DNA Barcoding and Fertilization Strategies in Sideritis syriaca subsp. syriaca, a Local Endemic Plant of Crete with High Medicinal Value.

Herein, we applied DNA barcoding for the genetic characterization of Sideritis syriaca subsp. syriaca (Lamiaceae; threatened local Cretan endemic plant) using seven molecular markers of cpDNA. Five fertilization schemes were evaluated comparatively in a pilot cultivation in Crete. Conventional inorganic fertilizers (ChFs), integrated nutrient management (INM) fertilizers, and two biostimulants were utilized (foliar and soil application). Plant growth, leaf chlorophyll fluorescence, and color were assessed and leaf content of chlorophyll, key antioxidants (carotenoids, flavonoids, phenols), and nutrients were evaluated. Fertilization schemes induced distinct differences in leaf shape, altering quality characteristics. INM-foliar and ChF-soil application promoted yield, without affecting tissue water content or biomass partitioning to inflorescences. ChF-foliar application was the most stimulatory treatment when the primary target was enhanced antioxidant contents while INM-biostimulant was the least effective one. However, when the primary target is yield, INM, especially by foliar application, and ChF, by soil application, ought to be employed. New DNA sequence datasets for the plastid regions of petB/petD, rpoC1, psbK-psbI, and atpF/atpH were deposited in the GenBank for S. syriaca subsp. syriaca while the molecular markers rbcL, trnL/trnF, and psbA/trnH were compared to those of another 15 Sideritis species retrieved from the GenBank, constructing a phylogenetic tree to show their genetic relatedness.

Chemometric Analysis Evidencing the Variability in the Composition of Essential Oils in 10 Salvia Species from Different Taxonomic Sections or Phylogenetic Clades.

Essential oil (EO) of Salvia spp. has been widely used for culinary purposes and in perfumery and cosmetics, as well as having beneficial effects on human health. The present study aimed to investigate the quantitative and qualitative variations in EOs in wild-growing and cultivated pairs of samples from members in four Salvia sections or three clades, namely S. argentea L. (Sect. Aethiopis; Clade I-C), S. ringens Sm. (Sect. Eusphace; Clade I-D), S. verticillata L. (Sect. Hemisphace; Clade I-B), S. amplexicaulis Lam., and S. pratensis L. (Sect. Plethiosphace; Clade I-C). Furthermore, the natural variability in EO composition due to different genotypes adapted in different geographical and environmental conditions was examined by employing members of three Salvia sections or two phylogenetic clades, namely S. sclarea L. (six samples; Sect. Aethiopis or Clade I-C), S. ringens (three samples; Sect. Eusphace or Clade I-D), and S. amplexicaulis (five samples; Sect. Plethiosphace or Clade I-C). We also investigated the EO composition of four wild-growing species of two Salvia sections, i.e., S. aethiopis L., S. candidissima Vahl, and S. teddii of Sect. Aethiopis, as well as the cultivated material of S. virgata Jacq. (Sect. Plethiosphace), all belonging to Clade I-C. The EO composition of the Greek endemic S. teddii is presented herein only for the first time. Taken together, the findings of previous studies are summarized and critically discussed with the obtained results. Chemometric analysis (PCA, HCA, and clustered heat map) was used to identify the sample relationships based on their chemical classes, resulting in the classification of two distinct groups. These can be further explored in assistance of classical or modern taxonomic Salvia studies.

Bioactivities and phenolic composition of Limonium boitardii Maire and L. cercinense Brullo & Erben (Plumbaginaceae): two Tunisian strict endemic plants.

Limonium genus is traditionally used in North Africa for disease treatment and in cosmetic. This study investigates for the first time the antioxidant, anti-inflammatory and enzyme inhibitory activities of two Tunisian endemic plants, L. boitardii and L. cercinense. Analysis of phenolic compounds was carried out by using RP-HPLC. Total phenolic and flavonoid contents, antioxidant activity (antiradical, reducing and total antioxidant activities), tyrosinase and α-glucosidase, collagenase inhibition activities were determined. Inhibiting of NO release in LPS-stimulated macrophages was assessed. L. cercinense exhibited strong antioxidant and anti-inflammatory effects. In the tyrosinase and α-glucosidase inhibitor activity tests, the two species were highly active, especially L. cercinense. High total phenolic and flavonoid contents were recorded in L. cercinense and myricitrin, myricetin, myricetin 3-O-ß-D-galatopyranoside, luteolin 7-O-glucoside and rutin were the main phenolics in both species. The results obtained render L. boitardii and L. cercinense as valuable new natural sources for cosmetic and pharmacological applications.

