Shoots and Turions of Aquatic Plants as a Source of Fatty Acids.

BACKGROUND: Fatty acids are essential for human health. Currently, there is a search for alternative sources of fatty acids that could supplement such sources as staple crops or fishes. Turions of aquatic plants accumulate a variety of substances such as starch, free sugars, amino acids, reserve proteins and lipids. Our aim is to see if turions can be a valuable source of fatty acids. METHODS: Overwintering shoots and turions of aquatic carnivorous plants were collected. The plant material was extracted with hexane. The oils were analyzed using a gas chromatograph with mass spectrometer. RESULTS: The dominant compound in all samples was linolenic acid. The oil content was different in turions and shoots. The oil content of the shoots was higher than that of the turions, but the proportion of fatty acids in the oils from the shoots was low in contrast to the oils from the turions. The turions of Utricularia species were shown to be composed of about 50% fatty acids. CONCLUSIONS: The turions of Utricularia species can be used to obtain oil with unsaturated fatty acids. In addition, the high fatty acid content of turions may explain their ability to survive at low temperatures.

Light-Emitting Diodes and Liquid System Affect the Caffeoylquinic Acid Derivative and Flavonoid Production and Shoot Growth of Rhaponticum carthamoides (Willd.) Iljin.

Plant in vitro cultures can be an effective tool in obtaining desired specialized metabolites. The purpose of this study was to evaluate the effect of light-emitting diodes (LEDs) on phenolic compounds in Rhaponticum carthamoides shoots cultured in vitro. R. carthamoides is an endemic and medicinal plant at risk of extinction due to the massive harvesting of its roots and rhizomes from the natural environment. The shoots were cultured on an agar-solidified and liquid-agitated Murashige and Skoog's medium supplemented with 0.1 mg/L of indole-3-acetic acid (IAA) and 0.5 mg/L of 6-benzyladenine (BA). The effect of the medium and different treatments of LED lights (blue (BL), red (RL), white (WL), and a combination of red and blue (R:BL; 7:3)) on R. carthamoides shoot growth and its biosynthetic potential was observed. Medium type and the duration of LED light exposure did not affect the proliferation rate of shoots, but they altered the shoot morphology and specialized metabolite accumulation. The liquid medium and BL light were the most beneficial for the caffeoylquinic acid derivatives (CQAs) production, shoot growth, and biomass increment. The liquid medium and BL light enhanced the content of the sum of all identified CQAs (6 mg/g DW) about three-fold compared to WL light and control, fluorescent lamps. HPLC-UV analysis confirmed that chlorogenic acid (5-CQA) was the primary compound in shoot extracts regardless of the type of culture and the light conditions (1.19-3.25 mg/g DW), with the highest level under R:BL light. BL and RL lights were equally effective. The abundant component was also 3,5-di-O-caffeoylquinic acid, accompanied by 4,5-di-O-caffeoylquinic acid, a tentatively identified dicaffeoylquinic acid derivative, and a tricaffeoylquinic acid derivative 2, the contents of which depended on the LED light conditions.

Advancing Simultaneous Extraction and Sequential Single-Particle ICP-MS Analysis for Metallic Nanoparticle Mixtures in Plant Tissues.

Engineered nanoparticles (ENPs) have been increasingly used in agricultural operations, leading to an urgent need for robust methods to analyze co-occurring ENPs in plant tissues. In response, this study advanced the simultaneous extraction of coexisting silver, cerium oxide, and copper oxide ENPs in lettuce shoots and roots using macerozyme R-10 and analyzed them by single-particle inductively coupled plasma-mass spectrometry (ICP-MS). Additionally, the standard stock suspensions of the ENPs were stabilized with citrate, and the long-term stability (up to 5 months) was examined for the first time. The method performance results displayed satisfactory accuracies and precisions and achieved low particle concentration and particle size detection limits. Significantly, the oven drying process was proved not to impact the properties of the ENPs; therefore, oven-dried lettuce tissues were used in this study, which markedly expanded the applicability of this method. This robust methodology provides a timely approach to characterize and quantify multiple coexisting ENPs in plants.

