Seasonal and Diurnal Variation in Leaf Phenolics of Three Medicinal Mediterranean Wild Species: What Is the Best Harvesting Moment to Obtain the Richest and the Most Antioxidant Extracts?

Mediterranean plants biosynthesize high amounts of polyphenols, which are important health-promoting compounds. Leaf polyphenolic composition changes according to environmental conditions. Therefore, it is crucial to know the temporal variation in their production. This study aimed to: i) evaluate the monthly and daily changes in polyphenols of Phyllirea latifolia, Cistus incanus, and Pistacia lentiscus to identify their best harvesting moment, ii) verify the possible correlations between phenolic production and temperature and irradiation, iii) evaluate their antioxidant capacity using 2,2-diphenyl-1-picrylhydrazyl (DPPH) and hydroxyl radical (OH)scavenging assays. The extracts of leaves harvested at 8:00, 13:00 and 18:00, in May, July, and October for two years were analysed by HPLC-DAD. Both "month" and "time of the day" affected the polyphenolic content in all species. July at 13:00 was the best harvesting moment for all polyphenolic classes of P. latifolia and only for some classes of C. incanus and P. lentiscus. Environmental parameters positively correlated with the polyphenols of C. incanus and P. latifolia, while the antioxidant capacity only varied in this last species, reaching the highest value in July. Results of the study allow to determine the balsamic time for each species. Moreover, the relationship between polyphenols and environmental data can be useful for the cultivation of these plants under controlled conditions.

Use of phytoremediated sediments dredged in maritime port as plant nursery growing media.

We evaluated the potential of a phytoremediated sediment (TR) dredged from maritime port as peat-free growth substrate for seven ornamental plants, in comparison with an untreated sediment (NT), in a greenhouse experiment. The studied plants were Quercus ilex, Photinia x fraseri, Viburnum tinus, Cistus albidus, Raphiolepis indica, Westringia fruticosa and Teucrium fruticans. Plant growth was monitored for ten months, and the changes in the physico-chemical properties, toxicity, microbial biomass and enzyme activities involved in the C, P and N cycles were also monitored during the plant growth period. The results showed that the studied ornamental plants could grow on both NT and TR sediments, but that the growth was higher on TR sediment. The plant growth induced changes in the sediment chemical functional groups, with clear separation between NT and TR sediments for each of the studied plant. Microbial biomass and enzyme activities significantly increased during the plant growth, more in TR than in NT sediment. Toxicity was detected in NT sediments during the plant growth whereas it was not observed in NT sediments during the whole growth period. We concluded that phytoremediation converted the dredged maritime sediments into suitable substrates for growing ornamental plants, and that the re-use by plant nursery industry can be a sustainable management and valorization for remediated sediments.

The physiological mechanisms underlying the ability of Cistus monspeliensis L. from São Domingos mine to withstand high Zn concentrations in soils.

Cistus monspeliensis L. is a species that grows spontaneously in contaminated mining areas from the Iberian Pyrite Belt. This species can have high concentrations of Zn in the shoots without visible signs of phytotoxicity. In order to understand the physiological mechanisms underlying this tolerance, C. monspeliensis was grown at several concentrations of Zn(2+) (0, 500, 1000, 1500, 2000µM) and the effects of this metal on plant development and on the defence mechanisms against oxidative stress were evaluated. Independently of the treatment, Zn was mainly retained in the roots. The plants with the highest concentrations of Zn showed toxicity symptoms such as chlorosis, low leaf size and decrease in biomass production. At 2000µM of Zn, the dry biomass of the shoots decreased significantly. High concentrations of Zn in shoots did not induce deficiencies of other nutrients, except Cu. Plants with high concentrations of Zn had low amounts of chlorophyll, anthocyanins and glutathione and high contents of H2O2. The highest concentrations of Zn in shoots of C. monspeliensis triggered defence mechanisms against oxidative stress, namely by triggering antioxidative enzyme activity and by direct reactive oxygen species (ROS) scavenging through carotenoids, that are unaffected by stress due to stabilisation by ascorbic acid.

Quantitative Variation of Flavonoids and Diterpenes in Leaves and Stems of Cistus ladanifer L. at Different Ages.

