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The present study aimed to investigate the reciprocal effects of Phoradendron perrottetii (mistletoe) and T. guianensis (host plant) regarding their polyphenol composition. Taking into account that tannins are important molecules in plant defense and their biosynthesis tends to be enhanced when a species is exposed to stress, we address the following questions: (1) Are the tannins found in our model species important in the interaction between host and mistletoe? (2) Does the presence of mistletoe induce changes in the content of tannins and other polyphenols in the host plant? (3) Do we find differences between the tannin sub-groups in the responses of the host plant to mistletoe? (4) Could the observed differences reflect the relative importance of one tannin group over another as chemical defense against the mistletoe? Using a polyphenol and tannin group-specific MRM methods we quantified four different tannin sub-groups together with flavonoid and quinic acid derivatives by ultra-performance liquid chromatography tandem mass spectrometry together with the oxidative and protein precipitation activities of leaves and branches of Tapirira guianensis and Phoradendron perrottetii. We selected leaves and branches of six non-parasitized trees of T. guianensis. Leaves and branches of nine individuals of T. guianensis parasitized by P. perrottetii were also sampled. For each parasitized tree, we sampled an infested branch and its leaves, as well as a non-infested branch and its leaves. Infested branches were divided into three groups: gall (the host-parasite interface), proximal, and distal region. Both proanthocyanidins and ellagitanins seem to be important for plant-plant parasitism interaction: host infested tissues (gall and surrounding regions) have clearly less tannin contents than healthy tissues. Mistletoe showed high levels of quinic acid derivatives and flavonoids that could be important during hastorium formation and intrusion on host tissues, suggesting a defense mechanism that could promote oxidative stress together with an inhibition of mistletoe seed germination, consequently avoiding secondary infestations. Polyphenol detected in T. guianensis-P. perrottetii interaction could play different role as plant-mistletoe strategies of survival.

Xyloglucan (XyG) is the major noncellulosic nonpectic matrixpolysaccharide in cell walls of most land plants. Initially thought tobe restricted to land plants, the last decade has seen the detection ofXyG and the discovery of synthesis and modification/degradationgenes in charophycean green algae (CGA). Recently, a totally newfunction of XyG was discovered as a potent soil aggregator releasedby roots and rhizoids of all major groups of land plants. In thisViewpoint, I show the presence of a complex XyG geneticmachinery in most CGA groups. I discuss the context of XyGevolution in light of the terrestrialization of early CGA that gave riseto embryophytes and its possible role in early soil formation. O xiloglucano (XyG) é o principal polissacarídeo de matriz não celulósica não-ótica nas paredes celulares da maioria das plantas terrestres. Inicialmente considerada restrita a plantas terrestres, a última década viu a detecção de XyG e a descoberta de genes de síntese e modificação / degradação em algas verdes carofíceas (CGA). Recentemente, uma função totalmente nova de XyG foi descoberta como um potente agregador de solo liberado por raízes e rizóides de todos os principais grupos de plantas terrestres. Neste artigo, mostro a presença de uma maquinaria genética XyG complexa na maioria dos grupos CGA. Discuto o contexto de XyGevolution à luz da terrestrialização do CGA inicial que deu origem a embriófitos e seu possível papel na formação inicial do solo.

[EN] Biochar and hydrochar are produced by charring organic matter through pyrolysis and hydrothermal carbonization, respectively. Biochar has been tested extensively in agricultural soils. Nevertheless, both materials have scarcely been tested as soilless growth medium constituents. We have conducted studies on the properties and behaviour of biochar [forest waste (BCH-FW) and olive mill waste (BCH-OMW)] and hydrochar [forest waste (HYD-FW)] as growth medium constituents under different culture conditions. We have grown ornamental, forest and vegetable species. We have conducted assays on seed germination, cutting rooting and plantlet growth under nursery conditions, and tomato growth under commercial conditions. The materials proved useful for growth medium formulation. However, they were not good performers as sole constituents, because they posed specific problems depending on the material. BCH-FW was alkaline and its major problem was the large particle size that some batches presented, which caused low retention of water and nutrients leading to low plant performance. Nevertheless, good results were obtained for seed germination, cutting rooting and plant survival. BCH-OMW was alkaline and highly saline, and as a result was phytotoxic to plants. This was a drawback for seed germination and young plant growth, but not for adult plants and saline-tolerant species such as tomato. HYD-FW was highly hygroscopic and showed high microbial activity, emitting large amounts of CO2. Under these conditions, the low O2 pressure in the rhizosphere led to a decrease in plant performance.

Fast urbanization has led to ozone (O3) being the main pollutant in summer in most of China. To assess future ground-level O3 effects on the service of urban greening species and clarify the underlying mechanism of O3 damage, four common urban greening species, Ailanthus altissima (AA), Fraxinus chinensis(FC), Platanus orientalis (PO) and Robinia pseudoacacia (RP) were exposed to non-filtered air (NF) and to elevated O3 (E-O3) in open-top chambers. E-O3 induced visible injury in all species as well as microscopic alterations such as collapse of the palisade parenchyma cells, callose accumulation, or chloroplast and mitochondrial accelerated senescence. E-O3 significantly reduced light-saturated CO2 assimilation (Asat),the maximum activity of Rubisco (Vcmax), the maximum electron transport rate (Jmax), and fluorescence parameters such as the quantum yield of noncyclic electron transport (�PSII), and the quenching of photochemical efficiency of PSII (qP). It also increased total antioxidant capacity, phenolics and ascorbate contents. No significant interaction between O3and species was found in photosynthetic performance and antioxidant systems, suggesting that the four species selected were sensitive to O3. Of all four species,AA was the most sensitive species due to a combination of earlier injury onset, anatomical features, lower antioxidant responses and higher stomatal conductance. The sensitivity of tree species to O3 is a factor to be considered for urban greening. Ozone may affect important urban forest ecosystem services by reducing CO2 assimilation, This study has been funded by the Hundred Talents Program, Chinese Academy of Sciences and State Key Laboratory of Urban and Regional Ecology. Collaboration between RCEES and Fundacion CEAM has been possible thanks to project AMIS (Fate and Impact of Atmospheric Pollutants, PIRSES-GA-2011-295132), and by the Chinese Academy of Sciences Visiting Professorships for Senior International Scientists (grant number: 2013T2Z0009). VC also acknowledges the support of PROMETEOII/2014/038 project (Generalitat Valenciana, G.V.), and FGB and JRA that of PROMETEOII2013/021 (G.V.), and CGL2012-40058-C02-01/02 (MINECO). We thank Mr. Yulong Zhang for the experimental management.

[EN] Gypsum areas are stressful environments inhabited by gypsophytes, plants that are exclusive for such habitats, and by plants that grow on gypsum but also on other soil types, the so-called gypsovags. To investigate possible differences between gypsovags and gypsophytes with respect to basic stress response mechanisms, two common osmolytes, glycine betaine and total soluble sugars, as well as monovalent (Na+ and K+) and bivalent (Ca2+ and Mg2+) cations, were quantified, under field conditions, in two Iberian endemic gypsophytes (Gypsophila struthium subsp. hispanica and Ononis tridentata) and two common Mediterranean gypsovags (Rosmarinus officinalis and Helianthemum syriacum). Their spatial variation according to a topographic gradient and their temporal variation over a period of three successive seasons were correlated with climatic data and soil characteristics. This analysis confirmed that water stress is the main environmental stress factor in gypsum habitats, whereas the percentage of gypsum in the soil does not seem to play any relevant role in the activation of stress responses in plants. Glycine betaine may contribute to stress tolerance in the gypsophytes, but not in the gypsovags, according to the close correlation found between the level of this osmolyte and the gypsophily of the investigated taxa. Cation contents in the plants did not correlate with those present in the soil, but the gypsophytes have higher levels of Ca2+ and Mg2+ than the gypsovags, under all environmental conditions, which may represent an adaptation mechanism to their specific habitat, This work has been supported by the Spanish Ministry of Science and Innovation (Project CGL2008-00438/BOS), with contribution from the European Regional Development Fund.

