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The aim of this study was to determine the chemical profile of the essential oils (EOs) of three Moroccan lavender species (Lavandula pedunculata, LP; Lavandula angustifolia, LA; and Lavandula maroccana, LM) and to investigate, for the first time, the synergistic effect of the optimal mixture of the EOs with conventional antibiotic ciprofloxacin against three pathogenic foodborne bacteria. Gas chromatography/mass spectrometry analysis showed that eucalyptol (39.05%), camphor (24.21%) and borneol (8.29%) were the dominant compounds of LA-EO. LP-EO was characterized by the abundance of camphor (74.51%) and fenchone (27.06%), whereas carvacrol (42.08%), camphor (17.95%) and fenchone (12.05%) were the main constituents of LM-EO. EOs alone or combined showed a remarkable antimicrobial activity against the tested bacteria with minimum inhibitory concentrations (MICs) ranging from 3.53 mg/mL to 15.96 mg/mL. The optimal mixture, calculated using a mixture design, corresponded to 19% LA, 38% LP and 43% LM. All combination of the EOs and the best EO mixture with ciprofloxacin exhibited a total synergism with fractional inhibitory concentration index (FICI) values ranging from 0.27 to 0.37. The best EO mixture showed the highest gain of 128 fold, especially against Salmonella spp., more than that found testing the EOs separately. These findings should be taken into consideration for a possible application in the pharmaceutical and food industries.

Lavandula pedunculata (Mill.) Cav. (LP) is one of lavender species traditionally used in Morocco to prevent or cure diabetes, alone or in the form of polyherbal preparations (PHP). Therefore, the primary objective of this study was to test the antihyperglycemic effect of the aqueous extract of LP, alone and in combination with Punica granatum L. (PG) and Trigonella foenum-graecum L. (FGK). The secondary objective was to explore some mechanisms of action on the digestive functions. The antihyperglycemic effect of the aqueous extract of LP, alone and in combination with PG and FGK, was studied in vivo using an oral glucose tolerance test (OGTT). In addition, LP extract was tested on the activities of some digestive enzymes (pancreatic α-amylase and intestinal α-glucosidase) in vitro and on the intestinal absorption of glucose ex vivo using a short-circuit current (Isc) technique. Acute and chronic oral administration of LP aqueous extract reduced the peak of the glucose concentration (30 min, p < 0.01) and the area under the curve (AUC, p < 0.01). The effect of LP + PG was at the same amplitude to that of the positive control Metformin (MET). LP aqueous extract inhibited the pancreatic α-amylase with an IC50 almost identical to acarbose (0.44 ± 0.05 mg/mL and 0.36 ± 0.02 mg/mL, respectively), as well as the intestinal α-glucosidase, (IC50 = 131 ± 20 µg/mL) and the intestinal glucose absorption (IC50 = 81.28 ± 4.01 µg/mL) in concentration-dependent manners. LP aqueous extract exhibited potent actions on hyperglycemia, with an inhibition on digestive enzymes and glucose absorption. In addition, the combination with PG and FGK enhanced oral glucose tolerance in rats. These findings back up the traditional use of LP in type 2 diabetes treatment and the effectiveness of the alternative and combinative poly-phytotherapy (ACPP).

Lavandula cinsi (Lamiaceae) ekonomik ve tıbbi açıdan büyük önem taşıyan uçucu yağ bitkilerinin yer aldığı önemli bir taksondur. Bu cinsin Türkiye’de doğal yayılış gösteren Lavandula stoechas L. subsp. stoechas ve L. pedunculata subsp. cariensis (Boiss.) Upson & S.Andrews olmak üzere iki türü mevcuttur. Bunlar “karabaşotu, karan, yalancı lavanta, karahan” gibi isimlerle bilinmekte ve halk arasında genellikle birbirinin yerine tıbbi amaçlarla yüzyıllardır kullanılmaktadır. İnfüzyonu ve distilasyonla elde edilen uçucu yağı, analjezik, antiseptik, yara iyi edici, balgam söktürücü, sinir ve kalp kuvvetlendirici gibi etkileri nedeniyle halk arasında kullanılmaktadır. Çalışmamızda, ülkemizin farklı bölgelerinden temin edilen karabaşotu çiçeklerinin uçucu yağ bileşimleri ile hazırlanan çaylarında Avrupa Gıda Güvenliği Otoritesi (EFSA) tarafından toksisite dozları yayımlanan kâfurun, kantitatif miktarı ortaya konmuştur. Gaz kromatografisi/kütle spektometresi ve gaz kromatografisi/alev iyonlaşma dedektörü kullanılarak yapılan analizlerde, tüm örneklerden elde edilen uçucu yağların ana bileşenleri olarak kâfur (%7-53) ve fenkon (%27-58) saptanırken, hazırlanan %5 infüzyonlarda ise 4,6-243 µg/mL aralığında kâfur tespit edilmiştir.