The impact of extraction processes on the physicochemical, functional properties and structures of bamboo shoot protein.

This study aimed to extract bamboo shoot protein (BSP) using different extraction approaches and compare their functional and physicochemical properties with commercial protein ingredients, including whey protein and soy protein isolates. The extraction methods including alkali extraction (AE), salt extraction (SE), and phosphate-aided ethanol precipitation (PE) were used. An enhanced solvent extraction method was utilized in combination, resulting in a significant improvement in the protein purity, which reached 81.59 %, 87.36 %, and 67.08 % respectively. The extraction methods had significant effects on the amino acid composition, molecular weight distribution, and functional properties of the proteins. SE exhibited the best solubility and emulsification properties. Its solubility reached up to 93.38 % under alkaline conditions, and the emulsion stabilized by SE with enhanced solvent extraction retained 60.95 % stability after 120 min, which could be attributed to its higher protein content, higher surface hydrophobicity, and relative more stable and organized protein structure. All three BSP samples demonstrated better oil holding capacity, while the SE sample showed comparable functional properties to soy protein such as foaming and emulsifying properties. These findings indicate the potential of BSP as an alternative plant protein ingredient in the food industry.

Comparative hepato-ameliorative effects of Bambusa nutans fresh and fermented shoot extracts on STZ induced diabetic LACA mice.

Bamboo shoots are nutritionally rich source of antioxidants and bioactive compounds with immense therapeutic potentials. The fresh shoot is acrid and needs to be processed to make it palatable. Fermentation is one the best processing methods for long term storage and make the shoot palatable and enhance taste. This study aims to assess the prophylactic hepatoprotective effects of fresh and fermented B. nutans shoot aqueous extract (200 mg/kg b.w.) in STZ induced diabetic LACA mice. Both extracts effectively improved body weight loss, hyperglycemia, and hepatomegaly. Fresh shoot reduced LDH activity and LPO level by 26.1% and 46.6%, while fermented shoot reduced them by 51.5% and 55.8%, respectively. The fermented shoot extract group demonstrated a noteworthy decrease in liver enzymes (SGPT, SGOT, ALP, and bilirubin levels) and an increase in albumin and A/G ratio, with more substantial improvements compared to the group treated with fresh extract. Additionally, the extracts enhanced antioxidant activities and showed histological improvements in hepatocytes and central vein structure. The findings indicate that both fresh and fermented B. nutans extracts are non-toxic and possess hepatoprotective potential in hyperglycaemic liver dysfunction, with fermented shoot extract exhibiting superior efficacy suggesting its potential as a therapeutic agent for hyperglycemic liver conditions.

Sensory properties of rehydrated Toona sinensis shoots after dehydrated by different drying methods and association with gamma-glutamyl transferase reaction.

Sulfur-containing compounds are responsible for the aroma of Toona sinensis shoot (TS). In this study, vacuum-freeze-drying (VFD), microwave-drying (MD), and hot-air-drying at 100 and 40 °C (HAD100 and HAD40, respectively), were applied to dehydrate perishable TS for preservation. VFD-TS retained most aroma of fresh/raw TS after rehydration. The content of sulfur-containing compounds reached to 118.00 µg/g with leading by methyl thiirane, (E,E)/(E,Z)/(Z,Z)-bis-(1-propenyl) disulfides, and (Z)/(E)-2-mercapto-3,4-dimethyl-2,3-dihydrothiophenes accounting for 86.33 %. They were undetected in the rehydrated MD-TS and HAD100-TS, as the indigenous enzymes in TS were deactivated under their dehydration conditions. Interestingly, the sulfur-containing compounds was restored by 77.47 % after the TS was treated by gamma-glutamyl transferase (GGT). Thus, the release of sulfur-containing compounds from TS could depend on GGT reaction. It was different from alliaceous vegetables relying on alliinase reaction. The results revealed the aroma formation in TS and provided an approach to enhance the aroma of TS dried by different methods.