Three tobacco types (Burley, Flue Cured, and Oriental) and eight cigarette brands were unequivocally identified using an electronic nose formed by only three sensors based on a single novel conducting polymer (PF-BTB) doped with different porphyrins (H2TPP, H2TPFP, and H2BTBOP). The synthesis and characterization of the polymer are also discussed. Small changes in the porphyrin structure caused significant changes in the electrical conductance response patterns of the sensors upon exposure to complex chemical matrixes, representing a novel approach for tuning the selectivity of chemiresistive sensors for e-nose application. This e-nose is fast, cheap, reliable, can be easily operated, and could be a valuable tool for border agents fighting cigarette smuggling around the world, helping them prevent losses of millions in tax revenues and sales.

Composition of essential oils of different populations of Thymus richardii grex of six localities from Bosnia-Herzegovina (Konjic, Borci), Spain (Majorca, Ibiza, Valencia) and Italy (Marettimo, Sicily) were determined by GC/FID and GC/MS. The main constituents in most of the samples were aromatic monoterpenes corresponding to non-phenolic cyclic compounds (p-cymene, gamma-terpinene). The highest monoterpene concentrations were found in the Bosnian samples (70%), and the lowest in samples from the Balearic Islands ( 50%) in samples from Majorca with beta-bisabolene (>40%) being the principal constituent. Discriminant analysis (LDA) shows the differentiation of two chemotypes: A (phenol chemotype), with p-cymene and gamma-terpinene as characteristic compounds and B, with beta-bisabolene and carvacrol, as major and significative compounds. The occurrence of the chemotypes was related to summer positive precipitation and to deep of soils.

Despite several pharmacological applications of Rosmarinus officinalis L. (Lamiaceae), studies on its analgesic and anti-inflammatory properties have been scarce.The aim of this work was to use in vivo models to evaluate the analgesic and anti-inflammatory activities of the aqueous extracts obtained from leaves (AEL) and stems (AES) of Rosmarinus officinalis, as well as its isolated compound--rosmarinic acid (RA). We also prepared and assessed the acetyl ester derivative of RA.The analgesic activity was evaluated using abdominal constriction and formalin tests. For the evaluation of the anti-inflammatory effects, carrageenin-induced paw edema in rats were used. The extracts were used at doses of 100, 200 and 400 mg kg⁻¹ compounds were tested at 10, 20 and 40 mg kg⁻¹.Orally administered AEL, AES and RA were not significantly active at any of the doses tested during the abdominal constriction test; the acetyl ester derivative of RA displayed significant analgesic activity. In the carrageenin-induced paw edema assay, the acetyl derivative of RA at all the tested doses produced significant anti-inflammatory effects and reduced the number of paw licks in the second phase of the formalin test.The results suggest that the analgesic effects of the acetyl derivative of RA operate via a peripheral-mediated mechanism. The acetyl ester derivative of RA is potentially applicable as a new lead compound for the management of pain and inflammation.

[EN] The feasibility of composted (C), composted plus vermicomposted (V1) and straight vermicomposted (V2) tomato crop waste as component of rooting media for Euonymus japonicus ‘Microphylla’ and Lavandula angustifolia vegetative propagation was studied. Mixes of C, V1 and V2 with coir fibre (CF) at the proportions 100:0, 75:25, 50:50, 25:75, 0:100 (v:v) were assayed. Physical, physico-chemical and nutritional characteristics of all materials and mixes were determined and correlated with cutting rooting and growth performances. The compost and the two vermicomposts were markedly different from CF. They had higher bulk density and lower total porosity than CF. Compost had lower water-holding capacity and shrinkage in response to drying than vermicomposts and CF. Compost and vermicomposts were alkaline materials whilst CF was almost neutral. Electrical conductivity (EC) was low in CF and vermicomposts, and high in compost due to the high mineral contents, mainly of K+, SO4 2– and Na+ in this material. EC and the ions contributing to it (K+, SO4 2–, Na+) showed highly significative inverse correlations with rooting percentage for the two species and with root and shoot growth but only for E. japonicus. Due to its high EC, compost C (average rooting = 22.5%) performed worse than vermicomposts V1 (av. rooting = 97%) and V2 (av. rooting = 98%) whilst the latter performed similarly to CF control (av. rooting = 100%). Thus vermicomposts appeared to be more appropriate than compost as rooting media constituent., Authors are deeply grateful to Dr. Maria Jose Lopez and Dr. Raquel Melgar for supervising the composting and vermicomposting procedures. This study was funded by the Institut Nacional de Investigacion y Tecnologia Agraria (INIA) of the Spanish Ministerio de Ciencia e Innovacion (Project Reference RTA2006-00008-C02-02).

[EN] Essential oil (EO) extracts coming from two representative populations of Mentha suaveolens Ehrh. subesp. suaveolens in Eastern Iberian Peninsula were analyzed by gas chromatography coupled with mass spectrometry and flame ion detector. Plant sampling was carried out in the morning and evening in order to study diurnal variation in EO profiles. Likewise, leaves and inflorescences were analyzed separately. Two chemotypes corresponding to each one of the populations were identified, with piperitenone oxide (35.2 - 74.3%) and piperitone oxide (83.9 - 91.3%), respectively, as major compounds. Once different chemotypes were identified, canonical correspondence analysis was employed to evaluate the effect of the bioclimatic and edaphic factors recorded in each location on the observed differences. Statistical analysis suggested that these chemotypes were closely related to specific environmental factors, mainly the bioclimatic ones. Concretely, piperitenone oxide chemotype can be associated to supramediterranean bioclimatic conditions and soils with major salinity and water field capacity. On the other hand, the most volatile fraction (hydrocarbon monoterpenes) reached its higher level in the morning; specifically, a noticeable amount of limonene was found in morning samples of flowers (4.8 - 10.6%). This fact can be related to ecological role of volatile compounds in order to attract pollinator insects.

[EN] Biomass waste from rice straw has many management problems, including field firing causing severe air pollution and natural organic decomposition resulting in methane emission. The conversion of this waste to ashes may offer the possibility of reusing them in cementing systems. For the first time ashes from different parts of the rice plant (Oryza sativa) were characterised from the chemical composition point of view: rice leaf ash (RLA), rice leaf sheath ash (RlsA) and rice stem ash (RsA). Microscopic studies on ashes revealed heterogeneity in the distribution of chemical elements in the remaining cellular structure (spodograms). The highest concentration of SiO2 was found in dumbbell-shaped phytoliths (%SiO2 > 78%). In the global chemical composition of ashes, SiO2 was also the main oxide present. According to Vassilev¿s classification of chemical composition, RLA belongs to the K-MA zone (medium acid), RlsA to the K-zone (low acid) and RsA to the S-zone (high acid). Calcination temperatures ¿550 ¿C completely removed organic matter from the straw and ashes underwent significant sinterisation by calcining at 650 ¿C due to the presence of potassium chloride. Here, ashes from global straw (rice straw ash, RSA) are characterised (via X-ray diffraction, Fourier transform infrared spectroscopy and thermogravimetry) and tested from a reactivity point of view (reaction towards calcium hydroxide) in order to assess the possibility for its reuse in cementing systems. Results from pastes made by mixing RSA and calcium hydroxide showed that the pozzolanic reactivity of the ashes is important (hydrated lime fixation of 82% for 7 days and 87% for 28 days in RSA:hydrated lime paste) and cementing C S H gel is formed after 7 and 28 days at room temperature. Compressive strength development of Portland cement mortars with 10% and 25% replacements by RSA yielded 107% and 98% of the strength of control mortar after 28 days of curing. Frattini test confirmed the pozzolanicity of the RSA blended cements. These reactivity results are very promising in terms of the potential reuse of ashes in cementing systems., The authors acknowledge the financial support of the Ministerio de Economia y Competitividad MINECO, Spain, and FEDER funding [Project: B1A2015-70107-R] The authors thank the Electron Microscopy Service of the Universitat Politecnica de Valencia (Spain).

