Search

Of the six juniper species found in the Bulgarian flora, three of the species have controversial taxonomic positions. Juniperus pygmaea K. Koch and J. sibirica Burgsd. exhibit similar morphological characteristics to J. communis L. in terms of leaves and female cones (galbuli). This is one of the reasons why, in the recent taxonomic developments, J. pygmaea and J. sibirica were united in a common variety of J. communis, namely, J. communis var. saxatilis. However, such a grouping of species in the Flora of Bulgaria has not been adopted. This study aimed to evaluate the degrees of similarity or difference in the structure of the leaves, galbuli, seeds, and pollen of J. communis, J. sibirica, and J. pygmaea using the methods of comparative anatomy by light microscope (LM) and scanning electron microscopy (SEM) observations and complex morphological measurements. The working hypothesis of this study was that the three species would show a different degree of similarity with each other, which would clarify their taxonomic rank. The morphological parameters revealed differences between the length/width ratio of galbuli and seed length of the three species, while leaf characteristics (length and width) showed a stronger resemblance between J. sibirica and J. pygmaea. Furthermore, a greater distinction between the leaves and galbili of J. communis and J. sibirica was found. The SEM analyses showed variations in the seed shape and spermoderm among the three species. The shape of J. communis seeds was oval and elongated, while J. pygmaea seeds were pear-shaped, and J. sibirica seeds were triangular-rhombic. The length and height of striations were diverse on seed spermoderm in the three species. The epicuticular waxes of leaves, located on the tips of the anticlinal walls of the elongated epidermal cells in J. pygmaea and J. communis, were oval, while they formed raised comb-like crystals in J. sibirica. The morphological, anatomical, and SEM analysis affirmed the accepted taxonomic status of J. communis and J. sibirica as independent species within the Bulgarian flora. Based on most of the analyzed parameters, J. pygmaea exhibits significant similarity with J. sibirica. Additionally, the similar habitats of these two species support the determination of J. pygmaea as a variety or form of J. sibirica rather than J. communis (J. sibirica forma pygmaea).

Pinus heldreichii Christ. (Bosnian pine), a Tertiary relict and Balkan sub-endemic, has not been comprehensively studied for its essential oil (EO) profile and bioactivity of its different plant parts. This study aimed to determine the EO yield, composition and antimicrobial activity from different parts of P. heldreichii at three different populations (mountains) in Bulgaria. Furthermore, the study assessed the antioxidant activities of plant tissue, including leaves (needles), twigs wood, male and female cones. The EOs yield from different plant parts ranged from 0.09 % (leaves) to 0.74 % (wood of twigs), with monoterpenes being the predominant class. Limonene, α-pinene, β-caryophyllene, germacrene D, β-pinene, and β-myrcene were detected in the EO extracted from all analyzed trees. However, these compounds were not found in the EO extracted from all plant parts of the same trees. Four chemical groups (chemotypes) were identified for EO from twigs, and three chemotypes were identified for EO from leaves. The chemotypes were based on the percent ratio of the main EO constituents (>5 %). Leaves tissue showed the highest values in terms of polyphenols and flavonoids, as well as higher ABTS radical scavenging activity, while the highest antimicrobial activity against Staphylococcus aureus subsp. aureus was seen in the EOs obtained from twigs. This is the first study to identify several chemotypes based on leaf and twigs EO of P. heldreichii distributed in Bulgarian flora. Furthermore, the EO of twigs tips (TT), male cones (MC), and wood of one-two-year-old twigs (WT) of the same trees were reported for the first time. The total polyphenol, flavonoid content, and radical scavenging activity of tissues of annual twigs wood and biennial twigs wood, leaf tissue, MC tissue, and the twigs tips tissue is also reported for the first time in the accessible literature. These findings highlight the potential of P. heldreichii to provide EOs with varying compositions and bioactivities, making them suitable for nutraceutical, pharmacological, and potentially food additive applications. Furthermore, the identification of chemotype accessions in this study suggests their selection for the development of new forest crop as a source for natural products with desirable composition and bioactivity.

As a staple crop, potatoes (Solanum tuberosum) play an important role in meeting daily caloric needs. To ensure adequate supplies for year-round consumption, potato quality must be maintained throughout lengthy storage periods. Towards this end, potato sprouting during storage must be minimized. Due to changing regulations regarding chemical means of potato sprout suppression, increased focus has turned to alternative products including essential oils (EO) as sprout suppressants in recent years. The complex composition of various EOs promises numerous options for sprout suppression. Furthermore, blends of several EOs may achieve enhanced sprout suppressant properties if synergistic interactions are present. We evaluated Syzygium aromaticum, Artemisia herba-alba, and Laurus nobilis EOs and blends thereof as sprout suppressants in potato cultivar Ranger Russet stored at room temperature and also tested for their antifungal activity against Colletotrichum fragariae, a causal organism of anthracnose disease in strawberries including other vegetables and fruits. A. herba-alba EO was an effective sprout suppressant when used alone and suppressed sprouting over the 90-day storage period. Interactions between A. herba-alba and S. aromaticum affected sprout length whereas interactions between A. herba-alba and L. nobilis EOs affected sprout number. An optimum blend of 50% - 82.31% A. herba-alba, 17.69% - 50% L. nobilis, and 0% - 1.01% S. aromaticum EOs could more effectively minimize tuber sprout length and number than any of the three whole EOs used alone. Among these three EOs, only S. aromaticum EO showed antifungal activity against C. fragariae in bioautography assay. These results exhibit the potential of EOs blends as a novel tactic in potato sprout suppression as well as potential natural product-based fungicides in managing C. fragariae.

Abstract Hemp (Cannabis sativa L.) synthesizes and accumulates a number of secondary metabolites such as terpenes and cannabinoids. They are mostly deposited as resin into the glandular trichomes occurring on the leaves and, to a major extent, on the flower bracts. In the last few years, hemp for production of high-value chemicals became a major commodity in the U.S. and across the world. The hypothesis was that hemp biomass valorization can be achieved through distillation and procurement of two high-value products: the essential oil (EO) and cannabinoids. Furthermore, the secondary hypothesis was that the distillation process will decarboxylate cannabinoids hence improving cannabinoid composition of extracted hemp biomass. Therefore, this study elucidated the effect of steam distillation on changes in the content and compositional profile of cannabinoids in the extracted biomass. Certified organic CBD-hemp strains (chemovars, varieties) Red Bordeaux, Cherry Wine and Umpqua (flowers and some upper leaves) and a T&H strain that included chopped whole-plant biomass, were subjected to steam distillation, and the EO and cannabinoids profile were analyzed by gas chromatography-mass spectrometry (GC–MS) and HPLC, respectively. The distillation of hemp resulted in apparent decarboxylation and conversion of cannabinoids in the distilled biomass. The study demonstrated a simple method for valorization of CBD-hemp through the production of two high-value chemicals, i.e. EO and cannabinoids with improved profile through the conversion of cannabidiolic acid (CBD-A) into cannabidiol (CBD), cannabichromenic acid (CBC-A) into cannabichromene (CBC), cannabidivarinic acid (CBDV-A) into cannabidivarin (CBDV), cannabigerolic acid (CBG-A) into cannabigerol (CBG), and δ-9-tetrahydrocannabinolic acid (THC-A) into δ-9-tetrahydrocannabinol (THC). In addition, the distilled biomass contained CBN while the non-distilled did not. Distillation improved the cannabinoids profile; e.g. the distilled hemp biomass had 3.4 times higher CBD in variety Red Bordeaux, 5.6 times in Cherry Wine, 9 times in variety Umpqua, and 6 times in T&H compared to the original non-distilled samples, respectively. Most of the cannabinoids remained in the distilled biomass and small amounts of CBD were transferred to the EO. The CBD concentration in the EO was as follows: 5.3% in the EO of Umpqua, 0.15% in the EO of Cherry Wine and Red Bordeaux and 0.06% in the EO of T&H. The main 3 EO constituents were similar but in different ratio; myrcene (23.2%), (E)-caryophyllene (16.7%) and selina-3,7(11)-diene (9.6%) in Cherry Wine; (E)-caryophyllene (~ 20%), myrcene (16.6%), selina-3,7(11)-diene (9.6%), α-humulene (8.0%) in Red Bordeaux; (E)-caryophyllene (18.2%) guaiol (7.0%), 10-epi-γ-eudesmol (6.9%) in Umpqua; and (E)-caryophyllene (30.5%), α-humulene (9.1%), and (E)-α-bisabolene (6.5%) in T&H. In addition, distillation reduced total THC in the distilled biomass. Scanning electron microscopy (SEM) analyses revealed that most of the glandular trichomes in the distilled biomass were not disturbed (remained intact); that suggest a possibility for terpenes evaporation through the epidermal membrane covering the glandular trichomes leaving the cannabinoids in the trichomes. This explained the fact that distillation resulted in terpene extraction while the cannabinoids remained in the distilled material.