Exploration of Global and Specialized Near-Infrared Calibrations for the Quantification of Nutritional Content in Grapevine Organs, Berry Phenological Stages, and Shoot Lignification.

Current infrared spectroscopy applications in the field of viticulture are moving toward direct in-field measuring techniques. However, limited research is available on quantitative applications using direct measurement of fresh tissue. The few studies conducted have combined the spectral data from various cultivars, growing regions, grapevine organs, and phenological stages during model development. The spectral data from these heterogeneous samples are combined into a single data set and analyzed jointly during quantitative analysis. Combining the spectral information of these diverse samples into a global data set could be an unsuitable approach and could yield less accurate prediction results. Spectral differences among samples could be overlooked during model development and quantitative analysis. The development of specialized calibrations should be considered and could lead to more accurate quantitative analyses. This study explored a model optimization strategy attempting global and specialized calibrations. Global calibrations, containing data from multiple organs, berry phenological, and shoot lignification stages, were compared to specialized calibrations per organ or stage. The global calibration for organs contained data from shoots, leaves, and berries and produced moderately accurate prediction results for nitrogen, carbon, and hydrogen. The specialized calibrations per organ yielded more accurate calibrations with a coefficient of determination in validation (R2val) at 90.65% and a root mean square error of prediction (RMSEP) at 0.32% dry matter (DM) for the berries' carbon calibrations. The leaves and shoots carbon calibrations had R2val and RMSEP at 84.99%, 0.34% DM, and 90.06%, 0.37% DM, respectively. The specialized calibrations for nitrogen and hydrogen showed similar improvements in prediction accuracy per organ. Specialized calibrations per phenological and lignification stage were also explored. Not all stages showed improvement, however, most stages had comparable or improved results for the specialized calibrations compared to the global calibrations containing all phenological or lignification stages. The results indicated that both global and specialized calibrations should be considered during model development to optimize prediction accuracy.

Multifunctional self-assembled adsorption microspheres based on waste bamboo shoot shells for multi-pollutant water purification.

In this study, multilayer self-assembled multifunctional bamboo shoot shell biochar microspheres (BSSBM) were prepared, in which bamboo shoot shell biochar was used as the carrier, titanium dioxide as the intermediate medium, and chitosan as the adhesion layer. The adsorption behavior of BSSBM on heavy metals Ag(I) and Pd(II), antibiotics, and dye wastewater was systematically analyzed. BSSBM shows a wide range of adsorption capacity. BSSBM is a promising candidate for the purification of real polluted water, not only for metal ions, but also for Tetracycline (TC) and Methylene Blue (MB). The maximum adsorption amounts of BSSBM on Pd(II), Ag(I), TC and MB were 417.3 mg/g, 222.5 mg/g, 97.2 mg/g and 42.9 mg/g, respectively.The adsorption of BSSBM on Pd(II), MB and TC conformed to the quasi-first kinetic model, and the adsorption on Ag(I) conformed to the quasi-second kinetic model. BSSBM showed remarkable selective adsorption capacity for Ag(I) and Pd(II) in a multi-ion coexistence system. BSSBM not only realized the high value-added utilization of waste, but also had the advantages of low cost, renewable and selective adsorption. BSSBM demonstrated its potential as a new generation of multifunctional adsorbent, contributing to the recovery of rare/precious metals and the treatment of multi-polluted water.

Stimulation of Lignan Production in Schisandra rubriflora In Vitro Cultures by Elicitation.

