Intraspecific Variability of Stinging Nettle (Urtica dioica L.).

The purpose of the work was to determine the intraspecific variability of the stinging nettle, in respect of the mass of leaves and their chemical composition, including the content of phenolic compounds and assimilative pigments. The objects of the study were 10 populations of nettle, originating from the eastern and southern part of Poland. The results obtained indicate a high level of variability between and within the populations investigated but not strictly related to their geographical locations. The mass of the leaves ranged from 0.19 to 0.28 kg dry weight (DW)/plant (Coefficient of variation (CV) = 16.33%). Using HPLC-DAD, four phenolic acids were detected, i.e., caffeoylmalic (570.97-1367.40 mg/100 g DW), chlorogenic (352.79-1070.83 mg/100 g DW), neochlorogenic (114.56-284.77 mg/100 g DW) and cichoric (58.31-189.52 mg/100 g DW) acids, with the last one differentiating populations to the highest degree (CV = 48.83%). All of the analyzed populations met the requirements of the European Pharmacopoeia (Ph Eur 10th) concerning the minimum content of caffeoylmalic and chlorogenic acids in nettle leaves (not less than 0.3%). Within the flavonoid fraction, two compounds were identified, namely rutoside (917.05-1937.43 mg/100 g DW, CV = 21.32%) and hyperoside (42.01-289.45 mg/100 g DW; CV = 55.26%). The level of chlorophyll a ranged from 3.82 to 4.49 mg/g DW, chlorophyll b from 1.59 to 2.19 mg/g DW, while the content of carotenoids varied from 2.34 to 2.60 mg/100 g DW. Given all the traits investigated, the level of a population's polymorphism (CV) was visibly higher within a population than between populations. Population no. 4 was distinguished by the highest mass of leaves, and the highest content of rutoside, while population no. 2 was distinguished by the highest content of hyperoside, caffeoylmalic and chlorogenic acid.

Chemical Diversity of Wild-Growing and Cultivated Common Valerian (Valeriana officinalis L. s.l.) Originating from Poland.

Common valerian is a medicinal plant. The underground organs of this species are used as a mild sedative and sleeping aid. Poland is one of the largest producers of this raw material in Europe, with local cultivar 'Lubelski' as a primary cultivated form. Although valerian is the subject of more or less deliberate selection carried out by farmers, it is still genetically unstable. The aim of this study was to determine the diversity of the 'Lubelski' cultivar originating from four regions of Poland (forms: L1-L4) in relation to wild-growing populations of the species. The plants were assessed in terms of the mass of underground organs and the content of valerenic acids and essential oils (EOs). The content of valerenic acids was determined using HPLC, whereas the content of EOs was determined using hydrodistillation. The composition of EOs was assessed using GC-MS GC-FID. The ploidy level of the analyzed objects was determined as well. Wild-growing populations (diploids) were characterized by lower masses of underground organs and lower contents of valerenic acid than cultivated forms (tetraploids). However, they produced higher contents of EOs. All the cultivated forms were strongly diversified with respect to the analyzed traits, including the mass of the roots (CV 49-75%), the content of valerenic acids (CV 18-55%), and the content of EOs (CV 28-57%). A total of 44 compounds were identified in the EOs. The dominant compound of both wild-growing populations and the 'Lubelski' forms were: α-fenchene, bornyl acetate, and valerenal. Among 'Lubelski' forms, the most interesting seems to be the L2 form, which was characterized by a relatively high yield and high content of valerenic acids and EOs. Thus, it appears to be a promising source of objects for further valerian cultivar improvement.

Intraspecific Variability of Wild-Growing Common Valerian (Valeriana officinalis L.).

