Floral Scent and Pollinators of Cypripedium calceolus L. at Different Latitudes

Floral scent is an important trait in plant–pollinator interactions. It not only varies among plant species but also among populations within species. Such variability might be caused by various non–selective factors, or, as has been shown in some instances, might be the result of divergent selective pressures exerted by variable pollinator climates. Cypripedium calceolus is a Eurasian deceptive orchid pollinated mainly by bees, which spans wide altitudinal and latitudinal gradients in mainly quite isolated populations. In the present study, we investigated whether pollinators and floral scents vary among different latitudes. Floral scents of three C. calceolus populations in the Southern Alps were collected by dynamic headspace and analyzed by gas chromatography coupled to mass spectrometry (GC/MS). These data were completed by previously published scent data of the Northern Alps and Scandinavia. The scent characteristics were compared with information on pollinators recorded for present study or available in the literature. More than 80 scent compounds were overall recorded from plants of the three regions, mainly aliphatics, terpenoids, and aromatics. Seven compounds were found in all samples, and most samples were dominated by linalool and octyl acetate. Although scents differed among regions and populations, the main compounds were similar among regions. Andrena and Lasioglossum species were the main pollinators in all three regions, with Andrena being relatively more abundant than Lasioglossum in Scandinavia. We discuss natural selection mediated by pollinators and negative frequency–dependent selection as possible reasons for the identified variation of floral scent within and among populations and regions.

Guide to Harvesting Heracleum Persicum: How Essential Oil Composition and Phenolic Acid Profile Fluctuate at Different Phenological Stages?

BACKGROUN: Heracleum persicum commonly named Golpar, is a principal native medicinal plant in Iran. Collecting H. persicum at the appropriate growing stage is the key factor to achieve the high phytochemical quality to meet consumer’s needs. In the present experiment, the aerial parts of this plant were harvested at up to six different developmental stages during the growing season to determine the phytochemical profiles. RESULTS: Our results indicated that the highest essential oil yield and extract were obtained in the mid-mature seed stage (3.5%),) and floral budding stage (10.40%). In the vegetative stage, limonene (18.05), in floral budding stage, caryophyllene (14.07), anethole (14.55%), and β-bisabolene (12.56%), in the full flowering stage, myristicin (15.02%), and hexyl butyrate (9.05%); in the early development of seeds stage, hexyl butyrate (32.08%), and octyl acetate (11.67%); in the mid-mature seeds stage hexyl butyrate (38.75%), octyl acetate (14.47%); in the late-mature/ripe seeds stage, hexyl butyrate (23.59%), and octyl acetate (10.48%) recorded as the main components. The analysis of phenolic acids demonstrated cinnamic acid, p-coumaric acid, p-hydroxybenzoic acid, ferulic acid, and rosmarinic acid as the main phenolic acids. The highest phenolic acids content was obtained in the floral budding stage (287.40 mg g –1 dry extract). Cinnamic acid was found as the major phenolic compound in the vegetative stage following by floral budding, the full flowering stage, the early development of seeds and late-mature/ripe seeds stages. P-coumaric acid was the most abundant phenolic compounds in the mid-mature seeds stage. The development stage has a significant impact on the content and composition of both essential oil and phenolic acid composition. CONCLUSION: In this regard, the harvest time of H. persicum aerial parts can be selected to achieve the highest secondary metabolites of interest. The results of this study can be used as a guideline for grower to obtain the highest amount of desirable metabolites, beneficial in the food and pharmaceutical industries as well as economic benefits.

Comparison of the Effects of Silver in Nanostructured and Ultrahigh Diluted Form on Growth and Volatile Compounds Produced by Escherichia coli and Staphylococcus aureus

Introduction: In this project, the growth and volatile metabolites profiles of Escherichia coli (E. coli ) and Staphylococcus aureus were monitored under the influence of silver base chemical, nanoparticle and ultra-highly diluted compounds. Materials & Methods: The treatments were done for 12000 life cycles using silver nanoparticles (AgNPs) as well as ultra-highly diluted Argentum nitricum (Arg-n). Volatile organic metabolites analysis was performed using gas chromatography mass spectrometry (GC-MS). The results indicated that AgNPs treatment made the bacteria resistant and adapted to growth in the nanoparticle condition. The use of ultra-highly diluted Arg-n initially increased growth but it decreased later. Also, with the continuous usage of these materials, no more bacterial growth was observed. Results: The most important compounds produced by E. coli are Acetophenone, Octyl acetate, Styrene, 1,8-cineole, 4-t-butyl-2-(1-methyl-2-nitroethyl)cyclohexane, hexadecane and 2-Undecanol. The main compounds derived from S. aureus are Acetophenone,1,8-cineole, Benzaldehyde, 2-Hexan-1-ol, Tridecanol, Dimethyl Octenal and tetradecane. Acetophenone and 1,8-cineole were common and produced by both organisms. Conclusion: Based on the origin of the produced volatiles, main volatiles percentage of untreated sample is hydrocarbon (>50%), while bacteria treatments convert the ratio in to aldehydes, ketones and alcohols in the case of AgNPs, (>80%) and aldehydes, ketones and terpenes in the case of Arg-n (>70%).

