Chemical profile of Juniperus excelsa M. Bieb. essential oil within and between populations and its weed seed suppression effect.

The aims of this study were to (1) establish the intrapopulation and seasonal variation of Juniperus excelsа essential oil (EO); (2) compare the J. excelsa concrete and resinoid composition with its EO composition; and (3) investigate the potential herbicidal activity of J. excelsa EO against seeds of Papaver rhoeas L., Consolida orientalis (J.Gay) Schrödinger, Anthemis arvensis L., Avena fatua L., and Agrostemma githago L. Four independent studies were performed to meet these objectives. Twenty-eight individual trees were analyzed from two populations to establish intrapopulation and interpopulation variability of EOs yield and composition. In the seasonal dynamic study of leaf EO, samples from the same three trees and in the same population were collected in January, March, May, July, October, and December and their EO yield and composition determined. The EOs (intrapopulation and seasonal) were extracted by hydrodistillation, while the EO for the herbicidal test was obtained by steam distillation in a semi-commercial (SCom) apparatus. Overall, the EO yield varied significantly from 0.93% to 2.57%. α-Pinene (8.85-35.94%), limonene (11.81-50.08%), and cedrol (3.41-34.29%) were the predominant EO compounds in all samples (intrapopulation variability); however, trans-2,4-decadienol and ß-caryophyllene were predominant in some individual trees. Four chemical groups were identified in the samples collected from two natural populations (intrapopulation). This is the first report on the compositions of J. excelsa concrete and resinoid. Cedrol (15.39%), 7-hydroxy-4-methyl-coumarin (17.63%), 1-octacosanol (36.85%), tritriacontane (16.08%), and tiacontanoic acid were the main compounds in the concrete and resinoid. Juniperus excelsa EO suppressed seed germination and seedling growth of P. rhoeas, C. orientalis, A. arvensis, A. fatua, and A. githago, demonstrating its potential to be used for the development of new biopesticides. The highest EO yield with high content of limonene and cedrol was obtained from samples harvested during the winter months (December, January, and March).

Plant In Vitro Culture Factories for Pentacyclic Triterpenoid Production.

Pentacyclic triterpenoids are a diverse subclass of naturally occurring terpenes with various biological activities and applications. These compounds are broadly distributed in natural plant resources, but their low abundance and the slow growth cycle of plants pose challenges to their extraction and production. The biosynthesis of pentacyclic triterpenoids occurs through two main pathways, the mevalonic acid (MVA) pathway and the 2-C-methyl-D-erythritol-4-phosphate (MEP) pathway, which involve several enzymes and modifications. Plant in vitro cultures, including elicited and hairy root cultures, have emerged as an effective and sustainable system for pentacyclic triterpenoid production, circumventing the limitations associated with natural plant resources. Bioreactor systems and controlling key parameters, such as media composition, temperature, light quality, and elicitor treatments, have been optimized to enhance the production and characterization of specific pentacyclic triterpenoids. These systems offer a promising bioprocessing tool for producing pentacyclic triterpenoids characterized by a low carbon footprint and a sustainable source of these compounds for various industrial applications.

Metabolomic Profile of Arthrospira platensis from a Bulgarian Bioreactor-A Potential Opportunity for Inclusion in Dietary Supplements.

The present study aims to elucidate the metabolomic profile of Arthrospira platensis grown in a bioreactor in Bulgaria. The results show that Arthrospira platensis has a high content of mannose, 137.02 mg g-1, and vitamin A (retinol)-10.3 µg/100 g. High concentrations of calcium, sulfur, and zinc distinguish its elemental composition. The freeze-dried powder contained 15.81 ± 0.45% dietary fiber, 50.16 ± 0.25% total protein content, and 1.22 ± 0.11% total fat content. Among the unsaturated fatty acids with the highest content is α-linolenic acid (25.28%), while among the saturated fatty acids, palmitic acid prevails (22.55%). Of the sterols in the sample, ß-sitosterol predominated. There is no presence of microcystins LR, RR, YR, and nodularin. Therefore, Arthrospira platensis grown in a Bulgarian bioreactor is safe for use in the pharmaceutical and food industries. Many of the organic compounds found have applications in medicine and pharmacology and play an important role in biochemical processes in the body. Therefore, Arthrospira platensis grown in Bulgaria has a high potential for use as an independent food supplement or in combination with other natural products.

