Growth and production of volatile compounds of yarrow (Achillea millefolium L.) under different irrigation depths.

Yarrow (Achillea millefolium L., Asteraceae) is an important medicinal plant used worldwide for its medicinal properties such as the analgesic, antioxidant and anti-inflammatory ones. The aim of this study was to evaluate the growth and production of photosynthetic pigments and of volatile constituents of Achillea millefolium L. under different irrigation depths. The treatments were the application of 55, 110, 220, 440 and 880 mm of water for a period of 110 days. Data were submitted to polynomial regression analysis at 5% probability, while the volatile constituents were analyzed by standard deviation. Different irrigation depths provided quadratic growth responses being the highest dry matter production at the depth of 440 mm. The contents of chlorophyll a, b, total and carotenoids were higher at the lower depth tested (55 mm). The major volatile compounds identified were sabinene, 1,8-cineol, borneol and ß-caryophyllene. Increased water availability reduced the complexity of the volatile fraction of essential oil. Thus, it is recommended that the species be cultivated at 440 mm irrigation depth to have a higher production of dry matter and lower variation in the volatile profile of the essential oil.

Variation in Essential Oil Composition, Bioactive Compounds, Anatomical and Antioxidant Activity of Achillea aucheri, an Endemic Species of Iran, at Different Phenological Stages.

In the present research, the essential oil composition, bioactive compounds, anatomical and antioxidant activity of Achillea aucheri were investigated at four phenological stages. The yield of essential oil ranged from 0.1 % (five leaves appearance stage) to 0.7 % (full flowering stage). So, the oils from the plants harvested at the full flowering stage provided higher essential oil contents than those at other stages. The analysis of the essential oils by GC/MS revealed the presence of a large number of components represented mainly by oxygenated monoterpenes (24.36-77.81 %). The compounds revealed high variation at different phenological stages. The major constituents of A. aucheri were carvacrol (1.07-19.17 %), camphor (5.44-9.99 %), δ-cadinene (0.20-13.88 %), thymol (2.15-7.80 %), germacrene D (4.50-7.24 %), 1,8-cineole (1.62-5.54 %) and α-terpineol (1.71-3.42 %). The total phenolic (51.63 mg tannic acid/g DW) and flavonoid (10.88 mg quercetin/g W) contents were more accumulated at the 50 % flowering stage than others. The results revealed that the accumulation of essential oil in A. aucheri increased as the plant grew from the five leaves appearance stage to the 100 % flowering stage with increasing the size and number of secretory structures. Finally, the extracts collected at the 50 % flowering stage showed the highest antioxidant activity as measured based on FTC and DPPH method with the IC50 values ranging from 719.65 µg/mL to 1039.17 µg/mL for the samples.

Herbicide Toxicity Testing with Non-Target Boreal Plants: The Sensitivity of Achillea millefolium L. and Chamerion angustifolium L. to Triclopyr and Imazapyr.

Terrestrial plant toxicity tests were conducted to determine the sensitivity of two boreal plants, yarrow (Achillea millefolium L.) and fireweed (Chamerion angustifolium L.), to the herbicides imazapyr and triclopyr. Both plants are common non-target species on northern powerline rights-of-way where the impacts of proposed herbicide applications are of concern. In the vegetative vigour test, triclopyr foliar spray caused extensive damage to A. millefolium at <50% of the maximum field application rate (inhibition concentration (IC)50 = 1443.8 g a.i. ha-1) and was lethal to C. angustifolium at the lowest dose tested (1210.9 g a.i. ha-1). Both species demonstrated extremely high sensitivity to imazapyr foliar spray: IC50s = 8.29 g a.i. ha-1 and 4.82 g a.i. ha-1 (<1.5% of the maximum field rate). The seedling emergence and seedling growth tests were conducted in the organic horizon of five boreal soils. Few differences in herbicide bioavailability between soils were detected. Triclopyr limited growth of A. millefolium, C. angustifolium and standard test species Calamagrostis canadensis at low levels (most IC50 estimates between 2-20 µg g-1). For imazapyr, IC50 estimates could not be calculated as there was >75% inhibition of endpoints at the lowest doses of ~2 µg g-1. A foliar application of triclopyr or imazapyr for woody species control would likely cause significant damage to boreal non-target plants. The high sensitivity of both species to herbicide residues in soil indicates long term impacts are dependent on herbicide degradation rates in northern conditions. A. millefolium performed well and is recommended for use in toxicity testing relevant to boreal regions.