Development and Validation of an HPLC Method for the Analysis of Flowers of Wild-Growing Primula veris from Epirus, Greece.

An HPLC-PDA method was developed for the determination of the flavonoids in the flowers of Primula veris from Epirus, Greece. The aim was to investigate the chemical content of the over-harvested P. veris populations of Epirus and to develop and optimize an extraction protocol to allow fast, exhaustive, and repeatable extraction. Qualitative analysis revealed that the P. veris flowers from Epirus were particularly rich in flavonoids, especially flavonol triglycosides including derivatives of quercetin, isorhamnetin, and kaempferol. A phytochemical investigation of a 70% hydromethanolic extract from the flowers afforded a new flavonoid, namely, isorhamnetin-3-Ο-ß-glucopyranosyl-(1 → 2)-ß-glucopyranosyl-(1 → 6)-ß-glucopyranoside, which is also the main constituent of the flower extracts. Its structure elucidation was carried out by means of 1D and 2D NMR and mass spectrometry analyses. The HPLC-PDA method was developed and validated according to the International Council for Harmonisation guidelines. Since the main flavonol glycoside of the plant is not commercially available, rutin was used as a secondary standard and the response correction factor was determined. Finally, the overall method was validated for precision (% relative standard deviation ranging between 1.58 and 4.85) and accuracy at three concentration levels. The recovery ranged between 93.5 and 102.1% with relative standard deviation values < 5%, within the acceptable limits. The developed assay is fast and simple and will allow for the quality control of the herbal drug.

Antibacterial Effects of Essential Oils of Seven Medicinal-Aromatic Plants Against the Fish Pathogen Aeromonas veronii bv. sobria: To Blend or Not to Blend?

Despite progress achieved, there is limited available information about the antibacterial activity of constituents of essential oils (EOs) from different medicinal-aromatic plants (MAPs) against fish pathogens and the complex interactions of blended EOs thereof. The present study aimed to investigate possible synergistic antimicrobial effects of EOs from seven Greek MAPs with strong potential against Aeromonas veronii bv. sobria, a fish pathogen associated with aquaculture disease outbreaks. The main objective was to evaluate whether blending of these EOs can lead to increased antimicrobial activity against the specific microorganism. A total of 127 combinations of EOs were prepared and their effect on A. veronii bv. sobria growth was tested in vitro. We examined both the inhibitory and bactericidal activities of the individual EOs and compared them to those of the blended EOs. The vast majority of the investigated combinations exhibited significant synergistic and additive effects, while antagonistic effects were evident only in a few cases, such as the mixtures containing EOs from rosemary, lemon balm and pennyroyal. The combination of EOs from Greek oregano and wild carrot, as well as the combinations of those two with Spanish oregano or savoury were the most promising ones. Overall, Greek oregano, savoury and Spanish oregano EOs were the most effective ones when applied either in pure form or blended with other EOs.

Latitudinal variation in soil nematode communities under climate warming-related range-expanding and native plants.

Current climate change has led to latitudinal and altitudinal range expansions of numerous species. During such range expansions, plant species are expected to experience changes in interactions with other organisms, especially with belowground biota that have a limited dispersal capacity. Nematodes form a key component of the belowground food web as they include bacterivores, fungivores, omnivores and root herbivores. However, their community composition under climate change-driven intracontinental range-expanding plants has been studied almost exclusively under controlled conditions, whereas little is known about actual patterns in the field. Here, we use novel molecular sequencing techniques combined with morphological quantification in order to examine nematode communities in the rhizospheres of four range-expanding and four congeneric native species along a 2,000 km latitudinal transect from South-Eastern to North-Western Europe. We tested the hypotheses that latitudinal shifts in nematode community composition are stronger in range-expanding plant species than in congeneric natives and that in their new range, range-expanding plant species accumulate fewest root-feeding nematodes. Our results show latitudinal variation in nematode community composition of both range expanders and native plant species, while operational taxonomic unit richness remained the same across ranges. Therefore, range-expanding plant species face different nematode communities at higher latitudes, but this is also the case for widespread native plant species. Only one of the four range-expanding plant species showed a stronger shift in nematode community composition than its congeneric native and accumulated fewer root-feeding nematodes in its new range. We conclude that variation in nematode community composition with increasing latitude occurs for both range-expanding and native plant species and that some range-expanding plant species may become released from root-feeding nematodes in the new range.

