Ageing kinetics of fern chlorophyllous spores during dry storage is determined by its antioxidant potential and likely induced by photosynthetic machinery.

Ageing in dry chlorophyllous propagules is leaded by photooxidation through the photosynthetic machinery, but why species differ in longevity and the ageing mechanisms of when light and oxygen are absent are unknown. We hypothesize that the cellular antioxidant capacity is key for the inter- and intra-specific differences in the ageing process. We have tested this hypothesis in chlorophyllous spores of two ferns. They were subjected to four different storage regimes resulting from light/dark and normoxia/hypoxia combinations. Lipophilic and hydrophilic antioxidants, reactive oxygen species (ROS), and photosynthetic pigments were analysed in parallel to germination and the recovery of Fv/Fm over a storage period of up to 22-months. We show that light and oxygen accelerate the ageing process, but their mechanisms (ROS, increase, antioxidant capacity decrease, loss of efficiency of the photosystem II, pigment degradation) appear the same under all conditions tested. The end of the asymptomatic phase of longevity, when a sudden drop of germination occurs, seems to be determined by a threshold in the depletion of antioxidants. Our results support the hypothesis that ageing kinetics in dry plant propagules is determined by the antioxidant system, but also suggests an active role of the photosynthetic machinery during ageing, even in darkness and hypoxia.

Unraveling metabolic mechanisms behind chloroplast desiccation tolerance: Chlorophyllous fern spore as a new promising unicellular model.

Fern spores are unicellular structures produced by the sporophyte generation that give rise to the haploid gametophyte. When released from the sporangium, spores are desiccation tolerant (DT) in the royal fern (Osmunda regalis) and contain fully developed chloroplasts. As a consequence, this type of spores is called chlorophyllous spores (CS). Upon transfer to germination conditions, CS initiate a process of imbibition that suppresses DT in 72 h, before the germination starts. In parallel to such change in DT, thylakoids undergo a profound remodelling in composition and function. Firstly, sustained quenching of chlorophyll fluorescence is relaxed, giving rise to photochemically active CS, while lipid composition shifts from that of a resting structure to a metabolically active cell. Basically trigalactolipids decreased in favour of monogalactolipids, with a parallel desaturation of fatty acids. Storage lipids such as triacylglycerol were quickly depleted. These results highlight the importance of the structure of thylakoids lipid as a key to protect membrane integrity during desiccation, together with the saturation of fatty acids and the constitutive chlorophyll quenching to prevent oxidative damage. The CS used here, in which the same cell shifts from DT to sensitive strategy in 72 h, reveal their potential as unicellular models for future studies on DT.

Does plant colour matter? Wax accumulation as an indicator of decline in Juniperus thurifera.

The photosynthesis in evergreen trees living in Mediterranean ecosystems is subjected to multiple climatic stresses due to water shortage and high temperatures during the summer and to low temperatures during the winter. Mediterranean perennials deploy different photoprotective mechanisms to prevent damage to the photosynthetic system. Wax accumulation in leaves is a primary response which by enhancing light scattering in the leaf surface reduces incident radiation in the mesophyll. The existence of high variability in wax accumulation levels between coexisting individuals of a species has a visual effect on colour that provides distinguishable green and glaucous phenotypes. We explored this variability in a Mediterranean evergreen tree Juniperus thurifera (L.) to evaluate the impact of epicuticular wax on optical and ecophysiological properties and on the abundance of photoprotective pigments throughout an annual cycle. Because of light attenuation by waxes, we expected that glaucous phenotypes would lower the need for photoprotective pigments. We evaluated the effect of phenotype and season on reflectance, defoliation levels, photochemical efficiency and photoprotective pigment contents in 20 green and 20 glaucous junipers. Contrary to our expectations, the results showed that glaucous trees suffered from a diminution in photochemical efficiency, but there was no reduction in photoprotective pigments. Differences between glaucous and green phenotypes were greater in winter, which is the most stressful season for this species. Glaucous individuals also showed the highest levels of leaf defoliation. The lower photochemical efficiency of glaucous trees, together with higher defoliation rates and equal or greater number of physiological photoprotective mechanisms, suggests that in spite of wax accumulation, glaucous trees suffer from more severe stress than green ones. This result suggests that changes in colouration in Mediterranean evergreens may be a decline indicator.

Antioxidant and photoprotective responses to elevated CO(2) and heat stress during holm oak regeneration by resprouting, evaluated with NIRS (near-infrared reflectance spectroscopy).