Satureja pilosa Velen., S. coerulea Janka and S. kitaibelii Wierzb. ex Heuff. are Balkan endemic species, and they are distributed in restricted territories, primarily found in dry grasslands, rocky slopes, and stony habitats. This study presents the results of the first embryological and micromorphological analyses of three Satureja species (S. pilosa, S. kitaibelii, and S. coerulea) from the Bulgarian flora. The aim of this study was to establish the features of the male and female reproductive sphere, as well as surface characteristics of leaves, stem, and calyx in order to understand the mode of reproduction, character, size and state of species populations and delimitation. For the embryological study, flowers and flower buds in different developmental stages were collected from plants of natural populations and treated with the classic paraffin method. Reproductive capacity was assessed using the following approaches: (1) acetocarmine test for pollen viability, (2) tetrazolium test (TTZ) for seed viability, and (3) germination test. The surfaces characteristics of leaves, stem, and calyx were analyzed by scanning electron microscopy (SEM). As a result, the study revealed the structures of the male (M) and female (F) generative spheres as well as the processes of gamete development, pollination, and endosperm and embryo formation. It was found that the three Satureja species exhibited a high pollen viability but low seed viability and germination. The SEM analysis showed both common and distinct micromorphology features regarding epidermis, calyx and stem surfaces among the three Satureja species. Notably, the S. coerulea surfaces (leaves, calyx, stem) were clearly distinguishable from the other two Satureja species. Regarding the nutlet surfaces, it was observed that the exocarp surfaces of S. coerulea and S. kitaibelii have a reticulate convex type surface and comprise two types of cells: (1) highly bulging, bubble-like cells; and (2) flat cells with numerous striations. On the other hand, the exocarp of S. pilosa displayed rectangular or polygonal shapes without bubble-like cells, and it had a tabular to slightly convex type surface. Additionally, nutlets (seeds) of both S. coerulea and S. pilosa exhibited distinct papilla formations resembling non-glandular trichomes seen on the ends of the nutlets for S. coerulea and over the entire surface for S. pilosa. The characteristics of the embryological structures and processes, along with the absence of apomixis, characterized the three studied Satureja species as sexually reproducing. The established balanced processes and stable structures contribute to their high reproductive potential and population stability. However, these traits may also decrease their adaptability to environmental changes.

Currently, there are increased interests in growing grain and fiber hemp (Cannabis sativa L.) as well as in large-scale hemp products. Cannabis has been grown/utilized for thousands of years as a fiber, grain, and drug/medicinal plant. However, the strict control of cannabis cultivation to combat illegal use, the spread of new yarns and oilseeds, and the advent of cheap synthetic fibers caused a decreased/eliminated hemp production. Hemp has been banned in most of the world for more than seven decades; it missed out on the Green Revolution and the adoption of new technologies and varieties, creating a knowledge gap. After the 2014 and 2018 Farm Bill in the USA, hemp became legal and the land grand universities launched research programs. The ability to utilize the entire plant for multiple purposes creates opportunity for the market to value hemp products. Hemp production technology varies depending on the type of hemp cultivated (grain, fiber, or cannabinoids), soil characteristics, and environmental factors. Hemp has the potential to be a very sustainable and ecologically benign crop. Hemp roots have a significant potential for absorbing and storing heavy metals such as lead, nickel, cadmium, and other harmful substances. In addition, hemp has been proven to be an excellent carbon trap and biofuel crop. Hemp has the ability to successfully suppress weeds, and it is generally regarded a pesticide-free crop. The purpose of this paper is to examine historic and recent industrial hemp (grain and fiber) literature, with a focus on hemp agronomy and utilization.

Abstract Previous studies in the long-term experiments at Pendleton, OR (USA), were focused on organic matter cycling, but the consequences of land management for nutrient status over time have received little attention. Soil and wheat (Triticum aestivum L.) tissue samples were analyzed to determine the macronutrient dynamics associated with residue management methods and fertilizer rate under a dryland winter wheat-fallow rotation. The treatments included: no burn residue incorporation with farmyard manure (FYM) or pea vines, no burn or spring burn with application of N fertilizer (0, 45, and 90 kg ha−1), and fall burn wheat residue incorporation. The results revealed no differences on the effect of residue burning on macronutrient concentration over time. After receiving the same treatments for 84 years, the concentrations of soil organic C, total N and S, and extractable Mg, K, P in the 0–10 cm depth significantly increased in FYM plots compared to the rest of the plots. The N fertilization rate of 90 kg ha−1 reduced the accumulations of P, K, and Ca in grain compared to the 0 and 45 kg N ha−1 applications. The results indicate that residue incorporation with FYM can play vital role in reducing the macronutrient decline over time.

The intensive use of lead (Pb)-based insecticides (lead arsenate, PbHAsO4) has led to Pb accumulation in agricultural soil, endangering human health through the possibility of transferring it to the food chain. The aim of this study was to evaluate the potential for the immobilization of Pb in the soil by applying organic (sludge, biocompost, yard compost, and peat) and inorganic (bonemeal, zeolite, lime, and wood ash) amendments, in combination with diammonium phosphate (DAP) in a greenhouse experiment. Two amendment rates were used: low and high, and three rates of DAP: 0 (zero), low (0.25 g of DAP/kg soil), and high (1.25 g DAP/kg soil). The results showed that the dry yield of carrot (Daucus carota susp. sativus) was the highest for the organic amendments in combination with the low rate of DAP. The high rate of inorganic amendments also increased the yield. Applications of inorganic bonemeal, inorganic lime, and inorganic wood ash yielded the lowest Pb tissue concentration (TC), and organic peat had the highest Pb TC. Inorganic bonemeal combined with DAP most effectively immobilized Pb in soil.

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.

Juniperus sabina L. var. balkanensis R.P. Adams & Tashev is a recently described endemic variety from the Balkan Peninsula. Its strong sprouting ability and fast vegetative propagation, on one hand, and fragmented distribution, on the other, can lead to lower genetic diversity in local populations and to the differentiation of populations. As there has been no detailed investigation of this variety, we studied Balkan natural populations using phytochemical and molecular markers. Leaf essential oils (EOs) were chosen based on their proven usability in the population studies of Juniperus taxa, while ISSRs (Inter Simple Sequence Repeats) have been used due to their high resolution. In addition, since this variety is best described using molecular markers, the chloroplast trnS-trnG region was amplified from individuals from different populations having different chemotypes. Based on the essential oil profile, three chemotypes could be identified with a difference in their distribution. The analysis of molecular variance showed moderate differentiation of populations and regions, attesting to the start of the separation of three regions in the Balkans: west, east and south. The bioclimatic and environmental parameters and sex of the individual did not influence the EO profile, although some of the compounds present in low-to-medium concentrations showed strong correlation with several bioclimatic parameters.

Our previous research has proven that the immortelle (Helichrysum italicum) essential oil (EO) grown in Serbia possesses respectable biological potential and desirable composition of volatile compounds with the potential for a wide range of applications in the food, cosmetics, and pharmaceutical industries. Within this study, the impact of seasonal variations (temperature, precipitation, and insolation) during three successive years (2017, 2018, and 2019), on the volatile profile of γ-curcumene + ar-curcumene immortelle chemotype was determined. Steam distillation was utilized to extract EO from the plant material, followed by chromatographic mass spectrometric analysis revealing 50 volatile compounds. A multiple linear regression model was developed, and principal component analyses were conducted to deliver detailed information regarding the prediction, component profile, and parallel contents of active compounds of the immortelle EO. Under Serbian agro-ecological conditions, with appropriate harvest method, immortelle can achieve two harvests per year: the first in July, and the second in August. The γ-curcumene + ar-curcumene chemotype usually occurs as the dominant chemotype in the region of the ex-Yugoslavia countries. This chemotype probably developed as an adaptation to climatic conditions, and spring and summer precipitation positively influenced curcumene accumulation in plants. Such a phenomenon was especially noticeable in the first harvest.

