Impact of silver nanoparticles on secondary metabolite composition and toxicity in anise (Pimpinella anisum L.) callus culture.

BACKGROUND: There are numerous challenges associated with producing desired amounts of secondary metabolites (SMs), which are mostly unique and cannot be chemically synthesized. Many studies indicate that nanoparticles (NPs) can boost the production of SMs. Still, the precise manner in which NPs induce metabolic changes remains unidentified. This study examines the influence of eco-friendly silver NPs (AgNPs) on the chemical makeup and toxicity of Pimpinella anisum L. (anise). RESULTS: AgNPs were introduced into anise callus cultures at different concentrations (0, 1.0, 5.0, 10, and 20 mg/L). The induced oxidative stress was tracked over intervals of 7, 14, 28, and 35 days. Chemical composition evaluations were carried out on the 35th day. Within the first 14 days, plant stress was evident, though the plant adapted to the stress later on. Notably, the plant showed high tolerance at 1 mg/L and 5 mg/L concentrations despite increased toxicity levels. However, relatively high toxicity levels were identified at 10 and 20 mg/L. The AgNP-induced stress significantly impacted anise SMs, particularly affecting fatty acid content. In the 10 and 20 mg/L AgNP groups, essential metabolites, including palmitic and linoleic acid, showed a significant increase. Polyunsaturated (omega) and monounsaturated fatty acids, vital for the food and pharmaceutical industries, saw substantial growth in the 1 and 5 mg/L AgNP groups. For the first time, vanillyl alcohol and 4-Hydroxybenzoic acid were detected along with various phenolic compounds, such as t-anethole, Salicylic acid, and Thiamazole. CONCLUSION: AgNPs can function as an elicitor to efficiently generate essential SMs such as omegas and phenolic compounds in anise callus culture. This study explores the application of AgNPs as plant elicitors in anise SM production, offering invaluable insight into potential uses.

Water deficit stress changes in physiological, biochemical and antioxidant characteristics of anise (Pimpinella anisum L.).

This study was designed to evaluate the impact of water deficit stress on the seed yield and its components, physiological functions, fatty acid content and compositions, essential oil (EO) content and compositions, phenolic acids and flavonoids amounts, and antioxidant activities of anise seeds. Plants evaluations were performed under well-watered (WW), moderate water deficit stressed (MWDS), and severe water deficit stressed (SWDS). The results revealed that SWDS significantly reduced seed yield, branch number per plant, seed number, umbel number, and thousand seed weight. Water deficit stress also caused a decrease in chlorophyll content, relative water content, quantum efficiency of photosystem II, and cell membrane stability, while increasing leaf temperature. The analysis of fatty acid composition indicated that petroselinic acid was the main fatty acid and its percentage increased by 8.75% and 14.60% under MWDS and SWDS, respectively. Furthermore, MWDS increased the EO content by 1.48 times, while it decreased by 41.32% under SWDS. The chemotype of EO was altered from t-anethole/estragole in WW seeds to t-anethole/ß-bisabolene in treated seeds. Higher levels of total phenolics were detected in stressed seeds. Water deficit stress increased the amount of the major class, naringin, by 1.40 and 1.26 times under MWDS and SWDS. The evaluation of antioxidant activity through reducing power, DPPH, and chelating ability assays indicated that stressed seeds exhibited the highest activity. The study's findings suggest that the application of drought stress before harvesting can regulate the production of bioactive compounds, which can affect the industrial and nutritional values of anise seeds.

Exploring the quantitative genetics of traits associated with drought tolerance and yield in Pimpinella anisum L. under different water regimes.

MAIN CONCLUSION: Genetic improvement of seed yield and drought resistance could be simultaneously gained in anise when breeding for drought resistance. Improving the water use efficiency of anise is a primary objective of anise breeding programs aimed at mitigating the impacts of drought stress. This study aimed to determine the predominant mechanisms involved in drought tolerance and investigate the genetic control of associated traits with drought tolerance and higher grain yield. According to these aims, 10 half-diallel hybrids and their five parents were evaluated in both field and greenhouse lysimetric experiments under well-watered and water deficit stress conditions. The results indicated that the inheritance of grain yield is complex and affected by water deficit stress. Similar heritability and genetic architecture were detected for flowering time and percentages of photosynthate partitioned to grain (PPPG) in both well-watered and water deficit stress treatments. Significant negative genetic correlations were observed between grain yield and flowering time, root dry mass, root diameter, root volume, root number, percentages of photosynthate partitioned to shoot, and percentages of photosynthate partitioned to root. Therefore, the selection of low values of these attributes can be used to improve grain yield under drought conditions. In contrast, a positive significant genetic linkage between grain yield and PPPG, chlorophyll content, cell membrane stability, and leaf relative water content reveal selection for high values of these attributes is favored. These attributes could be used as surrogate selection criteria in the early segregating generations. The P1 parent (early ripening parent) contained key genes associated with PPPG and drought escape. It was concluded that improvement of drought tolerance and grain yield could be simultaneously achieved in anise breeding programs.