Photosynthetic, photoprotective and antioxidant responses during high temperature stress were determined in leaves of evergreen holm oak (Quercus ilex L.), the main species in Mediterranean forests, during resprouting under elevated CO(2) (750 µl·l(-1) ). Leaf chemicals, chloroplast pigments and non-enzymatic antioxidants were quantified in a single measurement using NIRS (near-infrared spectroscopy), a rapid and suitable method for ecophysiological purposes. Resprouts from plants grown under elevated CO(2) (RE) showed photosynthetic down-regulation, higher starch content and lower stomatal conductance, but similar stomatal density, than plants grown under current CO(2) concentrations (350 µl·l(-1) ) (RA). The photosynthetic sink reduction and need for more antioxidants and photoprotection in RE were reflected in an increased concentration of ascorbate (Asc) and phenolic compounds and in the contribution of the xanthophyll (Z/VAZ) and lutein epoxide cycles to excess energy dissipation as heat, and also reflected in chlorophyll fluorescence measurements. CO(2) assimilation parameters were stable from 25 to 35 °C in RE and RA, declining thereafter in RA in spite of a 2.3 °C lower leaf temperature. RE showed a more marked decline in photorespiration above 35 °C and less sensitive stomatal responses to high temperature stress than RA. During heat stress, RE had higher Asc, Z/VAZ and phenolics content, together with delayed enhancement of chloroplast lipophilic antioxidant compounds (carotenes and tocopherols). The high contribution of photoprotective systems and high temperature tolerance in resprouts developed under elevated CO(2) would mitigate the effect of photosynthesis acclimation during the regeneration of Q. ilex plants under climate change.

Native plant communities in an abandoned Pb-Zn mining area of northern Spain: implications for phytoremediation and germplasm preservation.

Plants growing on metalliferous soils from abandoned mines are unique because of their ability to cope with high metal levels in soil. In this study, we characterized plants and soils from an abandoned Pb-Zn mine in the Basque Country (northern Spain). Soil in this area proved to be deficient in major macronutrients and to contain toxic levels of Cd, Pb, and Zn. Spontaneously growing native plants (belonging to 31 species, 28 genera, and 15 families) were botanically identified. Plant shoots and rhizosphere soil were sampled at several sites in the mine, and analyzed for Pb, Zn and Cd concentration. Zinc showed the highest concentrations in shoots, followed by Pb and Cd. Highest Zn concentrations in shoots were found in the Zn-Cd hyperaccumulator Thlaspi caerulescens (mean = 18,254 mg Zn kg(-1) DW). Different metal tolerance and accumulation patterns were observed among the studied plant species, thus offering a wide germplasm assortment for the suitable selection of phytoremediation technologies. This study highlights the importance of preserving metalliferous environments as they shelter a unique and highly valuable metallicolous biodiversity.

Dehydration-mediated activation of the xanthophyll cycle in darkness: is it related to desiccation tolerance?

The development of desiccation tolerance by vegetative tissues was an important step in the plants' conquest of land. To counteract the oxidative stress generated under these conditions the xanthophyll cycle plays a key role. Recent reports have shown that desiccation itself induces de-epoxidation of xanthophyll cycle pigments, even in darkness. The aim of the present work was to study whether this trait is a common response of all desiccation-tolerant plants. The xanthophyll cycle activity and the maximal photochemical efficiency of PS II (F(v)/F(m)) as well as ß-carotene and α-tocopherol contents were compared during slow and rapid desiccation and subsequent rehydration in six species pairs (with one desiccation-sensitive and one desiccation-tolerant species each) belonging to different taxa. Xanthophyll cycle pigments were de-epoxidised in darkness concomitantly with a decrease in F(v)/F(m) during slow dehydration in all the desiccation-tolerant species and in most of the desiccation-sensitive ones. De-epoxidation was reverted in darkness by re-watering in parallel with the recovery of the initial F(v)/F(m). The stability of the ß-carotene pool confirmed that its hydroxylation did not contribute to zeaxanthin formation. The α-tocopherol content of most of the species did not change during dehydration. Because it is a common mechanism present in all the desiccation-tolerant taxa and in some desiccation-sensitive species, and considering its role in antioxidant processes and in excess energy dissipation, the induction of the de-epoxidation of xanthophyll cycle pigments upon dehydration in the dark could be understood as a desiccation tolerance-related response maintained from the ancestral clades in the initial steps of land occupation by plants.