Chemical Composition and Antibacterial Activity of the Bulgarian Endemic Species Achillea thracica from its Natural Habitat, and in vitro Propagated and ex vitro Established Plants.

The present study reports the chemical composition of headspace volatiles (HS) and acetone extracts of the endemic Bulgarian species Achillea thracica Velen. from its natural habitat (N), in vitro propagated (IN) and ex vitro established (EX) plants. Additionally, acetone extracts were tested by a disk diffusion method for antibacterial activity. Irregular monoterpenes were the most abundant HS volatile constituents, while 0,0-dimethyl quercetin was the most abundant flavonoid in the acetone extracts. The secondary metabolites of A. thracica grown in its natural habitat (N), propagated in vitro (IN) and ex vitro established (EX) showed that the qualitative composition is mutually similar, but there are differences in the quantitative composition. Considering antibacterial activity, IN and EX samples showed moderate activity against Pseudomonas aeruginosa and Escherichia coli.

Pesticide residues in some herbs growing in agricultural areas in Poland.

The aim of this paper was to assess residue content of plant protection products in selected herbs: Achillea millefolium L., Cichorium intybus L., Equisetum arvense L., Polygonum persicaria L., Plantago lanceolata L., and Plantago major L. The study comprises herbs growing in their natural habitat, 1 and 10 m away from crop fields. The herbs, 30 plants of each species, were sampled during the flowering stage between 1 and 20 July 2014. Pesticide residue content was measured with the QuECHERS method in the dry matter of leaves, stalks, and inflorescence, all mixed together. Out of six herb species growing close to wheat and maize fields, pesticide residues were found in three species: A. millefolium L., E. arvense L., and P. lanceolata L. Most plants containing the residues grew 1 m away from the wheat field. Two active substances of fungicides were found: diphenylamine and tebuconazole, and one active substance of insecticides: chlorpyrifos-ethyl. Those substances are illegal to use on herbal plants. Samples of E. arvense L. and P. lanceolata L. contained two active substances each, which constituted 10% of all samples, while A. millefolium L. contained one substance, which is 6.6% of all samples.

A comparative study of antimony accumulation in plants growing in two mining areas in Iran, Moghanlo, and Patyar.

Antimony occurs locally at high concentrations in some mineralized soils. Very little is known about behavior of antimony in plants. In this study, we analyzed the soil and vegetation of two mining areas in Iran, Patyar, and Moghanlo. Total Sb concentrations in soil were 358-3482 mg/kg in Moghanlo and 284-886 mg/kg in Patyar. Corresponding Sb concentrations in plant shoots were 0.8-287 and 1.3-49 mg/kg, respectively. In both areas, foliar Sb concentrations increased with acid-extractable soil Sb, although the slope was about 2-fold steeper for Patyar than for Moghanlo. Regressing the foliar concentrations on water-soluble Sb yielded identical slopes for both areas, suggesting that the soluble fraction of Sb rather than total Sb is the direct determinant of foliar Sb accumulation. Both in Patyar and Moghanlo, only a minor part of the total variance of shoot Sb was explained by soluble Sb. The major part was explained by plant species, demonstrating that plant taxonomic identity is the most important determinant of foliar Sb accumulation capacity in both areas. The translocation factor (TF) was highly variable too, with species as the only significant variance component. Only four species were able to accumulate more than 100 mg/kg Sb in their leaves. Among these species, Achillea wilhelmsii and Matthiola farinosa were by far the best Sb accumulators, with, on average, 141 and 132 mg/kg Sb in their leaves. Of these two, only Matthiola farinosa consistently maintained TF values far above unity across the whole range of soluble Sb in Moghanlo.

Root foraging influences plant growth responses to earthworm foraging.

