Cyanobacterial elicitor enhances the biomass of Mentha piperita L. and improves the production of high-value rosmarinic acid under in vitro culture of apical meristem.

BACKGROUND: Rosmarinic acid (RA), like other phenolic compounds, is sources of antioxidants and anti-inflammatory agents in medicinal plants. In vitro culture of plants can improve the medicinal plants' metabolite profile and phenolic compound quantity. To date, various methods have been proposed to increase this medicinal metabolite in plants, among which the use of bioelicitors can be mentioned. In the present study, a native isolate of heterocystous cyanobacteria, Nostoc spongiaeforme var. tenue ISB65, was used to stimulate the production of biomass and content of RA in Mentha piperita L. (peppermint) grown in vitro from apical meristem. Mentha piperita L. explants were inoculated in half strength Murashige and Skoog (1/2 MS) medium containing cyanobacterial lysate (CL). After 50 days of culturing, the growth indices, the content of photosynthetic pigments, and RA in control and treated plants were measured. RESULTS: CL inoculation resulted in a significant enhancement in the vegetative growth indices of peppermint, including root and shoot length, plant biomass and leaf number. The content of photosynthetic pigments also increased in cyanobacteria-treated plants. Inoculation with CL increased the RA content by 2.3-fold, meaning that the plants treated with CL had the highest RA content (7.68 mg. g- 1 dry weight) compared to the control (3.42 mg. g- 1 dry weight). Additionally, HPLC analysis revealed the presence of several auxins in CL. CONCLUSIONS: The presence of auxins and the chemical content of CL such as K+ and Ca2+, as regulators of metabolic pathways and molecular activities of cells, may be responsible for the enhanced growth and phenolic compounds of plants under tissue culture conditions. An improvement in RA content in the tissue culture of medicinal plants treated with CL was reported for the first time in this investigation.

Application of insecticides on peppermint (Mentha × piperita L.) induces lignin accumulation in leaves by consuming phenolic acids and thus potentially deteriorates quality.

Irrational use of pesticides may lead to physiological and metabolic disorders in different crops. However, there are limited investigations on impacts of insecticides on physiology and biochemistry, secondary metabolic pathways, and associated quality of medicinal plants such as peppermint (Mentha × piperita L.). In this study, target metabolites in peppermint were monitored following foliar spraying of five insecticides: imidacloprid, pyriproxyfen, acetamiprid, chlorantraniliprole, and chlorfenapyr. Compared with the control, all insecticide treatments caused a significant loss of soluble protein (decreased by 22.3-38.7%) in peppermint leaves. Insecticides induced an increase in the levels of phytohormones jasmonic acid and abscisic acid in response to these chemical stresses. Among them, imidacloprid increased jasmonic acid by 388.3%, and pyriproxyfen increased abscisic acid by 98.8%. The contents of phenylpropanoid metabolites, including rutin, quercetin, apigenin, caffeic acid, 4-hydroxybenzoic acid, ferulic acid, syringic acid, and sinapic acid showed a decreasing trend, with pyriproxyfen decreasing the levels of quercetin and 4-hydroxybenzoic acid by 78.8% and 72.6%, respectively. Combined with correlation analysis, the content of lignin in leaves shows different degrees of negative correlations with several phenolic acids. It could be inferred that insecticides may trigger plant defense mechanisms that accumulate lignin (increased by 24.6-49.1%) in leaves by consuming phenolic acids to barricade absorption of insecticides. Through constructing networks between phytohormones and secondary metabolites, peppermint may regulate the contents of caffeic acid, 4-hydroxybenzoic acid, and sinapic acid by the antagonistic effect between salicylic acid and abscisic acid in response to insecticidal stresses. Principal component analysis and systemic cluster analysis revealed that the most pronounced changes in physiological indexes and metabolites were caused by the pyriproxyfen treatment. In conclusion, this study improves our understanding of the mechanism by which insecticides affect plant physiological and metabolic processes, thus potentially altering the quality and therapeutic value of peppermint as an example.