Phytochemical and biological investigations on Centranthus kellereri (Stoj., Stef. & T. Georgiev) Stoj. & Stef. and C. ruber (L.) DC. and their potential as new medicinal and ornamental plants.

INTRODUCTION: Centranthus kellereri is a Bulgarian endemic plant species, found only in two locations in the world: The Balkans Mountains (Stara Planina), above the town of Vratsa, and The Pirin Mountains, above the town of Bansko, Bulgaria. Being endemic and endangered species precluded any significant research on it. The hypothesis of this study was that the populations of C. kellereri may represent genetically, phytochemically, and morphologically distinct forms and these will differentiate from C. ruber. Furthermore, C. kellereri possibly imperfect embryology may preclude its more widespread distribution under natural conditions. RESULTS: This study revealed the phytochemical profile, antioxidant activity, embryology, surface microstructural morphological traits, and genetic differences between the C. kellereri plants from the only two natural populations and compares them to the ones of the related and better-known plant C. ruber. The essential oil (EO) content in aboveground plant parts and in roots was generally low and the EO composition varied significantly as a function of plant part, year of sampling, location, and species. Methylvaleric acid was a major EO constituent in the C. kellereri EO, ranging between 60.2% and 71.7% of the total EO. The EO included monoterpenes, sequiterpenes, long-chain alkanes and fatty acids. Phytochemical analyses of plant tissue revealed the occurrence of 32 compounds that were tentatively identified as 6 simple phenolics, 18 flavonoids, 1 quinone, 1 lipid, 1 alkaloid, 2 diterpenes, and 3 triterpenes. There were differences in detected compounds between the C. kellereri plants at the two locations and between the roots and shoots in both species. The total phenols and flavonoids varied between the two species but were also dissimilar between the plants from the two populations of C. kelleri. Free radical scavenging activity, measured with ABTS and DPPH in aqueous and methanol extracts, had similar values; however, overall, C. kellereri from Vratsa showed the highest antioxidant activity while C. ruber had the lowest activity. Genetic analyses showed a clear differentiation between C. kellereri and C. ruber, and between the two populations of C. kellereri. Embryological studies revealed the peculiarities of the male and female generative spheres of the two species that were defined as being sexually reproducing. The pollen had high viability; however, the low viability of seeds demonstrated possible high sensitivity of C. kellereri to the environmental conditions, perhaps the main factor modifying and restricting the population sizes. The SEM analyses exposed differences in surface microstructural traits between the species (C. kellereri and C. ruber) but also between the two populations of C. kellereri. The observed dissimilarities in genetic makeup, micromorphological characteristics, and phytochemical composition strongly indicate that the two populations can be classified as distinct subspecies or varieties of C. kellereri; var. pirinensis and var. balkanensis. Further research is needed to introduce C. kellereri into culture and develop it as a high-value specialty crop or ornamental in order to conserve C. kellereri natural populations. C. kellereri may be utilized as a source for phytochemicals of interest and as an ornamental plant like C. ruber; however, it may have a greater environmental plasticity and adaptation as evidenced by its current locations.

Transcriptional and Metabolic Profiling of Arabidopsis thaliana Transgenic Plants Expressing Histone Acetyltransferase HAC1 upon the Application of Abiotic Stress-Salt and Low Temperature.