Comparative Evaluation of Essential Oils from Medicinal-Aromatic Plants of Greece: Chemical Composition, Antioxidant Capacity and Antimicrobial Activity against Bacterial Fish Pathogens.

The administration of antibiotics in aquaculture has raised concern about the impact of their overuse in marine ecosystems, seafood safety and consumers' health. This "green consumerism" has forced researchers to find new alternatives against fish pathogens. The present study focused on 12 Mediterranean medicinal-aromatic plants as potential antimicrobials and antioxidant agents that could be used in fish aquaculture. In vitro assays showed that the essential oils (EOs) from all studied plants had anti-bacterial and antioxidant properties, with their efficacy being dependent on their chemical composition. More specifically, EOs rich in carvacrol, p-cymene and γ-terpinene exhibited not only the strongest inhibitory activity against the growth of bacterial pathogens (inhibitory concentration: 26-88 µg mL-1), but also the greatest total antioxidant capacity (ABTS: 2591-5879 µmole mL-1; CUPRAC: 931-2733 µmole mL-1). These compounds were mainly found in the EOs from Greek oregano (Origanum vulgare subsp. hirtum), Spanish oregano (Thymbra capitata) and savoury (Satureja thymbra) collected from cultivations in Greece. The specific EOs stand out as promising candidates for the treatment of bacterial diseases and oxidative stress in farmed fish. Further in vivo experiments are needed to fully understand the effects of EO dietary supplementation on fish farming processes.

Exploring the Ecological Preferences and Essential Oil Variability in Wild-Growing Populations of the Endangered Local Greek Endemic Thymus holosericeus (Lamiaceae).

Thymus holosericeus Celak. (Lamiaceae) is a taxonomically isolated and endangered local endemic thyme species which is geographically isolated in four Ionian Islands (West Greece). The present study investigated the essential oil (EO) composition, the ecological preferences, and their correlations in three T. holosericeus wild-growing populations from Zakynthos (ΤH-Z), Cephalonia (ΤH-C) and Lefkada (ΤH-L). The variations in essential oil yield and the composition of T. holosericeus populations were evaluated using hydrodistillation, GC/MS, TLC and NMR analysis. The climatic conditions of each sample were organized and analyzed in RStudio with the raster package and in SPSS with Pearson's Canonical Correlation Analysis (CCA), respectively. The aerial parts of the plants varied in EO yields from 1.92 to 2.28% w/v. The analysis of EO constituents revealed noteworthy qualitative and quantitative inter-population variation. The composition of EOs revealed the presence of linalool (82.77%) and borneol (5.95%) as major compounds in ΤH-Z, while carvacrol (35.34%), geraniol (23.98%), linalool (14.37%), borneol (5.66%), thymol (4.27%) and p-cymene (4.08%) were the main compounds in ΤH-C and linalool (40.37%), geraniol (39.42%) and borneol (5.20%) were dominant components in ΤH-L. The results of the gas chromatography procedure have also been confirmed by 1H and 13C-NMR and TLC analysis. The ecological profile showed an average annual precipitation of 942 ± 18.33 mm and the temperature limits in which T. holosericeus seems to adapt to are 6.80± 1.08 °C 27.70 ± 0.70 °C. Regarding the examined samples, TH-C was adapted to the driest summer and coldest winter conditions, TH-Z was adapted to the lowest annual precipitation with the most complex climatic conditions, and TH-L was adapted to the highest summer temperatures with the lowest precipitation in the wettest period of the year. For each sampled population, the CCA identified the association of the samples' EOs composition with the prevailing local environmental conditions.

Ecological Preferences and Diversity of Essential Oil Composition in Endangered Wild-Growing Populations of Sideritis sipylea Boiss. (Lamiaceae) of the East Aegean Islands (Greece): Evidencing Antioxidant Potential, Antimicrobial and Cytotoxic Activities.