Peppermint Essential Oil and Its Major Volatiles as Protective Agents against Soft Rot Caused by Fusarium sambucinum in Cera Pepper (Capsicum pubescens).

Cera pepper (Capsicum pubescens) is an exotic fruit considered as a rich source of nutraceuticals with known benefits for human health and also an economic resource for local producers in Mexico. The present investigation reports on the in vitro and in situ antifungal activity of the essential oil from Mentha piperita and its two major volatiles (menthol and menthone) against Fusarium sambucinum, which is a causal agent of soft rot in cera pepper. The application of these components in pepper fruits previously infected with F. sambucinum caused a significant delay (p<0.05) in the emergence of soft rot symptoms. This effect was reflected in the maintenance of pH and fruit firmness during a period of 10 days. The nutrimental content of the fruits (protein, fiber, fat and other proximate parameters) was conserved in the same period of time. The nutraceutical content of these fruits was estimated by the quantification of seven carotenoids (violaxanthin, cis-violaxanthin, luteoxanthin, antheraxanthin, lutein, zeaxanthin and ß-carotene), ascorbic acid and capsaicinoids (capsaicin and dihydrocapsaicin). According to our results, the essential oil from M. Piperita and its major volatiles exerted a preservative effect on these metabolites. Our findings demonstrated that the essential oil of M. Piperita and its major volatiles represent an ecological alternative for the control of fusariosis caused by F. sambucinum in cera peppers under postharvest conditions.

Peppermint extract inhibits protein aggregation.

The extracts of 7 herbs were screened and compared for their functional ability to inhibit the aggregation of trypsin as an appropriate model protein for in vitro fibrillation in aqueous ethanol at pH 7.0. Turbidity measurements, total phenolic content determination, aggregation kinetics, Congo red binding assay as well as transmission electron microscopy were used to analyse the inhibition of amyloid fibril formation. This correlated with the total phenolic content of the herb extracts. The peppermint extract proved to be the most potent anti-amyloidogenic agent. Results showed that the peppermint extract exerted dose-dependent inhibitory effect on trypsin fibril formation.

Does co-inoculation of mycorrhiza and Piriformospora indica fungi enhance the efficiency of chlorophyll fluorescence and essential oil composition in peppermint under irrigation with saline water from the Caspian Sea?

Symbiotic associations with endophytic fungi are ecologically important for medicinal and aromatic plants. Endophytic fungi highly affect the quantity and quality of herbal products. In this study, a pot experiment was carried out in the greenhouse to investigate the interactive effects of Piriformospora indica and arbuscular mycorrhizal (AMF) inoculation on the chlorophyll fluorescence, essential oil composition, and antioxidant enzymes of peppermint under saline condition. The results showed that Fo, YNPQ, YNO, and NPQ values were obviously increased under salinity conditions, while essential oil content, chlorophyll a and b, gs, Fm, Fv, ETR, ФPSII and Fv/Fm ratio decreased by increasing salinity. In addition, salt induced the excess Na+ uptake, whereas the opposite trend was observed for P and K+. The synergistic association of P. indica and AMF caused a considerable increase in the antioxidant ability, essential oil content, Fv/Fm ratio, ФPSII, and amount of P and K+ uptake in salt-stressed plants. The main peppermint oil constituents, menthol, menthone, and 1,8-cineole increased considerably in inoculated plants. Besides, the applied endophytic fungi positively enhanced the ability of peppermint to alleviate the negative effect of the salinity stress.

Antioxidant and enzyme-inhibitory activity of peppermint extracts and essential oils obtained by conventional and emerging extraction techniques.