This work consists of the comparison of the chemical composition, the antioxidant activity, and the antimicrobial effect of two species of the genus Lavandula essential oils (EOs) and extracts (hydroethanolic, aqueous and decocted), that were collected from the region of Ifrane in Morocco "Lavandula angustifolia and Lavandula peduncultata". Gas chromatographic analyzes coupled with mass spectrometry of the studied EOs showed that L. angustifolia contains camphor and linalool as majority compounds with 20,17% and 12,41% respectively, while camphor (41,15%) and fenchone (15,70%) are the major compounds of L. peduncultata. The dosage of hydroethanolic, aqueous and decocted extracts showed that the content of polyphenols and flavonoids is higher in the hydroethanolic extracts of L. pedunculata, while L. angustifolia contains the highest content of tannins. The determination of the antioxidant activity was carried out using FRAP and DPPH methods, the last method revealed that the EO of L.angustifolia is less effective (IC50=7,98 mg/ml) than that of L. pedunculata (EC50=7,47 mg/ml) while, there was no great divergence concerning the IC50 values of the hydroethanolic and decocted extracts of the two species. As for the aqueous extract, the value recorded by L. pedunculata was slightly higher than that of L. angustifolia. Finally, tests of the antimicrobial activity of EOs and extracts of the two species were carried out on six bacteria and five fungal strains whose antimicrobial power was remarkable. As a conclusion, the two Lavenders’extracts can be considered as alternative sources of powerful natural antioxidants, antifungals and antibiotics.

The present study aimed at evaluating the effectiveness of different natural deep eutectic solvents (NADES) on the extraction of phenolic compounds from Lavandula pedunculata subsp. lusitanica (Chaytor) Franco, on the antioxidant activity, and acetylcholinesterase (AChE), butyrylcholinesterase (BChE) and tyrosinase (Tyr) inhibitory capacities. Ten different NADES were used in this research and compared with conventional solvents. Ultrasound-assisted extraction (UAE) for 60 min proved to be the best extraction condition, and proline:lactic acid (1:1) and choline chloride:urea (1:2) extracts showed the highest total phenolic contents (56.00 ± 0.77 mgGAE/gdw) and antioxidant activity [64.35 ± 1.74 mgTE/gdw and 72.13 ± 0.97 mgTE/gdw in 2.2-diphenyl-1- picrylhydrazyl (DPPH) and 2.20 -azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) (ABTS) methods, respectively]. These extracts also exhibited enzymes inhibitory capacity particularly against Tyr and AChE. Even so, organic acid-based NADES showed to be the best extractants producing extracts with considerable ability to inhibit enzymes. Twenty-four phenolic compounds were identified by HPLC-HRMS, being rosmarinic acid, ferulic acid and salvianolic acid B the major compounds. The results confirmed that the combination of UAE and NADES provide an excellent alternative to organic solvents for sustainable and green extraction, and have huge potential for use in industrial applications involving the extraction of bioactive compounds from plants. Grant SFRH/BD/145243/2019, UIDB/05183/2020 info:eu-repo/semantics/publishedVersion