The compounds derived from secondary metabolism in plants perform a variety of ecological functions, providing the plant with resistance to biotic and abiotic factors. The basal levels of these metabolites for each organ, tissue or cell type depend on the development stage of the plant and they may be modified as a response to biotic and/or abiotic stress. As a consequence, the resistance state of a plant may vary in space and time. The secondary metabolites of Cistus ladanifer have been quantified in leaves and stems throughout autumn, winter, spring and summer, and at different ages of the plant. This study shows that there are significant differences between young leaves, mature leaves and stems, and between individuals of different ages. Young leaves show significantly greater synthesis of flavonoids and diterpenes than mature leaves and stems, with a clear seasonal variation, and the differences between leaves at different growth stages and stems is maintained during the quantified seasons. With respect to age, specimens under one year of age secreted significantly lower amounts of compounds. The variation in the composition of secondary metabolites between different parts of the plant, the season and the variations in age may determine the interactions of Cistus ladanifer with the biotic and abiotic factors to which it is exposed.

Perennially young: seed production and quality in controlled and natural populations of Cistus albidus reveal compensatory mechanisms that prevent senescence in terms of seed yield and viability.

The question of whether or not perennial plants senesce at the organism level remains unresolved. The aim of this study was to unravel whether or not plant age can influence the production and composition of seeds. Flower and seed production was examined in 3-, 8-, and 13-year-old Cistus albidus plants growing in experimental plots corresponding to the F2, F1, and F0 generations of the same population. Furthermore, the phytohormone, fatty acid, and vitamin E content of the seeds was evaluated, and their viability was examined. Whether or not age-related differences in seed quality were observed in a natural population in the Montserrat Mountains (NE Spain) was also tested. The results indicate that under controlled conditions, the oldest plants not only produced fewer flowers, but also had higher rates of embryo abortion in mature seeds. However, germination capacity was not negatively affected by plant ageing. Seeds of the oldest plants contained significantly higher salicylic acid, jasmonic acid, and vitamin E levels compared with those from younger plants. Despite vigour (in terms of plant growth) being severely reduced due to harsh environmental conditions in the natural population, the oldest individuals produced seeds with no decline in viability. Seed biomass was instead positively correlated with seed viability. In conclusion, increased plant size may explain the loss of seed viability in the experimental field, but older smaller individuals in natural populations can escape senescence in terms of seed viability loss.

Large and abundant flowers increase indirect costs of corollas: a study of coflowering sympatric Mediterranean species of contrasting flower size.

Large floral displays receive more pollinator visits but involve higher production and maintenance costs. This can result in indirect costs which may negatively affect functions like reproductive output. In this study, we explored the relationship between floral display and indirect costs in two pairs of coflowering sympatric Mediterranean Cistus of contrasting flower size. We hypothesized that: (1) corolla production entails direct costs in dry mass, N and P, (2) corollas entail significant indirect costs in terms of fruit set and seed production, (3) indirect costs increase with floral display, (4) indirect costs are greater in larger-flowered sympatric species, and (5) local climatic conditions influence indirect costs. We compared fruit set and seed production of petal-removed flowers and unmanipulated control flowers and evaluated the influence of mean flower number and mean flower size on relative fruit and seed gain of petal-removed and control flowers. Fruit set and seed production were significantly higher in petal-removed flowers in all the studied species. A positive relationship was found between relative fruit gain and mean individual flower size within species. In one pair of species, fruit gain was higher in the large-flowered species, as was the correlation between fruit gain and mean number of open flowers. In the other pair, the correlation between fruit gain and mean flower size was also higher in the large-flowered species. These results reveal that Mediterranean environments impose significant constraints on floral display, counteracting advantages of large flowers from the pollination point of view with increased indirect costs of such flowers.

A copal-8-ol diphosphate synthase from the angiosperm Cistus creticus subsp. creticus is a putative key enzyme for the formation of pharmacologically active, oxygen-containing labdane-type diterpenes.

The resin of Cistus creticus subsp. creticus, a plant native to Crete, is rich in labdane-type diterpenes with significant antimicrobial and cytotoxic activities. The full-length cDNA of a putative diterpene synthase was isolated from a C. creticus trichome cDNA library. The deduced amino acid sequence of this protein is highly similar (59%-70% identical) to type B diterpene synthases from other angiosperm species that catalyze a protonation-initiated cyclization. The affinity-purified recombinant Escherichia coli-expressed protein used geranylgeranyl diphosphate as substrate and catalyzed the formation of copal-8-ol diphosphate. This diterpene synthase, therefore, was named CcCLS (for C. creticus copal-8-ol diphosphate synthase). Copal-8-ol diphosphate is likely to be an intermediate in the biosynthesis of the oxygen-containing labdane-type diterpenes that are abundant in the resin of this plant. RNA gel-blot analysis revealed that CcCLS is preferentially expressed in the trichomes, with higher transcript levels found in glands on young leaves than on fully expanded leaves, while CcCLS transcript levels increased after mechanical wounding. Chemical analyses revealed that labdane-type diterpene production followed a similar pattern, with higher concentrations in trichomes of young leaves and increased accumulation upon wounding.