Plants from the genus Sideritis (Lamiaceae) have been widely used in folk medicine for a long time and consequently are a focus of the scientific community. Despite this interest, explicit data about the essential oils (EOs) of the Endangered Sideritis sipylea have not been readily available to date. In this study, we investigated the ecological preferences of Greek S. sipylea and the chemical composition of the essential oils of wild-growing S. sipylea populations from two Greek islands (S1: Samos, S2: Lesvos); we explored concomitant associations with environmental factors; and we assessed their (i) antioxidant potential (two tests), (ii) antimicrobial activity against six microbial and two fungal strains, and (iii) cytotoxic effect in two human and one murine cell lines. We compiled an ecological profile in R based on all known Greek localities of S. sipylea, outlining for the first time its preferences regarding temperature (3.48 ± 1.53 °C to 30.70 ± 1.11 °C) and the precipitation regimes (5.92 ± 2.33 mm to 136 ± 11.43 mm) shaping its natural occurrence in the wild. The chemical analysis (42 compounds in total) confirmed the domination of monoterpene hydrocarbons in both samples (with quantitative and qualitative differences) and identified 12 new constituents reported in S. sipylea for the first time (e.g., Bicyclogermacrene and Cumacrene). Dominant compounds in S1 (39 constituents) were ß-Myrcene (20.4%) followed by ß-caryophyllene (11.8%), bicyclogermacrene (7.1%), ß-pinene (6.3%), carvacrol (6.2%) and α-terpinene (6.1%), whereas in S2 (26 constituents) the main ones were α-pinene (37.3%), ß-pinene (15.1%) and sabinene (12.1%), followed by ß-caryophyllene (5.6%) and bicyclogermacrene (5.5%). The strong antioxidant capacity and cytotoxic activity of S. sipylea EOs are reported herein for the first time, while new insight is provided regarding their effect on bacterial and fungal strains (four ones originally tested herein). The biological activity analysis demonstrated variation among samples, with S2 being more potent than S1. Altogether, the results of the present study demonstrate the high biological potential of S. sipylea EOs with an interesting antioxidant capacity and antimicrobial and cytotoxic effects and reveal associations of natural chemodiversity with climatic factors.

Taxonomic Identification and Molecular DNA Barcoding of Collected Wild-Growing Orchids Used Traditionally for Salep Production.

Molecular DNA barcoding combined with botanical taxonomy can be used for the identification and conservation of collected Greek orchids used for salep production as well as in the regulation of fair salep trade. A modified CTAB protocol was used for DNA extraction, amplification of barcoding regions (ITS, matK, rbcL, trnH-psbA), and sequencing. Sequencing data were assembled using Bioedit software, and the BLAST algorithm was used on the NCBI database for species identification at the genus level. Molecular barcoding data based on genetic similarity identification was in full coherence with taxonomic classification based on morphological data. The combination of ITS and matK exhibited a greater capacity to identify a species among the Greek salep samples. Out of the 53 samples examined, 52.9% were classified as Dactylorhiza spp. and 33.3% as Anacamptis spp., whereas only 6 samples were identified as Orchis spp. (11.8%). Given that a superior-quality salep beverage comes from tubers of the latter, the number of samples classified as such in northwestern Greece is unexpectedly low. A database of 53 original reference sequences from wild-growing samples of Greek origin was generated, providing a valuable resource for the identification of other salep samples from different regions. The DNA barcoding results unveiled that salep samples from northwestern Greece are related to nine members of four different genera of Orchidaceae. All species are nationally protected and covered by the CITES convention, while many of these orchids are included in the EU Directive 92/43/EEC appendix as "Other Important Species". Thus, expedited coordinated management actions are needed to ensure their survival in the future.

The Invasion of Alien Populations of Solanum elaeagnifolium in Two Mediterranean Habitats Modifies the Soil Communities in Different Ways.