Some autochthones plant species with aromatic and medicinal properties are able to naturally colonize contaminated soils from mining areas from Iberian Pyrite Belt contributing to their rehabilitation. A study was carried out in order to characterize and valorise an autochthones species, which has adequate ecophysiological behaviours for phytostabilization of mining areas, as new sources of bioactive substances. The main aims of this study were to: i) characterise the phytochemical profile of the bioextracts from shoots of L. pedunculata growing in soils from São Domingos mining area and a control area; and ii) evaluate the influence of potentially hazardous elements (PHEs) accumulated in the shoots on the quality of the bioextracts.Composite samples of soils, developed on mine wastes and/or host rocks, as well as C Lavandula pedunculata shoots were collected in São Domingos mine (Iberian pyrite Belt, SE of Portugal) and in a reference area with non-contaminated soils and the same climatic conditions. Classical characterisation of soils and total concentrations of potentially hazardous elements in soils and plant shoots were determined. The bioextracts from Lavandula pedunculata shoots were obtained by an accelerated solvent extractor, and the compounds were analysed by GC-MS. Extracts were extracted with hexane and major components were quantified.The total concentrations of some potentially hazardous elements (e.g. As, Cu, Pb and Zn) were higher in soils from São Domingos than in reference area. However, soils from São Domingos are considered as contaminated with As, Cu, Pb and Sb for agriculture and residential/parkland uses. Concentrations of the PHEs (excepted Cr and Mn) in the shoots collected in São Domingos mine were higher than in the non-contaminated areaIn the L. pedunculata extracts, obtained in the single extraction with hexane, were identified 34 compounds accounting between 79 and 89 % of the total identified compounds. Camphor was the major component in all extracts but Fenchone, eucalyptol, verbenone, bornyl acetate, borneol and linalool oxide cis also showed considerable amounts. All these compounds present economic interest. Some variation was obtained in the qualitative composition of the L. pedunculata extracts but, in general, it was not clear the differentiation between populations and, consequently, soil contamination level and concentrations of the potentially hazardous elements in shoots. Environmental rehabilitation of mining areas from Iberian Pyrite Belt with this species can provide economic valorisation by the exploration of this plant-based product for fragrance and pharmaceutical industries.Acknowledgment: This research was supported by Portuguese funds, through Fundação para a Ciência e Tecnologia within the scope of the project UID/AGR/04129/202, and Xunta de Galicia (GRC2014/003).

Spontaneous lavender growing in uncultivated fields in Portugal have been used in traditional medicine for internal and external uses. The essential oils (EOs) of Lavandula stoechas subsp. luisieri are characterized by the presence of trans-α-necrodyl acetate and trans-necrodol. These EOs are able to prevent the generation and deposition of neurotoxic β-amyloid peptide in Alzheimer’s disease. The EOs also present antibacterial, anti-fungal, anti-Leishmania, antioxidant, anti-inflammatory and antifeedant effects. In the case of hydrodistillation, the predominant compound of Lavandula viridis EO was 1,8-cineole, nevertheless in the case of supercritical fluid extraction, the main constituent was camphor. In in vitro shoots EOs, 1,8-cineole and α-pinene were the most important compounds. The EOs presented anti-fungal activity particularly against Cryptococcus neoformans and dermatophytes. The antioxidant and anti-protozoal activities of L. viridis EOs were lower than L. stoechas subsp. luisieri EOs, with hydrodistillation being the best method for obtaining samples with higher antioxidant and anti-acetylcholinesterase activities. The presence of fenchone, 1,8-cineole and camphor was a common trace of the Lavandula pedunculata subsp. pedunculata EOs and in in vitro axillary shoots EOs. Lavandula pedunculata subsp. lusitanica EOs were predominantly constituted of fenchone and camphor. The antioxidant activity of L. pedunculata subsp. lusitanica EOs was poorer than other Lavandula EOs from Portugal.

This study aimed to: i) evaluate the accumulation and translocation patterns of potentially hazardous elements into the Lavandula pedunculata and their influence in the concentrations of nutrients; and ii) compare some physiological responses associated with oxidative stress (concentration of chlorophylls (Chla, Chlb and total), carotenoids, and total protein) and several components involved in tolerance mechanisms (concentrations of proline and acid-soluble thiols and total/specific activity of catalase (CAT) and superoxide dismutase (SOD)), in plants growing in soils with a multielemental contamination and non-contaminated. Composite samples of soils, developed on mine wastes and/or host rocks, and L. pedunculata (roots and shoots) were collected in Sao Domingos mine (SE of Portugal) and in a reference area with non-contaminated soils, Corte do Pinto, with the same climatic conditions. Sao Domingos soils had high total concentrations of several hazardous elements (e.g. As and Pb) but their available fractions were small (mainly