Peppermint is widely used medicinal plant with distinguished bioactive potential, therefore, the aim of present work was to develop novel peppermint extracts with high activity by application of traditional and emerging separation techniques. Conventional hydrodistillation and microwave-assisted hydrodistillation (MWHD) were applied for recovery of essential oil (EO), while organic solvent extraction using Soxhlet apparatus, microwave-assisted and ultrasound-assisted process and supercritical fluid extraction (SFE) were applied for non-selective recovery of peppermint lipophilic extracts. Extracts were characterized in terms of terpenoids profile with special emphasis on content of major compounds (mentol, menthone, isomenthol and eucalyptol). Antioxidant activity (DPPH, ABTS, CUPRAC, FRAP, chelating and phosphomolybdenum assay) and enzyme-inhibitory assays (acetylcholinesterase, butyrylcholinesterase, tyrosinase, amylase and glucosidase inhibition) were used for screening of peppermint bioactivity. MWHD was recognized as alternative for traditional process in EO recovery, while SFE extracts were useful for green production of solvent-free peppermint extracts rich in terpenoids and other lipophilic bioactives.

A simple method to determine antioxidant status in aromatic plants subjected to drought stress.

Drought is a major environmental stress factor that affects the growth and development of plants. All plants have to maintain the reactive oxygen species within certain levels for normal cellular homeostasis by means of their antioxidant systems, which can be classified as enzymatic and non-enzymatic. Plants under drought stress generate an excess production of reactive oxygen species. At high concentrations, this can be detrimental by producing damage to the protein structures and inhibiting enzymes, as well as oxidizing macromolecules, which may eventually lead to cell death. There has been increasing attention paid to the antioxidant capacity of aromatic/medicinal plants, with a high antioxidant content having been reported in some plant extracts, such as in Mentha piperita (peppermint). Peppermint plants cultivated under drought stress also present high levels of phenolic compounds, peroxidase enzyme activity and lipid peroxidation of membranes. A simple and inexpensive laboratory class is proposed for teaching some mechanisms that plants have evolved to avoid reactive oxygen species damage. The series of lab experiments described is aimed at demonstrating the antioxidant status in aromatic plants subjected to drought stress, by measuring total phenolic compound content (non-enzymatic antioxidant compound), peroxidase activity (enzymatic antioxidant) and malondialdehyde, as convenient biomarkers for lipid peroxidation. The proposed class will be carried out by undergraduate students of the advanced biochemistry course, as part of our biology and agronomy studies. The experiment presented is intended to be used as a vehicle to emphasize the concepts that students have learned in their lectures. This lab exercise to be carried out by the students has dual goals: to apply a methodology only learned superficially on previous courses, and also to increase their understanding of how plants developed resistance mechanisms in order to tolerate drought stress.

Changes in Ecophysiology, Osmolytes, and Secondary Metabolites of the Medicinal Plants of Mentha piperita and Catharanthus roseus Subjected to Drought and Heat Stress.

Global warming contributes to higher temperatures and reduces rainfall for most areas worldwide. The concurrent incidence of extreme temperature and water shortage lead to temperature stress damage in plants. Seeking to imitate a more natural field situation and to figure out responses of specific stresses with regard to their combination, we investigated physiological, biochemical, and metabolomic variations following drought and heat stress imposition (alone and combined) and recovery, using Mentha piperita and Catharanthus roseus plants. Plants were exposed to drought and/or heat stress (35 °C) for seven and fourteen days. Plant height and weight (both fresh and dry weight) were significantly decreased by stress, and the effects more pronounced with a combined heat and drought treatment. Drought and/or heat stress triggered the accumulation of osmolytes (proline, sugars, glycine betaine, and sugar alcohols including inositol and mannitol), with maximum accumulation in response to the combined stress. Total phenol, flavonoid, and saponin contents decreased in response to drought and/or heat stress at seven and fourteen days; however, levels of other secondary metabolites, including tannins, terpenoids, and alkaloids, increased under stress in both plants, with maximal accumulation under the combined heat/drought stress. Extracts from leaves of both species significantly inhibited the growth of pathogenic fungi and bacteria, as well as two human cancer cell lines. Drought and heat stress significantly reduced the antimicrobial and anticancer activities of plants. The increased accumulation of secondary metabolites observed in response to drought and/or heat stress suggests that imposition of abiotic stress may be a strategy for increasing the content of the therapeutic secondary metabolites associated with these plants.