[EN] Aloe vera gel, analysed as to its antifungal properties against six fungi causing plant diseases, was found to be most effective against Fusarium oxysporum. It was included in different ratios in starch based films plastiziced with glycerol to obtain antifungal films. These were characterized as to their physical (barrier and optical) and structural properties. Films containing the highest ratio of Aloe vera solids (1:1, with respect to starch) and different glycerol mass ratios with respect to starch (0.15 and 0.25) were analysed as to the water sorption and water plasticization effects, in order to discover the water sensitivity of the films in different RH ambients. Films with 0.15 g glycerol/g starch were more homogenous, with a lower degree of starch crystallization, reduced water vapour permeability and higher gloss and transparency. These films with the highest Aloe vera ratio were effective at controlling fungal decay and weight loss in cherry tomatoes. (C) 2017 Elsevier Ltd. All rights reserved., The authors acknowledge the financial support from the Spanish Ministerio de Economia y Competitividad throughout the project AGL2013-42989-R and AGL2016-76699-R. Rodrigo Ortega-Toro thanks Universidad Nacional Abierta y a Distancia from Colombia for their support through project PIE-01 2016. Authors also thank the Electron Microscopy Service of the Universitat Politecnica de Valencia for their technical assistance.

[EN] Commercial thyme and lavender essential oils were analysed by GC/MS. Sixty-six compounds accounting for 98.6¿99.6% of total essential oil were identified. Thymol (52.14 ± 0.21%), followed by pcymene (32.24 ± 0.16%), carvacrol (3.71 ± 0.01%) and ¿-terpinene (3.34 ± 0.02%), were the main compounds in thyme essential oil, while large amounts of oxygenated monoterpenes linalool acetate (37.07 ± 0.24%) and linalool (30.16 ± 0.06%) were found in lavender one. In vitro antifungal activity of the essential oils was evaluated at 200 and 300 ¿g/mL against 10 phytopathogenic and post-harvest fungi, which significantly affect agriculture. Micelial growth inhibition was calculated for each tested fungus and dose. Thyme essential oil showed satisfactory results with 90¿100% growth inhibition in almost all the assayed fungi at 300 ¿g/mL, while lavender essential oil showed no noteworthy inhibition data at either dose, and its growth was even enhanced. Thyme essential oil represents a natural alternative to control harvest and post-harvest fungi, and to extend the shelf-life of agriculture products., This study has been financed by the Spanish Ministry of Economy and Competitiveness, Programme-oriented Societal Challenges 2014-2017 [grantnumber AGL2013-42989-R-AR].

Comparative studies on the responses to salt stress of taxonomically related taxa should help to elucidate relevant mechanisms of stress tolerance in plants. We have applied this strategy to three Plantago species adapted to different natural habitats, P. crassifolia and P. coronopus both halophytes and P. major, considered as salt-sensitive since it is never found in natural saline habitats. Growth inhibition measurements in controlled salt treatments indicated, however, that P. major is quite resistant to salt stress, although less than its halophytic congeners. The contents of monovalent ions and specific osmolytes were determined in plant leaves after four-week salt treatments. Salt-treated plants of the three taxa accumulated Na+ and Cl- in response to increasing external NaCl concentrations, to a lesser extent in P. major than in the halophytes; the latter species also showed higher ion contents in the non-stressed plants. In the halophytes, K+ concentration decreased at moderate salinity levels, to increase again under high salt conditions, whereas in P. major K+ contents were reduced only above 400 mM NaCl. Sorbitol contents augmented in all plants, roughly in parallel with increasing salinity, but the relative increments and the absolute values reached did not differ much in the three taxa. On the contrary, a strong (relative) accumulation of proline in response to high salt concentrations (600 800 mM NaCl) was observed in the halophytes, but not in P. major. These results indicate that the responses to salt stress triggered specifically in the halophytes, and therefore the most relevant for tolerance in the genus Plantago are: a higher efficiency in the transport of toxic ions to the leaves, the capacity to use inorganic ions as osmotica, even under low salinity conditions, and the activation, in response to very high salt concentrations, of proline accumulation and K+ transport to the leaves of the plants., MAH was a recipient of an Erasmus Mundus pre-doctoral scholarship financed by the European Commission (Welcome Consortium). AP acknowledges the Erasmus mobility programme for funding her stay in Valencia to carry out her Master Thesis.

[EN] Charred organic matter is recently receiving attention for its potential use as soilless growth medium. However, depending on its origin and on the manufacturing technology, it can result toxic for plants. This fact implies that a detoxifying treatment ought to be devised in order to reclaim char in this way. We have studied three materials which combine these factors: two pyrolyzed biochars, one from forest waste (BCH-FW) and another from olive mill waste (BCH-OMW), and one hydrothermally carbonized hydrochar from forest waste (HYD-FW). These materials are suspicious of phytotoxicity due to their high pH, high salinity, or presence of organic toxics. For these new materials, it is mandatory to select fast and reliable bioassays to predict their potential phytotoxicity. In order to achieve this goal water extracts of the three chars were subjected to bioassays of seed germination and bioassays of seedling growth in hydroponic conditions. The biochar from olive mill waste and the hydrochar, but not the biochar from forest waste, showed considerable phytotoxicity as seed germination and plant growth were negatively affected (e.g. BCH-OMW reduced seed germination by 80% and caused early seedling death). In order to adjust pH and electrical conductivity for plant growth, treatments of acidification and salt leaching with optimal diluted HNO3 solutions (03 N, 0.2 N, and 0.75 N for BCH-OMW, BCH-FW, and HYD-FW, respectively) as calculated from titration curves, were conducted. The acid treatment reduced electrical conductivity in BCH-OMW (from 9.2 to 4.5 dS m(-1)), pH (maximum in BCH-FW from 9.6 to 6.2) and water soluble carbonaceous compounds (maximum in HYD-FW from 5969 to 2145 mg kg(-1)) in the three chars, and increased N content (maximum in BCH-OMW from 50 to 6342 mg kg(-1)) in the three chars. Bioassays on acid-treated chars demonstrated the absence of phytotoxicity and even stimulation of seedling growth over the control (increase of 86% and 56% for BCH-FW and HYD-FW, respectively). We conclude that acidification of chars with diluted HNO3 is a viable technique to conform chars to standards for plant growth purposes. (C) 2017 Elsevier Ltd. All rights reserved., This study was funded by the Universitat Politecnica de Val encia (SP20120433-PAID-05-12) (Proyectos de nuevas Líneas de Investigacion Multidisciplinares; PAID-05-12). We thank INGELIA S.L. for giving us the hydrochar for free.This study was funded by the Universitat Politecnica de Val encia (SP20120433-PAID-05-12) (Proyectos de nuevas Líneas de Investigacion Multidisciplinares; PAID-05-12). We thank INGELIA S.L. for giving us the hydrochar for free.