Common valerian (Valeriana officinalis L.) is an important medicinal plant revealing sedative, hypotensive, anti-spasmodic and anxiolytic activity. The purpose of the study was to determine the intraspecific variability of the common valerian growing wild in Poland and the 'Lubelski' landrace, as to their developmental traits, chemical composition and selected genetic parameters. Both wild-growing populations (19) and the landrace (1) were evaluated under ex situ conditions. Observations of the underground organs parameters, both developmental and chemical (according to the European Pharmacopoeia) were carried out in the first year of the plant's development, while the characteristics of the aboveground organs, followed by the sowing value of seeds (according to the International Seed Testing Association)-in the second year. The genetic analyses were performed using the NGS-DArT-seq method. Results indicate the presence of five different gene pools covering the regions of population's origin, with a gene flow within and between them. A high level of developmental and chemical variabilities among the wild-growing populations was noticed, however without a clear relation to the region of the origin. The mass of underground organs ranged from 107.4 to 403.6 g FW × plant-1 with the content of sesquiterpenic acids at the level of 0.004-0.094%. Population no 18 was distinguished by the highest content of sesquiterpenic acids and the relatively high mass of underground organs, followed by the admixture of the gene pool, typical for the 'Lubelski' landrace. Unlike the 'Lubelski' landrace, the wild-growing populations were characterized by a high amount of an essential oils (3.90 to 10.04 mL/kg), which may be promising from the perspective of their potential use. In turn, the sowing value of the seeds obtained from the populations, expressed as the germinability, was rather low (25.25-62.25%).

Antioxidant and Antibacterial Activity of Helichrysum italicum (Roth) G. Don. from Central Europe.

Helichrysum italicum (Roth) G. Don. is one of the most important cosmetic and medicinal plants originating from the Mediterranean region of Europe. The aim of this study was to assess the chemical profile as well as antioxidant and antibacterial potential of the species cultivated in the temperate climate of Central Europe. The analyses were carried out using herbs and inflorescences. The content of essential oil ranged from 0.25 g × 100 g-1 in the herb to 0.31 g × 100 g-1 in the inflorescences. Neryl acetate, accompanied by α-pinene in the herb (10.42%), and nerol in inflorescences (15.73%) were the dominants here. Rutoside, as well as rosmarinic, chlorogenic, neochlorogenic, isochlorogenic b and cichoric acids, were detected in both raw materials using HPLC-DAD. Within this group, cichoric acid was the dominant (2647.90 mg × 100 g-1 in the herb, 1381.06 mg × 100 g-1 in the inflorescences). The herb appeared to be more abundant in phenolics in comparison with the inflorescences. When given antioxidant activity (determined using DPPH and ABTS assays), both methanolic extract and essential oil obtained from the herb indicated higher potential than those originating from the inflorescences (74.72, 61.38 and 63.81, 58.59% in the case of DPPH, respectively). In turn, regarding antimicrobial activity, the essential oil from inflorescences was distinguished by stronger bacteriostatic power than the herb essential oil. Gram-positive bacteria were more sensitive to both essential oils in comparison with Gram-negative ones, with S. aureus ATCC 25923 as the most susceptible (MIC 1; MBC 16 mg × mL-1) among tested strains.

Antiviral Activity of Selected Essential Oils against Cucumber Mosaic Virus.

The aim of the study was to assess the antiviral activity of selected essential oils (EOs) against Cucumber mosaic virus (CMV), both in vitro and in vivo. The observations were made using Chenopodium quinoa as a local host. The EOs were obtained from Greek oregano, thyme, and costmary. Their chemical composition was determined using GC/FID followed by GC/MS. The dominant compound in oregano EO was carvacrol (59.41%), in thyme EO-thymol (59.34%), and in costmary EO-ß-thujone (90.60%). Among the analysed EOs, thyme EO exhibited the most promising effects against CMV. However, its activity was influenced by the time of application. In an in vivo experiment, thyme EO showed protective (pre-inoculation) rather than curative (post-inoculation) activity.

Antioxidant and Antibacterial Activity of Essential Oils and Hydroethanolic Extracts of Greek Oregano (O. vulgare L. subsp. hirtum (Link) Ietswaart) and Common Oregano (O. vulgare L. subsp. vulgare).

Greek oregano and common oregano were compared in respect of the antioxidant and antibacterial activity of the corresponding essential oils and hydroethanolic extracts in relation with their chemical profile. The chemical composition of essential oils was determined by GC-MS and GC-FID, while extracts (phenolic acids and flavonoids fractions) were analyzed by HPLC-DAD. Based on given volatiles, the investigated subspecies represented two chemotypes: a carvacrol/γ-terpinene/p-cymene type in the case of Greek oregano and a sabinyl/cymyl type rich in terpinen-4-ol in common oregano. Within non-volatile phenolic compounds, rosmarinic acid appeared to dominate in both subspecies. Lithospermic acid B, chlorogenic acid and isovitexin were present only in Greek oregano extracts. However, the total content of flavonoids was higher in common oregano extracts. The essential oil and extract of Greek oregano revealed visibly stronger antibacterial activity (expressed as MIC and MBC) than common oregano, whereas the antioxidant potential (determined by DPPH, ABTS and FRAP) of these extracts was almost equal for both subspecies. In the case of Origanum plants, the potential application of essential oils and extracts as antiseptic and antioxidant agents in the food industry should be preceded by subspecies identification followed by recognition of their chemotype concerning both terpene and phenolics composition.