The study investigated the effect of elicitation with: chitosan (CH) (200 mg/L), yeast extract (YeE) (3000 mg/L), ethephon (ETH) (25 µM/L), and methyl jasmonate (MeJA) (50 µM/L), on lignan accumulation in agitated and bioreactor (Plantform temporary immersion systems) microshoot cultures of female (F) and male (M) Schisandra rubriflora Rehd. et Wils. (Schisandraceae) lines. The elicitors were supplemented on the 10th day of culture. Biomasses were collected at 24 h and 48 h, and 4, 6, and 8 days after the addition of each elicitor. The 24 compounds from the dibenzocyclooctadiene, aryltetralin, dibenzylbutane, and tetrahydrofuran lignans and neolignans were determined qualitatively and quantitatively in biomass extracts using the UHPLC-MS/MS method. The highest total contents [mg/100 g DW] of lignans were: for CH-95.00 (F, day 6) and 323.30 (M, 48 h); for YeE 104.30 (F, day 8) and 353.17 (M, day 4); for ETH 124.50 (F, 48 h) and 334.90 (M, day 4); and for MeJA 89.70 (F, 48 h) and 368.50 (M, 24 h). In the biomass extracts of M cultures grown in bioreactors, the highest total lignan content was obtained after MeJA elicitation (153.20 mg/100 g DW). The maximum total lignan contents in the biomass extracts from agitated and bioreactor cultures were 3.29 and 1.13 times higher, respectively, than in the extracts from the non-elicited cultures. The poor understanding of the chemical composition and the lack of studies in the field of plant biotechnology of S. rubriflora emphasize the innovativeness of the research.

Antioxidant Potential and Enhancement of Bioactive Metabolite Production in In Vitro Cultures of Scutellaria lateriflora L. by Biotechnological Methods.

Studies carried out using three different in vitro assays and a biological setting (Escherichia coil) demonstrated the antioxidant activity of Scutellaria lateriflora microshoot extract. Moreover, the extract exhibited no toxicity in a brine shrimp lethality bioassay. These results indicated that microshoots are a rich, safe source of antioxidants, which encouraged us to enhance their production in vitro. In agar and agitated cultures, two biotechnological strategies were applied: feeding the cultures with the biogenetic precursors of the phenolics-phenylalanine and tyrosine, and eliciting them with methyl jasmonate. Specific Scutellaria flavonoids and verbascoside were analysed by HPLC. Feeding with precursors (1 g/L) in agar cultures decreased the production of the metabolites. In agitated cultures, different concentrations of precursors (1.0-2.5 g/L) and the elicitor (10; 50; 100 µM) were tested. Additionally, parallel feeding with the precursor and elicitor in a concentration of 50 µM were applied. The best strategy for total flavonoid and verbascoside production was phenylalanine feeding (1.5 g/L), max. 3765 and 475 mg/100 g DW, respectively, after 7 days. This is the first report documenting the high antioxidant production in S. lateriflora microshoots after feeding with phenylalanine. Moreover, for the first time, bioreactor cultures were successfully maintained, obtaining attractive results (max. total flavonoid content 2348 and verbascoside 485 mg/100 g DW).

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.

Shoot and root single cell sequencing reveals tissue- and daytime-specific transcriptome profiles.

Although several large-scale single-cell RNA sequencing (scRNAseq) studies addressing the root of Arabidopsis (Arabidopsis thaliana) have been published, there is still need for a de novo reference map for both root and especially above-ground cell types. As the plants' transcriptome substantially changes throughout the day, shaped by the circadian clock, we performed scRNAseq on both Arabidopsis root and above-ground tissues at defined times of the day. For the root scRNAseq analysis, we used tissue-specific reporter lines grown on plates and harvested at the end of the day (ED). In addition, we submitted above-ground tissues from plants grown on soil at ED and end of the night to scRNAseq, which allowed us to identify common cell types/markers between root and shoot and uncover transcriptome changes to above-ground tissues depending on the time of the day. The dataset was also exploited beyond the traditional scRNAseq analysis to investigate non-annotated and di-cistronic transcripts. We experimentally confirmed the predicted presence of some of these transcripts and also addressed the potential function of a previously unidentified marker gene for dividing cells. In summary, this work provides insights into the spatial control of gene expression from nearly 70,000 cells of Arabidopsis for below- and whole above-ground tissue at single-cell resolution at defined time points.

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.

Arbuscular mycorrhizal symbioses alleviating salt stress in maize is associated with a decline in root-to-leaf gradient of Na+/K+ ratio.