The photosynthetic responses to a flowering-inductive water-stress period and recovery were studied and compared in two Citrus species. Under greenhouse conditions, Fino lemon and Owari satsuma trees were subjected to moderate (-2 MPa at predawn) and severe (-3 MPa) water stress levels and were re-watered after 60 days. Vegetative growth was inhibited during the stress assays, and strong defoliation levels were reported, especially in Fino lemon. In both species, bud sprouting was induced after re-watering. Flowers and vegetative shoots developed in Owari satsuma after a drought period, and the development was independent of the stress level. In Fino lemon, vegetative shoots and flowers were primarily formed after moderate and severe stress, respectively. The photosynthetic rate and stomatal conductance were reduced by water stress, and a marked increase in water-use efficiency at the moderate water deficit level was observed. Nevertheless, the photosynthetic apparatus was not damaged, since the maximum quantum yield, photosynthetic pigment concentrations and Rubisco level and activity did not change. Furthermore, the measured malonyldialdehyde (MDA) and peroxidase activity indicated that oxidative stress was not specifically triggered by water stress in our study. Therefore, the gas exchange, fluorescence and biochemical parameters suggested that diffusional limitations to photosynthesis predominated in both of the studied Citrus species, and explained the rapid recovery of the photosynthetic parameters after rehydration. The net CO 2 fixation rate and stomatal conductance were recovered within 24 h in Fino lemon, whereas 3 days were required in Owari satsuma. This suggests the presence of some metabolic limitations in the latter species. Furthermore, the sensibility of the defoliation rates, the accumulation of proline and the stomatal behaviour in response to water stress indicated a higher drought tolerance of Fino lemon, according to its better acclimation to hot climates. © 2011 Springer-Verlag., The authors thank Dr. J. Moreno and co-workers from the Departamento de Bioquimica of the Universidad de Valencia for his help and support in the Rubisco assays, and Dr. F. Fornes, Dr. A. Calatayud and Dr. E. Primo-Millo for the critical review of the manuscript. This work was funded by the Universitat Politecnica de Valencia, Spain (Ayudas para primeros proyectos de investigacion PAID06-06).

[EN] Rice is exposed in the field and in stored conditions to a great variety of fungi that can cause a lot of diseases with potential risk to consumers. In the present study, the chemical composition of commercial Laurus nobilis L. and Syzygium aromaticum L. Merr. & Perry essential oils and antifungal activity against five pathogenic fungi isolated from Mediterranean rice grains has been investigated. Thirty-seven compounds accounting for more than 99.5% of the total essential oil were identified by GC and GC/MS. 1,8-Cineole (51.95%), alpha-terpinyl acetate (12.93%) and the monoterpene hydrocarbon sabinene (9.56%) were the main compounds in bay leaf essential oil, while the phenylpropanoid eugenol (88.58%), and the sesquiterpene hydrocarbons beta-caryophyllene (8.13%) and alpha-humulene (2.35%) were found in clove essential oil. Clove essential oils at 300 mu g/mL showed more antifungal effect than bay leaf essential oil against all tested strains. S. aromaticum essential oil showed the best antifungal activity towards Fusarium graminearum and similar antifungal activity compared to pure eugenol against all tested phytopathogenic fungi. In inoculated rice grain, clove essential oil significantly reduced the fungal infection in food so S. aromaticum essential oil could be a good alternative as preservative in stored rice grain. (C) 2015 Elsevier Ltd. All rights reserved., This study has been financed by the Ministerio de Economia y Competitividad; Subdireccion General de Proyectos de Investigacion; Convocatoria de Ayudas a Proyectos de I+D+i; Research Challenges; Programme-oriented Societal Challenges 2014-2017, reference number AGL2013-42989-R-AR.

[EN] The antimicrobial activity of essential oils (EO) is very well-known and it has been reported that incorporating them into edible films based on biopolymers extends the food's shelf-life. In this study, cinnamon, clove and oregano EO, at 25% with respect to the polymer, were incorporated into glycerol plasticized starch-gelatin blend films (ratio 1:1) in order to elucidate their effect on the physical (barrier, mechanical and optical), structural and antifungal properties of the films. Whereas EOs exhibited no significant effect on tensile behavior in the case of casting films conditioned at 53% relative humidity and 25 degrees C, the EO compounds did significantly reduce the water vapor and oxygen permeability of the films. Likewise, the EOs increased the films' transparency but reduced their gloss. Despite the fact that about 60% of the incorporated EOs were lost during the film drying step, they exhibited antifungal activity against the two tested fungal species, Colletotrichum gloesporoides (CG) and Fusarium oxysporum (FOG), as revealed by the in vitro agar diffusion method. (C) 2016 Elsevier Ltd. All rights reserved., The authors acknowledge the financial support provided by the Ministerio de Economia y Competitividad-Spanish Government (AGL2013-42989-R).

[EN] Recently, biochars and hydrochars have been proposed as soil amendments and, to a lesser degree, as substrate constituents for soilless cultivation. These materials are interesting because their use is a means to reclaim organic matter and because, in some instances, they have been proved to sequester CO2 from the atmosphere. Myrtle and mastic are native to Mediterranean ecosystems and are grown for their essential oil production. This study aims to assess the properties of two biochars from different feedstock and a hydrochar from the same feedstock as one of the biochars as substrate constituents, in order to partially substitute either coir or soil in the cultivation of myrtle and mastic. Although the physical properties of the four organic materials are reasonably adequate for their use in substrate mixes, the biochar of olive mill waste is extremely saline as a pure material and remains very saline in a 50% mix with coir. The hydrochar showed less easily available water for the plants and a lower pH than the others. In incubation experiments using mixes with soil, biochars absorbed more 02 than CO2 emitted, which suggests they were sequestering CO2. The hydrochar showed O-2 consumption and CO2 emission of one order of magnitude larger than the biochars. It also showed that an outstanding amount of organic carbon was being metabolized in anaerobic routes. The biochar of forest waste and the coir displayed N immobilization when mixed with soil, whilst the biochar of olive mill waste had little microbial activity. Although hydrochars had been proved not to immobilize N as pure materials, they did immobilize N when mixed with soil. When tried for plant growth, mixes of the local soil with the biochar of forest waste and with hydrochar produced the best mastic plants. However, unmixed soil gave the best results for myrtle. The biochar of olive mill waste proved phytotoxic for myrtle in mixes with coir, whilst the biochar of forest waste gave optimal results for both species. Depending on their production process and the feedstock from which they are produced, biochars and hydrochars can vary in their characteristics and therefore in their effect on plants. (C) 2016 Elsevier B.V. All rights reserved.

[EN] Different adverse environmental conditions cause oxidative stress in plants by generation of reactive oxygen species (ROS). Accordingly, a general response to abiotic stress is the activation of enzymatic and non-enzymatic antioxidant systems. Many phenolic compounds, especially flavonoids, are known antioxidants and efficient ROS scavengers in vitro, but their exact role in plant stress responses in nature is still under debate. The aim of our work is to investigate this role by correlating the degree of environmental stress with phenolic and flavonoid levels in stress-tolerant plants. Total phenolic and antioxidant flavonoid contents were determined in 19 wild species. Meteorological data and plant and soil samples were collected in three successive seasons from four Mediterranean ecosystems: salt marsh, dune, semiarid and gypsum habitats. Changes in phenolic and flavonoid levels were correlated with the environmental conditions of the plants and were found to depend on both the taxonomy and ecology of the investigated species. Despite species-specific differences, principal component analyses of the results established a positive correlation between plant phenolics and several environmental parameters, such as altitude, and those related to water stress: temperature, evapotranspiration, and soil water deficit. The correlation with salt stress was, however, very weak. The joint analysis of all the species showed the lowest phenolic and flavonoid levels in the halophytes from the salt marsh. This finding supports previous data indicating that the halophytes analysed here do not undergo oxidative stress in their natural habitat and therefore do not need to activate antioxidant systems as a defence against salinity., This work has been funded by the Spanish Ministry of Science and Innovation (Project CGL2008-00438/BOS), with contribution from the European Regional Development Fund. Thanks to Dr. Rafael Herrera for critical reading of the manuscript.

Chemical studies of green leaves of A. tetraphyllum afforded beta-sitosterol, a mixture containing the ethyl esters of long chain carboxylic acids, 30-normethyl-lupan-20-one, hopan-22-ol, phytol, phyten-3(20)-1,2-diol, quercetin and quercetin-3-O-beta-D-glucoside. The structures of the compounds were elucidated by spectroscopic and GC analysis. The allelopathic potentials of the crude ethanolic extract and fractions were evaluated against Lactuca sativa (letuce) and Allium cepa (onion) seeds.