Augmented knowledge of plant responses upon application of stress could help improve our understanding of plant tolerance under abiotic stress conditions. Histone acetylation plays an important role in gene expression regulation during plant growth and development and in the response of plants to abiotic stress. The current study examines the level of transcripts and free metabolite content in transgenic Arabidopsis thaliana plants expressing a gene encoding histone acetyltransferase from Medicago truncatula (MtHAC1) after its heterologous expression. Stable transgenic plants with HAC1 gain and loss of function were constructed, and their T5 generation was used. Transgenic lines with HAC1-modified expression showed a deviation in root growth dynamics and leaf area compared to the wild-type control. Transcriptional profiles were evaluated after the application of salinity stress caused by 150 mM NaCl at four different time points (0, 24, 48, and 72 h) in treated and non-treated transgenic and control plants. The content and quantity of free metabolites-amino acids, mono- and dicarbohydrates, organic acids, and fatty acids-were assessed at time points 0 h and 72 h in treated and non-treated transgenic and control plants. The obtained transcript profiles of HAC1 in transgenic plants with modified expression and control were assessed after application of cold stress (low temperature, 4 °C).

Chemical Composition of Essential Oils from Nepeta transcaucasica Grossh. and Nepeta cataria L. Cultivated in Bulgaria and Their Antimicrobial and Antioxidant Activity.

The genus Nepeta, belonging to the family Lamiaceae, includes about 300 species, most of which are used in folk medicine due to their pronounced biological properties. The aim of the present study was to evaluate the agrobiological characteristics of Nepeta transcaucasica (N. transcaucasica) Grossh. and Nepeta cataria (N. cataria) L., cultivated in Bulgaria, and obtain their essential oils and determine their antimicrobial and antioxidant activities. The agrobiological characteristics of the two species growing in Kazanlak were analyzed; therefore, high variability in the population of N. transcaucasica and comparative homogeneity in N. cataria was shown. The species N. transcaucasica contained 0.28% essential oil with main components ß-citronellol (52.05%), eucalyptol (7.34%), ß-citronellal (6.06%), germacrene D (5.45%), (Z)-ß-ocimene (5.14%), and ß-caryophyllene (3.06%). The species N. cataria consisted of 0.19% essential oil with main components ß-citronellol (26.31%), geraniol (15.92%), neral (11.45%), nerol (9.56%), carvacrol (6.04%), and ß-citronellal (5.35%). The antibacterial activity against Gram-positive bacteria Listeria monocytogenes and Staphylococcus aureus and Gram-negative bacteria Escherichia coli (E. coli) and Salmonella enterica subsp. enterica serovar Abony was determined. The essential oils showed antimicrobial activity only against E. coli. The diameters of the inhibition zones were found to be 26 mm for the species N. transcaucasica and 10 mm for the species N. cataria. The antioxidant activity of the two essential oils was also determined by four different methods, DPPH, ABTS, FRAP, and CUPRAC, with the highest values for the ABTS radical, for the species N. transcaucasica (48.72 µM TE/mL), and the species N. cataria (310 µM TE/mL).

Sambucus ebulus (Elderberry) Fruits Modulate Inflammation and Complement System Activity in Humans.

Sambucus ebulus (SE) fruits are used for immune stimulation and amelioration of gastrointestinal inflammatory conditions. Currently, there is no scientific evidence of their effects on various aspects of the immune response mechanisms in humans. The purpose of this study was to evaluate the immunomodulatory potential of SE fruit infusion intake in healthy humans. Anthocyanin content was determined with UPLC-ESI-MS/MS. Fifty-three volunteers enrolled in a 4-week SE infusion intake intervention. Blood count, serum total protein, Interleukin 1 beta (IL-1ß), Interleukin 6 (IL-6), Tumor Necrosis Factor Alpha (TNFα), High-sensitivity C-reactive protein (hs-CRP), C3, and C4 levels were measured on automatic analyzers, and Interleukin 8 (IL-8) was measured manually with an ELISA kit. Cyanidin-3-O-galactoside (48.15 mg/g DW), followed by cyaniding-3-sambubioside (43.41 ± 1.07 mg/g DW), were the most abundant anthocyanins in SE samples. A significant decrease in total protein (2.82%), IL-6 (20.15%), TNFα (5.38%), IL-8 (5.50%), C3 (4.16%), and C4 (14.29%) was established in the whole group. Total protein, IL-8, TNFα, and C4 decreased in women (3.11%, 4.76%, 5.09%, and 11.11%), and IL-6 decreased (40.61%) in men. Hb (1.20%) and hematocrit (1.55%) levels decreased in the whole group and in the women group (1.61% and 2.20%). SE fruits exert immune-modulatory activity as revealed by decreased pro-inflammatory status and complement activity markers in healthy volunteers after a 4-week intervention.