In order to restore the plant cover, improve ecosystem services and decrease the environmental risk of two mine wastes (gossan and sulfide-rich wastes), an integrated biotechnology was tested at long-term and greenhouse conditions. This integrated biotechnology involves the natural isolation of sulfide-rich wastes through an alkaline barrier, covered by designed Technosols (both of them derived from mining and agro-industrial wastes) and a plant cover with Lavandula pedunculata and Cistus ladanifer. Technosols allowed significant germination (L. pendunculata: 16-18%; C. ladanifer: 5-11%) and biomass production of both species (g FW/pot; Roots: 16.3-30.9, Shoots: 41.2-76.4 depending on species and Technosol). In the control was reached the lowest germination (3%) and seedlings died after 40 days, so the improvement of the chemical characteristics of the surface layer, i.e. the implementation of the designed Technosols, is essential to ensure good vegetative development. No visual symptoms of nutritional deficiency and phytotoxicity neither element concentrations above hazardous levels for domestic animals intake were observed in those plants species. The alkaline barrier's components stabilise the sulfide-rich wastes by decreasing the oxidation and capillary rise of acid solutions that are rich in metals/metalloids. The limestone gravel showed, in some places, a thin layer of salts from alunite-jarosite group and metal-oxyhydroxides. As an outcome, the biotechnology was efficient and sustainable allowing the combined rehabilitation of both mine wastes at long-term.

Lavenders ( Lavandula species) are aromatic plants of great economic value for fragrance, pharmaceutical and cosmetic industries. Their biological activities and application on therapy can be compromised due to bioactive compounds physicochemical instability. Therefore, nanotechnology can be used as a way of achieving this stabilization. The antioxidant profile of different extracts from Lavandula stoechas ssp. luisieri and L . pedunculata were established by lipid peroxidation inhibition and the antioxidant activity confirmed by the free radical scavenging method. Methanol extracts of L . stoechas ssp. luisieri and L . pedunculata , due to their high antioxidant activity, phenolic content (1387.21 and 1044.19 mg gallic acid equivalents g sample −1 , respectively) and flavonoid amount (482.4 and 360.0 mg rutin equivalents g sample −1 ) were selected for encapsulation. The produced polymeric poly (lactic-co-glycolic) acid (PLGA) nanoparticles observed by scanning electron microscopy (SEM) showed a well-defined spherical shape and a high encapsulation efficiency (>96%) when the concentration of the extracts main component, rosmarinic acid, was used as indicator of encapsulation efficiency. The e pidermal permeation of both extracts through human epidermis and their in vitro cytotoxicity in human keratinocytes studies were suggestive of low risks of toxicity. In conclusion, the current study provides data for promising new cosmetic or dermatological formulations for the pharmaceutical industry, as anti-aging and antioxidant agents.

In the present study, the effect of S-cyclodextrin (beta-CD) and 2-hydroxypropyl-beta-cyclodextrin (HP-beta-CD) on the antioxidant activity and storage stability of the essential oils (EOs) from Lavandula viridis, Lavandula pedunculata subsp. Iusitanica and Thymus lotocephalus (Lamiaceae) was investigated. The presence of both cyclodextrins (CDs) was found to produce a remarkable enhancement on the antioxidant activity as assessed by the oxygen radical absorbance capacity (ORAC) assay. The highest free radical-scavenging effect was observed in the EO from T. lotocephalus, particularly at the highest concentration of HP-beta-CD (i.e., 1900 mu moI(TE)/g(EO)). Moreover, the antioxidant activity of the EO/HP-beta-CD complexes was found to be considerably stable over 30 days, at 4 degrees C, possibly due to the higher solubility of the modified beta-CD. The results obtained suggest that these systems can be a very interesting alternative as a natural source of antioxidants for different applications such as in the food preservation field.

We investigated the metabolic profile and biological activities of the essential oil and polar extracts of Lavandula pedunculata subsp. lusitanica (Chaytor) Franco collected in south Portugal. Gas chromatography-mass spectrometry (GC-MS) analysis revealed that oxygen-containing monoterpenes was the principal group of compounds identified in the essential oil. Camphor (40.6%) and fenchone (38.0%) were found as the major constituents. High-performance liquid chromatography with diode array detection (HPLC-DAD) analysis allowed the identification of hydroxycinnamic acids (3-O-caffeoylquinic, 4-O-caffeoylquinic, 5-O-caffeoylquinic and rosmarinic acids) and flavones (luteolin and apigenin) in the polar extracts, with rosmarinic acid being the main compound in most of them. The bioactive compounds from L. pedunculata polar extracts were the most efficient free-radical scavengers, Fe2+ chelators and inhibitors of malondialdehyde production, while the essential oil was the most active against acetylcholinesterase. Our results reveal that the subspecies of L. pedunculata studied is a potential source of active metabolites with a positive effect on human health. © 2013 Elsevier Ltd. All rights reserved.