Alkanna tinctoria (L.) Tausch Boraginaceae is a medicinal plant whose root is used for its antimicrobial and anti-inflammatory properties. A. tinctoria roots have been subject to numerous studies. However, the aerial parts have been explored less. The objective of the present study was to compare the chemical profile of aerial parts and roots as well as the total alkannin content in roots of 11 populations of the species from different floristic regions of Bulgaria. Methanolic extracts from 22 samples were analyzed by GC/MS. Phenolic, fatty, and organic acids, sterols, polyols, fatty alcohols, and sugars were identified. Ononitol (4-O-methyl-myo-inositol) was found as the main compound in the aerial parts. The total alkannin content in the roots was evaluated by the spectrophotometric method and compared with that of the commercial product. Populations with high alkannin content and rich in other bioactive compounds were identified. A relatively low genetic diversity in the studied populations was observed. The present study is the first comprehensive study on metabolite profiles and genetic diversity of the Bulgarian populations of A. tinctoria. The occurrence of ononitol in the aerial parts of the species is reported for the first time, as well as the phenolic acid profiles of the species in both aerial parts and roots. The results showed that aerial parts of the plant are also promising for use as a source of valuable biologically active substances.

The prospect of replacing traditional chemical fertilization with organic and microorganism-based fertilization meets the current demand for more sustainable cropping systems and healthy food. In this respect, research was carried out to evaluate the effects of the factorial combination between four basil cultivars (‘Aromat de Buzau’, ‘Macedon’, ‘Cuisoare’ and ‘Serafim’) and three types of fertilization, namely chemical fertilization (with a solid chemical fertilizer), organic fertilization (with chicken manure formulate) and microorganisms’ fertilization (with microorganisms formulate), on basil yield, biochemical and physiological parameters and essential oil composition. The results showed that the biometric parameters (plant height, number of stems and leaves and leaf area) were significantly influenced by the cultivar; ‘Macedon’ obtained the highest values of plant height (64.7 cm) and number of stems (20.33) and leaves (618.3) and ‘Serafim’ the largest leaf area (4901.7 cm2 per plant), while the type of fertilization did not affect these parameters. Regarding the biomass, the influence of the cultivar was not significant on fresh biomass but was significant on dry biomass, with ‘Macedon’ showing the highest value (56.4 g·plant−1 dry biomass). The mentioned parameters were significantly influenced by the type of fertilization, with the highest values recorded with chemical fertilization. Both the cultivar and the fertilization type significantly influenced the physiological parameters (the total content of assimilatory pigments and photosynthesis). Five phenolic compounds were quantified from leaf extracts by HPLC-MS (caffeic acid, hyperoside, isoquercitrin, rutin and quercitrin). Hyperoside was identified only in ‘Macedon’, while the rest of the compounds were found in all the cultivars and varied depending on the cultivar and fertilization type. Regarding the composition of the essential oil, variation was found depending on the cultivar and fertilization type. In ‘Aromat de Buzau’, the main compounds were methyl chavicol and β-linalool; in ‘Macedon’, geranial and neral; and in ‘Cuisoare’ and ‘Serafim’, β-linalool. Moreover, the PCA showed that the ‘Serafim’ cultivar has exclusive properties compared to the other cultivars. Our results highlight that identifying the most effective interaction between genotype and fertilization type allows to optimize yield and quality targets for sweet basil.

The composition, herbicidal, larvicidal, and toxic activities of Juniperus horizontalis Moench essential oil and sabinene, its main component, were evaluated. The seed germination percentage and root length of eight different plant species (crops and weeds) were measured for in vitro herbicidal activity tests. Different doses (100, 50, 10, 5, 1, 0.5 µg/mL) of the samples were applied to seeds for 120 h. The same doses were applied to test the toxicity of the samples on Tubifex tubifex (sludge worm) and Chironomus aprilinus (blood worm). Four doses (435, 652.5, 870, and 1740) of samples were in a larvicidal test on Tenebrio molitor L. (mealworm), and bioassays were checked after 24 and 48 h. The analysis of the oil allowed for the identification of dominant components as sabinene (38.7%), α-pinene (10.0%), elemol (8.6%), γ-terpinene (8.3%), limonene (7.8%) and α-thujene (5.3%). The results showed that the effect of oil on root length inhibition was significant in all crop species. The doses which affected root growth were not toxic to Tubifex tubifex and Chironomus aprilinus. Finally, the obtained results in the larvicidal bioassay indicate that the potential of J. horizontalis in yellow mealworm survivorship limitation can be a starting point for future research.

Chlorpropham (CIPC) has been the dominant method of chemical sprout suppression for the last half-century. However, stricter regulations including outright bans on its use in several countries has prompted investigation into alternative products to replace it. Growing interest in organic foods has increased focus on the use of biopesticides, including essential oils (EOs), as potential sprout suppressants in stored potato. We evaluated the potential of ten EOs for sprout suppression in potato cultivar Ranger Russet at room temperature. Treatment with Cymbopogon citratus EO was found to be the most effective sprout suppressant, completely suppressing sprouting over the 90-day storage period. The EOs of Myrtus communis and Melaleuca quinquenervia significantly reduced sprout length relative to the control but did not have any effect on sprout number. These findings demonstrate the potential of select EOs as effective potato sprout suppressants that could replace CIPC use in this industry while also giving more power to organic potato producers and processors to control sprouting in their operations.

As a global staple, potato plays an important role in meeting human dietary needs and alleviating malnutrition. Potato sprouting during storage is a major issue that threatens food security by increasing food waste and must therefore be controlled. Biopesticides, including essential oils (EOs), have a history of use as potato sprout suppressants, and interest in their use has been renewed in response to stricter regulations on CIPC, the dominant chemical sprout suppressant over the last half-century. We evaluated twenty-one EOs as potential sprout suppressants in cv. Ranger Russet potatoes at room-temperature storage. Treatment with Artemisia herba-alba EO was the most effective at suppressing both sprout length and sprout number over a 90-day storage period. GC—MS—FID analysis of A. herba-alba EO revealed the presence of α-thujone, hexadecenoic acid, β-thujone, camphor, sabinene, and camphene at amounts >1%. Cistus ladanifer, Ocimum basilicum, Ormenis mixta, and Salvia sclarea EOs significantly reduced sprout length for shorter storage periods, whereas Cinnamomum zeylanicum (bark) and Laurus nobilis EOs also significantly reduced sprout number. Syzygium aromaticum (clove) EO did not significantly suppress sprouting at room temperature. These results indicate the potential of certain EOs to be used as sprout suppressants for room-temperature potato storage, providing needed alternatives for both organic and conventional potato industries.

Winter savory (Satureja montana L.) is a well-known spice and medicinal plant with a wide range of activities and applications. Two subspecies of S. montana, subsp. montana and subsp. variegata, were used for the preparation of seven different extracts: steam distillation (essential oil (EO) and hydrolate (HY)), subcritical water (SWE), ultrasound-assisted (UAE-MeOH and UAE-H2O), and microwave-assisted (MAE-MeOH and MAE-H2O) extraction. The obtained EOs, HYs, and extracts were used for an in vitro evaluation of the antioxidant activity (DPPH, ABTS, reducing power, and superoxide anion methods) and in vitro antimicrobial activity against Bacillus cereus, Staphylococcus aureus, Enterococcus faecalis, Escherichia coli, Salmonella Typhimurium, Saccharomyces cerevisiae, and Candida albicans. The antimicrobial screening was conducted using disk-diffusion assessment, minimal inhibitory concentration, time–kill kinetics modeling, and pharmacodynamic study of the biocide effect. The total phenolic content (TPC) was highest in EO, followed by SWE, MAE, and UAE, and the lowest was in HY. The highest antimicrobial activity shows EO and SWE for both varieties, while different UAE and MAE extracts have not exhibited antimicrobial activity. The natural antimicrobials in the S. montana extract samples obtained by green extraction methods, indicated the possibility of ecologically and economically better solutions for future in vivo application of the selected plant subspecies.