The Influence of Two-Component Mixtures from Spanish Origanum Oil with Spanish Marjoram Oil or Coriander Oil on Antilisterial Activity and Sensory Quality of a Fresh Cut Vegetable Mixture.

The aim of this study was to evaluate two-component mixtures of essential oils (EOs) in sub-inhibitory concentrations for antilisterial protection of minimally processed vegetables. Spanish origanum oil (SOO), Spanish marjoram oil (SMO) and coriander oil (CO) and a mixture of fresh leafy vegetables with red beet were used. The chemical compositions of selected EOs were analyzed by gas chromatography. Carvacrol was the major component of SOO. The dominant active component of SMO was 1,8-cineole, while the major constituent in CO was linalool. The study shows that essential oils in combination, namely SOO + SMO and SOO + CO in a » MIC + » MIC (Minimal Inhibitory Concentration), have a synergistic effect against L. monocytogenes. The reduction of L. monocytogenes in vegetable mixtures treated by EOs in these selected combinations after 24 h of storage was approximately equal to the limit of detection. Furthermore, it was found that the vegetable mixture treated with SOO + SMO had the highest-rated sensorial quality and could therefore meet consumer expectations.

The Quality of Greek Oregano (O. vulgare L. Subsp. hirtum (Link) Ietswaart) and Common Oregano (O. vulgare L. Subsp. vulgare) Cultivated in the Temperate Climate of Central Europe.

The purpose of the study was to determine the differences between two subspecies: O. vulgare L. subsp. hirtum (Link) Ietswaart (Greek oregano) and O. vulgare L. subsp. vulgare (common oregano) growing in cultivation conditions within temperate climate of Central Europe. The characteristic of the subspecies was undertaken in terms of selected morphological parameters and the quality of the raw material. The herb of both subspecies was evaluated on the content and composition of essential oil by hydrodistillation followed by GC-MS and GC-FID (gas chromatography coupled with mass spectrometry and flame ionization detector), the total content of phenolic acids (according to PP 6th ed.) and the content of rosmarinic acid (by HPLC). The sensory evaluation (QDA) was performed, as well. Greek oregano was distinguished by visibly higher number of glandular trichomes on the leaves (up to 4.85 per 1 mm2) followed by higher content of essential oil in the herb (up to 3.36 g × 100 g-1 DW) in comparison to common oregano. Based on the essential oil composition, Greek oregano was classified as mixed carvacrol/γ-terpinene chemotype, while common oregano as mixed sabinyl/cymyl type rich in sesquiterpenes. Greek oregano was also characterized by higher total content of phenolic acids (up to 6.16 g × 100 g-1 DW) and rosmarinic acid (up to 6787.2 mg × 100 g-1 DW) than common oregano. Essential oil content reached the maximum at the beginning of blooming (common oregano) and at the full blooming stage (Greek oregano). In turn, the amount of phenolic acids followed by rosmarinic acid was the highest at the beginning of seed-setting stage, in the case of both subspecies. The differences between subspecies concerning chemical composition (especially essential oil) were reflected in the sensory attributes, where both odor and taste notes were found at higher level for Greek oregano. Results of our work indicate that Greek oregano is well adapted to grow in the temperate zone conditions. Such adaptation was reflected mainly in the satisfied yield and maintaining characters typical for the Mediterranean plant, e.g., a high essential oil content followed by high carvacrol share, traits the most important from practice viewpoint.

Chemical Diversity of Bastard Balm (Melittis melisophyllum L.) as Affected by Plant Development.