In dry-summer seasonal climates, the beginning of the rainy season can prompt germination under different temperatures, depending on altitude. Understanding germination responses to temperature with altitude is important in fire-prone environments for species regenerating after fire from seeds (seeders), particularly under changing climate. Here we investigated the role of temperature in four Mediterranean seeder shrubs from Central Spain. Seeds from 17 sites (285–1,253 m altitude), of two hard-seeded nanophanerophytes (Cistus ladanifer and C. salviifolius) and two soft-seeded chamaephytes (Lavandula pedunculata and Thymus mastichina) were investigated. Intact and heat shock treated seeds were set to germinate under four temperature regimes, including a treatment simulating future warming. GLM with binomial or gamma functions were used to test treatment effects using altitude as a covariate. Altitude was a significant covariate only in L. pedunculata. Temperature did not affect final germination in either Cistus, but it significantly affected T. mastichina, and interacted with altitude in L. pedunculata, whereby the higher the altitude the less it germinated with decreasing temperature. Germination speed (T50) was lower at colder temperatures in all but C. salviifolius that was insensitive to our treatments. Heat shock significantly increased final germination in both Cistus and T. mastichina, but did not interact with temperature or altitude. We conclude that germination response to temperature, including varying sensitivity with altitude, differed among these species; thus, changes in the timing of the onset of the rainy season will diversely affect populations at various altitudes. We discuss our results in a context of changing climate and fire.

Lavanders (Lavandula spp.) are aromatic plants with high industrial and commercial value, used in food industry, perfumery and pharmaceutical preparations. Micropropagation can be used for large-scale multiplication of essential oil producing plants thus avoiding an overexploitation of natural resources. This work aims to develop a reliable protocol for the in vitro propagation of Lavandula pedunculata and to evaluate the potential of these plants for essential oil production with industrial application. In vitro cultures were established on media with different concentrations of benzyladenine. Trichome morphology was examined by SEM and their secretion analyzed by histochemical tests. Essential oils were isolated by hydrodistillation and analyzed by GC and GC-MS. The essential oils and trichomes of in vitro plants were compared with those of field-growing plants. Best propagation rates were achieved with 0.25 mg/l BA. Rooting occurred without treatment with auxins thus preventing callus growth. Trichomes and essential oils of the in vitro propagated plants were similar to those of the parent plants. Two chemotypes were characterized: 1,8-cineole/camphor type and fenchone type. Since trichomes and essential oils of L. pedunculata micropropagated plantlets are identical to those produced by field-growing plants, in vitro cultures can be used for essential oil production without affecting natural resources. Moreover, the quality of the essential oils is assured and this protocol can be further applied to the propagation of selected chemotypes for industrial purposes.

The aim of this study was to characterize the phenolic composition and evaluate the bioactivity of several samples of Lavandula pedunculata (Mill.) Cav, and to compare aqueous and hydroethanolic extracts. Plant materials were obtained by growing some accessions (seed samples) of various wild populations from different regions of Portugal conserved at the Portuguese Genebank in Braga. Phenolic compounds were analised by HPLC-DAD-ESI/MSn, antioxidant potential through in vitro assays (DPPH radical scavenging activity, reducing power and inhibition of lipid peroxidation), cytotoxicity on tumor cells (MCF-7, NCI-H460, HeLa and HepG2) and non-tumor (PLP2) cells, anti-inflammatory activity in rat RAW 264.7 macrophages, by the ability to inhibit NO production and antimicrobial potential by the microdilution method with INT dye (iodonitrotetrazolium chloride). Thirteen compounds were identified, being salvianolic acid B, rosmarinic acid and luteolin-7-O-glucuronide, the main compounds present, with values ranging between 44.3⁻582, 50.9⁻550, and 24.36⁻101.5 mg/g extract, respectively. L. pedunculata aqueous extract revealed a higher antioxidant potential (EC50 values between 14 to 530 μg/mL), which could be related to its higher concentration in phenolic compounds; however, the hydroethanolic extract showed a higher anti-inflammatory (lower EC50 values than 124 μg/mL) potential and antiproliferative capacity (lower GI50 values than 34 μg/mL). Thus, this study highlights the bioactive effects of this species and opens up possibilities of uses in food and pharmaceutical formulations. However, there are potential differences in such properties according to geographical origin of plant material, as in general, the samples from Alentejo presented higher results in all the bioactivities, compared with Trás-os-Montes samples. The authors are grateful to the Foundation for Science and Technology (FCT, Portugal) and FEDER under Program PT2020 for financial support to CIMO (UID/AGR/00690/2013), L. Barros and R. Calhelha contracts. The authors are grateful to FEDER-Interreg España-Portugal programme for financial support through the project 0377_Iberphenol_6_E. info:eu-repo/semantics/publishedVersion