Laurel (Laurus nobilis L.) is a plant species from Lauraceae family, and is native to the Mediterranean region. The goal of this study was to compare chemical composition of laurel leaves and antibacterial activity of its essential oil (EO) from wild-grown trees in Greece and Georgia. The laurel leaves from the two native habitats had dissimilar concentrations of phenolic acids. Of the conjugated flavonols and flavons, kaempferol (1981.3 μg/g) and apigenin (1433.6 μg/g) were the major representatives in the leaves from Greece, while luteolin (839.1 μg/g) and kaempferol (688.1 μg/g) were the major ones in the leaves from Georgia, respectively. The EO content was 1.42% and 4.54% in the leaves from Greece and Georgia, respectively. The main EO constituents of the Greek laurel plants were 1,8-cineole (30.8%), α-terpinyl acetate (14.9%), α-terpineol (8.0%), sabinene (7.9%), and terpinen-4-ol (6.0%). The main EO constituents of the Georgian laurel plants were 1,8-cineole (29.2%), α-terpinyl acetate (22.6%), sabinene (12.2%), and methyleugenol (8.1%). The EO antimicrobial activities against 20 microorganisms were determined. Among the Gram-positive bacteria, the Enterococcus faecalis strain was the most sensitive, followed by Staphylococcus aureus ATCC 6538. Among the Candida species, C. albicans ATCC 10231 was the most sensitive to the laurel leaf EOs.

Satureja pilosa and S. kitaibelii (Lamiaceae) are Balkan endemic plant species, and the composition of their essential oil (EO) is highly variable. The aim of the present study was to establish: (1) the EO variability in two populations of S. pilosa (the intrapopulation), and (2) the EO variation in S. kitaibelii between nine populations (interpopulation) from Bulgaria and two from Serbia. The EOs of two Satureja species were obtained from aboveground plant parts by hydrodistillation and were analyzed by GC/MS/FID. Overall, the EO yield on the intrapopulation level of S. pilosa varied from 0.54% to 2.15%, while the EO of S. kitaibelii varied from 0.04% to 0.43% (interpopulation). The EO of S. pilosa was found to contain thymol and carvacrol as the main constituents, with other major constituents being p-cymene and γ-terpinene. S. pilosa samples in both studied populations formed six chemical groups. The major constituents (p-cymene, terpinen-4-ol, bornyl acetate, γ-muurolene, endo-borneol, cis-β-ocimene, trans-β-ocimene, carvacrol, α-pinene, thymoquinone, geranial, geranyl acetate, spathulenol, and caryophyllene oxide) of S. kitaibelii EO were considered for grouping the populations into ten chemotypes. The present study is the first report on the interpopulation diversity of S. kitaibelii EOs in Bulgaria. It demonstrated variability of the EOs between and within the populations of S. kitaibelii from Bulgaria. This study identified promising genetic material that could be further propagated and developed into cultivars for commercial production of S. kitaibelii and S. pilosa, thereby reducing the impact of collection on wild populations.

Sagebrush (Artemisia spp.) are dominant wild plants in large areas of the U.S., Canada and Mexico, and they include several species and subspecies. The aim was to determine if there are significant differences in essential oil (EO) yield, composition, and biological activity of sagebrush within the Bighorn Mountains, U.S. The EO yield in fresh herbage varied from 0.15 to 1.69% for all species, including 0.25–1.69% in A. tridentata var. vaseyana, 0.64–1.44% in A. tridentata var. tridentata, 1% in A. tridentata var. wyomingensis, 0.8–1.2% in A. longifolia, 0.8–1% in A. cana, and 0.16% in A. ludoviciana. There was significant variability in the EO profile between species, and subspecies. Some EO constituents, such as α-pinene (0–35.5%), camphene (0–21.5%), eucalyptol (0–30.8%), and camphor (0–45.5%), were found in most species and varied with species and subspecies. The antioxidant capacity of the EOs varied between the species and subspecies. None of the sagebrush EOs had significant antimicrobial, antimalarial, antileishmanial activity, or contained podophyllotoxin. Some accessions yielded EO with significant concentrations of compounds including camphor, eucalyptol, cis-thujone, α-pinene, α-necrodol-acetate, fragranol, grandisol, para-cymene, and arthole. Therefore, chemotypes can be selected and possibly introduced into culture and be grown for commercial production of these compounds to meet specific industry needs.

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.

Steam distillation was used for the isolation of Dracocephalum moldavica L. (Moldavian dragonhead) essential oil (DMEO). This aromatic herbaceous plant is widespread across the Northern Hemisphere regions and has been utilized in health-improving studies and applications. In addition to the DMEO, the hydrolate (DMH), a byproduct of the distillation process, was also collected. The DMEO and DMH were analyzed and compared in terms of their chemical composition, as well as their in vitro biological activities. The main component in DMEO was geranyl acetate, while geranial was dominant in DMH. The DMEO demonstrated better antioxidant and antimicrobial activities compared with the DMH against Staphylococcus aureus, Escherichia coli, Salmonella Typhimurium, and Listeria monocytogenes, which represent sources of food-borne illness at the global level. The DMEO and DMH show promise as antioxidant and antimicrobial additives to various products.

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 β-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%), β-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.

Periodic removal of sediment from aquaculture ponds is practiced to maintain their productivity and animal welfare. The recovery of sediment as a plant fertilizer could alleviate the costs of sediment removal. The objective of this study was to test the effects of a dried sediment, extracted from an aquaculture pond used for common carp cultivation, on the growth and physiology of potted wheat grass and the quality of the juice obtained from wheat grass. The results showed that sediment application did not produce significant morphological changes, although the values for plant height (16.94–19.22 cm), leaf area (19.67–139.21 mm2), and biomass (3.39–4.26 g/plant) were higher in sediment-grown plants. However, at a physiological level, the effect was negative, decreasing photosynthesis (0.82–1.66 μmol CO2 m2s−1), fluorescence ΦPSII (0.737–0.782), and chlorophyll content (1.40–1.83 CCI). The juice yield was reduced in the sediment treatments (46–58 g/100 g), while the quality was improved by increasing the content of phenols (2.55–3.39 µg/mL gallic acid equivalent), flavonoids (1.41–1.85 µg/mL quercetin equivalent), and antioxidant activity (47.99–62.7% inhibition of; 2,2-diphenyl-1-picrylhydrazyl). The positive results obtained in this study can be attributed to the moderate nutrient content of the sediment and a negligible concentration of heavy metals.

Genus Galanthus (Amaryllidaceae) includes 19 species in Europe and the Middle East. The Flora of Bulgaria recognizes two species: G. nivalis L. and G. elwesii Hook. Galanthus elwesii is characterized by relatively high morphological variability, leading some authors to identify some populations as G. gracilis Celak. However, the occurrence of G. gracilis in the Bulgarian flora is disputed. The hypothesis was that populations previously identified as G. gracilis belong indeed to a separate species. Therefore, the objective of this study was to compare G. nivalis and G. elwesii with plants from populations previously identified as G. gracilis. Morphological, DNA, embryological and anatomical analyzes were conducted to meet the objective. The morphological characteristics and DNA dendrogram revealed that G. gracilis and G. elwesii were situated in the same cluster and had significant morphological similarity, whereas plants from populations identified as G. nivalis were dissimilar in morphology and situated in a separate cluster. The revealed features of the generative sphere showed similarities across the species. Scanning electron microscopy (SEM) analyses of the surface revealed that the anticillinal walls of G. elwesii and G. gracilis were straight, while those of G. nivalis were wavy. This research demonstrated that the plants of G. elwesii and those from populations identified as G. gracilis are morphologically, embryologically and genetically similar, thus refuting the hypothesis. This study did not provide sufficient evidence to support the claim of the existence of G. gracilis in the Bulgarian flora; the populations identified as G. gracilis in Bulgaria may be forms of G. elwesii.

