatile compounds of this plant in vitro and ex vivo was undertaken. Thus, the chemical volatile compounds excreted by the studied plant to attract insects were recognized with the application of the headspace solid-phase microextraction (HS-SPME) coupled with the GC-MS technique. As the major volatile compounds (Z)-3-hexen-1-ol (16.48% ± 0.31), (E)-3-hexen-1-ol acetate (19.99% ± 0.01) and ß-caryophyllene (11.30% ± 0.27) were identified. Further, both the chemical assumed to be responsible for attracting insects, i.e., pyridine (3.10% ± 0.07), and whole plants were used in in vivo bioassays with two insect species, namely Drosophila hydei and Acyrthosiphon pisum. The obtained results bring a new perspective on the possibilities of cultivating rare carnivorous plants in vitro since they are regarded as a valuable source of bioactive volatile compounds, as including ones with repellent or attractant activity.

In Vitro Phytotherapeutic Properties of Aqueous Extracted Adenia viridiflora Craib. towards Civilization Diseases.

Adenia viridiflora Craib. is an indigenous edible plant that became an endangered species due to limited consumption of the local population with unknown reproduction and growth conditions. The plant is used as a traditional herb; however, its health applications lack scientific-based evidence. A. viridiflora Craib. plant parts (old leaves and young shoots) from four areas as Kamphaeng Phet (KP), Muang Nakhon Ratchasima (MN), Pakchong Nakhon Ratchasima (PN), and Uthai Thani (UT) origins were investigated for phenolic compositions and in vitro health properties through the inhibition of key enzymes relevant to obesity (lipase), diabetes (α-glucosidase and dipeptidyl peptidase-IV), Alzheimer's disease (cholinesterases and ß-secretase), and hypertension (angiotensin-converting enzyme). Phenolics including p-coumaric acid, sinapic acid, naringenin, and apigenin were detected in old leaves and young shoots in all plant origins. Old leaves exhibited higher total phenolic contents (TPCs) and total flavonoid contents (TFCs), leading to higher enzyme inhibitory activities than young shoots. Besides, PN and MN with higher TPCs and TFCs tended to exhibit greater enzyme inhibitory activities than others. These results will be useful to promote this plant as a healthy food with valuable medicinal capacities to support its consumption and agricultural stimulation, leading to sustainable conservation of this endangered species.

Widely Targeted Metabolomics Analysis of Different Parts of Salsola collina Pall.

Salsola collina Pall has a long history of being used as a traditional medicine to treat hypertension, headache, insomnia, constipation and vertigo. However, only a few biologically active substances have been identified from S. collina. Here, the shoots and roots of S. collina, namely L-Sc and R-Sc, were studied. The primary and secondary metabolites were investigated using ultrahigh-performance liquid chromatography-electrospray ionization-tandem mass spectrometry (UPLC-ESI-MS/MS). A total of 637 putative metabolites were identified and these metabolites were mainly classified into ten different categories. Correlation analysis, hierarchical clustering analysis, principal component analysis and orthogonal partial least squares discriminant analysis of metabolites showed that the L-Sc samples could be clearly separated from the R-Sc samples. Differential accumulated metabolite analysis revealed that most of differential primary metabolites were significantly lower in the L-Sc than in the R-Sc. Conversely, the major differential secondary metabolites had higher levels in the L-Sc than in the R-Sc. Further analysis indicated that the flavonoids were the major putative antioxidant components and most of putative antioxidant components exhibited higher relative concentrations in the L-Sc than the R-Sc. These results improve our understanding of metabolite accumulation and provide a reference for the study of medicinal value in S. collina.

Chemical and Nutritional Compounds of Different Parts of Lemongrass (Cymbopogon citratus (DC) Stapf.) Cultivated in Temperate Climate of Poland.

Lemongrass (Cymbopogon citratus (DC) Stapf.) is a perennial plant indigenous to semi-tropical regions of Asia and cultivated in other semi-tropical countries. The present study aimed to examine the key chemical constituents of various parts of lemongrass cultivated in the temperate climate of Poland. The content of essential oil and its composition were determined in 4 plant parts: leaves (part C), overground shoots (part B), underground shoots (part A), and roots (part R). Moreover, the content of dry weight, chlorophyll, polyphenols and macro- and microelements was determined in the edible parts (excluding roots). The essential oil from the aerial part predominantly contained neral (> 30%) and geranial (> 40%), which is consistent with the data reported in literature; the main component of essential oil (EO) from the underground part was elemol (65%); interestingly, such a high concentration of it was found for the first time. The concentration of chlorophyll was found to be higher in leaves, as compared to parts B and A. The highest level of potassium, magnesium, zinc and sodium was found in part A while of calcium and copper in leaves. The quality of lemongrass raw materials grown in temperate climates did not differ significantly from those obtained in warmer regions. The study findings confirmed the usefulness of leaves as a raw material for the preparation of infusions (higher concentration of pigments, polyphenols and EO) and of near-ground parts of a plant as a culinary material (a higher content of macroelements at lower concentrations of green pigments and dry weight).