ANTIMICROBIAL AND ANTIRADICAL ACTIVITY OF INDIVIDUAL FRACTIONS OF ESSENTIAL OIL FROM SEEDS OF HERACLEUM DISSECTUM LEDEB. OF SIBERIAN REGION

By the method of exhaustive hydroponically obtained essential oil from beans of Heracleum dissectum Ledeb., growing in the Krasnoyarsk region. Separate fractions of oil were obtained: the first after 45 minutes from the beginning of distillation, the second – after 2 hours, the third-after 5 hours, the fourth fraction was collected after the end of hydro-distillation. The component composition of both whole essential oil and its separate fractions was studied. The main components are octyl acetate (60.0%), octyl-2-methylpropanoate (10.2%), n-hexyl-2-methylbutanoate (9.0%). The main amount of octyl acetate (64.7%) is concentrated in the first fraction of the oil. The antimicrobial activity of various fractions of essential oil of borscht dissected against strains of opportunistic microorganisms: Escherichia coli, Pseudomonas aeruginosa, Klebsiella pneumoniae, Staphylococcus aureus 209p, MRSA, Proteus vulgaris. It was found that, depending on the duration of isolation, the antimicrobial activity of essential oil fractions in relation to Staphylococcus aureus 209p, MRSA and Pseudomonas aeruginosa decreases, and in relation to Escherichia coli, Klebsiella pneumoniae and Proteus vulgaris increases. The most pronounced inhibitory effect of the third and fourth fractions of essential oil against Klebsiella pneumonia. The antiradical activity of all studied samples of borscht essential oil dissected in reaction with stable free 2,2-diphenyl-1-picrylhydrazyl radical was established. The first fraction showed minimal antiradical activity (15.1%), the fourth – maximum (49.2%).

Purification and Properties of an Esterase from Bacillus licheniformis and it’s Application in Synthesis of Octyl Acetate

Background: Esterase plays a major role in the degradation of natural materials, industrial pollutants and also provides an immense contribution to the eco-friendly approaches in various industrial applications. Objective: In the present study, extracellular esterase from bacterial isolate Bacillus licheniformis was purified, characterized and used in the synthesis of octyl acetate. Methods: Purification of esterase from Bacillus licheniformis was achieved using Sephadex G-75 column chromatography. Gas chromatography was used to analyze the octyl acetate synthesis. Results: The enzyme was salted out using ammonium sulphate precipitation and 60-70% saturation gave maximum specific activity of the enzyme during precipitation. A purification fold of 6.46 and yield of 9.69% was achieved when esterase from Bacillus licheniformis was purified using Sephadex G-75 column chromatography. Native as well as SDS-PAGE analysis gave a single band of 42 kDa. This showed that the enzyme was purified to homogeneity and it was a monomer with molecular weight of 42 kDa. Biochemical characterization of the enzyme revealed that it had optimum temperature of 45°C in 0.1 M Tris-HCl buffer of pH 8.0. On optimizing different parameters, such as molar ratio of reactants, incubation time, temperature, and amount of protein, the % yield of octyl acetate was found to be 77.3%. Conclusion: In this work, simple method was used to purify esterase and the enzyme was further used in producing esters/products of commercial value within a reasonably short period of 12 h with a maximum yield of 77.3%.

Protective Effect of Boswellic Resin Against Memory Loss and Alzheimer’s Induced by Aluminum Tetrachloride and D-Galactose (Experimental study in Mice)

Nowadays, because of the industrialization, a lot of contaminant were available ; the consequences of this availability are apparition of diseases including neurodegeneration. Neurodegenerative diseases of the human brain comprise a variety of disorders that affect an increasing percentage of the population. This study is based on the effect of the Boswellic resin, which is from a medicinal plant and known for its antioxidant effects on nerve cell damage. The objective of this work was to evaluate the in vitro and in vivo effects of the Boswellic resin on anticholinesterase activity and Alzheimer’s disease (AD) induced by D-galactose and aluminum tetrachloride in Swiss mice. Chemical composition of the resin essential oil was identified by the CG-MS analysis. The antioxidant activity was also assessed by the DMPD and metal chelation methods. In order to understand the mechanism of memory improvement, the acetylcholinesterase, AChE, and butyrylcholinesterase, BChE, inhibitory assays were performed. In vivo part of the study was achieved on Swiss mice divided into four groups: control, AD model, treated AD, and treated control group. The identification of chemical composition by CG-MS reach the 89.67% of the total extract compounds presented some very important molecules (p-Cymene, n-Octyl acetate, α-Pinene…). The present study proves that Boswellic resin improves memory and learning in treated Alzheimer’s group, modulates the oxidative stress and be involved in the protective effect against amyloid deposition and neurodegeneration, and stimulates the immune system in mice’s brain.