Effect of Heat Stress on Yield, Monoterpene Content and Antibacterial Activity of Essential Oils of Mentha x piperita var. Mitcham and Mentha arvensis var. piperascens.

Heat stress affects the yield of medicinal plants and can reduce biomass and/or metabolite production. In order to evaluate the effect of heat-induced stress on the essential oil production in Mentha x piperita L. var. Mitcham (Mitcham mint) and Mentha arvensis var. piperascens Malinv. ex L. H. Bailey (Japanese mint), we studied the chemical composition of the oils of the two mint species under different heat shock stresses in growth chambers. The antibacterial activity of the essential oils was also evaluated; microscopic observation (fluorescence and electron transmission) was used to assess the effect of the tested samples on bacterial growth. The results obtained shed light on the mint essential oils composition and biological activity in relation to heat stress.

Elevated CO2 induces a global metabolic change in basil (Ocimum basilicum L.) and peppermint (Mentha piperita L.) and improves their biological activity.

Many studies have discussed the influence of elevated carbon dioxide (eCO2) on modeling and crop plants. However, much less effort has been dedicated to herbal plants. In this study, a robust monitoring for the levels of 94 primary and secondary metabolites and minerals in two medicinal herbs, basil (Ocimum basilicum L.) and peppermint (Mentha piperita L.), grwon under both ambient (aCO2, 360 ppm) and eCO2 (620 ppm) was performed. We also assessed how the changes in herbal tissue chemistry affected their biological activity. Elevated CO2 significantly increased herbal biomass, improved the rates of photosynthesis and dark respiration, and altered the tissue chemistry. Principal Component Analysis of the full data set revealed that eCO2 induced a global change in the metabolomes of the two plants. Moreover, Hierarchical Clustering Analyses showed quantitative differences in the metabolic profiles of the two plants and in their responsiveness to eCO2. Out of 94 metabolites, 38 and 31 significantly increased in basil and peppermint, respectively, as affected by eCO2. Regardless of the plant species, the levels of non-structural carbohydrates, fumarate, glutamine, glutathione, ascorbate, phylloquinone (vitamin K1), anthocyanins and a majority of flavonoids and minerals were significantly improved by eCO2. However, some metabolites tended to show species specificity. Interestingly, eCO2 caused enhancement in antioxidant, antiprotozoal, anti-bacterial and anticancer (against urinary bladder carcinoma; T24P) activities in both plants, which was consequent with improvement in the levels of antioxidant metabolites such as glutathione, ascorbate and flavonoids. Therefore, this study suggests that the metabolic changes triggered by eCO2 in the target herbal plants improved their biological activities.

Bio-mediated synthesis, characterization and cytotoxicity of gold nanoparticles.

We report here a "green" approach for the synthesis of gold nanoparticles (GNPs) in which the Mentha piperita extract was applied for the bioreduction of chloroauric acid and the stabilization of the formed nanostructures. The obtained GNPs were characterized by UV-Vis absorption spectroscopy and transmission electron microscopy (TEM). The reduction of gold ions with the plant extract leads to the production of nanoparticles with various shapes (spherical, triangular and hexagonal) and sizes (from 10 to 300 nm). The kinetics of the reaction was monitored and various conditions of the synthesis were investigated. As a result, we established protocols optimized towards the synthesis of nanospheres and nanoprisms of gold. The cytotoxic effect of the obtained gold nanoparticles was studied by performing MTT assay, which showed lower cytotoxicity of the biosynthesized GNPs compared to gold nanorods synthesized using the usual seed-mediated growth. The results suggest that the synthesis using plant extracts may be a useful method to produce gold nanostructures for various biological and medical applications.

Development of an experimental apparatus and protocol for determining antimicrobial activities of gaseous plant essential oils.