Essential Oil Composition of Seven Bulgarian Hypericum Species and Its Potential as a Biopesticide.

Hypericum species and especially H. perforatum L. are well known for their therapeutic applications. The present study assessed the essential oil (EO) composition, and antifungal and aphid suppression activity of seven Bulgarian Hypericum species. The EOs were analyzed by GC-MS-FID. Two experiments were conducted. In the first experiment, H. perforatum, H. maculatum, and H. hirsutum were used. Additionally, the EO composition of H. perforatum extracted via hydrodistillation (ClevA) and via commercial steam distillation (Com) were compared. The second experiment compared the EOs of H. perforatum, H. cerastoides, H. rumeliacum, H. montbretii, and H. calycinum (flowers and leaves) extracted via hydrodistillation and collected with n-hexane. Overall, the EO constituents belonged to four classes, namely alkanes, monoterpenes, sesquiterpenes, and fatty acids. The main class for compounds in H. maculatum and H. perforatum (section Hypericum) were sesquiterpenes for both experiments except for H. perforatum (Com). Hypericum montbretii (section Drosocarpium) EO had monoterpenes (38.09%) and sesquiterpenes (37.09%) as major groups, while H. hirsutum EO (section Taeniocarpium) contained predominately alkanes (67.19%). Hypericum hirsutum EO contained cedrol (5.04%), found for the first time in Hypericum species. Fatty acids were the main compounds in H. cerastoides (section Campylopus), while monoterpenes were the most abundant class in H. rumeliacum and H. calycinum EOs. α-Pinene and germacrene D were the major EO constituents of all analyzed Hypericum species except for H. hirsutum and H. cerastoides. Hypericum perforatum EO (Com) had significant repellent and insecticidal activity against two aphid species, Rhopalosiphum padi (Bird Cherry-oat aphid) and Sitobion avenae (English grain aphid) at concentrations of 0%, 1%, 2.5%, 3.5%, 4.5%, and 5%. The tested EOs did not show significant activity against selected economically important agricultural fungal pathogens Fusarium spp., Botrytis cinerea, Colletotrichum spp., Rhizoctonia solani, and Aspergillus sp. The EO of the Hypericum species found in the Bulgarian flora could be utilized for the development of new biopesticides for aphid control.

Valorization of Peels of Eight Peach Varieties: GC-MS Profile, Free and Bound Phenolics and Corresponding Biological Activities.

Sustainability, becoming essential for food processing and technology, sets goals for the characterization of resources considered as food waste. In this work, information about the GC-MS metabolites of peach peels was provided as a tool that can shed more light on the studied biological activities. In addition, distribution patterns and contribution of the chemical profile and free and bound phenolic compounds as antioxidant, antimicrobial, and enzymatic clusters in peach peels of different varieties of Bulgarian origin were studied. The two applied techniques (alkaline and acid hydrolysis) for releasing the bound phenolics reveal that alkaline hydrolysis is a better extraction approach. Still, the results indicate the prevalence of the free phenolics in the studied peach peel varieties. Total phenolics of peach wastes were positively correlated with their antioxidant activity. The antioxidant activity results certainly defined the need of an individual interpretation for each variety, but the free phenolics fractions could be outlined with the strongest potential. The limited ability of the peels' extracts to inhibit α-amylase and acetylcholinesterase, and the moderate antimicrobial activity, on the other hand, indicate that the potential of peach peels is still sufficient to seek ways to valorize this waste. Indeed, this new information about peach peels can be used to characterize peach fruits from different countries and/or different food processes, as well as to promote the use of this fruit waste in food preparation.