The presence and extent of genetic variation in saffron crocus are still debated, as testified by several contradictory articles providing contrasting results about the monomorphism or less of the species. Remarkably, phenotypic variations have been frequently observed in the field, such variations are usually unstable and can change from one growing season to another. Considering that gene expression can be influenced both by genetic and epigenetic changes, epigenetics could be a plausible cause of the alternative phenotypes. In order to obtain new insights into this issue, we carried out a molecular marker analysis of 112 accessions from the World Saffron and Crocus Collection. The accessions were grown for at least three years in the same open field conditions. The same samples were analysed using Amplified Fragment Length Polymorphism (AFLP) and Methyl Sensitive AFLP in order to search for variation at the genetic (DNA sequence) and epigenetic (cytosine methylation) level. While the genetic variability was low (4.23% polymorphic peaks and twelve (12) effective different genotypes), the methyl sensitive analysis showed the presence of high epigenetic variability (33.57% polymorphic peaks and twenty eight (28) different effective epigenotypes). The pattern obtained by Factorial Correspondence Analysis of AFLP and, in particular, of MS-AFLP data was consistent with the geographical provenance of the accessions. Very interestingly, by focusing on Spanish accessions, it was observed that the distribution of the accessions in the Factorial Correspondence Analysis is not random but tends to reflect the geographical origin. Two clearly defined clusters grouping accessions from the West (Toledo and Ciudad Real) and accessions from the East (Cuenca and Teruel) were clearly recognised.

[EN] The localized application of the synthetic cytokinin CPPU ((2-chloro-4-pyridyl)-N-phenyl urea) to ovaries at flower opening was as effective as free pollination in setting parthenocarpic fruit in the triploid watermelon cultivar ' Reina de Corazones', and increased yield per unit land area by at least 50%, simply due to the lack of requirement for diploid pollen producing plants within the orchard. The application of the synthetic auxin 2,4-D (2,4-dichlorophenoxy acetic acid) as a full coverage spray, was also effective in setting fruit; total yield was however 10% smaller than in the CPPU-treated plots, but the cost of application was much less expensive. These applications had no adverse effect on fruit quality, and their effectiveness in commercial watermelon production was evaluated over 4 years. Localized applications of 2,4-D to ovaries were less effective in setting fruit, and increased hollow fruit.

[EN] An open-top chamber study was conducted to investigate the tissue and cellular-level foliar effects of ozone (O-3) on a Mediterranean evergreen species, the mastic plant (Pistacia lentiscus L.). Plants were exposed at three different O-3 levels, and leaf samples were collected periodically from the beginning of the exposure. Although no visible foliar injury was evident, alterations of the plastids and vacuoles in the mesophyll were observed. Senescence processes were accelerated with an anomalous stacking of tannin vacuoles, and a reduction in the size and number of the chloroplasts. Overall, most of the modifications induced by O-3 were consistent with previously reported observations on deciduous broadleaf species, with the exception of alterations in the cells covering the secretory channels, reported here as a new finding. Comments on the feasibility of using microscopy to validate O-3 related field observations and subtle foliar injury are also given. (C) 2004 Elsevier Ltd. All rights reserved.

[EN] Two biochars, one from forest waste (BCH-FW) and the other from olive mill waste (BCH-OMW), and one hydrochar from forest waste (HYD-FW) were analyzed from the horticultural and environmental point of view. Several incubation assays were performed to foresee the material evolution when used as soilless growth media. HYD-FW was slightly alkaline and rich in nutrients and showed high microbial respiration Moreover, it did not caused N immobilization but N mineralization which contradicts previous reports. Biochars were highly alkaline. BCH-FW was poor in nutrients whilst BCH-OMW was saline and rich in soluble cations. Both materials had low respiratory activity, similar to that of coir fibre (CF). Whilst BCH-FVV and CF immobilize N, BCH-OMW did not. Both biochars captured CO2 from the atmosphere, particularly BCH-OMW, which concomitantly showed a decrease in soluble cations. We conclude that pure hydrochar does not immobilize N and that alkaline biochars act as CO2 traps. (C) 2014 Elsevier Ltd. All rights reserved., This study was funded by the Universitat Politecnica de Valencia (Proyectos de nuevas Lineas de Investigacion Multidisciplinares; SP 20120433-PAID-05-12). We thank INGELIA S.L. for giving the hydrochar for free and to Mr Manuel Sanchez-Perales and Mrs Joana Oliver for their technical assistance.

Essential oils of bay leaf, cinnamon, clove and oregano were tested in vitro and oregano essential oil in vivo, against two foodborne fungi belonging to the dominant mycobiota of stored rice, Fusarium culmorum and Fusarium verticillioides, collected from the Albuferarice-producing Mediterranean area near Valencia (Spain). Chemical composition was identified by gas chromatography-mass spectrometry. Essential oils presented a high percentage of oxygenated components: 78.8 % in bay leaf (eucalyptol 51 %); 90.3 % in clove (eugenol 89.8 %); 92 % in cinnamon (eugenol 60 % and eugenyl acetate18.3 %); 71.8 % in oregano (carvacrol 49.6 % and thymol 21.2 %). Monoterpenes and sesquiterpenes were: 18 % in bay leaf, 9 % in clove, 5 % in cinnamon, 25 % in oregano. This research showed that essential oils have a great potential to control both fungal pathogens. In the in vitro test, the essential oils of cinnamon, clove and oregano reduced fungal growth by 90 % and almost 100 %, being oregano the most effective essential oil to inhibit fungal growth. The effect of the oregano essential oil on fungal development in inoculated rice grains demonstrated its effectiveness., This study has been financed by the Vice-Chancellor for Research at the Polytechnic University of Valencia as part of its Support Programme to Research and Development, New Lines of Multidisciplinary Research (PAID-05-10), reference number 2644.

[Background] In saffron (Crocus sativus), new corms develop at the base of every shoot developed from the maternal corm, a globular underground storage stem. Since the degree of bud sprouts influences the number and size of new corms, and strigolactones (SLs) suppress growth of pre-formed axillary bud, it was considered appropriate to investigate SL involvement in physiology and molecular biology in saffron. We focused on two of the genes within the SL pathway, CCD7 and CCD8, encoding carotenoid cleavage enzymes required for the production of SLs., [Results] The CsCCD7 and CsCCD8 genes are the first ones isolated and characterized from a non-grass monocotyledonous plant. CsCCD7 and CsCCD8 expression showed some overlapping, although they were not identical. CsCCD8 was highly expressed in quiescent axillary buds and decapitation dramatically reduced its expression levels, suggesting its involvement in the suppression of axillary bud outgrowth. Furthermore, in vitro experiments showed also the involvement of auxin, cytokinin and jasmonic acid on the sprouting of axillary buds from corms in which the apical bud was removed. In addition, CsCCD8 expression, but not CsCCD7, was higher in the newly developed vascular tissue of axillary buds compared to the vascular tissue of the apical bud., [Conclusions] We showed that production and transport of auxin in saffron corms could act synergistically with SLs to arrest the outgrowth of the axillary buds, similar to the control of above-ground shoot branching. In addition, jasmonic acid seems to play a prominent role in bud dormancy in saffron. While cytokinins from roots promote bud outgrowth. In addition the expression results of CsCCD8 suggest that SLs could positively regulate procambial activity and the development of new vascular tissues connecting leaves with the mother corm., The laboratory is funded by the Spanish Ministerio de Ciencia e Innovación (BIO2009-07803) and participates in the IBERCAROT network (112RT0445). Dr. Ahrazem was funded by FPCYTA through the INCRECYT Programme.