Unravelling the roles of desiccation-induced xanthophyll cycle activity in darkness: a case study in Lobaria pulmonaria.

Desiccation-tolerance ability in photosynthetic organisms is largely based on a battery of photoprotective mechanisms. Xanthophyll cycle operation induced by desiccation in the absence of light has been previously proven in the desiccation-tolerant fern Ceterach officinarum. To understand the physiological function of xanthophyll cycle induction in darkness and its implication in the desiccation tolerance in more detail, we studied its triggering factors and its photochemical effects in the lichen Lobaria pulmonaria. We found that both the drying rate and the degree of desiccation play a crucial role in the violaxanthin de-epoxidase activation. De-epoxidation of violaxanthin to zeaxanthin (Z) occurs when the tissue has lost most of its water and only after slow dehydration, suggesting that a minimum period of time is required for the enzyme activity induction. Fluorescence analysis showed that Z, synthesised during tissue dehydration in the absence of light, prevents photoinhibition when rewatered tissues are illuminated. This is probably due to Z implication in both non-photochemical quenching and/or antioxidative responses.

Phytoextraction potential of two Rumex acetosa L. accessions collected from metalliferous and non-metalliferous sites: effect of fertilization.

Metal tolerance and phytoextraction potential of two common sorrel (Rumex acetosa L.) accessions, collected from a Pb/Zn contaminated site (CS, Lanestosa) and an uncontaminated site (UCS, Larrauri), were studied in fertilized and non-fertilized pots prepared by combining soil samples from both sites in different proportions (i.e., 0%, 33%, 66% and 100% of Lanestosa contaminated soil). The original metalliferous mine soil contained 20480, 4950 and 14 mg kg(-1) of Zn, Pb and Cd, respectively. The microcosm experiment was carried out for two months under greenhouse controlled conditions. It was found that fertilization increased mean plant biomass of both accessions as well as their tolerance. However, only the CS accession survived all treatments even though its biomass decreased proportionally according to the percentage of contaminated mine soil present in the pots. This metallicolous accession would be useful for the revegetation and phytostabilization of mine soils. Due to its high concentration and bioavailability in the contaminated soil, the highest values of metal phytoextracted corresponded to Zn. The CS accession was capable of efficiently phytoextracting metal from the 100% mine soil, indeed reaching very promising phytoextraction rates in the fertilized pots (6.8 mg plant(-1) month(-1)), similar to the ones obtained with hyperaccumulator plants. It was concluded that fertilization is certainly worth being considered for phytoextraction and revegetation with native plants from metalliferous soils.

Short- and long-term modulation of the lutein epoxide and violaxanthin cycles in two species of the Lauraceae: sweet bay laurel (Laurus nobilis L.) and avocado (Persea americana Mill.).

Short- and long-term responses of the violaxanthin (V) and lutein epoxide (Lx) cycles were studied in two species of Lauraceae: sweet bay laurel (Laurus nobilis L.) and avocado (Persea americana L.). The Lx content exceeded the V content in shade leaves of both species. Both Lx and V were de-epoxidised on illumination, but only V was fully restored by epoxidation in low light. Violaxanthin was preferentially de-epoxidised in low light in L. nobilis. This suggests that Lx accumulates with leaf ageing, partly because its conversion to lutein is limited in shade. After exposure to strong light, shade leaves of avocado readjusted the total pools of alpha- and beta-xanthophyll cycles by de novo synthesis of antheraxanthin, zeaxanthin and lutein. This occurred in parallel with a sustained depression of F(v)/F(m). In Persea indica, a closely related but low Lx species, F(v)/F(m) recovered faster after a similar light treatment, suggesting the involvement of the Lx cycle in sustained energy dissipation. Furthermore, the seasonal correlation between non-reversible Lx and V photoconversions and pre-dawn F(v)/F(m) in sun leaves of sweet bay supported the conclusion that the Lx cycle is involved in a slowly reversible downregulation of photosynthesis analogous to the V cycle.

Plasticity of photoprotective mechanisms of Buxus sempervirens L. leaves in response to extreme temperatures.