New Insights into the Research of Bioactive Compounds from Plant Origins with Nutraceutical and Pharmaceutical Potential.

Plant bioactive compounds are essential for human health due to their multiple biological effects, such as antioxidant, anticarcinogenic, antiallergenic, anti-inflammatory, antimutagenic, and antimicrobial activities, which can have beneficial effects on various noncommunicable diseases, such as autoimmune, inflammatory, cardiovascular, cancer, metabolic, and neurodegenerative diseases [...].

Pattern Recognition of Varieties of Peach Fruit and Pulp from Their Volatile Components and Metabolic Profile Using HS-SPME-GC/MS Combined with Multivariable Statistical Analysis.

A fruit's aroma profile, composed of a complex mixture of volatile organic compounds, is among the core attributes related to the overall taste and consumer preference. Prunus persica L. is a preferred summer fruit with a distinct, favorable olfactory characteristic. The volatile compositions of both peach fruits and fruit pulps from eight peach cultivars (four native and four introduced) was investigated to compare their composition and assess flavor-contributing compounds. In total, 65 compounds were profiled after a HS-SPME-GC-MS analysis: 16 esters, 14 aldehydes, 5 alcohols, 7 hydrocarbons, 7 ketones, 8 acids, and 8 terpenes. The most common compounds were esters, acids, and aldehydes. Although the same compounds were identified in both fruit and pulp, their %TIC (total ion current) differed in favor of the whole fruit. Following the metabolic profiling of the whole fruit and fruit pulp, a total of 44 compounds were identified from the studied varieties. Among them, amino acids, organic acids, sugar alcohols, saccharides, fatty acids, and phenolic acids were identified as existing groups. According to the provided principal component analysis (PCA) and hierarchical cluster analysis (HCA), the relative %TIC of the identified volatile compounds fluctuated depending on the studied cultivar. No differences were visible in the PCA biplots, which suggested that the polar and lipid metabolites do not provide significant variations when considering different parts of the fruit, contrary to the volatile compounds. The obtained results could successfully be applied in the metabolic chemotaxonomy of peaches and the differentiation of the metabolites present in different parts of the peach.

Comparative Phytochemical Analysis of Aronia melanocarpa L. Fruit Juices on Bulgarian Market.

Aronia melanocarpa L. (black chokeberry), belonging to the Rosaceae family, contains high amounts of polyphenolics and therefore exhibits one of the highest antioxidant and anti-inflammatory activities among berry fruits. Chokeberries are used in the food industry for juice, nectar, and wine production and as colorants. We aimed to compare the phytochemical composition of three chokeberry juices commercially available in the local market as sources of beneficial phytochemicals. Using GC-MS and LC-MS/MS, we performed the identification and quantitation of polar compounds and polyphenolics. The concentrations of 13 amino acids, including 6 essential amino acids, 10 organic acids, 20 sugar alcohols and derivatives, 14 saccharides, 12 fatty acids and esters, and 38 polyphenols, were estimated. One of the analyzed juices had the highest polyphenolic content (5273.87 ± 63.16 µg/mL), possibly due to 2.9 times higher anthocyanin concentration compared to anthocyanins in other tested juices. This study provides new data concerning phytochemical composition in terms of amino acids, organic acids, sugar acids, fatty acids and their esters, and polyphenols as phytocomponents of commercially available chokeberry juices. Results show that after all processing techniques and possibly different plant growth conditions, chokeberry juices are a valuable source of health-promoting phytochemicals such as phenolic acids, pro-anthocyanins, and anthocyanins, thus considering them as functional foods. We demonstrated a diversity of the active substances in bioactive foods marketed as "same"; therefore, the standardized therapeutic effect could be expected only by the utilization of food supplements with guaranteed constant content.