We aimed to explore how the invasion of the alien plant Solanum elaeagnifolium affects soil microbial and nematode communities in Mediterranean pines (Pinus brutia) and maquis (Quercus coccifera). In each habitat, we studied soil communities from the undisturbed core of both formations and from their disturbed peripheral areas that were either invaded or not by S. elaeagnifolium. Most studied variables were affected by habitat type, while the effect of S. elaeagnifolium was different in each habitat. Compared to maquis, the soil in pines had higher silt content and lower sand content and higher water content and organic content, supporting a much larger microbial biomass (PLFA) and an abundance of microbivorous nematodes. The invasion of S. elaeagnifolium in pines had a negative effect on organic content and microbial biomass, which was reflected in most bacterivorous and fungivorous nematode genera. Herbivores were not affected. In contrast, in maquis, organic content and microbial biomass responded positively to invasion, raising the few genera of enrichment opportunists and the Enrichment Index. Most microbivores were not affected, while herbivores, mostly Paratylenchus, increased. The plants colonizing the peripheral areas in maquis probably offered a qualitative food source to microbes and root herbivores, which in pines was not sufficient to affect the much larger microbial biomass.

Adaptability of Wild-Growing Tulips of Greece: Uncovering Relationships between Soil Properties, Rhizosphere Fungal Morphotypes and Nutrient Content Profiles.

Wild-growing Greek tulips are protected plants but almost nothing is known about their natural nutrient status and rhizosphere fungal morphotypes in the wild, thus no insight is currently available into their growth and adaptation to their natural environment or artificial settings. To this end, several botanical expeditions were conducted with a special collection permit, and 34 tulip and soil samples were collected, representing 13 species from two phytogeographical regions of Greece (North Aegean Islands, Crete Island) and seven regions of mainland Greece. The tulips' content in essential macro- and micro-nutrients, respective physicochemical soil properties, and rhizosphere fungal morphotypes were assessed across samples, and all parameters were subjected to appropriate statistical analysis to determine their interrelationships. The results showed that soil variables played a significant role in shaping tulips' nutrient content, explaining up to 67% of the detected variability as in the case of phosphorus (P) in the above-ground plant tissue. In addition, significant correlations were observed (with an r value of up to 0.65, p < 0.001) between essential nutrients in the tulips, such as calcium (Ca) and boron (B). The principal component analysis (PCA) revealed that between the three spatial units examined, the total variability of tulips' nutrient content produced a clear distinction among sampled species, while the first two PCA axes managed to explain 44.3% of it. This was further confirmed by the analysis of variance (ANOVA) results which showed corresponding significant differences (at p < 0.05) in both the tulips' nutrient content and the studied soil properties as well (mean values of N, P, and K in the North Aegean Islands tulips' nutrient content, up to 53%, 119%, and 54% higher compared to those of the Crete Island, respectively). Our study sheds light on Greek tulips' adaptability and resilience in their original habitats, facilitating at the same time the undertaken efforts regarding their conservation and potential domestication in artificial settings.

Exploring the Medicinal Potential of Achillea grandifolia in Greek Wild-Growing Populations: Characterization of Volatile Compounds, Anti-Inflammatory and Antioxidant Activities of Leaves and Inflorescences.

Various species of the genus Achillea L. (Asteraceae) are traditionally used worldwide for wound healing against diarrhea, flatulence, and abdominal pains, as diuretic and emmenagogue agents. In the present study, the essential oils (EOs) obtained separately from the leaves and inflorescences of wild-growing Achillea grandifolia Friv. from Mt. Menoikio and Mt. Pelion (Greece) were analyzed by Gas Chromatography-Mass Spectrometry. The major compounds found in EOs of A. grandifolia inflorescences from Mt. Menoikio were as follows: cis-thujone (36.9%), 1,8-cineole (11.9%), camphor (10.0%), ascaridole (7.3%), α-terpinene (6.4%), sabinene (4.1%), trans-thujone (3.6%), and cis-jasmone (3.4%). In leaves from Mt. Menoikio, they were as follows: cis-thujone (50.8%), 1,8-cineole (20.0%), trans-thujone (5.5%), camphor (5.5%), borneol (3.6%), and α-terpineol (3.1%). In inflorescences from Mt. Pelion, they were as follows: camphor (70.5%), camphene (5.9%), cis-jasmone (3.2%), bornyl acetate (3.2%). In leaves from Mt. Pelion, they were as follows: camphor (83.2%), camphene (3.9%), and borneol (3.7%). Subsequently, the samples were first time tested for their antioxidant activities with the interaction of EOs with DPPH (2,2-diphenyl-1-picrylhydrazyl) and their inhibition of lipid peroxidation, as well as for their anti-inflammatory activity through the soybean LOX (lipoxygenase) inhibition. All of the examined samples were found effective. A. grandifolia leaves presented the highest antioxidant potential according to the DPPH method, and the highest percentage of LOX inhibition. The study herein investigated for the first time the leaves and the inflorescences of A. grandifolia separately, and the results generally align with similar studies from neighboring countries (Turkey and Serbia) in terms of the yields and categorization of main EO compounds (oxygenated monoterpenes). However, the findings were not in agreement with previously studied Greek material, as a higher amount of cis-thujone and lower antioxidant activity are reported herein. Both the EOs of inflorescences and the leaves of the wild-growing population collected from Mt. Menoikio were characterized by a high quantity of cis-thujone (36.9% and 50.8%, respectively).