The chemical composition and antifungal activity of the essential oils of Lavandula pedunculata (Miller) Cav., harvested in North and Central Portugal, were investigated. The essential oils were isolated by hydrodistillation and analyzed by GC and GC/MS. The minimal-inhibitory concentration (MIC) and the minimal-lethal concentration (MLC) of the essential oils and of their major constituents were used to evaluate the antifungal activity against different strains of fungi involved in candidosis, dematophytosis, and aspergillosis. The oils were characterized by a high percentage of oxygenated monoterpenes, the main compounds being 1,8-cineole (2.4-55.5%), fenchone (1.3-59.7%), and camphor (3.6-48.0%). Statistical analysis differentiated the essential oils into two main types, one characterized by the predominance of fenchone and the other one by the predominance of 1,8-cineole. Within the 1,8-cineole chemotype, two subgroups were well-defined taking into account the percentages of camphor. A significant antifungal activity of the oils was found against dermatophyte strains. The essential oil with the highest content of camphor was the most active with MIC and MLC values ranging from 0.32-0.64 microl/ml.

International audience; Lavandula pedunculata (Mill.) Cav. subsp. lusitanica, L. stoechas L. subsp. stoechas and L. viridis l'Hér. are three lavender taxa that belong to the botanical section Stoechas and are widely used as aromatherapy, culinary herb or folk medicine in many Mediterranean regions. The analysis of their bioactive volatile constituents revealed the presence of 124 substances, the most abundant being the bicyclic monoterpenes fenchone, camphor and 1,8-cineole that give these three species their respective chemotypes. Most noteworthy was fenchone which, with its reduced form fenchol, made 48% of the total volatile constituents of L. pedunculata whilst present at 2.-% in L. stoechas and undetectable in L. viridis. In order to provide a molecular explanation to the differences in volatile compounds of these 3 species, two monoterpene synthases (monoTPS) and one sesquiterpene synthase (sesquiTPS) were cloned in L. pedunculata and functionally characterized as fenchol synthase (LpFENS), α-pinene synthase (LpPINS) and germacrene A synthase (LpGEAS). The two other lavender species contained a single orthologous gene for each of these three classes of TPS with similar enzyme product specificities. Expression profiles of FENS and PINS genes matched the accumulation profile of the enzyme products unlike GEAS. This study provides one of the rare documented cases of chemotype modification during plant speciation via changes in the level of plant TPS gene expression, and not functionality.

In this study the characterization of a total of 60 honey samples with Protected Denomination of Origin (PDO) collected over three harvests (2009–2011, inclusive), from the Northeast of Portugal was carried out based on the presence of pollen, physicochemical and microbiological characteristics. All samples were found to meet the European Legislation, but some didn’t meet the requirements of the PDO specifications. Concerning the floral origin of honey, our results showed the prevalence of rosemary (Lavandula pedunculata) pollen. The microbiological quality of all the analyzed samples was satisfactory, since fecal coliforms, sulfite-reducing clostridia and Salmonella were absent, and molds and yeasts were detected in low counts. Significant differences between the results were studied using one-way analysis of variance (ANOVA), followed by Tukey’s HSD test. The samples were submitted to discriminant function analysis, in order to determine which variables differentiate between two or more naturally occurring groups (Forward Stepwise Analysis). The variables selected were in this order: diastase activity, pH, reducing sugars, free acidity and HMF. The pollen spectrum has perfect discriminatory power. This is the first study in which a honey with PDO was tested, in order to assess its compliance with the PDO book of specifications X.F. would also like to thank the Xunta de Galicia (Isidro Parga Pondal Program for young researchers, Grant No. IPP-020) SI