Metabolic Profiling of Bulgarian Potato Cultivars.

Potatoes (Solanum tuberosum L.) are the fourth most economically important crop in the world. They have a short period of vegetation and are an excellent source of carbohydrates, amino acids, vitamins, organic acids, minerals and phenolics as antioxidant substances. Potato can be a major dietary source of various bioactive compounds. In this study, we applied gas chromatography coupled with mass spectrometry (GC-MS) metabolite profiling to classify eight Bulgarian potato cultivars bred in the Maritsa Vegetable Crops Research Institute (VCRI), Plovdiv, according to their metabolite contents. Altogether, we determine their flavonoids/phenolics to evaluate their nutritive quality for the breeding program with the target of determining strong health-promoting compounds. The "Kalina" cultivar is highlighted as the best one with the highest number of metabolites, containing 14 out of the 26 evaluated; it was selected as the highest-quality cultivar, compared with the other seven cultivars studied. According to the grouping of the cultivars in principal component analysis PCA, their positive distribution is explained mainly by them having the highest contents of aminobutyric and isocitric acids, methionine and alanine and lower levels of fumaric acid, pyroglutamic acid and glycine, in contrast to the cultivars distributed negatively, which had high contents of carbohydrates and relatively low contents of most of the amino acids. The highest number of amino acids was found in the cultivar "Kalina", followed by "Perun" and "Bor". The highest number of carbohydrates was found in "Pavelsko" and "Iverce", while the prominent accumulation of organic acids was found in "Kalina", "Bor" and "Rozhen". The highest number of flavonoids in the flesh of the tubers was found in the cultivars "Nadezhda" and "Pavelsko", followed by "Bor". The highest ratio of flavonoids/phenolics in the flesh was found in "Pavelsko" and in "Nadezhda", followed by "Iverce".

Chemical Profile and Antimicrobial Activity of the Essential Oils of Helichrysum arenarium (L.) Moench. and Helichrysum italicum (Roth.) G. Don.

This study compared the essential oils (EO) composition of Helichrysum arenarium (Bulgarian populations) with that of the cultivated species H. italicum. The EO composition of H. arenarium and H. italicum were analyzed via gas chromatography. In general, 75 components were identified in H. arenarium EO and 79 in H. italicum EO. The predominant constituents in H. arenarium EO were α-pinene (34.64-44.35%) and sabinene (10.63-11.1%), which affirmed the examined population as a new chemical type. Overall, the main EO constituents of H. italicum originating in France, Bosnia and Corsica were neryl acetate (4.04-14.87%) and ß-himachalene (9.9-10.99%). However, the EOs profile of H. italicum introduced from the above three countries differed to some extent. D-limonene (5.23%), italicene, α-guaiene and neryl acetate (14.87%) predominated in the H. italicum introduced from France, while α-pinene (13.74%), δ-cadinene (5.51%), α-cadinene (3.3%), ß-caryophyllene (3.65%) and α-calacorene (1.63%) predominated in plants introduced from Bosnia. The EOs of the plants introduced from France and Corsica had similar chemical composition and antimicrobiological activity.

Wheat and Barley Grass Juice Addition to a Plant-Based Feed Improved Growth and Flesh Quality of Common Carp (Cyprinus carpio).