Summer 2003 was unusually hot in Western Europe, with local droughts and an intense heatwave, that led to a massive damage in vegetation. Since high temperatures are supposed to generate photooxidative stress, we analysed photoprotective responses in leaves of the evergreen boxtree (Buxus sempervirens L.) during summer 2003. All the photoprotective compounds analysed (alpha-tocopherol, beta-carotene, and xanthophylls cycle pigments) were simultaneously induced in parallel with a reduction in photochemical efficiency (Fv/Fm). To characterise these responses, we compared these data with other data obtained during cold stress periods (2003, 2005) and with an unstressful summer (2002). Photoprotective responses observed during the heatwave were also induced by low temperature stress, and in both situations, this effect was exacerbated by light. In parallel with such induction the accumulation of red retro-carotenoids and xanthophyll esters was also observed under unfavourable conditions, suggesting a photoprotective role for both groups of carotenoids. This is the first report showing that in any species (Buxus sempervirens L.), the same retro-carotenoids can be induced in response to winter and summer stress. Present results demonstrate that the same mechanisms are induced as response to sub- and supraoptimal temperatures and the plasticity of such responses plays a critical role in plant acclimation to extreme temperatures, an ability that is specially important in the context of any future climate warming.

Synthesis of low molecular weight thiols in response to Cd exposure in Thlaspi caerulescens.

In this study, we investigated the accumulation of phytochelatins (PCs) and other low molecular weight (LMW) thiols in response to Cd exposure in two contrasting ecotypes differing in Cd accumulation. Using a root elongation test, we found that the highly accumulating ecotype Ganges was more tolerant to Cd than the low Cd-accumulation ecotype Prayon. L-buthionine-(S,R)-sulphoximine (BSO), a potent inhibitor of the gamma-glutamylcysteine synthetase gamma-ECS) (an enzyme involved in the PC biosynthetic pathway), increased the Cd sensitivity of Prayon, but had no effect on Ganges. Although PC accumulation increased in response to Cd exposure, no significant differences were observed between the two ecotypes. Cd exposure induced a dose-dependent accumulation of both Cys and a still unidentified LMW thiol in roots of both ecotypes. Root accumulation of Cys and this thiol was higher in Ganges than in Prayon; the ecotypic differences were more pronounced when the plants were treated with BSO. These findings suggest that PCs do not contribute to the Cd hypertolerance displayed by the Ganges ecotype of Thlaspi caerulescens, whereas Cys and other LMW thiols might be involved.

Role of red carotenoids in photoprotection during winter acclimation in Buxus sempervirens leaves.

The red leaf coloration of several plant species during autumn and winter is due to the synthesis of phenolic compounds such as anthocyanins or red carotenoids. The latter occur very rarely and are non-ubiquitous and taxonomically restricted compounds. The present study shows that the leaves of common box ( Buxus sempervirens L.) accumulate red carotenoids (eschscholtzxanthin, monoanhydroeschscholtzxanthin, anhydroeschscholtzxanthin) as a response to photoinhibitory conditions during winter acclimation. These compounds are produced in a coordinated manner with the operation of other photoprotective systems: accumulation and sustained deepoxidation of VAZ pigments with a concomitant decrease in maximal photochemical efficiency, accumulation of alpha-tocopherol and a gradual decrease on chlorophyll content. All these processes were reversed when the photosynthetic tissues were transferred from photoinhibitory winter conditions to room temperature for 9 days. Buxus leaves showed a large degree of phenotype variation in the degree of reddening, ranging from green to orange. The differences in colour pattern were mainly due to differences in the accumulation of red carotenoids and xanthophyll esters. Red pigments were mainly anhydroeschscholtzxanthin and esters of eschscholtzxanthin. Conversely to fruit or petal chromoplasts, the plastids of red leaves in this species are not the terminal differentiated state but are able to redifferentiate again to chloroplasts. Their photoprotective role during winter as a light screen system or as antioxidants, in a similar way to other red pigments, and their implications on the wide ecological tolerance of this evergreen species are discussed.

Estado clínico y psicosocial de reclusos en tratamiento con antirretrovirales / Clinical and psychosocial state of inmates in treatment with antiretrovirals