We studied the water relations of 6 shrub and 3 tree species typical of the mediterranean climate region of central Spain to identify differential responses to water stress between and within species, and to determine if free proline concentration in leaves could be used as a water stress indicator. Predawn and midday water potentials (Ψw) on a seasonal basis, relative water content (RWC), leaf mass per area, foliar nitrogen and free proline concentrations were measured. The lowest water potentials were observed at the end of the summer, with recovery to higher water potentials in the fall and winter seasons. Species differed regarding the annual Ψw fluctuation. Thymus zygis, Halimium viscosum, Genista hirsuta and Juniperus oxycedrus exhibited the most negative midday and predawn Ψw (both less than -6 MPa) with a large magnitude of response to changing conditions in soil moisture of the upper horizon of the soil. Lavandula pedunculata and Cistus ladanifer showed a moderate response. Quercus rotundifolia, Quercus faginea and Retama sphaerocarpa showed a modest response. The Ψw of different size individuals of Quercus rotundifolia and Cistus ladanifer were compared. The annual Ψw fluctuation was greater in small individuals as compared to large individuals. In every species, there was an increase in proline concentration of bulk leaf tissues when predawn Ψw dropped below -5 MPa. Small plants of Cistus ladanifer reached lower water potentials and also higher concentration of proline than bigger plants. Proline could possibly be used as a drought stress indicator in every species except Q. rotundifolia. It is suggested that in addition to water stress avoidance due to deep root systems, some mechanisms of water stress tolerance may operate among shrub and tree species of central Spain.

Essential oil, ethanolic extract and decoction of 10 plant species from interior Portugal were analyzed for their activity towards acetylcholinesterase (AChE) enzyme and their antioxidant activity. Of these, Melissa officinalis, Paronychia argentea, Sanguisorba minor, Hypericum undulatum and Malva silvestris are used in herbal medicine, Laurus nobilis and Mentha suaveolens as condiments, and Salvia officinalis, Lavandula angustifolia and Lavandula pedunculata also as aromatics. Melissa officinalis and Mentha suaveolens showed AChE inhibitory capacity higher then 50% in the essential oil fraction. Laurus nobilis, Hypericum undulatum, and Sanguisorba minor showed a high inhibition value of AChE in the ethanolic fraction, 64% (1 mg ml(-1)) 68% (0.5 mg ml(-1)), and 78% (1 mg ml(-1)), respectively. Higher values of AChE inhibitory activity were found using decoctions of Lavandula pedunculata, Mentha suaveolens and Hypericum undulatum, 68, 69 and 82% (at a concentration of 5mg dry plant ml(-1) of assay), respectively. The free radical scavenger activity was higher for the polar extracts. In the water extracts most of the plants showed values around 90%. When antioxidant activity was measured with the beta-carotene-linoleic acid assay high activity (65-95%) was also found in the water extracts. Hypericum undulatum, Melissa officinalis and Laurus nobilis showed both high AChE inhibitory capacity and antioxidant activity.

The selection of native Lavandula species and their economic exploitation have increased in the last few years. Micropropagation techniques have been used as an alternative for vegetative propagation allowing the multiplication of selected genotypes and chemotypes. Our previous studies showed that the essential oils of Lavandula pedunculata have an important antifungal activity against dermatophyte strains. Therefore, a new line of investigation concerning the in vitro culture of this species is justified. In the present study we compare the morphology of the leaf trichomes and the chemical composition of their essential oils in both field-growing and in vitro propagated plants.

Juvenile plants of Thymus mastichina, Thymus zygis, Lavandula pedunculata, Genista hirsuta, and Cistus ladanifer were transplanted from the field to the greenhouse; the soil used was the same in which they had grown at their natural site. The plants were grown to maturity to determine comparative growth, water status, and mycorrhizal colonization under greenhouse conditions and to determine the influence of drought on the symbiosis. After 24 months, L. pedunculata showed the most developed root system; G. hirsuta showed the lowest root/shoot ratio and the lowest percentage of mycorrhizal colonization. This percentage ranged from 32.0% to 82.4% for G. hirsuta and L. pedunculata, respectively. There was a positive relationship between root biomass and root/shoot ratio with mycorrhization percentage, and a positive response of water potential to that percentage also occurred. After a moderate drought treatment over 4 weeks, root and shoot biomass and root/shoot ratio as well as mycorrhizal colonization decre...