There is a growing interest in the use of naturally-occurring antimicrobial agents such as plant essential oils (EOs) to inhibit the growth of hazardous and spoilage microorganisms in foods. Gaseous EOs (EO gases) have many potential applications in the food industry, including use as antimicrobial agents in food packaging materials and sanitizing agents for foods and food-contact surfaces, and in food processing environments. Despite the potentially beneficial applications of EO gases, there is no standard method to evaluate their antimicrobial activities. Thus, the present study was aimed at developing an experimental apparatus and protocol to determine the minimal inhibitory concentration (MIC) and minimal lethal concentration (MLC) of EO gases against microorganisms. A sealed experimental apparatus was constructed for simultaneous evaluation of antimicrobial activities of EO gases at different concentrations without creating concentration gradients. A differential medium was then evaluated in which a color change allowed for the determination of growth of glucose-fermenting microorganisms. Lastly, an experimental protocol for the assessment of MIC and MLC values of EO gases was developed, and these values were determined for 31 EO gases against Escherichia coli O157:H7 as a model bacterium. Results showed that cinnamon bark EO gas had the lowest MIC (0.0391 µl/ml), followed by thyme-thymol EO gas (0.0781 µl/ml), oregano EO gas (0.3125 µl/ml), peppermint EO gas (0.6250 µl/ml), and thyme-linalool EO gas (0.6250 µl/ml). The order of the MLC values of the EO gases against the E. coli O157:H7 was thyme-thymol (0.0781 µl/ml)

Salicylic acid elicitation during cultivation of the peppermint plant improves anti-diabetic effects of its infusions.

Peppermint (Mentha piperita) infusions represent an important source of bioactive compounds with health benefits, which can be enhanced by applying salicylic acid (SA) during plant cultivation. The aim of this study was to evaluate the effect of SA (0, 0.5 and 2 mM) during peppermint cultivation on the chemical profile of saponins and alkaloids, as well as the anti-diabetic properties of the resulting infusions. The results showed that a 2 mM SA treatment significantly improved the chemical profiles of the infusions. Furthermore, the administration of 2 mM SA-treated peppermint infusions for 4 weeks to a high-fat diet/streptozotocin-induced diabetic rats decreased serum glucose levels (up to 25%) and increased serum insulin levels (up to 75%) as compared with the diabetic control. This can be related to the observed protection on pancreatic ß-cells. Furthermore, 0.5 and 2 mM SA-treated peppermint infusions decreased LDL (24 and 47%, respectively) and increased HDL levels (18 and 37%, respectively). In addition, all groups treated with peppermint infusions had lower serum and liver triglyceride contents, where 2 mM SA peppermint infusion showed the highest effect (44% and 56%, respectively). This is probably caused by its higher capacity to inhibit pancreatic lipase activity and lipid absorption. Moreover, SA-treated peppermint infusions improved the steatosis score in diabetic rat liver and decreased serum transaminase levels, probably as a result of the increase in steroidal saponins and alkaloids, such as trigonellin. Therefore, the application of 2 mM SA during cultivation of peppermint could be used to improve the anti-diabetic properties of peppermint infusions.

Metabolite profile, antioxidant capacity, and inhibition of digestive enzymes in infusions of peppermint (Mentha piperita) grown under drought stress.

Peppermint (Mentha piperita) infusions represent an important source of antioxidants, which can be enhanced by inducing abiotic stress in plants. The aim of this study was to evaluate the effect of drought stress on peppermint cultivation as well as the metabolite profile, antioxidant capacity, and inhibition of digestive enzymes of resulting infusions. At 45 days after planting, irrigation was suppressed until 85 (control), 65, 35, 24, and 12% soil moisture (SM) was reached. The results showed that 35, 24, and 12% SM decreased fresh (20%) and dry (5%) weight. The 35 and 24% SM treatments significantly increased total phenolic and flavonoid contents as well as antioxidant capacity. Coumaric acid, quercetin, luteolin, and naringenin were detected only in some drought treatments; however, in these infusions, fewer amino acids and unsaturated fatty acids were identified. The 24 and 12% SM treatments slightly improved inhibition of pancreatic lipase and α-amylase activity. Therefore, induction of moderate water stress in peppermint is recommended to enhance its biological properties.