Rehabilitation of mining areas through integrated biotechnological approach: Technosols derived from organic/inorganic wastes and autochthonous plant development.

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.

Effect of forest fire prevention treatments on bacterial communities associated with productive Boletus edulis sites.

Cistus ladanifer scrublands, traditionally considered as unproductive, have nonetheless been observed to produce large quantities of king bolete (Boletus edulis) fruitbodies. These pyrophytic scrublands are prone to wildfires, which severely affect fungi, hence the need for fire prevention in producing C. ladanifer scrublands. In addition, B. edulis productions have severely decreased in the last years. A deeper understanding of the B. edulis life cycle and of biotic and abiotic factors influencing sporocarp formation is needed to implement management practices that facilitate B. edulis production. For example, some bacteria likely are involved in sporocarp production, representing a key part in the triple symbiosis (plant-fungus-bacteria). In this study, we used soil DNA metabarcoding in C. ladanifer scrublands to (i) assess the effect of site history and fire prevention treatment on bacterial richness and community composition; (ii) test if there was any correlation between various taxonomic groups of bacteria and mycelial biomass and sporocarp production of B. edulis; and to (iii) identify indicator bacteria associated with the most productive B. edulis sites. Our results show that site history drives bacterial richness and community composition, while fire prevention treatments have a weaker, but still detectable effect, particularly in the senescent plots. Sporocarp production correlated positively with genera in Verrucomicrobia. Several genera, e.g. Azospirillum and Gemmatimonas, were identified as indicators of the most productive sites, suggesting a potential biological role in B. edulis fructification. This study provides a better understanding of the triple symbiosis (plant-fungus-bacteria) involved in C. ladanifer-B. edulis systems.

Meristem aging is not responsible for age-related changes in growth and abscisic acid levels in the Mediterranean shrub, Cistus clusii.

To obtain new insights into the mechanisms underlying aging in perennials, we measured abscisic acid levels, growth and other stress indicators in leaves of Cistus clusii Dunal plants of different ages grown under Mediterranean field conditions. Recently emerged leaves from 9-year-old plants were compared to those of 1-year-old plants (obtained from cuttings from 9-year-old plants) to evaluate the effects of meristem aging on plant aging. Rooting and successful establishment of the cuttings allowed us to compare the physiology of plants with old meristems, but of different size. Plants obtained from cuttings were rejuvenated, with new leaves displaying a higher leaf area and chlorophyll content, but smaller leaf mass per unit area ratios and endogenous abscisic acid levels than those of 9-year-old plants. A comparative study in 1-, 4- and 9-year-old plants revealed that abscisic acid levels increase during the early stages of plant life (with increases of 90% between 1- and 4-year-old plants), but then remain constant at advanced developmental stages (between 4- and 9-year-old plants). Although leaf biomass was 53% smaller in 9-year-old compared to 4-year-old plants, the dry matter produced per shoot apical meristem was equivalent in both plant groups due to an increased number of leaves per shoot in the former. It is concluded that (i) C. clusii plants maintain the capacity to rejuvenate for several years; (ii) newly emerged leaves accumulate higher amounts of abscisic acid during early stages of plant life, but the levels of this phytohormone later remain constant; and (iii) although plant aging leads to the production of smaller leaves, the amount of biomass produced per shoot apical meristem remains constant at advanced developmental stages.

Seed germination of Cistus creticus L. and Cistus laurifolius L. as influenced by dry-heat, soaking in distilled water and gibberellic acid.