Investigation of Different Selenium Sources and Supplying Methods for Selenium Enrichment of Basil Vegetable (A Case Study under Calcareous and Non-calcareous Soil Systems).

BACKGROUND: Selenium (Se) is a crucial component of selenoaminoacids and selenoproteins. Therefore, Se-enriched agricultural products can reduce health complications induced by Se deficiency. OBJECTIVE: This research was carried out to investigate the effects of Se bio-enrichment on Basil grown in calcareous and non-calcareous soil systems and also to evaluate the changes in Se concentration in the soil after harvesting. METHODS: The experiment executed in two calcareous and one non-calcareous soil systems, and different Se application methods (control, soil application, seed inoculation, foliar application, and soil + foliar application) were administered. Selenobacteria, a plant growth-promoting rhizobacteria (PGPR), derived from the soil was used as a biofertilizer, compared to the other Se sources. RESULTS: The results showed that both soil types and the methods of Se application had significant effects (P ˂ 0.01) on root and shoot dry weights and concentrations of P, K, Zn, Fe, and Se in both of the root and shoot. Shoot dry weight of plants treated with foliar Se was maximum in the calcareous soil. Compared to the control treatment, foliar application of Se increased shoot Se content in both calcareous and non-calcareous soils by 242% and 204%, respectively. Furthermore, the increase in shoot Se concentration in calcareous soil induced by Se application increased the concentration of other nutrients in the shoot and root. Plant growth parameters and concentrations of nutrients were significantly increased by using selenobacter inoculum. CONCLUSION: The application of Se-containing compounds can improve vegetable quality. Considering the daily requirement of the human body for minerals and nutrients, enriching basil with Se can play an important role in community health. Moreover, some patents have reported the effectiveness of endophyte bacteria.

Cytotoxic, Apoptosis-Inducing Activities, and Molecular Docking of a New Sterol from Bamboo Shoot Skin Phyllostachys heterocycla var. pubescens.

Phytochemical screening of nonpolar fractions from the methanol extract of the Bamboo shoot skin Phyllostachys heterocycla var. pubescens resulted in the isolation of a new sterol-glucoside-fatty acid derivative (6'-O-octadeca-8'',11''-dienoyl)-sitosterol-3-O-ß-d-glucoside (1), together with six known compounds. The chemical structures of the pure isolated compounds were deduced based on different spectral data. The isolated compounds were assessed to determine their cytotoxic activity, and the results were confirmed by determining their apoptotic activity. Compound 1 was more cytotoxic against the MCF-7 cells (IC50 = 25.8 µM) compared to Fluorouracil (5-FU) (26.98 µM), and it significantly stimulated apoptotic breast cancer cell death with 32.6-fold (16.63% compared to 0.51 for the control) at pre-G1 and G2/M-phase cell cycle arrest and blocked the progression of MCF-7 cells. Additionally, RT-PCR results further confirmed the apoptotic activity of compound 1 by the upregulation of proapoptotic genes (P53; Bax; and caspases 3, 8, and 9) and downregulation of the antiapoptotic genes (BCL2). Finally, the identified compounds, especially 1, were found to have high binding affinity towards both tyrosine-specific protein kinase (TPK) and vascular endothelial growth factor receptor (VEGFR-2) through the molecular docking studies that highlight its mode of action.

1Selenium supply alters the subcellular distribution and chemical forms of cadmium and the expression of transporter genes involved in cadmium uptake and translocation in winter wheat (Triticum aestivum).