Kinetic modeling of plant metabolism and its predictive power: peppermint essential oil biosynthesis as an example.

The integration of mathematical modeling with analytical experimentation in an iterative fashion is a powerful approach to advance our understanding of the architecture and regulation of metabolic networks. Ultimately, such knowledge is highly valuable to support efforts aimed at modulating flux through target pathways by molecular breeding and/or metabolic engineering. In this article we describe a kinetic mathematical model of peppermint essential oil biosynthesis, a pathway that has been studied extensively for more than two decades. Modeling assumptions and approximations are described in detail. We provide step-by-step instructions on how to run simulations of dynamic changes in pathway metabolites concentrations.

Physiological indexese macro- and micronutrients in plant tissue and essential oil of Mentha piperita L. grown in nutrient solution with variation in N, P, K and Mg levels / Índices fisiológicos, macro e micronutrientes no tecido vegetal e óleo essencial de Mentha piperita L. cultivada em solução nutritiva com variação de N, P, K e Mg.

Mentha piperita L. is an aromatic and medicinal species of the family Lamiaceae, known as mint or peppermint, and its leaves and branches produce essential oil rich in menthol. This study aimed to evaluate physiological indexes, macro- and micronutrients inthe shootsand essential oil of Mentha piperita L. grown in nutrient solution number 2 of Hoagland and Arnon (1950) with different N, P, K and Mg levels. Shoot length, dry mass of the different organs, total dry mass, leaf area, essential oil yield and composition, and macronutrient (N, P, K, Mg, Ca, S) and micronutrient (Mn, Cu, Fe, Zn) contents in the shoot were evaluated. Plants treated with 65%N/50%P/25%K/100%Mg had a tendency towards longer shoot, greaterroot and leaf blade dry masses, higher essential oil yield, higher menthol levels and lower menthone levels. The results showed that Mentha can be grown in nutrient solution by reducing 65% N, 50% P, 25% K and 100% Mg. This solution had better development compared to the other tested treatments. Therefore,we recommendMentha piperita L. to be grown with such nutrient levels.

Mentha piperita L., é uma espécie aromática e medicinal pertencente à família Lamiaceae, conhecida como menta ou hortelã-pimenta que produz em sua parte aérea óleo essencial rico em mentol. O objetivo do trabalho foi avaliar os índices fisiológicos, macro e micronutrientes na parte aérea e óleo essencial de Mentha piperita L. cultivada em solução nutritiva numero Hoagland e Arnon (1950) com variação dos níveis de N, P, K e Mg. Foram avaliados comprimento da parte aérea, massa seca dos diferentes órgãos e total, área foliar, rendimento e composição do óleo essencial, teor de macronutrientes (N, P, K, Mg, Ca e S), e de micronutrientes (Mn, Cu, Fe e Zn) na parte aérea. As plantas nutridas com 65% de N, 50% de P, 25% de K, e 100% de Mg, apresentaram tendência de maior comprimento de parte aérea e massa seca de raízes e de lâminas foliares; maior rendimento de óleo essencial e, em média, maiores teores de mentol e menores teores de mentona. O resultados permitem concluir que a Mentha pode ser cultivada em solução nutritiva reduzindo-se 65% de N, 50% de P, 25% de K e 100% de Mg. Esta solução apresentou melhor desenvolvimento em relação aos outros tratamentos. Recomenda-se, portanto, o cultivo da Mentha piperita L. com tais níveis de nutrientes.