Interactions among the foraging behaviours of co-occurring animal species can impact population and community dynamics; the consequences of interactions between plant and animal foraging behaviours have received less attention. In North American forests, invasions by European earthworms have led to substantial changes in plant community composition. Changes in leaf litter have been identified as a critical indirect mechanism driving earthworm impacts on plants. However, there has been limited examination of the direct effects of earthworm burrowing on plant growth. Here we show a novel second pathway exists, whereby earthworms (Lumbricus terrestris L.) impact plant root foraging. In a mini-rhizotron experiment, roots occurred more frequently in burrows and soil cracks than in the soil matrix. The roots of Achillea millefolium L. preferentially occupied earthworm burrows, where nutrient availability was presumably higher than in cracks due to earthworm excreta. In contrast, the roots of Campanula rotundifolia L. were less likely to occur in burrows. This shift in root behaviour was associated with a 30% decline in the overall biomass of C. rotundifolia when earthworms were present. Our results indicate earthworm impacts on plant foraging can occur indirectly via physical and chemical changes to the soil and directly via root consumption or abrasion and thus may be one factor influencing plant growth and community change following earthworm invasion. More generally, this work demonstrates the potential for interactions to occur between the foraging behaviours of plants and soil animals and emphasizes the importance of integrating behavioural understanding in foraging studies involving plants.

Species-specific responses to community density in an unproductive perennial plant community.

Most studies of density dependent regulation in plants consider a single target species, but regulation may also occur at the level of the entire community. Knowing whether a community is at carrying capacity is essential for understanding its behaviour because low density plant communities may behave quite differently than their high density counterparts. Also, because the intensity of density dependence may differ considerably between species and physical environments, generalizations about its effects on community structure requires comparisons under a range of conditions. We tested if: (1) density dependent regulation occurs at the level of an entire plant community as well as within individual species; (2) the intensity (effect of increasing community density on mean plant mass) and importance (the effect of increasing density, relative to other factors, on mean plant mass) of competition increases, decreases or remains unchanged with increasing fertilization; (3) there are species-specific responses to changes in community density and productivity. In 63 1 m2 plots, we manipulated the abundance of the nine most common species by transplanting or removing them to create a series of Initial Community Densities above and below the average natural field density, such that the relative proportion of species was consistent for all densities. Plots were randomly assigned to one of three fertilizer levels. At the community level, negative density dependence of mean plant size was observed for each of the 4 years of the study and both the intensity and importance of competition increased each year. At the species level, most species' mean plant mass were negatively density dependent. Fertilizer had a significant effect only in the final year when it had a negative effect on mean plant mass. Our data demonstrate a yield-density response at the entire community-level using perennial plant species in a multi-year experiment.

Effect of Light Environment on Growth and Phenylpropanoids of Yarrow (Achillea collina cv. SPAK) Grown in the Alps.

A 2-year field study on the effect of different light environments, obtained by using cladding materials (polyethylene films and shade net) able to cut off specific regions of the photosynthetically active radiation and ultraviolet wavebands, on the growth and phenylpropanoids content of Achillea collina grown in the Alps was conducted. Overall the plant growth was strongly enhanced in the second growing season irrespective of radiation treatment. The light environment did not affect total biomass accumulation, but only carbon allocation to leaves or inflorescences. Indeed the phenylpropanoid levels in inflorescences appeared to be more sensitive to the light environment than leaves as the latter showed high constitutive amounts of these compounds. However, the use of polyethylene films improved to some extent the content of caffeic acid derivatives in leaves. Our results showed that yarrow production, in the alpine situation considered, is influenced by the growing season and the light environment, providing a basis to optimize its quality, depending on the concentration of bioactive compounds, by means of proper agronomic practices.

Involvement of bradykinin and prostaglandins in the diuretic effects of Achillea millefolium L. (Asteraceae).

ETHNOPHARMACOLOGICAL RELEVANCE: Achillea millefolium L. (Asteraceae), popularly known as "mil-folhas", is well recognized and widely used in Brazilian folk medicine to treat heart and kidney disorders. Among its popularly described effects are diuretic and hypotensive actions. AIM OF THE STUDY: The diuretic activity of Achillea millefolium L. extracts and its semi-purified fractions, as well as the mechanisms involved, were evaluated in male Wistar rats. MATERIAL AND METHODS: An aqueous extract (AEAM, 125-500 mg/kg), hydroethanolic extract (HEAM, 30-300 mg/kg), dichloromethane subfractions (DCM-2, 10 and 30 mg/kg), or hydrochlorothiazide (10mg/kg), were orally administered and the animals were kept in metabolic cages for 8h for urine collection. To evaluate the involvement of bradykinin and prostaglandins in the diuretic action of Achillea millefolium, selected groups of rats received HOE-140 (1.5mg/kg, i.p.) or indomethacin (5mg/kg, p.o.), before treatment with a DCM-2 subfraction (30 mg/kg). The urinary volume, conductivity, pH, density and electrolyte excretion were measured. RESULTS: Similar to hydrochlorothiazide, both HEAM and DCM-2, but not AEAM, increased urinary volume and the excretion of Na(+) and K(+) when compared with the control group (vehicle). The diuretic effect of DCM-2 was abolished by HOE-140 (a bradykinin B2 receptor antagonist), as well as by indomethacin (a cyclooxygenase inhibitor). CONCLUSION: The present study reveals that extracts obtained from Achillea millefolium are able to effectively increase diuresis when orally administered in rats. This effect depends on both the activation of bradykinin B2 receptors and the activity of cyclooxygenases.