BACKGROUND: Cadmium (Cd) accumulation in crops affects the yield and quality of crops and harms human health. The application of selenium (Se) can reduce the absorption and transport of Cd in winter wheat. RESULTS: The results showed that increasing Se supply significantly decreased Cd concentration and accumulation in the shoot and root of winter wheat and the root-to-shoot translocation of Cd. Se application increased the root length, surface area and root volume but decreased the average root diameter. Increasing Se supply significantly decreased Cd concentration in the cell wall, soluble fraction and cell organelles in root and shoot. An increase in Se supply inhibited Cd distribution in the organelles of shoot and root but enhanced Cd distribution in the soluble fraction of shoot and the cell wall of root. The Se supply also decreased the proportion of active Cd (ethanol-extractable (FE) Cd and deionized water-extractable (FW) Cd) in root. In addition, the expression of TaNramp5-a, TaNramp5-b, TaHMA3-a, TaHMA3-b and TaHMA2 significantly increased with increasing Cd concentration in root, and the expression of TaNramp5-a, TaNramp5-b and TaHMA2 in root was downregulated by increasing Se supply, regardless of Se supply or Cd stress. The expression of TaHMA3-b in root was significantly downregulated by 10 µM Se at both the 5 µM and 25 µM Cd level but upregulated by 5 µM Se at the 25 µM Cd level. The expression of TaNramp5-a, TaNramp5-b, TaHMA3-a, TaHMA3-b and TaHMA2 in shoot was downregulated by increasing Se supply at 5 µM Cd level, and 5 µM Se upregulated the expression of those genes in shoot at 25 µM Cd level. CONCLUSIONS: The results confirm that Se application limits Cd accumulation in wheat by regulating the subcellular distribution and chemical forms of Cd in winter wheat tissues, as well as the expression of TaNramp5-a, TaNramp5-b and TaHMA2 in root.

Phytochemical and Biological Activity Studies on Nasturtium officinale (Watercress) Microshoot Cultures Grown in RITA® Temporary Immersion Systems.

The main compounds in both extracts were gluconasturtiin, 4-methoxyglucobrassicin and rutoside, the amounts of which were, respectively, determined as 182.93, 58.86 and 23.24 mg/100 g dry weight (DW) in biomass extracts and 640.94, 23.47 and 7.20 mg/100 g DW in plant herb extracts. The antioxidant potential of all the studied extracts evaluated using CUPRAC (CUPric Reducing Antioxidant Activity), FRAP (Ferric Reducing Ability of Plasma), and DPPH (1,1-diphenyl-2-picrylhydrazyl) assays was comparable. The anti-inflammatory activity of the extracts was tested based on the inhibition of 15-lipoxygenase, cyclooxygenase-1, cyclooxygenase-2 (COX-2), and phospholipase A2. The results demonstrate significantly higher inhibition of COX-2 for in vitro cultured biomass compared with the herb extracts (75.4 and 41.1%, respectively). Moreover, all the studied extracts showed almost similar antibacterial and antifungal potential. Based on these findings, and due to the fact that the growth of in vitro microshoots is independent of environmental conditions and unaffected by environmental pollution, we propose that biomass that can be rapidly grown in RITA® bioreactors can serve as an alternative source of bioactive compounds with valuable biological properties.

Biosynthesis and characterization of silver nanoparticles using Ochradenus arabicus and their physiological effect on Maerua oblongifolia raised in vitro.

Silver nanoparticles (AgNPs) are presently the most commonly generated engineered nanomaterials and are found in a wide range of agro-commercial products. The present study was designed to synthesize AgNPs biologically using Ochradenus arabicus leaves and investigate their effect on the morphophysiological properties of Maerua oblongifolia raised in vitro. Physicochemical methods (ultraviolet-visible spectroscopy, Fourier transform infrared spectroscopy, and transmission electron microscopy were performed for characterization and for obtaining microphotographs of the AgNPs. Shoots of M. oblongifolia (2-3 cm) grown in Murashige and Skoog medium supplemented with different concentrations of AgNPs (0, 10, 20, 30, 40, or 50 mg L-1) were used. Following 6 weeks of in vitro shoot regeneration, the shoot number, shoot length, leaf number, fresh weight, dry weight, chlorophyll content, total protein, proline level, and antioxidant enzyme activities of the plants were quantified. We found that 20 mg L-1 AgNPs increased the shoot number, shoot length, fresh weight, dry weight, and chlorophyll content of the plants. The maximum total protein was recorded in plants that were administered the lowest dose of AgNPs (10 mg L-1), while high concentrations of AgNPs (40 and 50 mg L-1) increased the levels of proline and the enzymes superoxide dismutase and catalase. Our results indicate that green-synthesized AgNPs may be of agricultural and medicinal interest owing to their effects on plants in vitro.

Transformed Shoots of Dracocephalum forrestii W.W. Smith from Different Bioreactor Systems as a Rich Source of Natural Phenolic Compounds.

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