The phytochemical diversity of Melittis melissophyllum was investigated in terms of seasonal changes and age of plants including plant organs diversity. The content of phenolics, namely: coumarin; 3,4-dihydroxycoumarin; o-coumaric acid 2-O-glucoside; verbascoside; apiin; luteolin-7-O-glucoside; and o-coumaric; p-coumaric; chlorogenic; caffeic; ferulic; cichoric acids, was determined using HPLC-DAD. Among these, luteolin-7-O-glucoside, verbascoside, chlorogenic acid, and coumarin were the dominants. The highest content of flavonoids and phenolic acids was observed in 2-year-old plants, while coumarin in 4-year-old plants (272.06 mg 100 g-1 DW). When considering seasonal changes, the highest content of luteolin-7-O-glucoside was observed at the full flowering, whereas verbascoside and chlorogenic acid were observed at the seed-setting stage. Among plant organs, the content of coumarin and phenolic acids was the highest in leaves, whereas verbascoside and luteolin-7-O-glucoside were observed in flowers. The composition of essential oil was determined using GC-MS/GC-FID. In the essential oil from leaves, the dominant was 1-octen-3-ol, whilst from flowers, the dominant was α-pinene.

Evaluation of Annona muricata Acetogenins as Potential Anti-SARS-CoV-2 Agents Through Computational Approaches.

Annona muricata, a tropical plant which has been extensively used in ethnomedicine to treat a wide range of diseases, from malaria to cancer. Interestingly, this plant has been reported to demonstrate significant antiviral properties against the human immunodeficiency virus, herpes simplex virus, human papilloma virus, hepatitis C virus and dengue virus. Additionally, the bioactive compounds responsible for antiviral efficacy have also shown to be selectively cytotoxic while inhibiting tumorigenic cell growth without affecting the normal cell growth. Annonaceous Acetogenins are a class of bioactive compounds exclusive to the Annonaceae family at which the plant A. muricata belongs. In the current study, we have created a library of Acetogenins unique to the plant, comprising of Annomuricin A, Annomuricin B, Annomuricin C, Muricatocin C, Muricatacin, cis-Annonacin, Annonacin-10-one, cis-Goniothalamicin, Arianacin and Javoricin, for in silico and theoretical evaluations against the SARS-CoV-2 spike protein in an attempt toward promotion of plant based drug development for the current pandemic of coronavirus disease 2019 (COVID-19). We found that all the Acetogenins showing in silico spike protein significantly docking with good binding affinities. Moreover, we envision A. muricata Acetogenins can be further studied by in vitro and in vivo models to identify potential anti-SARS-CoV-2 agents.

Antioxidant and Antibacterial Activity of Roseroot (Rhodiola rosea L.) Dry Extracts.

Roseroot (Rhodiola rosea L.) belongs to plants revealing adaptogenic properties, which are attributed to the presence of specific phenolic compounds and are reflected mainly as antioxidant activity. The aim of the present study was to determine the antioxidant and antibacterial activity of various products obtained from R. rosea (underground organs as well as their aqueous and ethanolic dry extracts) in relation to the chemical profiles of phenolic and essential oil compounds. The chemical profiles were determined by High-performance Liquid Chromatography with a diode-array detector (HPLC-DAD) and Gas chromatography-mass spectrometry (GC-MS), antioxidant activity by (1,1-Diphenyl-2-picrylhydrazyl) Scavenging Capacity Assay (DPPH), (2,2'-Azino-bis(3-ethylbenzothiazoline-6-sulphonic acid)) Scavenging Capacity Assay (ABTS) and Ferric Reducing Antioxidant Power Assay (FRAP) and antimicrobial properties were expressed as minimum inhibitory concentration (MIC) and minimum bacterial concentration (MBC) values following the broth microdilutions method. The results show that the investigated samples differed in terms of their chemical compositions and biological activities. The extracts were more abundant in phenolic compounds (salidroside, tyrosol, and rosavin derivatives) in comparison to dried underground organs. The content of the determined phenolics in the analyzed extracts was affected by the solvent used for extraction. The ethanolic extract was characterized by the highest content of these substances in comparison to the aqueous one and the dried raw material, especially with regard to rosavin (969.71 mg/100 g). In parallel, this extract showed the strongest antioxidant and antibacterial activity. However, dried R. rosea underground organs also revealed strong antibacterial effects against, for example, Staphylococcus strains.

Phenolics in Primula veris L. and P. elatior (L.) Hill Raw Materials.