DNA binding with One Finger (DOF) transcription factors are involved in multiple aspects of plant growth and development but their precise roles in abiotic stress tolerance are largely unknown. Here we report a group of five tomato DOF genes, homologous to Arabidopsis Cycling DOF Factors (CDFs), that function as transcriptional regulators involved in responses to drought and salt stress and flowering-time control in a gene-specific manner. SlCDF1–5 are nuclear proteins that display specific binding with different affinities to canonical DNA target sequences and present diverse transcriptional activation capacities in vivo. SlCDF1–5 genes exhibited distinct diurnal expression patterns and were differentially induced in response to osmotic, salt, heat, and low-temperature stresses. Arabidopsis plants overexpressing SlCDF1 or SlCDF3 showed increased drought and salt tolerance. In addition, the expression of various stress-responsive genes, such as COR15, RD29A, and RD10, were differentially activated in the overexpressing lines. Interestingly, overexpression in Arabidopsis of SlCDF3 but not SlCDF1 promotes late flowering through modulation of the expression of flowering control genes such as CO and FT. Overall, our data connect SlCDFs to undescribed functions related to abiotic stress tolerance and flowering time through the regulation of specific target genes and an increase in particular metabolites., This work was supported by grants from Instituto Nacional de Investigación y Tecnología Agraria y Alimentaria (INIA; project numbers: 2009-0004-C01, 2012-0008-C01), the Spanish Ministry of Science and Innovation (project number: BIO2010-14871), and the MERIT Project (FP7 ITN2010-264474). ARC was supported by a pre-doctoral fellowship from the INIA.

Crocins, the glucosides of crocetin, are present at high concentrations in saffron stigmas and accumulate in the vacuole. However, the biogenesis of the saffron chromoplast, the changes during the development of the stigma and the transport of crocins to the vacuole, are processes that remain poorly understood. We studied the process of chromoplast differentiation in saffron throughout stigma development by means of transmission electron microscopy. Our results provided an overview of a massive transport of crocins to the vacuole in the later developmental stages, when electron dense drops of a much greater size than plastoglobules (here defined “crocinoplast”) were observed in the chromoplast, connected to the vacuole with a subsequent transfer of these large globules inside the vacuole. A proteome analysis of chromoplasts from saffron stigma allowed the identification of several well-known plastid proteins and new candidates involved in crocetin metabolism. Furthermore, expressions throughout five developmental stages of candidate genes responsible for carotenoid and apocarotenoid biogenesis, crocins transport to the vacuole and starch metabolism were analyzed. Correlation matrices and networks were exploited to identify a series of transcripts highly associated to crocetin (such as 1-Deoxy-d-xylulose 5-phosphate synthase (DXS), 1-Deoxy-d-xylulose 5-phosphate reductoisomerase (DXR), carotenoid isomerase (CRTISO), Crocetin glucosyltransferase 2 (UGT2), etc.) and crocin (e.g., ζ-carotene desaturase (ZDS) and plastid-lipid-associated proteins (PLAP2)) accumulation; in addition, candidate aldehyde dehydrogenase (ADH) genes were highlighted., This work was supported by the Spanish Ministerio de Economía y Competitividad (BIO2013-44239-R), participates in the IBERCAROT network (112RT0445) and benefited from the networking activities within the European Cooperation in Science and Technology Action CA15136 (EUROCAROTEN). The proteomics analysis LC-MS/MS by LTQ Orbitrap Velos was carried out in the Proteomics and Genomics Facility (CIB-CSIC), a member of ProteoRed-ISCIII networ

[EN] The essential oil composition of Cistus ladanifer grown in central Spain was analysed by gas chromatography and gas chromatography/mass spectrometry, revealing it to be rich in oxygenated compounds, with trans-pinocarveol (20.00%), bornyl acetate (7.03%) and terpinen-4-ol (6.37%) as the main monoterpene compounds. Viridiflorol (13.59%) and ledol (4.36%) were the main constituents of the oxygenated sesquiterpene fraction. Large amounts of-pinene (4.70%) were found in the hydrocarbonated fractions. To study its possible use as a natural herbicide, the oil was tested invitro against Amaranthus hybridus, Portulaca oleracea, Chenopodium album, Conyza canadensis and Parietaria judaica, completely inhibiting A. hybridus germination, and nearly blocking C. canadensis and P. judaica germination at all concentrations assayed. In P. oleracea, the oil showed less effect, inhibiting its germination at the higher doses studied. It did not control C. album germination. Regarding seedling length, the oil had strong phytotoxic activity, being effective at all concentrations tested. © 2012 Taylor & Francis.

[EN] Changes in physical and chemical characteristics of an organic waste (tomato crop waste) throughout composting (COM), vermicomposting (VER) and the combination of both processes (C + V) were assessed at five selected stages. COM consisted of a combination of the static Rutgers system with forced aeration and pile turning. For VER Eisenia worms were fed with the raw material. For C + V pre-composted material was added to the worms. Particle size decreased during COM, yet it increased during VER and C + V due to the amalgamation of small particles. pH was alkaline throughout the processes. Heavy irrigation during vermicomposting resulted in greater decrease of EC and greater leaching of organic matter, total nitrogen and most macronutrients in VER and C + V than in COM. Final materials were not phytotoxic but compost could have salinity related problems. Thus, COM, VER and C + V produced treated organic material, which could be suitable for horticultural purposes. (C) 2012 Elsevier Ltd. All rights reserved., Authors are deeply grateful to Dr. Maria Jose Lopez and Dr. Raquel Melgar for supervising the composting and vermicomposting procedures. They would also like to thank Professor Sebastia Balasch for his advice on statistics and Dr. Bruce Pearce for reviewing the English. This study was funded by the Instituto Nacional de Investigacion y Tecnologia Agraria (INIA) of the Spanish Ministerio de Ciencia e Innovacion (Project Reference RTA2006-00008-C02-02).

13C nuclear magnetic resonance (NMR) spectroscopy was explored as a simple and efficient technique for the quantitative analysis of nonpolar metabolites in plants. The method was first optimized with a mixture of known metabolites and then applied to the nonpolar leaf extracts of plants harvested in the Valencian community (eastern Spain) belonging to three different genera: Euphorbia (Euphorbiaceae), Araujia (Apocynaceae), and Morus (Moraceae). Furthermore, an exhaustive analysis of Euphorbia characias leaf and stem extracts from different geographic locations allowed that quantitative 13C NMR spectroscopy is a suitable tool for metabolic profiling purpose. © 2011 American Chemical Society., We thank the Spanish Ministerio de Ciencia e Innovacion of Spain (MICINN, SAF2008 01845) and the Centro de Investigacion Principe Felipe for their economic support We also thank Cemex Sustainable Chair in Spain for the predoctoral fellowship to P V E

Fruits from many citrus cultivars develop depressed areas in the flavedo (outer part of the peel) and albedo (inner part) following detachment. Although ultrastructural analysis may provide important information about multiple plant responses to stresses and external stimuli at the cell and tissue levels, and despite the proved efficacy of ethylene in reducing peel damage in citrus fruit, cytological responses of this horticultural crop to protective ethylene concentrations have not yet been reported. We show that applying high ethylene levels (2 mu L L(-1) for 14 days) causes sublethal stress as it favored the alteration of cuticle, vacuole, middle lamella and primary wall, especially in the albedo cells, but reduced peel collapse in detached mature "Navelate" oranges (C. sinensis, L. Osbeck) held under nonstressful environmental conditions (22 degrees C and 90-95% RH). Ethylene did not induce relevant changes in lignification but favored the deposition of pectic exudates and the release of sugars from degradation of cell polysaccharides including starch, cellulose, and pectins. In contrast, inhibiting ethylene perception by applying 1-methylcyclopropene (1-MCP) reduced these ethylene-related responses and favored degradation of cell membranes and peel damage. The overall results reflect that mature oranges tolerate high ethylene levels that might favor the activation of defense responses involving oxidative-stress related mechanisms and recycling of nutrients and carbon supply to enable cells to sustain respiration and cope with carbon deprivation stress caused by detachment. Microsc. Res. Tech. 74:970-979, 2011. (C) 2011 Wiley-Liss, Inc., Contract grant sponsor: Comision Interministerial de Ciencia y Tecnologia (CICYT), Spain; Contract grant number: AGL2002-1727; Contract grant number: AGL2009-11969; Contract grant sponsor: Conselleria D'Educacio Generalitat Valenciana, Spain; Contract grant number: PROMETEO/2010/010; Contract grant sponsor: SUPERA Programme, Mexico