Isolation of Secondary Metabolites from Achillea grandifolia Friv. (Asteraceae) and Main Compounds' Effects on a Glioblastoma Cellular Model.

This study aims at the isolation and structural determination of the secondary metabolites of the herbaceous perennial plant Achillea grandifolia Friv. (Asteraceae). The examination of the non-volatile content of the leaves and flowers of A. grandifolia afforded the isolation of sixteen secondary metabolites. On the basis of NMR spectra, the identified compounds included ten sesquiterpene lactones; three guaianolides-rupicolin A (1), rupicolin B (2), and (4S,6aS,9R,9aS,9bS)-4,6a,9-trihydroxy-9-methyl-3,6-dimethylene-3a,4,5,6,6a,9,9a,9b-octahydro-3H-azuleno [4,5-b]furan-2-one (3); two eudesmanolides-artecalin (4) and ridentin B (5); two sesquiterpene methyl esters-(1S,2S,4αR,5R,8R,8αS)-decahydro-1,5,8-trihydroxy-4α,8-dimethyl-methylene-2-naphthaleneacetic acid methylester (6) and 1ß, 3ß, 6α-trihydroxycostic acid methyl ester (7); three secoguaianolides-acrifolide (8), arteludovicinolide A (9), and lingustolide A (10); and an iridoid-loliolide (11). Moreover, five known flavonoids, i.e., apigenin, luteolin, eupatolitin, apigenin 7-O-glucoside, and luteolin 7-O-glucoside (12-16) were also purified from the aerial parts of the plant material. We also investigated the effect of rupicolin A (1) and B (2) (main compounds) on U87MG and T98G glioblastoma cell lines. An MTT assay was performed to define cytotoxic effects and to calculate the IC50, while flow cytometry was employed to analyze the cell cycle. The IC50 values of reduced viability during the 48 h treatment for compound (1) and (2) were 38 µM and 64 µM for the U87MG cells and 15 µM and 26 µM for the T98G cells, respectively. Both rupicolin A and B induced a G2/M cell cycle arrest.

From the Wild to the Field: Documentation, Propagation, Pilot Cultivation, Fertilization, and Phytochemical Evaluation of the Neglected and Underutilized Amelanchier ovalis Medik. (Rosaceae).

The snowy Mespilus, or serviceberry (Amelanchier ovalis Medik., Rosaceae) represents a neglected and underutilized small fruit tree species with high nutritional value. In this work, we present the results of a long-term study facilitating the sustainable exploitation of A. ovalis as a new germplasm resource from the Greek flora. Ten wild-growing population samples of A. ovalis have been collected from natural habitats in northern Greece. Asexual propagation trials on these materials delivered successful propagation (83.3% rooting) on a selected genotype via leafy cuttings of young, primary, non-lignified soft wood with the application of the rooting hormone. The ex situ cultivation potential of the selected genotype has been evaluated under distinct fertilization regimes in a pilot field trial. Three-year results of this ongoing trial have shown that A. ovalis does not require external nutrient enhancement to be established during its early stages since plant growth rates between conventional fertilization and control plants were similar for the first two years and higher compared to organic fertilization. Conventional fertilization delivered higher fresh fruit production in the third year, with higher fruit number and fruit size compared to organic fertilization and control plants. The phytochemical potential of the cultivated genotype was assessed via the total phenolic content and radical scavenging activity of separate extracts from leaves, twigs, flowers, and young fruits, which revealed that individual plant organs have strong antioxidant activity despite their moderate total phenolic content. The multifaceted approach applied herein has provided novel data that may set the framework for further applied research toward the sustainable agronomic exploitation of Greek A. ovalis as a diversified superfood crop.