Primula veris L. and Primula elatior (L.) Hill represent medicinal plants used for the production of herbal teas and preparations with antioxidant and expectorant activity. Flowers and roots of both species possess the same biological activity. In the presented study, raw materials of wild growing P. veris and P. elatior were compared in terms of the content and composition of phenolic compounds using a fast and simple HPLC-DAD method. The study showed that flowers of both species were rich in flavonoids. However, P. veris flowers were characterized with a distinctly higher content of isorhamnetin-3-O-glucoside, astragalin, and (+)-catechin, whereas P. elatior occurred to be a richer source of rutoside and isorhamnetin-3-O-rutinoside. Hyperoside was found exclusively in P. elatior flowers. Phenolic glycosides (primverin and primulaverin) were identified only in the roots. Their content was about ten times higher in P. veris in comparison with P. elatior underground organs. The obtained results clearly show that both Primula species differ distinctly in terms of the content and composition of phenolic compounds. The compounds differentiating both species to the highest degree (hyperoside, in flowers, as well as primverin and primulaverin, in the roots) may be useful chemical markers in the identification and evaluation of both species.

Preservation of Brussels Sprouts by Pullulan Coating Containing Oregano Essential Oil.

In this study, the effectiveness of pullulan (a fungal polysaccharide) film containing oregano essential oil (OEO) at 1.0 to 10.0% was evaluated against bacteria, yeasts, and molds. The quality of the sprouts, as determined by weight loss, color, and appearance, was monitored during storage at 2 and 16°C. An organoleptic evaluation of odor preference and odor acceptability of OEO on the Brussels sprouts was also conducted. The antimicrobial activity of pullulan films with OEO increased significantly with the increase in OEO concentration (1 to 10%). Pullulan films with OEO were more effective for inhibiting the growth of yeasts and molds than for inhibiting gram-positive and gram-negative bacteria. Pullulan with 1.0% OEO was an effective combination and was used subsequently as the base coating for maintaining the safety and quality of fresh Brussels sprouts stored at 16°C for 14 days. The pullulan coating containing 1.0% OEO reduced Aspergillus niger populations by 2 log CFU/g. This coating also reduced weight loss in the sprouts. Compared with uncoated Brussels sprouts, the percent weight loss after 14 days was reduced in samples coated with pullulan and with pullulan plus 1% OEO by 3.81 and 6.06%, respectively, after storage at 2°C and by 8.04 and 9.30%, respectively, after storage 16°C. The coating also significantly reduced changes in general appearance and color during storage. Evaluation of the organoleptic properties indicated that pullulan containing OEO had only a slight detrimental effect on odor properties. Incorporating OEO into a delivery system for antimicrobial compounds in pullulan coatings extended the microbiological shelf life of Brussels sprouts.

Antimicrobial activity of a pullulan-caraway essential oil coating on reduction of food microorganisms and quality in fresh baby carrot.

This research evaluated the antimicrobial efficacy of pullulan films containing caraway essential oil (CEO). The films were prepared from a 10% of pullulan, containing from 0.12% to 10.0% of CEO. The composition of the CEO was analyzed with the use of gas chromatography. The antimicrobial activity of the CEO was evaluated with the method of serial microdilutions, and the films containing CEO-with the agar diffusion method against selected Gram-negative, Gram-positive bacteria, and fungi. The structure of the film surface and its cross-section were analyzed using a scanning electron microscope (SEM). Analyses were also carried out to determine the efficacy of a pullulan coating with 10% CEO on baby carrots experimentally inoculated with Salmonella enteritidis, Staphylococcus aureus, Saccharomyces cerevisiae, or Aspergillus niger and stored at a room temperature for 7 d. At a concentration of 0.12%, CEO inhibited the growth of all the tested microorganisms. Pullulan films containing 8% to 10% of CEO were active against all tested microorganisms. Populations of S. aureus on carrot samples were reduced by approximately 3 log CFU/g, while those of A. niger and S. cerevisiae by, respectively, 5 and 4 log CFU/g, after 7 d of storage. S. enteritidis was the most resistant among the tested species, since it was not significantly reduced after 7 d of storage. At the end of storage, samples treated with pullulan-caraway oil coating maintained better visual acceptability than control samples. Results of this study suggest the feasibility of applying a pullulan film with incorporated CEO to extend the microbiological stability of minimally processed foods.