Marrubium friwaldskyanum Boiss (Lamiaceae) is a Bulgarian endemic species. Overall, the essential oil (EO) composition of M. friwaldskyanum was different from that of the other Marrubium species reported in the literature. The main EO constituents of M. friwaldskyanum were (E)-caryophyllene, germacrene D, and caryophyllene oxide. The effect of the harvest stage was significant only on α-copaene, (E)-caryophyllene, caryophyllene oxide, and τ-muurolol. The concentration of α-copaene (1.26–1.83% range of the total oil), (E)-caryophyllene (31–41%), caryophyllene oxide (6.4–11.8%), and τ-muurolol (1.3–2.8%) were the highest at 2–3 pair of leaves or before flowering and lower at flowering. The harvest stage did not significantly affect the concentrations of the other six identified EO compounds β-bourbonene (1.1%), α-humulene (2.8%), germacrene D (23.3%), bicyclogermacrene (2.85%), δ-cadinene (1.1%), and spathulenol (2.8%). In a separate experiment, grinding of the biomass prior to EO extraction had a significant effect only on the concentrations of D-limonene (0.24–3.3%) and bicyclogermacrene (3.6–9.1%). Grinding in water or without water, maceration, and addition of Tween®20 had rather small effects on the EO profile. The identified EO constituents and their mean concentrations in this experiment were (E)-caryophyllene (25.4%), germacrene D (17.6%), caryophyllene oxide (9.1%), spathulenol (6.5%), τ-muurolol (5.0%), carvacrol (3.9%), α-copaene (2.5%), β-bourbonene (2.5%), δ-cadinene (2.4%), α-humulene (1.8%), and Z-β-farnesene (1.3%). Embryological studies observed anther and the development of the male gametophyte and ovule and development of the female gametophyte of M. friwaldskyanum. Furthermore, pollen and seed viability assays were conducted, and mass spectrometry-based metabolomics analysis of an extract from shoots revealed the presence of 45 natural products, identified as flavonoids, phenolic acids, and (tri)terpenoids. Overall, the phytochemistry and some of the microscopic analyses distinguished this endemic species from other species in Marrubium.

In this study, we evaluated the allelopathic effects of essential oils (EOs) from six different plant species, namely, lavender (Lavandula angustifolia), hyssop (Hyssopus officinalis), English thyme (Thymus vulgaris), lovage (Levisticum officinale), costmary (Chrysanthemum balsamita), and cumin (Cuminum cyminum), on seed germination and seedling growth of barley (Hordeum vulgare) and wheat (Triticum aestivum). The main constituents of the EOs of L. angustifolia were 47.0% linalool acetate and 28.4% linalool; H. officinalis’ main constituents were 39.8% cis-pinocamphone, 9.8% trans-pinocamphone, 11.4% β-pinene, and 7.5% β-phellandrene; T. vulgaris’ were 38.2% para-cymene, 25.6% thymol, and 13.6% γ-terpinene; L. officinale’s were 64.8% α-terpinyl acetate and 14.7% β-phellandrene; C. balsamita’s were 43.7% camphor, 32.4% trans-thujone, and 11.6% camphene; C. cyminum’s were 49.6% cumin aldehyde, 10.4% para-cymene, 11.6% α-terpinen-7-al, and 9.1% β-pinene. All six EOs exhibited an allelopathic effect and suppressed the seed germination and seedling development of wheat and barley; however, the concentrations that exhibited a suppressing effect were different among the plants. C. cyminum EO completely suppressed both barley and wheat germination at 10-, 30-, and 90-µL application rates, making it the most effective treatment among the tested EOs. C. balsamita’s and H. officinalis’ EOs at 30 and 90 µL application rates completely suppressed barley and wheat radicles per seed, radicle length (mm), seedling height (mm), and germination (%). L. angustifolia’s EOs at 30- and 90-µL and T. vulgaris’ EO at 90 µL application rates also completely suppressed barley and wheat radicles per seed, radicle length (mm), seedling height (mm), and germination (%). C. balsamita’s, H. officinalis’, L. angustifolia’s, and T. vulgaris’ EOs at a 10 µL application rate reduced barley radicle length, seedling height, and % germination relative to the control. Wheat seed germination % was completely suppressed by the application of L. angustifolia’s and T. vulgaris’ EOs at 30 and 90 µL, while T. vulgaris’ EO at 10 µL rate reduced the germination relative to the control. Interestingly, C. balsamita and H. officinalis at 10 µL did not reduce wheat germination; however, they did reduce the number of radicles per seed, radicle length (mm), seedling height (mm), germination (%), and vigor index. Furthermore, L. officinale’s EO reduced the measured indices (radicles per seed, radicle length, seedling height, and vigor index) at the 10, 30, and 90 µL application rates relative to the non-treated control; however, none of the application rates of L. officinale’s EO had a suppression effect on wheat germination. This study demonstrated the allelopathic effects of the EOs of six different herbal plant species on seed germination of barley and winter wheat. The results can be utilized in the development of commercial products for controlling pre-harvest sprouting of wheat and barley. Further research is needed to verify the results under field conditions.

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, β-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.

Juniperus excelsa M. Bieb and J. sabina L. contain essential oil (EO), while J. sabina also contains podophyllotoxin, which is used as a precursor for anti-cancer drugs. Two studies were conducted. The first assessed the variability in the EO profile and podophyllotoxin concentration of the two junipers, depending on the location and tree gender. The main EO constituents of J. excelsa were α-cedrol, α-limonene and α-pinene, while the constituents in J. sabina were sabinene, terpinen-4-ol, myrtenyl acetate and α-cadinol. The podophyllotoxin yield of 18 J. sabina accessions was 0.07–0.32% (w/w), but this was not found in any of the J. excelsa accessions. The second study assessed the effect of hydrodistillation (Clevenger apparatus) and steam distillation (in a semi-commercial apparatus) on the EO profile and bioactivity. The extraction type did not significantly alter the EO composition. The EO profiles of the two junipers and their accessions were different and may be of interest to the industry utilizing juniper leaf EO. Breeding and selection programs could be developed with the two junipers (protected species) in order to identify chemotypes with (1) a high EO content and desirable composition, and (2) a high concentration of podophyllotoxin in J. sabina. Such chemotypes could be established as agricultural crops for the commercial production of podophyllotoxin and EO.

H. italicum essential oil (EO) is one of the most popular ingredients utilized by the cosmetic industry, and it is also used as natural antioxidant and as a value-added ingredient in food products. The chemical composition of the EO H. italicum cultivated in Serbia was analyzed by gas chromatography–mass spectrometry. The quantitative structure–retention relationship was used to predict the retention indices of the EO constituents acquired by GC-MS data, applying five molecular descriptors selected by factor analysis and a genetic algorithm. Also, antimicrobial activity, and biological activity by four common antioxidant tests (DPPH and ABTS assays, reducing power, and β-carotene bleaching test), and in vitro antihyperglycemic and anti-inflammatory capacities were evaluated. A total of 70 EO constituents were detected, of which 17 (8.5%) could not be identified. The H. italicum EO in this study belonged to γ-curcumene chemotype. The coefficients of determination reached the value of 0.964, demonstrating that this model could be used for prediction purposes. All applied tests showed that H. italicum EO possesses good biological activity and an interesting chemical composition. Therefore, the EO of H. italicum grown in Serbia has a potential to be used in food, cosmetic, and pharmaceutical products.

The development of organic farming as a result of increasing consumer preference for organic food has led to the development and registration of new pest-control products for certified organic production. In this study, the effects of three biocontrol products containing spores and mycelium of Arthrobotrys oligospora—Artis®, Beauveria bassiana—Bora®, and Coniothyrium minitans—Öko-ni® were tested on four basil (Ocimum basilicum L.) cultivars: ‘Aromat de Buzau’, ‘Serafim’, ‘Macedon’ and ‘Cuisoare’. The application of Öko-ni® increased basil yields by 8% relative to Control. The application of Bora® increased chlorophyll content of basil leaves by 2% and the activity of photosynthesis by 66% relative to the Control. Basil essential oil (EO) content was increased by 18% with the application of Artis® and by 34% with the application of Bora® and Öko-ni®, respectively. The content of phenolic compounds analyzed by HPLC varied; caffeic acid concentration was higher in the plants treated with Öko-ni®, hyperoside, isoquercitrin and rutin concentrations were higher in those treated with Artis®, while the quercitrin content was higher in Bora®-treated plants. The two main EO constituents that were identified were linalool and methyl chavicol in ‘Aromat de Buzau’, linalool and eugenol in ‘Serafim’, neral and geranial in ‘Macedon’, also linalool and eugenol in ‘Cuisoare’. The investigated myco-biocontrol products had positive effects on basil fresh biomass and EO content and also influenced the content of phenolic compounds.