Mechanisms for success after long-term nutrient enrichment in a boreal forest understory.

Global levels of reactive nitrogen are predicted to rise in the coming decades as a result of increased deposition from the burning of fossil fuels and the large-scale conversion of nitrogen into a useable form for agriculture. Many plant communities respond strongly to increases in soil nitrogen, particularly in northern ecosystems where nitrogen levels are naturally very low. An experiment in northern Canada that was initiated in 1990 has been investigating the effects of long-term nutrient enrichment (fertilizer added annually) on a boreal forest understory community. We used this experiment to investigate why some species increase in abundance under nutrient enrichment whereas others decline. We focused on four species that differed in their responses to fertilization: Mertensia paniculata and Epilobium angustifolium increased in abundance, Achillea millefolium remained relatively constant and Festuca altaica declined. We hypothesized that the two species that were successful in the new high-nutrient, light-limited environment would be taller, have higher specific leaf area, change phenology by growing earlier in the season and be more morphologically plastic than their less successful counterparts. We compared plant height, specific leaf area, growth spurt date and allocation to leaves in plants grown in control and fertilized plots. We demonstrated that each of the two species that came to dominate fertilized plots has a different combination of traits and responses that likely gave them a competitive advantage; M. paniculata has the highest specific leaf area of the four species whereas E. angustifolium is tallest and exhibits morphological plasticity when fertilized by increasing biomass allocation to leaves. These results indicate that rather than one strategy determining success when nutrients become available, a variety of traits and responses may contribute to a species' ability to persist in a nutrient-enriched boreal forest understory.

Chemical composition and free radical scavenging activity of the essential oil of Achillea ligustica growing wild in Lipari (Aeolian Islands, Sicily).

In the present study the chemical composition of the essential oils from aerial parts and flowers of Achillea ligustica All., collected in Lipari (Aeolian Islands) was evaluated by GC and GC-MS. (Z)-Chrysanthenyl acetate was the most abundant component of both oils (29.6% in A1 and 27.8% in F1), followed by viridiflorol (16.8% in A1 and 21.6% in F1), bornyl acetate (8.7% in A1 and 11.6% in F1) and 1,8-cineole (7.4% in A1 and 9.3% in F1). A comparison was made of the composition of the different populations studied so far. Futhermore, the free radical scavenging activity of the oil was determined by DPPH and ABTS methods.

Context dependence in foraging behaviour of Achillea millefolium.

Context-dependent foraging behaviour is acknowledged and well documented for a diversity of animals and conditions. The contextual determinants of plant foraging behaviour, however, are poorly understood. Plant roots encounter patchy distributions of nutrients and soil fungi. Both of these features affect root form and function, but how they interact to affect foraging behaviour is unknown. We extend the use of the marginal value theorem to make predictions about the foraging behaviour of roots, and test our predictions by manipulating soil resource distribution and inoculation by soil fungi. We measured plant movement as both distance roots travelled and time taken to grow through nutrient patches of varied quality. To do this, we grew Achillea millefolium in the centers of modified pots with a high-nutrient patch and a low-nutrient patch on either side of the plant (heterogeneous) or patch-free conditions (homogeneous). Fungal inoculation, but not resource distribution, altered the time it took roots to reach nutrient patches. When in nutrient patches, root growth decreased relative to homogeneous soils. However, this change in foraging behaviour was not contingent upon patch quality or fungal inoculation. Root system breadth was larger in homogeneous than in heterogeneous soils, until measures were influenced by pot edges. Overall, we find that root foraging behaviour is modified by resource heterogeneity but not fungal inoculation. We find support for predictions of the marginal value theorem that organisms travel faster through low-quality than through high-quality environments, with the caveat that roots respond to nutrient patches per se rather than the quality of those patches.