Phytogenics are plant extract additives used for their bioactive properties. The objective of this study was to assess the effect of plant extracts, wheat grass juice (WGJ) and barley grass juices (BGJ) addition to fish diet on growth and meat quality of common carp. Fish (51 ± 33 g initial weight) were fed for four weeks with three plant-based diets: (1) control feed (Con), (2) control feed supplemented with 2% WGJ (Con+WGJ), and (3) control feed supplemented with 2% BGJ (Con+BGJ). The results showed that the inclusion of the two juices in the plant-based feed stimulated the growth and improved meat quality by lowering the fat and ash content. Feed conversion ratio and condition factor were not affected. There were no significant differences in Fe and Zn contents of meat; however, Cu decreased, while Mn was lower in the Con+WGJ group and higher in the Con+BGJ group compared to Con. A high content of unsaturated fatty acids (FA, oleic acid and linoleic acid) and desirable ratios of saturated/unsaturated FA (0.27-0.29) and Ω6/Ω3 (2.5-2.78) were found in all groups. The juices were characterized in terms of lipid profile and polar compounds by GC-MS technique. The observed positive effects can be attributed to the rich composition of juices that included unsaturated FA, amino acids, sugars and organic compounds.

HS-SPME-GC-MS Volatile Profile Characterization of Peach (Prunus persica L. Batsch) Varieties Grown in the Eastern Balkan Peninsula.

The volatile compounds of eight peach varieties (Prunus persica L.)-"Filina", "Gergana", "Ufo-4", "July lady", "Laskava", "Flat Queen", "Evmolpiya", and "Morsiani 90"-growing in Bulgaria were analyzed for the first time. Gas chromatography-mass spectrometry (GC-MS) analysis and the HS-SPME technique revealed the presence of 65 volatile compounds; the main identified components were aldehydes, esters, and fatty acids. According to the provided principal component analysis (PCA) and hierarchical cluster analysis (HCA), the relative quantities of the identified volatile compounds depended on the studied peach variety. The results obtained could be successfully applied for the metabolic chemotaxonomy of peaches.

A common F-box gene regulates the leucine homeostasis of Medicago truncatula and Arabidopsis thaliana.

The F-box domain is a conserved structural protein motif that most frequently interacts with the SKP1 protein, the core of the SCFs (SKP1-CULLIN-F-box protein ligase) E3 ubiquitin protein ligases. As part of the SCF complexes, the various F-box proteins recruit substrates for degradation through ubiquitination. In this study, we functionally characterized an F-box gene (MtF-box) identified earlier in a population of Tnt1 retrotransposon-tagged mutants of Medicago truncatula and its Arabidopsis thaliana homolog (AtF-box) using gain- and loss-of-function plants. We highlighted the importance of MtF-box in leaf development of M. truncatula. Protein-protein interaction analyses revealed the 2-isopropylmalate synthase (IPMS) protein as a common interactor partner of MtF-box and AtF-box, being a key enzyme in the biosynthesis pathway of the branched-chain amino acid leucine. For further detailed analysis, we focused on AtF-box and its role during the cell division cycle. Based on this work, we suggest a mechanism for the role of the studied F-box gene in regulation of leucine homeostasis, which is important for growth.

High resolution LC-MS/MS characterization of polyphenolic composition and evaluation of antioxidant activity of Sambucus ebulus fruit tea traditionally used in Bulgaria as a functional food.

The purpose of this study was to perform phytochemical analysis of tea from Sambucus ebulus fruits concerning hydroxycinnamic acids, flavonol glucosides, stilbenes and proanthocyanidin mono-, di- and trimers content. In total, 33 compounds were identified and quantified using UPLC-DAD-ESI/MS/MS system and the results are presented in mg/g dry weight (DW). Among analyzed hydroxycinnamic acids, 5-Caffeoylquinic acid (114.17 mg/g) was most abundant, followed by 3-p-Coumaroylquinic acid (50.33 mg/g) and 3-p-Feruloylquinic acid, p-Coumaric acid glucoside and 4-p-Coumaroylquinic acid (31.36 mg/g, 29.78 mg/g and 27.70 mg/g, respectively). Flavonol glucosides were represented predominantly by Quercetin-3-O-galactoside, Quercetin-3-O-rhamnosyl-galactoside Quercetin-3-O-glucoside and Quercetin-3-O-rhamnosyl-glucoside (3.68 mg/g, 3.22 mg/g, 2.87 mg/g and 2.56 mg/g, respectively). trans-Resveratrol-3-O-glucoside, epicatechin (40.62 mg/g) and proanthocyanidin di- and -trimers (19.90 mg/g - 31.42 mg/g) also were present in the tea. ABTS cation decolorization assay revealed 1.248 mM UAE activity and the percent of DPPH radical scavenging was 14.25%, corresponding to 39.07 µM Trolox equivalents.