BACKGROUND: Inoculation of arbuscular mycorrhizal (AM) fungi has the potential to alleviate salt stress in host plants through the mitigation of ionic imbalance. However, inoculation effects vary, and the underlying mechanisms remain unclear. Two maize genotypes (JD52, salt-tolerant with large root system, and FSY1, salt-sensitive with small root system) inoculated with or without AM fungus Funneliformis mosseae were grown in pots containing soil amended with 0 or 100 mM NaCl (incrementally added 32 days after sowing, DAS) in a greenhouse. Plants were assessed 59 DAS for plant growth, tissue Na+ and K+ contents, the expression of plant transporter genes responsible for Na+ and/or K+ uptake, translocation or compartmentation, and chloroplast ultrastructure alterations. RESULTS: Under 100 mM NaCl, AM plants of both genotypes grew better with denser root systems than non-AM plants. Relative to non-AM plants, the accumulation of Na+ and K+ was decreased in AM plant shoots but increased in AM roots with a decrease in the shoot: root Na+ ratio particularly in FSY1, accompanied by differential regulation of ion transporter genes (i.e., ZmSOS1, ZmHKT1, and ZmNHX). This induced a relatively higher Na+ efflux (recirculating) rate than K+ in AM shoots while the converse outcoming (higher Na+ influx rate than K+) in AM roots. The higher K+: Na+ ratio in AM shoots contributed to the maintenance of structural and functional integrity of chloroplasts in mesophyll cells. CONCLUSION: AM symbiosis improved maize salt tolerance by accelerating Na+ shoot-to-root translocation rate and mediating Na+/K+ distribution between shoots and roots.

Biocatalytic Production of Aldehydes: Exploring the Potential of Lathyrus cicera Amine Oxidase.

Aldehydes are a class of carbonyl compounds widely used as intermediates in the pharmaceutical, cosmetic and food industries. To date, there are few fully enzymatic methods for synthesizing these highly reactive chemicals. In the present work, we explore the biocatalytic potential of an amino oxidase extracted from the etiolated shoots of Lathyrus cicera for the synthesis of value-added aldehydes, starting from the corresponding primary amines. In this frame, we have developed a completely chromatography-free purification protocol based on crossflow ultrafiltration, which makes the production of this enzyme easily scalable. Furthermore, we determined the kinetic parameters of the amine oxidase toward 20 differently substituted aliphatic and aromatic primary amines, and we developed a biocatalytic process for their conversion into the corresponding aldehydes. The reaction occurs in aqueous media at neutral pH in the presence of catalase, which removes the hydrogen peroxide produced during the reaction itself, contributing to the recycling of oxygen. A high conversion (>95%) was achieved within 3 h for all the tested compounds.

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.

Effect of Elicitation with (+)-Usnic Acid on Accumulation of Phenolic Acids and Flavonoids in Agitated Microshoots of Eryngium alpinum L.

The present work was aimed at studying the potential of elicitation on the accumulation of phenolic compounds in in vitro shoot cultures of Eryngium alpinum L., a protected plant from the Apiaceae family. The study examined the influence of (+)-usnic acid on the biomass growth as well as on the biosynthesis of the desired flavonoids and phenolic acids in the cultured microshoots. The phenolic compound content was determined by HPLC-DAD. The flavonoid of the highest concentration was isoquercetin, and the phenolic acids of the highest amount were rosmarinic acid, caffeic acid and 3,4-dihydroxyphenylacetic acid, both in the non-elicited and elicited biomass. Isoquercetin accumulation was efficiently increased by a longer elicitation with a lower concentration of lichenic compound (107.17 ± 4.67 mg/100 g DW) or a shorter elicitation with a higher concentration of acid (127.54 ± 11.34 and 108.37 ± 12.1 mg/100 g DW). Rosmarinic acid production generally remained high in all elicited and non-elicited microshoots. The highest content of this acid was recorded at 24 h of elicitation with 3.125 µM usnic acid (512.69 ± 4.89 mg/100 g DW). The process of elicitation with (+)-usnic acid, a well-known lichenic compound with allelopathic nature, may therefore be an effective technique of enhancing phenolic compound accumulation in alpine eryngo microshoot biomass.