Introducción: La mayoría de los estudios realizados en prisiones se han centrado en el estudio de la infección por VIH, la adherencia o la TBC. Sin embargo, llama la atención la escasez de estudios sobre las características psicosociales de esta población, como son por ejemplo la morbilidad psíquica, el apoyo social o el consumo de drogas. Objetivos: Describir el estado clínico y psicosocial de los reclusos en tratamiento con antirretrovirales en tres prisiones andaluzas (Córdoba, Granada y Huelva). Material y métodos: Mediante una encuesta transversal, con un cuestionario administrado por entrevistador, se entrevistaron a todos los reclusos en tratamiento con antirretrovirales de los tres centros. Las variables estudiadas fueron: datos sociodemográficos, variables del medio penitenciario, variables sobre la adherencia al tratamiento, variables de salud y variables de apoyo. Resultados: La frecuencia de morbilidad psíquica fue del 42% y el 64,4% refirió tener algún tipo de enfermedad crónica; de éstos, el 12,7% afirmó padecer tuberculosis. El 46,6% afirmaron contar con apoyo social dentro de la prisión, recibiéndolo de otros internos el 45% y de los profesionales de la prisión el 36,6%. El 54,8% fueron considerados no adherentes al tratamiento con antirretrovirales. El 89% consideraba que la comida era mala, y sólo al 29,9% se le facilitaban suplementos de comida. Por último, el 33,8% refería no tener flexibilidad para que le abrieran la celda en caso de olvido del tratamiento. El 31% refería peor salud autopercibida (AU)

Introduction: Most studies conducted in prisons have focused on the study of HIV infection, adherence to treatment or TBC. However, there is a surprisingly small number of studies on the psychosocial characteristics of this population, such as psychic morbidity, social support or drug use. Objective: To describe the clinical and psychosocial status of inmates in treatment with antiretrovirals in three Andalusian prisons (Córdoba, Granada and Huelva). Materials and method: All inmates in treatment with antiretrovirals in the three correctional facilities were interviewed using a heteroadministered questionnaire, with a cross-sectional design. The sociodemographic data, penitentiary, adherence to treatment, health and social support variables were studied. Results: The frequency of psychic morbidity was 42%, and 64,4% of the interviewed sample mentioned having some kind of chronic disease; of these, 12,7% admitted to suffering from tuberculosis. 46,6% stated that they had social support within the prison, receiving it from other inmates in 45% of the cases and 36,6% from professionals of the prison. 54,8% were considered non-adherent to antiretroviral treatment. 89% of the sample considered that the food was of bad quality, and only 29,9% were provided with food supplements. Finally, 33,8% stated that they were not able to have their cells opened if their medication was forgotten. 31% mentioned worse self-perceived health when compared with the previous year (AU)

Glyphosate effects on phenolic metabolism of nodulated soybean (Glycine max L. merr.).

Glyphosate is a herbicide that blocks the shikimic acid pathway. Three Bradyrhizobium japonicum strains with different sensitivities to glyphosate were used to test the effect of this herbicide on the phenolic metabolism of nodulated soybeans and on the bacteroid nitrogenase activity. Glyphosate caused an inhibition in the bacteroid nitrogenase activity that was related with the sensitivity of the nodule-forming strains. Both leaves and nodules accumulated huge amounts of shikimate and phenolic acids (mainly protocatechuic acid), indicating that the herbicide was translocated to the nodule and disturbed phenolic metabolism. However, this accumulation was not clearly related to the sensitivity of the different strains. Bacteroids from control plants were incubated with the same concentration of shikimate, and phenolic acid accumulated in glyphosate-treated plants. Despite the high levels found in nodules, they were not responsible for the decrease of the nitrogenase activity. Glyphosate by itself caused a small inhibition of the bacteroid nitrogenase activity.

Diurnal changes in photoprotective mechanisms in leaves of cork oak (Quercus suber) during summer.

Daily variations in photoprotective mechanisms were studied in sun and shade leaves of 40-year-old cork oak (Quercus suber L.) trees during early summer in Portugal. Although trees were not severely water stressed because predawn leaf water potentials remained high, photosynthesis and stomatal conductance decreased at midday. The midday depression in gas exchange was not reversed by short-term exposure to "optimal" conditions of temperature, light and vapor pressure deficit. Chlorophyll a fluorescence, maximum photochemical yield of photosystem II and the quantum yield of noncyclic electron transport showed midday depressions, but recovered by the evening. Both short-term changes in the components of the xanthophyll cycle (reversible de-epoxidation of violaxanthin during the day) as well as long-term changes (higher xanthophyll content in sun compared with shade leaves) were detected and may play a role in the dissipation of excess energy at midday. Because the activities of enzymes of the antioxidant system, superoxide dismutase and ascorbate peroxidase, were high enough to cope with the increase in oxygen reactive species likely to arise under the stressful conditions of midday, we conclude that these enzymes may provide an additional mechanism for energy dissipation.