HPLC- and NMR-Based Chemical Profiling, Wound-Healing Potential, Anti-Inflammatory and Antibacterial Activities of Satureja pilosa (Lamiaceae), a Neglected Medicinal-Aromatic Herb.

Satureja pilosa Velen. (Lamiaceae) is a perennial and melliferous aromatic-medicinal subshrub which is range-restricted in adjacent parts of Greece and Bulgaria and locally in Italy, known in Northern Greece as wild oregano ("agriorigani") and traditionally collected from the wild for culinary purposes. Since the ethnopharmacological data and modern biological activities of Satureja spp. suggest promising applications in skin conditions, the present study aimed to investigate the hitherto unknown phenolic content of cultivated S. pilosa and its potential biological activities, focusing mainly on wound-healing and anti-inflammatory effects. An HPLC-PDA-MS-targeted phytochemical investigation, along with NMR, allowed for the isolation and characterization of the main constituents, resulting in 18 compounds. Representative extracts and purified compounds were tested for wound-healing activity on NIH/3T3 fibroblasts. The butanol extract exhibited a significantly higher cell migration rate (73.4%) compared to aqueous (50.6%) and methanolic (49.6%) ones, enhancing the cell migration more rapidly at both concentration levels, whilst rosmarinic acid was the most potent among the isolated compounds, with a migration rate of 64.0% at the concentration level of 10-5 mg/mL, followed by 3,4-dihydrophenyllactic acid (54.7%). Moreover, potential effects on endothelial activation processes were explored, including the leukocyte-endothelial cell interaction during inflammatory processes and the migratory capacity during angiogenic actions, since these processes are commonly associated with skin diseases. Finally, extracts and purified compounds demonstrated weak antibacterial potential against two important pathogens (Staphylococcus aureus and Pseudomonas aeruginosa), suggesting that further investigation is warrented.

Bitter Is Better: Wild Greens Used in the Blue Zone of Ikaria, Greece.

The current study reports an ethnobotanical field investigation of traditionally gathered and consumed wild greens (Chorta) in one of the five so-called Blue Zones in the world: Ikaria Isle, Greece. Through 31 semi-structured interviews, a total of 56 wild green plants were documented along with their culinary uses, linguistic labels, and locally perceived tastes. Most of the gathered greens were described as bitter and associated with members of Asteraceae and Brassicaceae botanical families (31%), while among the top-quoted wild greens, species belonging to these two plant families accounted for 50% of the wild vegetables, which were consumed mostly cooked. Cross-cultural comparison with foraging in other areas of the central-eastern Mediterranean and the Near East demonstrated a remarkable overlapping of Ikarian greens with Cretan and Sicilian, as well as in the prevalence of bitter-tasting botanical genera. Important differences with other wild greens-related food heritage were found, most notably with the Armenian and Kurdish ones, which do not commonly feature many bitter greens. The proven role of extra-oral bitter taste receptors in the modulation of gastric emptying, glucose absorption and crosstalk with microbiota opens new ways of looking at these differences, in particular with regard to possible health implications. The present study is also an important attempt to preserve and document the bio-cultural gastronomic heritage of Chorta as a quintessential part of the Mediterranean diet. The study recommends that nutritionists, food scientists, and historians, as well as policymakers and practitioners, pay the required attention to traditional rural dietary systems as models of sustainable health.

Influence of Temperature on Seed Germination of Five Wild-Growing Tulipa Species of Greece Associated with Their Ecological Profiles: Implications for Conservation and Cultivation.