Apomixis is not prevalent in subnival to nival plants of the European Alps.

BACKGROUND AND AIMS: High alpine environments are characterized by short growing seasons, stochastic climatic conditions and fluctuating pollinator visits. These conditions are rather unfavourable for sexual reproduction of flowering plants. Apomixis, asexual reproduction via seed, provides reproductive assurance without the need of pollinators and potentially accelerates seed development. Therefore, apomixis is expected to provide selective advantages in high-alpine biota. Indeed, apomictic species occur frequently in the subalpine to alpine grassland zone of the European Alps, but the mode of reproduction of the subnival to nival flora was largely unknown. METHODS: The mode of reproduction in 14 species belonging to seven families was investigated via flow cytometric seed screen. The sampling comprised 12 species typical for nival to subnival plant communities of the European Alps without any previous information on apomixis (Achillea atrata, Androsace alpina, Arabis caerulea, Erigeron uniflorus, Gnaphalium hoppeanum, Leucanthemopsis alpina, Oxyria digyna, Potentilla frigida, Ranunculus alpestris, R. glacialis, R. pygmaeus and Saxifraga bryoides), and two high-alpine species with apomixis reported from other geographical areas (Leontopodium alpinum and Potentilla crantzii). KEY RESULTS: Flow cytometric data were clearly interpretable for all 46 population samples, confirming the utility of the method for broad screenings on non-model organisms. Formation of endosperm in all species of Asteraceae was documented. Ratios of endosperm : embryo showed pseudogamous apomixis for Potentilla crantzii (ratio approx. 3), but sexual reproduction for all other species (ratios approx. 1·5). CONCLUSIONS: The occurrence of apomixis is not correlated to high altitudes, and cannot be readily explained by selective forces due to environmental conditions. The investigated species have probably other adaptations to high altitudes to maintain reproductive assurance via sexuality. We hypothesize that shifts to apomixis are rather connected to frequencies of polyploidization than to ecological conditions.

Competition for patchy soil resources reduces community evenness.

The presence of small-scale patches of soil resources has been predicted to increase competition, because multiple species will proliferate roots in the same small area, and therefore decrease plant diversity. I tested whether such patches reduced species evenness in a community of four old-field species, both with and without interspecific interactions. In species mixtures, patches reduced evenness, while in "communities" constructed via combined monocultures, in which species did not compete, patches increased evenness. Therefore, the reduction in evenness in response to patches was due to changes in competition. Community-level changes may be attributable to plant foraging traits-in species with low foraging precision, competition reduced abundance much more in patchy soils than in even soils, while in species with high root foraging precision, the effect of competition was similar in patchy and even soils.

Influence of environmental factors on composition of phenolic antioxidants of Achillea collina Becker ex Rchb.

Effects of environmental growth conditions on the antioxidant capacity, total phenolic content and composition of Achillea collina Becker ex Rchb. were investigated. Methanol extracts and infusions obtained from leaves and inflorescences of plants cultivated in the Italian Alps at two different altitudes (600 and 1050 m a.s.l.) were evaluated. Infusions exhibited the highest antioxidant capacity (1/IC(50) values from 4.35 ± 0.72 to 8.90 ± 0.74), total phenolic content (from 31.39 ± 4.92 to 49.36 ± 5.70 mg gallic acid equivalents (GAE) g(-1) DW), chlorogenic acid (from 9.21 ± 1.52 to 31.27 ± 6.88 mg g(-1) DW), 3,5-di-O-caffeoylquinic acid (from 12.28 ± 3.25 to 25.13 ± 1.99 mg g(-1) DW) and 4,5-di-O-caffeoylquinic acid (from 7.38 ± 1.01 to 12.78 ± 2.61 mg g(-1) DW) content. Climate (as influenced by altitude) was shown to be the main environmental factor influencing yarrow composition and properties. Leaf extracts from the higher experimental site showed a 2-4-fold increase of chlorogenic acid level. Achillea collina can be considered as a very good source of bioactive phenolic compounds, and growing it at high altitude may constitute an effective way to significantly enhance its quality for both medicinal and nutritional uses.

Plant root growth and the marginal value theorem.