Photosynthesis down-regulation due to an imbalance between sources and sinks in Citrus leaves could be mediated by excessive accumulation of carbohydrates. However, there is limited understanding of the physiological role of soluble and insoluble carbohydrates in photosynthesis regulation and the elements triggering the down-regulation process. In this work, the role of non-structural carbohydrates in the regulation of photosynthesis under a broad spectrum of source-sink relationships has been investigated in the Salustiana sweet orange. Soluble sugar and starch accumulation in leaves, induced by girdling experiments, did not induce down-regulation of the photosynthetic rate in the presence of sinks (fruits). The leaf-to-fruit ratio did not modulate photosynthesis but allocation of photoassimilates to the fruits. The lack of strong sink activity led to a decrease in the photosynthetic rate and starch accumulation in leaves. However, photosynthesis down-regulation due to an excess of total soluble sugars or starch was discarded because photosynthesis and stomatal conductance reduction occurred prior to any significant accumulation of these carbohydrates. Gas exchange and fluorescence parameters suggested biochemical limitations to photosynthesis. In addition, the expression of carbon metabolism-related genes was altered within 24 h when strong sinks were removed. Sucrose synthesis and export genes were inhibited, whereas the expression of ADP-glucose pyrophosphorylase was increased to cope with the excess of assimilates. In conclusion, changes in starch and soluble sugar turnover, but not sugar content per se, could provide the signal for photosynthesis regulation. In these conditions, non-stomatal limitations strongly inhibited the photosynthetic rate prior to any significant increase in carbohydrate levels. © The Author 2011. Published by Oxford University Press. All rights reserved., This work was supported by the Conselleria de Cultura, Educacio i Esport de la Generalitat Valenciana [GV/2007/213 and GV/2009/034].

[EN] The essential oil composition of Peumus boldus and Drimys winterii was analyzed by means of capillary GC-FID and GC-MS. More than 96% of the total oil components (43 and 54 compounds, respectively) were identified, with ascaridole (51.17 ± 9.51), p-cymene (16.31 ± 2.52) and 1,8-cineole (14.45 ± 2.99) as the main compounds in P. boldus and ¿-eudesmol (21.65 ± 0.41), followed of elemol (12.03 ± 0.34) and terpinen-4-ol (11.56 ± 1.06) in D. winterii. The herbicidal activity was tested against Amaranthus hybridus and Portulaca oleracea. P. boldus essential oil was the most phytotoxic against both weeds, inhibiting seed germination and seedling growth at all concentrations assayed (0.125-1 ¿L/mL). D. winterii essential oil did not show any effect on A. hybridus germination and only affected P. oleracea germination at the highest concentration. The results suggest the possible use of the essential oil from P. boldus as a natural herbicide. © 2010 by the authors., Research performed under an International Cooperation Project (number A/024753/09), financed by Agencia Espanola de Cooperacion Internacional para el Desarrollo (AECID).

[EN] Responses to increasing salinity, during seed germination and vegetative plant growth, were studied in two related species of Juncus, J. maritimus and J. acutus. In both species, germination was optimal in the absence of salt, reduced by about 50% in the presence of 200 mM NaCl, and completely inhibited by NaCl concentrations above 300 mM. Previous exposure of the seeds to salt, up to 500 mM NaCl, did not affect the germination capacity in J. acutus, and clearly enhanced it in J. maritimus. A concentration-dependent inhibition of plant growth was observed in the presence of NaCl for both species, together with the parallel accumulation of sodium ions in the leaves, as determined by cation exchange HPLC. Regarding the levels of divalent cations, in J. acutus Ca2+ and Mg2+ increased up to about two-fold in plants treated with 500 mM NaCl, as compared to control plants, whereas in J. maritimus they were three-to four-fold higher than in J. acutus in the absence of salt, and did not change significantly with increasing NaCl concentrations. These results suggest that Ca2+ and Mg2+ participate in defence mechanisms against salt stress, which would be constitutive in J. maritimus and salt-inducible in J. acutus.

Ozone effects on the perennial forb Lamottea dianae were studied in an open-top chamber experiment. Ozone was found to induce reductions in CO 2 assimilation and water use efficiency in the leaves of this species. These reductions were mainly related to a decline in the in vivo CO 2 fixation capacity of Rubisco (V c,max), rather than to stomatal limitations or photoinhibitory damage (F v:F m). In addition to chloroplast degeneration, other observed effects were callose accumulation, formation of pectinaceous wart-like cell wall exudates and phloem alterations. Moreover, ozone exposure significantly reduced root dry biomass. The possible relevance of these adverse effects for Mediterranean forbs is commented. These results show that endemic plants can be very sensitive to ozone, suggesting that risks associated with this pollutant should be taken into account for conservation purposes. © 2011 Elsevier Inc., We thank the Conselleria de Medi Ambient, Aigua, territory i Habitatge and FEDER funds (project VegetPollOzone, Interreg IIIb, Meddoc) for supporting parts of this study. The Instituto Universitario CEAM-UMH is partly supported by Generalitat Valenciana, Fundacion Bancaja, and benefits form the projects CONSOLIDER-INGENIO 2010 (GRACCIE) and Prometeo Program (Generalitat Valenciana). Emilio Laguna and Mari Carmen Escriva are also thanked for providing us the plants and, together with Joan Perez, for their comments on aspects of the biology of this species. Carmen Martin is thanked for taking care of the plants. Four anonymous referees provided very useful comments for improving the manuscript.

The application of the cryo-scanning electron microscopy and light microscopy for the study of the interactions at different environmental conditions between Penicillium oxalicum and Fusarium verticillioides is described. A dual microculture was developed for the light microscopy analysis of the interaction. The microscope and macroscopic examinations were compared. Analysis of Petri plates revealed that F. verticillioides was a competitor for space and nutrients while P. oxalicum was a mycoparasite under the microscopic observations.

[EN] Potato plants were grown in open-top chambers under three ozone concentrations during two complete cropping seasons (93 and 77 d in 2004 and 2005, respectively). The effects of chronic exposure to ozone on leaf anatomy, cell ultrastructure and crop yield were studied. Severe cell damage was found, even at ambient ozone levels, mainly affecting the spongy parenchyma and areas near the stomata. Damage to the cell wall caused loss of cell contact, and loss of turgor pressure due to tonoplast disintegration, contributed to cell collapse. Phloem sieve plates were obstructed by callose accumulation, and damaged mesophyll cells increased their starch stores. Tuber yield fell sharply (24–44%), due to the biggest tubers becoming smaller, which affected commercial yield. These anatomical findings show the mechanisms of ozone effect on assimilate partitioning, and thus crop yield decrease, in potato. Further implications of ozone causing reductions in belowground biomass are also discussed., The authors thank Prof Secundino del Valle (Valencia University, Spain) for his helpful comments. We are also grateful to Mr Duncan Gates for revising the English style of the text. AAF was supported by a grant from the Generalitat Valenciana's FPI programme (Government of Valencia, Spain).