Biological Activity of Essential Oils of Four Juniper Species and Their Potential as Biopesticides.

The objective of this study was to assess the biological activity of essential oils (EOs) of four Juniperus species obtained via two different distillation methods and their potential as biopesticides. The studied factors were juniper species (Juniperus communis L., J. oxycedrus L., J. pygmaea C. Koch., and J. sibirica Burgsd), plant sex (male (M) and female (F)), and distillation method (hydrodistillation via a standard Clevenger apparatus (ClevA) and semi-commercial (SCom) steam distillation). The hypothesis was that the EO will have differential antioxidant, antimicrobial, and insecticidal activities as a function of plant species, plant sex, and distillation method. The two distillation methods resulted in similar EO composition within a given species. However, there were differences in the EO content (yield) due to the sex of the plant, and also differences in the proportions of some EO components. The concentration of α-pinene, ß-caryophyllene, δ-cadinene and δ-cadinol was dissimilar between the EO of M and F plants within all four species. Additionally, M and F plants of J. pygmaea, and J. sibirica had significantly different concentrations of sabinene within the respective species. The EOs obtained via ClevA extraction showed higher antioxidant capacity within a species compared with those from SCom extraction. All of the tested EOs had significant repellent and insecticidal activity against the two aphid species Rhopalosiphum padi (bird cherry-oat aphid) and Sitobion avenae (English grain aphid) at concentrations of the EO in the solution of 1%, 2.5%, and 5%. The tested EOs demonstrated moderate activity against selected pathogens Fusarium spp., Botrytis cinerea, Colletotrichum spp., Rhizoctonia solani and Cylindrocarpon pauciseptatum. The results demonstrate that the standard ClevA would provide comparable EO content and composition in comparison with SCom steam distillation; however, even slight differences in the EO composition may translate into differential bioactivity.

Phytochemical Composition, Anti-Inflammatory and ER Stress-Reducing Potential of Sambucus ebulus L. Fruit Extract.

Sambucus ebulus L. (SE) fruits are used for their immunostimulation, hematopoietic and antiviral potential. Recently, we focused on analyzing the mechanism underlying SE fruit aqueous extract's (FAE) immunomodulation and anti-inflammatory activities, with attention to its endoplasmic reticulum (ER) stress-reducing potential. J774A.1 macrophages were treated with SE FAE alone or in conditions of lipopolysaccharides (LPS) stimulation. Using GC-MS and LC-MS/MS, its phytochemical composition was analyzed. To measure transcription and protein levels, we used qPCR and Western blot, respectively. The prevailing phytochemicals in SE FAE were hydroxycinnamic acids, proanthocyanidins and anthocyanins. The content of some amino acids, organic acids, alcohols, fatty acids and esters were newly reported. Extracts exerted an immunostimulation potential by stimulating IL-6, TNFα, Ccl2, COX2 and iNOS transcription, without inducing ER stress. SE FAE suppressed the LPS-induced transcription of inflammation related genes (IL-1ß, IL-6, TNFα, Ccl2, Icam-1, Fabp4, COX2, iNOS, Noxo1, IL-1ra, Sirt-1) and reduced the protein levels of iNOS, peIF2α, ATF6α and CHOP. The effects were comparable to that of salicylic acid. SE suppresses LPS-stimulated inflammatory markers on the transcription and translation levels. Targeting ER stress is possibly another mechanism underlying its anti-inflammatory potential. These findings reveal the potential of SE fruits as a beneficial therapeutic of inflammation and ER stress-related pathological conditions.