The objective of this study was to evaluate wild growing Satureja kitaibelii, Thymus serpyllum, Origanum vulgare, Achillea millefolium and Achillea clypeolata with respect to their essential oil (EO) content, composition and antimicrobial activity. The five species were collected at Mt. Rtanj and the village of Sesalac, Eastern Serbia. The main EO constituents of Lamiaceae plants were p-cymene (24.4%), geraniol (63.4%) and germacrene D (21.5%) in Satureja kitaibelii, Thymus serpyllum and Origanum vulgare ssp. vulgare, respectively. A. millefolium EO had multiple constituents with major ones being camphor (9.8%), caryophyllene oxide (6.5%), terpinen-4-ol (6.3%) and 1,8-cineole (5.6%), while the main EO constituents of A. clypeolata were 1,8-cineole (45.1%) and camphor (18.2%). Antimicrobial testing of the EO showed that Staphylococcus aureus (Gram-positive) was more sensitive to all of the tested EOs than Escherichia coli (Gram-negative). S. kitaibelii EO showed the highest antimicrobial activity against both tested bacterial strains. This is the first study to characterize the EO composition and antimicrobial activity of these five medicinal species from Eastern Serbia in comparison with comprehensive literature data. The results can be utilized by the perfumery, cosmetics, food and pharmaceutical industries, but also for healing purposes in self-medication.

Hemp (Cannabis sativa L.) is currently one of the most controversial and promising crops. This study compared nine wild hemp (C. sativa spp. spontanea V.) accessions with 13 registered cultivars, eight breeding lines, and one cannabidiol (CBD) hemp strain belonging to C. sativa L. The first three groups had similar main essential oil (EO) constituents, but in different concentrations; the CBD hemp had a different EO profile. The concentration of the four major constituents in the industrial hemp lines and wild hemp accessions varied as follows: β-caryophyllene 11–22% and 15.4–29.6%; α-humulene 4.4–7.6% and 5.3–11.9%; caryophyllene oxide 8.6–13.7% and 0.2–31.2%; and humulene epoxide 2, 2.3–5.6% and 1.2–9.5%, respectively. The concentration of CBD in the EO of wild hemp varied from 6.9 to 52.4% of the total oil while CBD in the EO of the registered cultivars varied from 7.1 to 25%; CBD in the EO of the breeding lines and in the CBD strain varied from 6.4 to 25% and 7.4 to 8.8%, respectively. The concentrations of δ9-tetrahydrocannabinol (THC) in the EO of the three groups of hemp were significantly different, with the highest concentration being 3.5%. The EO of wild hemp had greater antimicrobial activity compared with the EO of registered cultivars. This is the first report to show that significant amounts of CBD could be accumulated in the EO of wild and registered cultivars of hemp following hydro-distillation. The amount of CBD in the EO can be greater than that in the EO of the USA strain used for commercial production of CBD. Furthermore, this is among the first reports that show greater antimicrobial activity of the EO of wild hemp vs. the EO of registered cultivars. The results suggest that wild hemp may offer an excellent opportunity for future breeding and the selection of cultivars with a desirable composition of the EO and possibly CBD-rich EO production.

Grecian juniper (Juniperus excelsa M. Bieb.) is an evergreen tree and a rare plant found in very few locations in southern Bulgaria. The aim of this study was to evaluate the phytochemical content and antioxidant potential of J. excelsa unripe and ripe galbuli from three different locations in Bulgaria. The essential oil content ranged between 1.9% and 5.1%, while the lipid fraction yield was between 4.5% and 9.1%. The content of total chlorophyll was 185.4–273.4 μg/g dw. The total carotenoid content ranged between 41.7 and 50.4 μg/g dw of ripe galbuli, and protein content was between 13.6% and 16.4%. Histidine (5.5 and 8.0 mg/g content range) and lysine (4.0 and 6.1 mg/g) were the major essential amino acids. The antioxidant potential of the 95% and 70% ethanol extracts was analyzed using four different methods. A positive correlation between the antioxidant potential and phenolic content of the galbuli was found. The results obtained in this study demonstrated the differences in phytochemical composition and antioxidant capacity of J. excelsa galbuli as a function of maturity stage and collection locality.

Industrial hemp (Cannabis sativa L., Cannabaceae) is an ancient cultivated plant originating from Central Asia and historically has been a multi-use crop valued for its fiber, food, and medicinal uses. Various oriental and Asian cultures kept records of its production and numerous uses. Due to the similarities between industrial hemp (fiber and grain) and the narcotic/medical type of Cannabis, the production of industrial hemp was prohibited in most countries, wiping out centuries of learning and genetic resources. In the past two decades, most countries have legalized industrial hemp production, prompting a significant amount of research on the health benefits of hemp and hemp products. Current research is yet to verify the various health claims of the numerous commercially available hemp products. Hence, this review aims to compile recent advances in the science of industrial hemp, with respect to its use as value-added functional food ingredients/nutraceuticals and health benefits, while also highlighting gaps in our current knowledge and avenues of future research on this high-value multi-use plant for the global food chain.

The hypothesis of this study was that we can modify the essential oil (EO) profile of hemp (Cannabis sativa L.) and obtain fractions with differential composition and antimicrobial activity. Therefore, the objective was to evaluate the effects of grinding of hemp biomass before EO extraction and fractionation during distillation on EO profile and antimicrobial activity. The study generated a several EO fractions with a diversity of chemical profile and antimicrobial activity. The highest concentrations of β-pinene and myrcene in the EO can be obtained in the 5–10 min distillation time (DT) of ground material or in the 80–120 min DT of nonground material. High δ-3-carene and limonene EO can be obtained from 0–5 min DT fraction of nonground material. High eucalyptol EO can be sampled either in the 0–5 min DT of the ground material or in the 80–120 min of nonground material. Overall, the highest concentrations of β-caryophyllene, α-(E)-bergamotene, (Z)-β-farnesene, α-humulene, caryophyllenyl alcohol, germacrene D-4-ol, spathulenol, caryophyllene oxide, humulene epoxide 2, β-bisabolol, α-bisabolol, sesquiterpenes, and cannabidiol (CBD) can be obtained when EO is sampled in the 80–120 min DT and the material is nonground. Monoterpenes in the hemp EO can be increased twofold to 85% by grinding the material prior to distillation and collecting the EO in the first 10 min. However, grinding resulted in a slight but significant decrease in the CBD concentration of the EO. CBD-rich oil can be produced by collecting at 120–180 min DT. Different EO fractions had differential antimicrobial activity. The highest antimicrobial activity of EO fraction was found against Staphylococcus aureus subsp. aureus. THC-free EO can be obtained if the EO distillation is limited to 120 min. The results can be utilized by the hemp processing industry and by companies developing new hemp EO-infused products, including perfumery, cosmetics, dietary supplements, food, and pharmaceutical industries.