Qualidade de folhas e rendimento de óleo essencial em hortelã pimenta (Mentha x Piperita L. ) submetida ao processo de secagem em secador de leito fixo / Quality of leaves and essential oil yield of peppermint (Mentha x piperita L. ) submitted to the drying process in fixed-bed dryer

O uso de plantas medicinais e seus derivados para o tratamento de doenças é uma prática antiga e se percebe, atualmente, uma crescente procura por produtos naturais, incluindo medicamentos, produtos alimentícios, e cosméticos. Hortelã pimenta (Mentha x Piperita L), além de ser uma planta medicinal, pode ser utilizada para obtenção de aromatizantes, infusões, e temperos. O processo de secagem se faz necessário para aumentar o tempo de conservação e a vida útil do produto facilitando seu transporte, manuseio, e armazenamento. Para que os produtos derivados da hortelã pimenta tenham qualidade é necessário estudos sobre o processo de pré e de pós-colheita. Objetivou-se neste estudo identificar a temperatura de secagem, em duas velocidades de ar circulante, que minimize a degradação da cor das folhas e permita obter maior rendimento de óleo essencial. Folhas da hortelã foram colhidas manualmente no horário entre 7:15 e 8:00, e submetidas a secagem em duas velocidades do ar (0,3 e 0,5 m.s-1) e em cinco temperaturas (30, 40, 50, 60 e 70 °C). O material seco foi analisado quanto à cor utilizando-se colorímetro com escala do sistema CIELab baseado em coloração dentro dos padrões da Norma DIN 6174 (1979). A quantificação do óleo essencial foi realizada por hidrodestilação em aparelho Clevenger. Observou-se que temperaturas superiores a 50 °C reduzem o rendimento. A cor das folhas submetidas à secagem diferencia-se da cor das folhas frescas. De acordo com a Norma DIN 6174, a cor das folhas submetidas a secagem com temperatura até 40 °C são "Facilmente distinguíveis" e para as folhas submetidas à secagem a temperaturas superior a 50 °C, a diferenciação é "Muito grande". Conclui-se que para obter o máximo de rendimento do óleo essencial e o mínimo de degradação da cor, a secagem deve ser realizada a temperatura de até 50 ºC. A velocidade do ar de secagem, na faixa de 0,3 a 0,5 m.s-1, não afetou os parâmetros avaliados.

The use of medicinal plants and their derivatives for the treatment of diseases is an ancient practice. Currently, there is a growing demand for natural products, not only medicines, but also food and cosmetics. The peppermint (Mentha x piperita L.), in addition to being a medicinal plant, can be used for obtaining flavorings, spices and tea infusions. The drying process is necessary to increase the shelf life of the product and to facilitatg itr transport, handling and storage. For peppermint derivatives of sufficient quality, it is necessary studies on the pre- and post-harvest. The objective of this study is to identify the temperature and air velocity that minimizs the degradation of the color of the leaves with higher oil yield. PepperMint leaves were harvested manually from 7:15 to 8:00 a.m. and dried in trays in a completely randomized design in a 2 x 5 factorial design, being two air velocities (0.3 and 0.5 m.s-1) and five temperatures (30, 40, 50, 60 and 70 °C) with three replications. The dried material was analyzed for color using a colorimeter with the CIELab scale system based on a color within the DIN 6174 standards (1979). The quantification of the essential oil was performed by hydrodistillation in a Clevenger-type apparatus. The analyzes were performed in triplicate. Ws observed that the yield of essential oil of the dry leaves was highertwhan compared to the yield presented by the fresh leaves. This effect was attributed to the fact that the lower water content in the leaves enables the vapor stream generated in the extractor to promots a more effective drag of the volatile oilcompounds stored in the cells; however, a temperature exceeding 50 °C promotes a reduction in yield. The color of the leaves submitted to drying is different from the color of fresh leaves. According to the DIN 6174 standard, the color of the leaves subjected to drying at the temperature up to 40 °C is "easily distinguishable" and, for the leaves submitted to drying at temperatures higher than 50 °C, the differentiation is "too large". We conclude that to get the maximum yield of essential oil and minimal color degradation, drying should be carried out at a temperature of up to 50 °C. The rate of the drying processes in the rangeofrom 0.3 to 0.5 m.s-1 did not affect the parameters evaluated.

Intrinsic isotopic 13C labelling of polyphenols.