8 pages, 6 figures, 1 table. -- Available online 15 September 2009., The effect of ethylene (2 μL L−1) on total and soft epicuticular wax content and wax morphology has been investigated in mature ‘Navelate’ (Citrus sinensis, L. Osbeck) oranges held under non-stressful environmental conditions (22 °C and constant high relative humidity (90–95% RH)). In addition, the objective of the study was to understand whether the ethylene-induced changes in epicuticular wax might participate in the beneficial effect of ethylene reducing non-chilling peel pitting, by modifying peel water, osmotic or turgor potential, or disease incidence caused by Penicillium digitatum (Pers.:Fr.) Sacc. Ethylene increased total and soft epicuticular wax content in ‘Navelate’ fruit and induced structural changes in surface wax that might be related to the formation of new waxes. Changes in epicuticular wax morphology, but not in its content, might be involved in the protective role of ethylene reducing non-chilling peel pitting, although the beneficial effect of the hormone is not related to water stress. Cell water and turgor potentials in freshly harvested fruit and fruit stored in air under non-stressful conditions suggest that water stress is not a limiting factor leading to the development of this physiological disorder. In addition, the results indicated that formation of new waxes in fruit treated with ethylene may partially cover stomata, cracks or areas lacking wax occurring in stored fruit and is likely to improve physical barriers to P. digitatum penetration., This work was supported by research grant AGL2002-1727 from the Comisión Interministerial de Ciencia y Tecnología (CICYT), Spain. Cajuste was the recipient of a fellowship from the Program SUPERA, México.

The effect on fruit quality of grafting and fruit set treatments, was determined in the triploid watermelon (Citrullus lanatus (Thun.) Matsum. and Nakai) cultivar 'Reina de Corazones' grown in a heated greenhouse. Grafting the scion onto 'Shintoza' (Cucurbita maxima X Cucurbita moschata) rootstock and setting fruit through application of the synthetic cytokinin CPPU [1-(2-chloro-4-pyridyl)-3-phenylurea] retarded the accumulation of sugar and development of the flesh colour of the fruit, compared to fruit from non-grafted, pollinated plants. The effects of grafting and of CPPU application on sugar accumulation were additive. At commercial maturity, the fruit of grafted and/or CPPU-treated plants had lower total soluble solids and sugar concentrations, and lower sucrose to hexose ratios, than fruit from control plants. Fruit from CPPU-treated plants also had a weaker red colour than fruit set by pollination. Despite this loss of quality, fruit from grafted plants, set with CPPU and grown in a heated greenhouse, had a total soluble solids concentration similar to fruit from pollinated, non-grafted plants, grown outdoors.

[EN] Background Reactive oxygen species (ROS) are normally produced in respiratory and photosynthetic electron chains and their production is enhanced during desiccation/rehydration. Nitric oxide (NO) is a ubiquitous and multifaceted molecule involved in cell signaling and abiotic stress. Lichens are poikilohydrous organisms that can survive continuous cycles of desiccation and rehydration. Although the production of ROS and NO was recently demonstrated during lichen rehydration, the functions of these compounds are unknown. The aim of this study was to analyze the role of NO during rehydration of the lichen Ramalina farinacea (L.) Ach., its isolated photobiont partner Trebouxia sp. and Asterochloris erici (Ahmadjian) Skaloud et Peksa (SAG 32.85 = UTEX 911). Results Rehydration of R. farinacea caused the release of ROS and NO evidenced by the fluorescent probes DCFH2-DA and DAN respectively. However, a minimum in lipid peroxidation (MDA) was observed 2 h post-rehydration. The inhibition of NO in lichen thalli with c-PTIO resulted in increases in both ROS production and lipid peroxidation, which now peaked at 3 h, together with decreases in chlorophyll autofluorescence and algal photobleaching upon confocal laser incidence. Trebouxia sp. photobionts generate peaks of NO-endproducts in suspension and show high rates of photobleaching and ROS production under NO inhibition which also caused a significant decrease in photosynthetic activity of A. erici axenic cultures, probably due to the higher levels of photo-oxidative stress. Conclusions Mycobiont derived NO has an important role in the regulation of oxidative stress and in the photo-oxidative protection of photobionts in lichen thalli. The results point to the importance of NO in the early stages of lichen rehydration., This project was funded by the Spanish Ministry of Education and Science [project numbers CGL2006 12917 C02 0 and CGL2009 13429 C02 01], project Prometeo 2008/1/4 of the Generalitat Valenciana and the project AECID PCI/A/024755/09 of the Spanish Ministry of Foreign Affaires. We are grateful to F. Gasulla, J. Gimeno-Romeu, E. Barreno, (ICBIBE, University of Valencia) and A. Guera (Plant Biology, University of Alcala) for communicating unpublished data, to Dr. R. Catala (CIB, Madrid), Dr. P. D'Ocon (UVEG, Valencia) and Dr. J. Medina (INIA, Madrid) for critical revision of the manuscript, and J. L. Rodriguez Gil for MDA protocol optimization. English revision was done by Wendy Ran.

[EN] Two chitinases induced in tomato leaves (Lycopersicon esculentum Mill cv. Rutgers) by citrus exocortis viroid (CEV) infection were purified. Their molecular masses determined by sodium dodecyl sulfate-gel electrophoresis were 32 kDa and 34 kDa and by filtration through Sephadex G-100 were 23 kDa and 25 kDa, respectively. These chitinases (P32 and P34) have been shown to be identical to the tomato pathogenesis-related proteins C7 and C6. They were purified in three stages: ammonium sulphate fractionation, chitin affinity chromatography and CM-Sepharose chromatography. The characterization of P32, the major component of the CEV-induced chitinase activity, revealed a basic protein (pI, 8·5), with a low optimum pH of 5·2 for hydrolytic activity, a high thermal stability and resistance to proteolytic degradation. These tomato chitinases have been shown to be serologically related to the tobacco chitinases. Our results lend further support to the hypothesis that enhanced chitinase activity is a component of a general mechanism of response to pathogens and other stress agents.

The chemical composition of essential oils from Satureja cuneifolia growing in east Spain was analyzed by GC, GC/MS. Forty-five compounds accounting for 99.1% of the total oil were identified. Camphor (47.6%), followed by camphene (13.6%) were the main compounds. Their herbicidal and antifungal activity was tested in vitro against three weeds (Amaranthus hybridus, Portulaca oleracea and Conyza canadensis) and eleven common pathogenic or saprophytic fungi (Phytophthora citrophthora, P. palmivora, Pythium litorale, Verticillium dahlia, Rhizoctonia solani, Penicillium hirsutum, Colletotrichum gloeosporioides, Phaeoacremonium aleophilum, Phaemoniella chlamydospora, Cylindrocarpon liriodendri and C. macrodidymum). The essential oil was very active against A. hybridus and C. canadensis significantly inhibiting their germination and seedling growth. Minor activity was shown against P. oleracea, depending on the concentration applied. P. palmivora, P. citrophthora and Pa. chlamydospora were the most sensitive fungi to the treatment with the essential oil, whereas R. solani showed no inhibition. Results showed that S. cuneifolia essential oil could be used for biocontrol of weeds and fungal plant diseases.

Carob seedlings were grown at different proportions of nitrate NO3) and ammonium (NH4) supplied at a concentration of 3.5 mM nitrogen (N). The effect of five different combinations of NH4- and NO3-N on growth rates, N and carbon (C) content of the plant and dry matter distribution were studied. The presence of NH4 in the nutrient solution enhanced plant growth rate, although net photosynthesis rates per unit area were lower than in NO3-fed plants. Nitrate –fed plants exbibited the highest rates of carbon dioxide (CO2) assimilation, water use, and N-use efficiency. Nevertheless, plants growing only with NO3 were more fragile and susceptible to fungus infections. The role of cotyledons on the regulation of the supply of photosynthates from leaves to roots in the different combinations of NO3- and NH4-N is described. Results suggest that the growth rates of carob may be improved using suitable combinations of NO3- and NH4-N. Equal concentrations of these two N sources allows good development and adequate allocation of low C and N to different developing sinks of the carob seedlings.