All organisms must find and consume resources to live, and the strategies an organism uses when foraging can have significant impacts on their fitness. Models assuming optimality in foraging behavior, and which quantitatively account for the costs, benefits, and biological constraints of foraging, are common in the animal literature. Plant ecologists on the other hand have rarely adopted an explicit framework of optimality with respect to plant root foraging. Here, we show with a simple experiment that the marginal value theorem (MVT), one of the most classic models of animal foraging behavior, can provide novel insights into the root foraging behavior of plants. We also discuss existing data in the literature, which has not usually been linked to MVT to provide further support for the benefits of an optimal foraging framework for plants. As predicted by MVT, plants invest more time and effort into highly enriched patches than they do to low-enriched patches. On the basis of this congruency, and the recent calls for new directions in the plant foraging literature, we suggest plant ecologists should work toward a more explicit treatment of the idea of optimality in studies of plant root foraging. Such an approach is advantageous because it forces a quantitative treatment of the assumptions being made and the constraints on the system. While we believe significant insight can be gained from the use of preexisting models of animal foraging, ultimately plant ecologists will have to develop taxa-specific models that account for the unique biology of plants.

Biological activities of yarrow species (Achillea spp.).

The genus Achillea consists of about 140 perennial herbs native to the Northern hemisphere. Traditional indications of their use include digestive problems, liver and gall-bladder conditions, menstrual irregularities, cramps, fever, wound healing. The Commission E approves its internal use for loss of appetite and dyspeptic ailments (gastric catarrh, spastic discomfort), externally it is used in form of sitz bath or as a compress against skin inflammation, slow healing wounds, bacterial or fungal infections. In the last decades, pharmacological studies became intensive, although human clinical investigations are still rare. Recent findings have confirmed several traditional uses. The largest number of data accumulated for antioxidant and anti-inflammatory effects. There are positive results on the analgesic, anti-ulcer, choleretic, hepatoprotective and wound healing activities. First results on other interesting therapeutical areas - antihypertensive, antidiabetic, antitumor, antispermatogenic activities -need confirmation. Yarrow can be used also as an insect repellent. Contact dermatitis as adverse effect may be connected to sesquiterpenes. The diversity and complexity of the effective compounds of yarrow species explains the broad spectrum of their activity. According to the literature the pharmacological effects are mainly due to the essential oil, proazulenes and other sesquiterpene lactones, dicaffeoylquinic acids and flavonoids. Synergistic actions of these and other compounds are also supposed. Achillea species have different chemical and therapeutical values. Despite of numerous data, correct evaluation of the results is difficult because of missing generally accepted taxonomical nomenclature. The used chemical-analytical methods and bio-assays are utmost diverse, making the comparison complicated. Further research on the activity is needed using exactly defined plant material, standardized methods and chemical analysis.

Geographical variation in selection, from phenotypes to molecules.

Molecular technologies now allow researchers to isolate quantitative trait loci (QTLs) and measure patterns of gene sequence variation within chromosomal regions containing important polymorphisms. I develop a simulation model to investigate gene sequence evolution within genomic regions that harbor QTLs. The QTLs influence a trait experiencing geographical variation in selection, which is common in nature and produces obvious differentiation at the phenotypic level. Counter to expectations, the simulations suggest that selection can substantially affect quantitative genetic variation without altering the amount and pattern of molecular variation at sites closely linked to the QTLs. Even with large samples of gene sequences, the likelihood of rejecting neutrality is often low. The exception is situations where strong selection is combined with low migration among demes, conditions that may be common in many plant species. The results have implications for gene sequence surveys and, perhaps more generally, for interpreting the apparently weak connection between levels of molecular and quantitative trait variation within species.

Physiological and biochemical stress responses in grassland species are influenced by both early-season ozone exposure and interspecific competition.

The effects of two-year early season ozone exposure on physiological and biochemical stress response were investigated in model plant communities. Achillea millefolium and Veronica chamaedrys target plants were grown in monocultures and in mixed cultures with Poa pratensis (phytometer) and exposed in open-top chambers over two years for five weeks to charcoal-filtered (CF) air plus 25 nl l(-1) O3 (control) and non-filtered (NF) air plus 50 nl l(-1) O3. Significant O3 effects were detected in different physiological and biochemical parameters, evidencing interspecific differences in metabolic stress responses and a strong influence of the competition factor. O3 induced strong oxidative effects in Achillea irrespective to the different growth modality. Veronica showed less O3-induced effects in monoculture than when grown in competition with the phytometer. Poa exhibited a different behaviour against O3 depending on the species in competition, showing an overall higher sensitivity to O3 when in mixture with Achillea.