The intrinsic isotopic labelling of plants with (13)CO2 is an effective method to generate highly labelled compounds using photosynthesis and avoiding labour-intensive complex organic syntheses. In this study, the intrinsic isotopic labelling of polyphenols in parsley, spinach and peppermint is shown for the first time. The plants were grown in an atmosphere where (12)CO2 was replaced by (13)CO2, in order to generate highly labelled compounds. The total content of (13)C as well as the individual polyphenols were analysed by Isotopic Ratio-MS and HPLC-Iontrap-MS(n). After 34 days of plant growth under (13)CO2, degree of labelling was found to be higher than 90 atom% (13)C for most polyphenols, predominantly consisting of highly and fully labelled isotopomers; the total plant material contained more than 88 atom% (13)C. Such highly labelled compounds can be used in future studies to dissect both metabolism and bioavailability of polyphenols in humans.

Mentha x piperita L. y su potencial en el control del vector del dengue (Aedes aegypti L.). Revisión / Mentha x piperita L. and its potential in controlling the dengue vector (Aedes aegypti L.). Review

El dengue es una de las enfermedades tropicales más fatales, afectando a 50-100 millones de personas al año. Endémico en más de cien países y transmitido a los humanos por la picadura subcutánea del mosquito Aedes aegypti L., puede desencadenar los síntomas de una gripe normal, episodios de fiebre hemorrágica y shock. Las medidas de erradicación del vector adoptadas por las autoridades sanitarias mundiales, como la eliminación del hábitat y uso de repelentes y/o insecticidas) no logran frenar la propagación del dengue, ya extendido por las áreas no endémicas, como Portugal. Se sabe que el mosquito es vulnerable a algunos productos vegetales y varios estudios los han considerado como alternativas potenciales a los productos químicos sintéticos, a la que el mosquito se ha vuelto resistente. Algunas de esas plantas pertenecen a la flora nativa, espontánea y subespontánea, estando accesibles a un gran número de personas. Es el caso de la Mentha x piperita., elegida como ejemplo por las propiedades insecticidas y repelentes de su aceite esencial (AU)

Dengue is one of the most fatal tropical diseases which affect 50-100 billion people/ year. It is endemic in more than 100 countries and transmitted to humans by the subcutaneous bite of the Aedes aegypti L., it can trigger different symptoms like normal flu, hemorrhagic fever and shock. The eradication of the vector adopted by global health authorities, like the elimination of its habitat, the use of repellents and/or insecticides, fail to curb the spread of the disease, which are expanding into non-endemic areas (Portugal). Vector is vulnerable to some herbal products and scientific studies have considered them as a potential alternative to the synthetic ones in use, to which the mosquito may become resistant. Some of these herbs belong to native, spontaneous and subspontaneous flora, resulting accessible to the population. Mentha x piperita L., was chosen as example due to insecticide and repellent properties of it essential oil (AU)

The roles of a flavone-6-hydroxylase and 7-O-demethylation in the flavone biosynthetic network of sweet basil.

Lipophilic flavonoids found in the Lamiaceae exhibit unusual 6- and 8-hydroxylations whose enzymatic basis is unknown. We show that crude protein extracts from peltate trichomes of sweet basil (Ocimum basilicum L.) cultivars readily hydroxylate position 6 of 7-O-methylated apigenin but not apigenin itself. The responsible protein was identified as a P450 monooxygenase from the CYP82 family, a family not previously reported to be involved in flavonoid metabolism. This enzyme prefers flavones but also accepts flavanones in vitro and requires a 5-hydroxyl in addition to a 7-methoxyl residue on the substrate. A peppermint (Mentha × piperita L.) homolog displayed identical substrate requirements, suggesting that early 7-O-methylation of flavones might be common in the Lamiaceae. This hypothesis is further substantiated by the pioneering discovery of 2-oxoglutarate-dependent flavone demethylase activity in basil, which explains the accumulation of 7-O-demethylated flavone nevadensin.