Ultrasound-assisted preparation of anise extract nanoemulsion and its bioactivity against different pathogenic bacteria.

The present work aimed to characterize the nanoemulsion of anise seed extract and to compare its efficacy with the bulk extract against pathogenic bacteria. The anise seeds extract was prepared by cold solvent extraction method using ethanol. Nanoemulsion of anise extract was formulated using ultrasound assisted method and analyzed using high-performance liquid chromatography (HPLC), fourier transform infrared spectroscopy (FTIR) and UV-visible spectrophotometry. The antimicrobial activity of the nanoemulsion was tested against seven foodborne pathogenic bacterial species. Results showed that the extract contained anethole (37%), naringenin (21%), and taxifolin (13%) as the major phytochemical components. The average droplet size of the nanoemulsion droplets was measured by dynamic light scattering (DLS) and confirmed by transmission electron microscope to be about 400 nm. Anise extract nanoemulsion showed higher antimicrobial activity against most of the tested pathogens. Anise extract nanoemulsion performed better than bulk extract as an antimicrobial agent against some foodborne pathogenic bacteria.

Composition, antibacterial and antioxidant activities of Pimpinella saxifraga essential oil and application to cheese preservation as coating additive.

The effect of Pimpinella saxifraga essential oil (PSEO) addition (1-3%) in sodium alginate coating on the bacterial and oxidative stability of cheese was studied during refrigerated storage. The GC-HRMS analysis of PSEO showed that anethole, pseudoisoeugenol and p-anisaldehyde were the main components. The PSEO exhibited strong in vitro DPPH radical scavenging activity (IC50 = 6.81 µg/mL), ß-carotene bleaching inhibition (IC50 = 206 µg/mL), ferric reducing power (EC50 = 35.20 µg/mL), total antioxidant activity (213.96 ±â€¯11.12 µmol/mL α-tocopherol equivalent) and notable DNA protection potential. Additionally, PSEO displayed potent antibacterial activity against 3 Gram-positive and 3 Gram-negative bacteria (MICs = 0.78-3.12 mg/mL). The acute toxicity of PSEO was determined using mice model (LD50 = 976.2 mg/kg). The enrichment of sodium alginate coating with PSEO, particularly at 3%, improved cheese preservation by reducing the weight loss, preserving the pH and color and enhancing oxidative and bacterial stability without unpleased flavor for consumers.

Extracellular chromone derivatives in cell cultures of Pimpinella anisum. Influence of elicitation with methyl jasmonate and 2ß-methyl cyclodextrins.

OBJECTIVES: To explore the potentiality of undifferentiated Pimpinella anisum L. cell cultures for the production of secondary metabolites by means of elicitation. RESULTS: Two chromone compounds were secreted to the medium of undifferentiated cultures of P. anisum: 4-methoxyfuro[3,2-g]chromen-7-one, known as bergapten, which is constitutive to anise, and 5-hydroxy-7-methoxy-2-methylchromen-4-one, the rare chromone eugenin, not yet described in P. anisum. Caffeoyl quinic acid species were also identified in the biomass. Elicitation with methyl jasmonate enhanced chromone accumulation in the medium and stimulated phenolic acid metabolism in the biomass (11 mg caffeoyl quinic acids g-1 DW cells). The application of 2,6-dimethyl-ß-cyclodextrins to cultures led to an intense accumulation of chromones, with nearly 10 mg l-1 bergapten and 150 mg l-1 eugenin being accumulated extracellularly after optimal elicitation conditions. CONCLUSIONS: The significant amounts of eugenin obtained in the anise cultures and the stability of production over long periods of time can be of interest for its biotechnological production and for future studies on biosynthesis regulation.

Characterization and Influence of Green Synthesis of Nano-Sized Zinc Complex with 5-Aminolevulinic Acid on Bioactive Compounds of Aniseed.