The genus Nicotiana (Solanaceae) includes over 70 species, with a long history of traditional use; many of them are nowadays used in bioengineering, biosynthesis, molecular biology, and other studies, while common tobacco, N. tabacum L., is one of the most economically important industrial crops worldwide. Although Nicotiana species have been extensively investigated, relatively less research has focused on flowers, especially research related to obtaining aromatic products for cosmetic and perfumery use. On the other hand, there is evidence that Nicotiana flowers accumulate various secondary metabolites with a distinct aroma and biological activities, and the flowers represent a biomass available in sufficient quantities. Therefore, this study aimed to determinate the chemical composition (by GC-MS) and the olfactory profiles of a specific type of natural aromatic product (concrete), obtained from the flowers of four Nicotiana species, in a direct comparison between them. The yields of extracted concrete were sufficiently high, varying between the species, 1.4% (N. rustica L.), 2.5% (N. glutinosa L.), 1.6% (N. alata Link&Otto genotype with white flowers), 2.7% (N. alata genotype with pink flowers), 3.2% (N. tabacum, Oriental type), and 5.2% (N. tabacum, Virginia type). The major components of the obtained concretes belonged to different chemical classes: N. rustica and N. tabacum (OR), the hydrocarbons n-tetratriacontane (14.5%; 15.0%) and n-triacontane (12.1%; 13.3%), and 3-methyl-pentanoic acid (11.1%; 12.2%); N. glutinosa, the diterpenes sclareol (25.9%), 3-α-hydroxy-manool (16.3%), and 13-epimanool (14.9%); N. alata (WF), the phenylpropanoid terephthalic acid and di(2-ethylhexyl) ester (42.9%); N. alata (PF), the diterpene tributyl acetylcitrate (30.7%); and N. tabacum (FCV), the hydrocarbons n-hexacosane (12.9%) and n-pentacosane (12.9%). Each of the flower concretes revealed a characteristic odor profile. This is the first report about Nicotiana species as a source for obtaining flower concretes; these initial results about the concrete yield, olfactory profile, and chemical composition are a prerequisite for the possible processing of Nicotiana flowers into new aromatic products for use in perfumery and cosmetics. The study provides new data in favor of the potential of the four Nicotiana species as aromatic plants, as well as a possible alternative use of flowers, a valuable, but discarded, plant material in other applications.

For the first time the volatile fraction of coffee silverskin has been studied focusing on odor-active compounds detected by gas chromatography-olfactometry/flame ionization detector (GC-O/FID) system. Two approaches, namely headspace (HS) analysis by solid-phase microextraction-gas chromatography-mass spectrometry (SPME-GC-MS) and odor-active compounds analysis by gas chromatography-olfactometry/flame ionization detector (GC-O/FID), have been employed to fully characterize the aroma profile of this by-product. This work also provided an entire characterization of the bioactive compounds present in coffee silverskin, including alkaloids, chlorogenic acids, phenolic acids, flavonoids, and secoiridoids, by using different extraction procedures and high performance liquid chromatography-tandem mass spectrometry (HPLC-MS/MS) system. Coffee silverskin was shown to be a good source of caffeine and chlorogenic acids but also of phenolic acids and flavonoids. In addition, the fatty acid composition of the coffee silverskin was established by GC-FID system. The results from this research could contribute to the development of innovative applications and reuses of coffee silverskin, an interesting resource with a high potential to be tapped by the food and nutraceutical sector, and possibly also in the cosmetics and perfumery.

Studies on the impact of agricultural practices on plant diversity provide important information for policy makers and the conservation of the environment. The aim of the present work was to evaluate wild plant diversity across the agroecosystems in two contrasting regions of Bulgaria; Pazardzhik-Plovdiv (representing agroecosystems in the lowlands) and Western Stara Planina (the Balkan Mountains, representing agroecosystems in the foothills of the mountains). This study conducted a two-year assessment of plant diversity in different types of agricultural and forest ecosystems, representing more than 30 land use types. Plant diversity, measured by species number, was affected by the land use type only in Pazardzhik-Plovdiv region. More pronounced was the effect of the groups of land use types on the diversity, measured by the mean species number per scoring plot. Climatic conditions, measured by 19 bioclimatic variables, were the most important factor affecting plant species diversity. Six bioclimatic variables had a significant effect on the plant diversity, and the effect was more pronounced when the analysis considered pooled data of the two regions. The highest plant diversity was found on grazing land with sparse tree cover, while the lowest one was in the land use types representing annual crops or fallow. The study also established a database on weed species, relevant to agriculture. A number of common weeds were found in the Pazardzhik-Plovdiv region, while the most frequent species in the Western Stara Planina region were indigenous ones. Overall, the natural flora of Western Stara Planina was more conserved; eleven orchid species with conservation significance were found in the pastures and meadows in that region. The present study is the first attempt in Bulgaria to characterize the plant diversity across diverse agroecosystems representing many different land use types and environmental conditions. The results can contribute to nature conservation, biodiversity, and the sustainable use of plant resources.

The intent of this study was to utilize distillation timeframes (DT) of nutmeg (Myristica fragrans) essential oil (EO) to generate fractions with differential chemical compositions and bioactivity. Ten fractions were captured at the following distillation timeframes: 0.0−0.5, 0.5−1.0, 1.0−2.5, 2.5−5.0, 5.0−10, 10−30, 30−60, 60−90, 90−120, and 120−240 min. In addition, a control EO was collected from a straight 0−240 min non-stop distillation. ANOVA and advanced regression modeling revealed that the produced EO fractions possess substantial variation in the concentration of potentially desired compounds. The concentrations (%) of α-phellandrene, 3-carene, p-cymene, limonene, α-thujene, α-pinene, camphene, sabinene, β-pinene, and myrcene decreased, while the concentrations (%) of α-terpinene, γ-terpinene, terpinolene, and myristicin increased in later DT fractions. Nutmeg EO showed some antimalarial activity against Plasmodium falciparum D6, but did not exhibit significant antifungal activity. In general, nutmeg seed oil yields increased with an increase of DT. These results may be utilized by industries using nutmeg EO.

The aim of this study was to establish the kinetics regression models for yield and composition of Ruta graveolens fruit and Hyssopus officinalis subsp. aristatus aboveground biomass essential oil (EO), collected at different time intervals during the hydrodistillation process. The hypothesis was that collecting the EO fractions during specific time frames may result in EOs with dissimilar composition that may have differential use by the industry. Furthermore, we calculated the kinetics regression models for the composition of EO, isolated by hydrodistillation in a Clevenger-type apparatus and characterized by GC-MS and GC-FID analyses. The EO yield of R. graveolens fruits was 0.39% (relative area % of GC-FID chromatogram), with major constituents in the Control fraction (0−90 min) being 2-nonanone, 2-undecanone, and 2-undecanol, representing 65% of the total oil. The highest concentration of 2-nonanone (60%) was found in the 30−60 min oil fraction, the concentration of 2-undecanone (35%) was highest in the Control (0−90 min) fraction, and the concentration of eucalyptol (19%) was highest in the 5−10 min fraction. The EO yield of H. officinalis subsp. aristatus dried biomass was 1.12%. The major constituents in the Control fraction (0−90 min) of H. officinalis biomass were eucalyptol, α-pinene, sabinene, β-pinene, and cis-3-pinanone, representing 86% of the total. Eucalyptol (58%) was the highest in the 0−5 min fraction. The highest β-pinene (15%) and cis-3-pinanone (20%) contents were found in the 20−40 min fraction. The kinetics regression models that were developed for EO composition of R. graveolens were second-order polynominal, Michaelis−Menten, and Exponential decay, while for EO composition of H. officinalis subsp. aristatus biomass were Exponential decay and Power. The results from this study could benefit the EO industry.

Currently, wastewater treatment plants produce large amounts of sewage sludge. Due to the rich content of organic matter and minerals, sewage sludge can be used as soil amendments for eroded soils. The aim of this work was to assess sewage sludge (SS) in combination with an eroded soil (ES) collected from the North Eastern Romania as growth substrate for sweet basil, and their effect on basil growth and physiology. The experiment was conducted in a greenhouse under controlled environment conditions. The tested substrates were: (1) eroded soil, ES; (2) mixture of eroded soil (15%) + sewage sludge, ES + SS (85%); and (3) sewage sludge, SS (100%). Three types of parameters were studied: morphological traits, physiological, and biochemical parameters. The maximum quantum yield of Photosystem II Fv/Fm was reduced in basil leaves grown on eroded soil (0.80) and was close to the normal value in ES + SS (0.83). Chlorophyll a and the carotenoids content were higher for plants grown on SS and significantly higher for those grown in ES + SS compared with the one of plants grown on ES. The fresh biomass yield and height of basil increased with 44% and 34.5% under ES + SS over ES. Total phenolic content was higher in plants grown on ES (7.34 mg/g dry weight Gallic acid equivalent), which also led to an increased antioxidant activity (44.4%) evaluated by the DPPH (2,2-diphenyl-1-picrylhydrazyl) method. Fourier-Transform Infrared (FT-IR) (4000–400 cm−1) spectra of basil did not show significant qualitative differences among the plants from different treatments. The results of this study demonstrated that SS application led to the improvement of the basil morpho-physiological parameters, allowing the growth of basil on ES + SS.