The effects of dry-heat and seed soaking in distilled water or in gibberellic acid on germination performance of Cistus creticus L. and C. laurifolius L. were studied in the present study Germination percentages of two Cistus species were low due to dormancy Soaking in distilled water for 24 hr resulted in 28% germination in C. creticus and 43% in C. laurifolius. Gibberellic acid applications (20, 100 and 250 mg l(-1)) for 24 hr gave 32, 30 and 23% germination, respectively in Cistus creticus and 33, 37 and 28% germination, respectively in C. laurifolius. Dry-heat pretreatments at 50, 80 and 100 degrees C for several times (1-60 min) also significantly increased germination percentage in two species. The highest germination in C. creticus was obtained with dry-heat at 100 degrees C for 1 or 5 min (80% and 83%, respectively) and in C. laurifolius at 100 degrees C for 5 min (87%). A significant increase in germination rate was also achieved under different pretreatments. The physiological dormancy caused by an impermeable seed coat can be overcome by dry-heat pretreatments in two Cistus species.

Cistus monspeliensis L. as a potential species for rehabilitation of soils with multielemental contamination under Mediterranean conditions.

The Iberian Pyrite Belt (IPB; SW of the Iberian Peninsula) is one of the most important volcanogenic massive sulphide ore deposits in the world. Cistus monspeliensis L. is a native woody shrub that grows spontaneously in non-contaminated soils as well as in soils with multielemental contamination from the IPB. In this study, different ecophysiological parameters of C. monspeliensis growing in soils with different levels of metal(loid)s were evaluated to assess the potential of this species for revegetation of degraded areas. Composite samples of plants and rhizosphere soils were sampled in São Domingos and Lousal mines and in a reference area without soil contamination (Pomarão, Portugal) (Portuguese sector of IPB). Classical characterisation of the soils and quantification of their total and available metal(loid) concentrations were done. Multielemental concentration was determined in plants (shoots and roots). Ecophysiological parameters were also determined in shoots: concentrations of pigments (chlorophylls, anthocyanins and carotenoids), antioxidants (glutathione and ascorbate) and hydrogen peroxide as well as activities of several antioxidative enzymes. Although mining soils present high total concentrations of potentially hazardous elements, their available fractions were low and similar among studied areas. Soil pH as well as concentrations of extractable P, total concentrations of As, Cd and Ni and concentrations of Cu, Cr, Ni, Pb and Sb in the soil available fraction differentiate the studied areas. Only concentrations of Cd, Pb and Sb in roots and shoots were explained by the concentrations of the same elements in the soil available fraction. Although the majority of elements were translocated from roots to shoots, the shoots concentrations were below the toxic values for domestic animals and only As, Mn and Zn reached phytotoxic concentrations. Ecophysiological parameters were similar independently of the studied area. Due to its adaptability, tolerance and standard plant features, C. monspeliensis is a good choice for rehabilitation of soils with multielemental contamination under similar climatic characteristics.

Morphology and biochemistry of non-glandular trichomes in Cistus salvifolius L. leaves growing in extreme habitats of the Mediterranean basin.

Here the functional roles of stellate and dendritic trichomes in Cistus salvifolius L leaves were studied by analysing i) both leaf surface and trichome morphology using scanning electron and light microscopy; and ii) the composition and localisation of polyphenols by coupling liquid chromatography with fluorescence spectroscopy and fluorescence microimaging. Red-coloured compounds were detected in the stalk cells and the channel in the trichome arm, and appeared to be released at the tip end of the trichome branch. We identified such metabolites as ellagitannins, namely punicalagin and two galloyl derivatives of punicalagin. These ellagitannins accounted for 4.3 % of leaf dry weight and their concentration in the leaf leachate averaged 289.4 mg L (-1). The trichome arms exhibited an appreciable orange-red autofluorescence (centred at 620 nm) when excited with UV light (at 365 nm) or emitted in the yellow waveband (peak centred at 566 nm) when stained with the Naturstoff reagent, and excited at 488 nm. The fluorescence signatures of the trichome arms were consistent with the presence of mono-hydroxy B-ring substituted flavonoids, which were identified as the mono- and di-coumaroyl derivative of a kaempferol 3-O-glycoside. Our data may provide some insights on the functional roles of stellate and dendritic trichomes in the response mechanisms of C. salvifolius to Mediterranean-type climate, based upon (i) the potential effect of released ellagitannins on the soil nitrogen dynamic and (ii) the ability of acylated kaempferol 3-O-glycosides to effectively absorb both the UV-B and UV-A wavelengths.

Leaf exchange in a Mediterranean shrub: water, nutrient, non-structural carbohydrate and osmolyte dynamics.