Although tulips are famous worldwide as ornamental plants, the knowledge about the seed germination of wild-growing species remains limited. The aim of the present study was to investigate the effect of temperature on seed germination of the local, wild-growing Greek endemics Tulipa bakeri and T. goulimyi and the sub-Balkan endemic T. undulatifolia, which are threatened with extinction, as well as the Mediterranean T. australis and the Asiatic T. clusiana naturalized on Chios Island (Greece). The germination responses at five constant temperatures (5, 10, 15, 20, and 25 °C) were assessed for all studied species in growth chambers under a 12:12 light-dark photoperiod. The ecological profile for each species was developed in R using open-source bioclimatic data; this was built to illustrate the abiotic environmental conditions of their wild habitats, to facilitate the examination of temperature effects on seed germination, and to facilitate their cultivation in artificial environments. The results indicated that the seed germination requirements of the studied species had a range-specific temperature dependence, reflecting their natural adaptation to local ecological conditions. Seed germination of T. bakeri, T. australis, and T. clusiana was observed only in a narrow range of very low temperatures (5-10 °C), whereas germination of T. undulatifolia and T. goulimyi occurred at temperatures between 5 and 15 °C. A temperature increase to 20 or 25 °C resulted in the absence of seed germination for all five Greek tulip species. The germinated seeds were planted in pots and bulblets were developed under greenhouse conditions. Seeds and bulblets constitute valuable genetic materials for the cultivation and ex situ conservation of these five Greek tulip species, three of which are threatened with extinction.

Effect of Temperature on Breaking of Morphophysiological Dormancy and Seed Germination Leading to Bulblet Production in Two Endemic Tulip Species from Greece.

Due to botanical tulips' economic interest coupled with limited information regarding their seed germination, we investigated the effect of temperature on dormancy release and germination in two endangered local endemic tulip species of Greece (Tulipa hageri Heldr., T. orphanidea Heldr.). Their germination responses at five constant temperatures (5, 10, 15, 20, and 25 °C) were evaluated in growth chambers, while the type of seed dormancy and the temperature effect on its release were determined based on open-sourced, R-derived species-specific ecological profiles illustrating abiotic conditions of their wild habitats. The results indicated a range-specific temperature dependence in seed germination for both studied species with seed germination observed only in very low temperatures (5-10 °C). The seeds of both species after dispersal had an underdeveloped embryo. The existence of a complex morphophysiological seed dormancy was confirmed in both species by the significant embryo development only at 5 and 10 °C (almost doubled after 30 days) coupled with observed delay in germination only at low temperatures. Furthermore, to facilitate their cultivation and ex situ conservation, the germinated seeds were planted in pots to develop bulblets in greenhouse conditions resulting in bigger T. orphanidea bulblets compared to T. hageri.

A Multifaceted Evaluation Approach for Greek Native Neglected and Underutilized Forest Fruit Trees and Shrubs as Natural Sources of Antioxidants: Consolidating the Framework for Their Sustainable Agronomic Exploitation.

Fruits from wild forest trees and shrubs represent a natural source of antioxidants against oxidative stress and a growing market for novel minor crops. This study presents a multifaceted approach which sets the basis for sustainable agronomic exploitation of selected Greek native germplasm of four traditional but neglected and underutilized forest fruit trees and shrubs, namely Amelanchier ovalis Medik., Cornus mas L., Rosa canina L., and Sambucus nigra L. The studied plant species are traditionally used in Greek ethnobotany but are currently neglected and underutilized in commercial terms, thus characterized as neglected and underutilized plant species (NUPs). The investigation includes new information on the evaluation of the ex situ cultivation of the Greek germplasm (three of the four focal NUPs), thus supplementing respective full datasets for their comparative evaluation based on four evaluation axes (documentation and molecular authentication of genotypes, phytochemical evaluation, asexual propagation via rooting of cuttings, and ex situ cultivation) after multi-year and multifaceted groundwork data previously acquired. Consecutively, the work includes feasibility and readiness timescale evaluation for the sustainable exploitation of each focal species based on existing literature and extant research experience. The feasibility for sustainable exploitation and readiness timescale evaluation results were very encouraging, showing high exploitation feasibility with an already achieved readiness timescale for R. canina and S. nigra, whereas C. mas and A. ovalis showed an achievable readiness in the short term. The comparative evaluation of the Greek native focal NUPs outlined the excellent potential of R. canina, S. nigra, and A. ovalis, and the high potential of C. mas. The results herein illustrate the very high fruit antioxidant potential (free radical scavenging activity) of all focal species, the diverse but effective asexual propagation capacity via cuttings at the species level, and summarize the results of a pilot cultivation trial set up in 2020 (still ongoing) outlining tree growth rates and the onset of fruit production among genotypes and species. Overall, the meta-analysis of previously published data in conjunction with new data generated herein may serve the sustainable exploitation of the studied NUPs.