Tobacco (Nicotiana tabacum L.) aroma is an important attribute of tobacco quality and is influenced by a variety of minor chemical components, including carotenoid degradation products. The objectives of this work were to determine the content of the most important fragrance-shaping carotenoid degradation products in the essential oils (EOs) of the three types of Bulgarian tobacco—Oriental (OR), flue-cured Virginia (FCV), and Burley (BU)—and to compare them with other aromatic products from tobacco. The content of total carotenoids and β-carotene was highest in BU tobacco (22.23 and 20.34 mg/100 g DW, respectively), followed by OR (13.60 and 12.09 mg/100 g DW in variety “Plovdiv 7” (Pd7); 6.27 and 5.45 mg/100 g DW in “Krumovgrad” (Kr), and FCV (5.93 and 3.73 mg/100 g DW). EOs were obtained by hydrodistillation in an acidified medium, and the main aroma-impact compounds from carotenoid degradation (identified by GC-MS) were as follows: FCV-α-ionone (0.61 mg/100 g DW), dihydro-β-ionone (0.96 mg/100 g DW), β-damascenone (1.26 mg/100 g DW); BU-α-ionone (0.73 mg/100 g DW), dihydro-β-ionone (1.19 mg/100 g DW), β-damascenone (1.35 mg/100 g DW); OR(Kr)-α-ionone (0.20 mg/100 g DW), β-ionone (1.08 mg/100 g DW), dihydro-β-ionone (1.34 mg/100 g DW), β-damascenone (0.36 mg/100 g DW); OR(Pd7)-α-ionone (1.43 mg/100 g DW), dihydro-β-ionone (1.73 mg/100 g DW), β-damascenone (1.23 mg/100 g DW). Ionone and its derivatives were not identified in the aroma extraction products concrete, resinoid, or absolute. The results suggest that temperature, pH of the medium, process duration, and possibly other unknown factors affect carotenoid transformation. The study provides insight into the composition of tobacco EOs and may be of interest to the fragrance industry.

Crop residues are an important source of plant nutrients. However, information on the various methods of residue management on micronutrients in soil and wheat (Triticum aestivum L.) over time is limited. A long-term (84-year) agroecosystem experiment was assessed to determine the impact of fertilizer type and methods of crop residue management on micronutrients over time under dryland winter wheat-fallow rotation. The treatments were: no N application with residue burning in fall (FB), spring (SB), and no residue burn (NB); 45 kg N ha−1 with SB and NB; 90 kg N ha−1 with SB and NB; pea vines; and farmyard manure (FYM) and a nearby undisturbed grass pasture (GP). Wheat grain, straw, and soil samples from 1995, 2005, and 2015 were used to determine tissue total and soil Mehlich III extractable Mn, Cu, B, Fe, and Zn, and soil pH. After 84 years, extractable Mn and B in the top 10 cm of soil decreased in all plots, except for B in FYM and SB. The FYM plots had the highest extractable Mn (114 mg kg−1) in the top 10 cm soil; however, it declined by 33% compared to the GP (171 mg kg−1). Extractable Zn in the top 10 cm of soil increased with FYM while it decreased with inorganic N application in 2015; however, total Zn in grain increased by 7% with inorganic N (90 kg ha−1) application compared to FYM application. The results suggest that residue management had similar impact on soil micronutrients. Inorganic N and FYM application can be integrated to reduce micronutrient losses from cultivation.

Tillage plays a major role in nutrient dynamics under dryland cropping systems, but there remains uncertainty regarding the long-term impacts of tillage on nutrient availability. The objective of this study was to examine the influence of tillage intensity and timing on micronutrient concentration of soils and winter wheat (Triticum aestivum L.) under dryland winter wheat−pea (Pisum sativum L.) or WW-P rotation. The treatments included moldboard tillage in fall (FT) and spring (ST), disk/chisel tillage (DT), and no-tillage (NT). The concentrations of Mehlich III extractable boron, manganese, zinc, copper, and iron in soil were unaffected by the tillage methods; however, a significant decline in extractable zinc in the top 10 cm soil was observed compared to an adjacent undisturbed grass pasture (GP) (NT: 2.3 mg kg−1 vs. GP: 6.0 mg kg−1). In the upper 10 cm soil surface, NT (123 mg kg−1) maintained the extractable manganese concentration with GP (175 mg kg−1) whereas FT (97 mg kg−1), ST (92 mg kg−1), and DT (113 mg kg−1) had lower manganese than GP. Soil pH declined in the upper 10 cm under NT more than in the rest of the WW-P treatments. The results suggest NT can play a vital role in sustaining micronutrient availability due to decreased soil pH and the greater amount of organic matter within the surface soil of NT compared to other tillage methods.

Coal-bed methane (CBM), an important energy source, coproduces a vast amount of saline-sodic wastewater, CBM water (CBMW), with environmental and economic disposal issues. This research evaluated under a greenhouse production system the influence of gypsum (CaSO4·2H2O) and CBMW on yields, essential oil (EO) content and composition in lemongrass (Cymbopogon flexuosus) and palmarosa (C. martinii), and on growth medium pH and available nutrients. CBMW treatments had higher pH than tap water treatment. Gypsum reduced pH in all CBMW treatments but did not affect pH in tap water treatment. While CBMW may increase the available Cu and Fe in growth medium, the application of gypsum may negate this effect. CBMW significantly increased growth medium Na. Gypsum increased growth medium S, and CBMW increased S in the high gypsum treatments. Palmarosa height, fresh weight, geranyl acetate, and isoneral in lemongrass EO were reduced, while geraniol in palmarosa EO increased with CBMW relative to tap water. In distillation waste plant tissue, CBMW increased Na in lemongrass and palmarosa by almost eight times; increased total P, S, and Mn in palmarosa; and reduced total N, S, Ca, and Mg in lemongrass and Ca in palmarosa, relative to tap water. This study demonstrated that CBMW may be used for greenhouse production of high-value crops, but it may affect the yields and oil content of some crops and growth medium characteristics.

The insights gained from the long-term impacts of tillage and N fertilization on soil fertility are crucial for the development of sustainable cropping systems. The objectives of this study were to quantify the effects of 75 years of tillage and N fertilization on macronutrients in soil and wheat (Triticum aestivum L.) tissues grown in a winter wheat–summer fallow rotation. The experiment included three types of tillage (disc, DP; sweep, SW; and moldboard, MP) and five N application rates (0, 45, 90, 135, and 180 kg ha−1). Soil and tissue samples were analyzed for the concentration of total N, S, and C, Mehlich III extractable P, K, Mg, Ca in the soil, and the total concentration of the same nutrients in wheat tissue. Soil N concentration was significantly greater under DP (1.10 g kg−1) than under MP (1.03 g kg−1). The P concentration in upper 20 cm soil depth increased with increased N rates. Comparison of experiment plots to a nearby undisturbed pasture revealed a decline of P (32%), SOC (34%), Mg (77%), and Ca (86%) in the top 10 cm soil depth. The results suggest that DP with high N rates could reduce the macronutrient decline in soil and plant over time.

The chemical profile and antioxidant capacity of Juniperus virginiana, J. excelsa, and J. sabina essential oil (EO) fractions as a function of time was the subject of this study. The hypothesis was that, capturing EO in sequential timeframes during hydrodistillation would generate fractions containing unique compositions and antioxidant capacity. In J. virginiana, the highest limonene (43%) was found in the 0–5 min oil fraction, with safrole (37%) being highest in the 10–20 and 20–40 min fractions, and elemol (34%) being highest in the 160–240 min fraction. In J. excelsa, α-pinene (34-36%) was the highest in the 0–5 min fraction and in the control (non-stop 0–240 min distillation) oil, limonene (39%) was the highest in the 0–10 min fractions and cedrol (50-53%) was the highest in the 40–240 min fractions. In J. sabina, sabinene (80%) was highest in the 0–3 min fraction. The highest antioxidant capacity of J. virginiana was demonstrated by the 5–10 min fraction; the one in J. sabina by the 3–10 min fraction; and, the one in J. excelsa, by the control. The kinetics regression models that were developed can predict EO composition of the three juniper species eluted at different timeframes. Various industries could benefit from the results from this study.