Leaf exchange is an abrupt phenological event that drastically modifies the morphology and physiology of the aerial portion of the plant. We examined if water and osmolyte differences between old leaves and new organs trigger leaf exchange, and whether the differences are closely linked to the resource resorption process in senescing leaves. We monitored concentrations of osmolyte, water, non-structural carbohydrate, nitrogen and potassium in senescing leaves and in emerging new leaves and inflorescences of a Mediterranean leaf exchanger (Cistus laurifolius L.) growing in NE Spain. Old leaves rehydrated markedly during most of the senescence process, which co-occurred with the extension of new shoots, suggesting the lack of a clear-cut switch in water supply from old to new organs. The accumulation of osmolytes in the early stage of leaf senescence might account for this rehydration. Osmolyte dynamics in old leaves depended largely on the progression of resource resorption from senescing organs but were mostly unrelated to water content during late senescence. We conclude that dehydration of old leaves is not a prerequisite for the triggering of leaf exchange. The finding that most nutrients and carbohydrates accumulated in new organs before senescing leaves massively exported resources, and the absence of relevant differences between the dynamics of old leaves at the base of inflorescences and those at the base of vegetative shoots, indicate that the nutrient and carbohydrate demands of new organs do not trigger leaf exchange.

Season-dependent and independent responses of Mediterranean scrub to light conditions.

Semi-arid plant species cope with excess of solar radiation with morphological and physiological adaptations that assure their survival when other abiotic stressors interact. At the leaf level, sun and shade plants may differ in the set of traits that regulate environmental stressors. Here, we evaluated if leaf-level physiological seasonal response of Mediterranean scrub species (Myrtus communis, Halimium halimifolium, Rosmarinus officinalis, and Cistus salvifolius) depended on light availability conditions. We aimed to determine which of these responses prevailed independently of the marked seasonality of Mediterranean climate, to define a leaf-level strategy in the scrub community. Thirty six leaf response variables - involving gas exchange, water status, photosystem II photochemical efficiency, photosynthetic pigments and leaf structure - were seasonally measured in sun exposed and shaded plants under field conditions. Physiological responses showed a common pattern throughout the year, in spite of the marked seasonality of the Mediterranean climate and of species-specific differences in the response to light intensity. Variables related to light use, CO2 assimilation, leaf pigment content, and LMA (leaf mass area) presented differences that were consistent throughout the year, although autumn was the season with greater contrast between sun and shade plants. Our data suggest that in Mediterranean scrub shade plants the lutein pool could have an important role in the photoprotection of the photosynthetic tissues. There was a negative linear correlation between the ratio lutein/total chlorophylls and the majority of leaf level variables. The combined effect of abiotic stress factors (light and drought or light and cold) was variable-specific, in some cases enhancing differences between sun and shade plants, while in others leading to unified strategies in all scrub species.

Cistus ladanifer L. Shrub is Rich in Saturated and Branched Chain Fatty Acids and their Concentration Increases in the Mediterranean Dry Season.

The Cistus ladanifer L. shrub is a widespread species of the Mediterranean region that is available as a feed source for ruminants all the year round, constituting a source of energy and nutrients when most of the vegetation is dry. However, there is no trustworthy information about the fatty acid composition of C. ladanifer, as well as no information about the seasonal and age related changes in their fatty acid composition. Thus, we collected the aerial parts of C. ladanifer plants of two age groups [young vs. older ones (2-6 years old)] during four consecutive seasons to characterize their fatty acid composition. The fatty acid composition of C. ladanifer is dominated by saturated fatty acids including the occurrence of two methyl branched chain fatty acids (BCFA), the iso-19:0 and iso-21:0, which as far as we know were detected for the first time in shrubs. Also, we demonstrated that several labdane type compounds might interfere with the fatty acid analysis of C. ladanifer. Marked seasonal changes in BCFA and polyunsaturated fatty acids (PUFA) were found, suggesting that BCFA can replace PUFA in plant lipids at high environmental temperatures.

Zeatin modulates flower bud development and tocopherol levels in Cistus albidus (L.) plants as they age.

In a previous study we showed that Cistus albidus (L.) experiences an age-dependent decay in flower vigour correlated with a decline in trans-zeatin (tZ) levels. In the present study we aimed to establish a causal relationship between these two phenomena. Exogenous tZ applied to plants grown under semi-controlled conditions did not rescue flower vigour; however, it accelerated flower development, but only in younger individuals. Older plants showed lower tocopherol levels in flower buds, which were restored by exogenous tZ, suggesting that a loss of antioxidant defences may underlie the age-dependent decay in flower vigour. We conclude that declining tZ levels may not be directly responsible for the age-associated loss of floral vigour; that tZ modulates the speed of flower development as plants age; and that flower buds alter their sensitivity to tZ as plants age.