A new water soluble zinc-aminolevulinic acid nano complex (n[Zn(ALA)2 ]), which was characterized by TEM, IR, and EDX spectra, has been prepared via sonochemical method under green conditions in water. In the current study, the effectiveness of foliar Zn amendment using synthetic Zn-ALA nano complex, as a new introduced Zn-fertilizer here, was evaluated. As the model plant, Pimpinella anisum, the most valuable spice and medicinal plant grown in warm regions, was used. By using zinc nano complex, further twenty compounds were obtained in the essential oil of anise plants. Application of 0.2% (w/v) Zn-ALA nano complex increased the levels of (E)-anethole, ß-bisabolene, germacrene D, methyl chavicol, and α-zingiberene in the essential oil. Nano Zn complex at the rate of 0.2% induced considerable high phenolic compounds and zinc content of shoots and seeds. Chlorogenic acid had the highest level between four detected phenolic compounds. The maximum antioxidant activity was monitored through the application of Zn nano complex. According to the results, nanoscale nutrients can be provided with further decreased doses for medicinal plants. Using Zn-ALA nano complex is a new and efficient method to improve the pharmaceutical and food properties of anise plants.

Antioxidant activity, polyphenolic contents and essential oil composition of Pimpinella anisum L. as affected by zinc fertilizer.

BACKGROUND: The antioxidant activity and essential oil content of plants may vary considerably with respect to environmental conditions, especially nutrient availability. Among micronutrients, zinc (Zn) is needed by plants in only small amounts but is crucial to plant development. This study aimed to evaluate the effects of Zn fertilization on the antioxidant activity, polyphenolic contents and essential oil composition of Pimpinella anisum fruit. RESULTS: Foliar application of Zn fertilizer considerably increased the number of detected essential oil components from 27 to 45. Zinc application at a rate of 0.2% (w/v) significantly enhanced the levels of ß-bisabolene, germacrene D, n-decane and α-zingiberene, whereas the opposite trend was observed for (E)-anethole and geijerene. Application of 0.2% Zn considerably increased the levels of phenolic compounds, with chlorogenic acid showing the highest content among eight phenolic compounds detected in treated plants. The maximum antioxidant activity was achieved through application of 0.2% Zn fertilizer. CONCLUSION: These findings indicated that the quality and quantity of anise fruit essential oil components were significantly altered by application of low levels of Zn. After foliar application of Zn, polyphenolic contents as well as antioxidant activity of anise fruit increased. Using Zn fertilizer is an efficient method to improve the pharmaceutical and food properties of anise fruit. © 2017 Society of Chemical Industry.

Seed-litter-position drives seedling establishment in grassland species under recurrent drought.

Changes in land use and climate interfere with grassland ecosystem processes. Here I experimentally investigated the combined effects of land-use change related litter cover and contrasting water supply on seedling emergence. In this context, the role of the initial relative position of seeds, i.e. seeds on top of the litter versus seeds beneath the litter in interaction with water supply has not been investigated so far. I hypothesised that facilitative effects of litter on seedling emergence occur when seeds are covered by litter and deteriorate when litter covers the ground and seeds fall on it (seeds on top of the litter). Further, I hypothesised that the importance of seed position for seedling emergence will increase under conditions of recurrent drought. I performed a controlled pot experiment on seedling emergence of three common European grassland species (Pimpinella saxifraga, Leontodon autumnalis, Sanguisorba officinalis) by experimental manipulations of litter and water availability. Seedling emergence under moist conditions showed no significant differences between each litter position compared to the control across species. In contrast, under recurrent drought, seedling emergence was significantly higher below the litter compared to seeds on top of the litter and the control (i.e. no litter). In abandoned land, seedling emergence may be limited when seeds fall on ground-covering litter. In contrast, in grasslands with regular low-intensity land use, seedling emergence may be enhanced when a moderate level of litter covers seeds at the end of the growing season. Protective mechanisms that occur with seeds positioned beneath litter are particularly important under recurrent drought.

Effect of gamma irradiation on microbial load and sensory characteristics of aniseed (Pimpinella anisum).

Seeds of anise (Pimpinella anisum) were exposed to doses of 0, 5, 10, 15 and 20kGy in a (60)Co package irradiator. Irradiated and unirradiated samples were stored at room temperature. Microbial populations on seeds, total and inorganic soluble solids in water extract and sensory properties of the latter were evaluated after 0, 6 and 12 months of storage. Results indicated that gamma irradiation reduced the aerobic plate counts of aniseed. Immediately after irradiation, the total soluble solids in an extract of irradiated seeds were greater than those of unirradiated ones. The total soluble solids in an extract of irradiated and un-irradiated seeds increased after 6 and 12 months of storage. There were no significant differences (p>0.05) in inorganic soluble solids between the water extract of irradiated and unirradiated aniseeds. Sensory evaluation indicated that gamma irradiation improved sensory characteristics of aniseed water extract tested immediately after irradiation; however, after 12 months of storage, no significant differences (p>0.05) were found in color, taste or flavor between extract of irradiated and unirradiated seeds.