Enhanced photo- and antioxidative protection, and hydrogen peroxide accumulation in drought-stressed Cistus clusii and Cistus albidus plants.

Mechanisms of drought stress resistance were studied in Cistus clusii Dunal and Cistus albidus L., two native Mediterranean shrubs that can withstand severe summer drought. While water deficit, solar radiation and temperature increased from winter to summer in the field, C. clusii and C. albidus reduced leaf area, increased root mass per leaf area, and showed diurnal changes in stomatal conductance to minimize water loss. In both species, the consequent reductions in CO2 assimilation were accompanied by reduced efficiency of photosystem II photochemistry, and protection against stress was afforded by enhanced de-epoxidation of violaxanthin in the xanthophyll cycle and increases in alpha-tocopherol and beta-carotene. In addition, hydrogen peroxide (H2O2) accumulation was observed in mesophyll cell walls of both species during the first stages of drought, although no accumulation of H2O2 was observed in chloroplasts or other organelles during the study. Despite these common responses, C. albidus and C. clusii differed in the extent of photo- and antioxidative protection. In response to drought, C. clusii showed a higher de-epoxidation state of the xanthophyll cycle and higher alpha-tocopherol and beta-carotene concentrations than C. albidus. We conclude that several structural and biochemical mechanisms underlie stress resistance in C. clusii and C. albidus, and are indicative of the different degrees of stress resistance of these shrubs.

Combined effects of clay immobilized Azospirillum brasilense and Pantoea dispersa and organic olive residue on plant performance and soil properties in the revegetation of a semiarid area.

The reestablishment of autochthonous shrubs species is an essential strategy for recovering degraded soils under semiarid Mediterranean areas. A field experiment was carried out to assess the effectiveness of an immobilized microbial inoculant (Azospirillum brasilense and Pantoea dispersa) and the addition of organic olive residue (alperujo), for plant growth promotion of Cistus albidus L. and enhancement of soil properties. Sixteen months after planting, the microbial inoculant and organic residue combined treatment was the most effective for stimulating the root dry weight of C. albidus (by 133% with respect to control plants) and microbial inoculant was the most effective treatment for increasing the shoot dry weight of plants (by 106% with respect to control plants). Available phosphorus and potassium content in the amended soils was about 100 and 70% respectively higher than the non-amended soil. Total C, total organic C and microbial biomass C content and enzyme activities (dehydrogenase, urease and protease) of the rhizosphere of C. albidus were increased by microbial inoculant and organic residue combined, but not by the microbial inoculation and organic residue applied independently. The combined treatment, involving microbial inoculant and the addition of the organic residue, had an additive effect improving the biochemical and microbiological quality of the soil.

Environmental control of terpene emissions from Cistus monspeliensis L. in natural Mediterranean shrublands.

The large amount of volatile organic compound (VOC) emitted by vegetation modifies air quality contributing to both tropospheric ozone and secondary organic aerosol production. A better understanding of the factors controlling VOC emissions by vegetation is mandatory in order to improve emission estimates derived from tropospheric chemistry models. Although the Mediterranean shrublands are particularly abundant and rich in emitting species, their emission potential is poorly known. Focusing on a VOC-emitting shrub species widespread in the Mediterranean area (Cistus monspeliensis L.), we measured and analysed its emissions of terpenes taking into account the age of individuals, the season of sampling and the soil type. Sampling was done under natural environmental conditions. Species of the genus Cistus are frequently reported to be storing species, although we found only one stored monoterpene and three sesquiterpenes in very low amount. Major emitted compounds were alpha-pinene and beta-myrcene. Total terpene emissions were not influenced by plant age but emission of some individual terpenes was positively correlated with age. A strong seasonal effect was evidenced. A larger amount of terpenes was emitted during spring and summer than during fall and winter. Summer emission rates were nearly 70 times higher than winter emission rates. Total and individual terpene emissions were influenced by soil type; emissions on siliceous substrate were ca. seven times higher than those on calcareous substrate. In conclusion, it appears clearly that environmental factors such as soil nature and season should be taken into account in order to achieve improved modelling of terpene emissions by shrub species.