Lichen rehydration in heavy metal polluted environments: Pb modulates the oxidative response of both Ramalina farinacea thalli and its isolated microalgae

Lichens are adapted to desiccation/rehydration and accumulate heavy metals, which induce ROS especially from the photobiont photosynthetic pigments. Although their mechanisms of abiotic stress tolerance are still to be unravelled, they seem related to symbionts' reciprocal upregulation of antioxidant systems. With the aim to study the effect of Pb on oxidative status during rehydration, the kinetics of intracellular ROS, lipid peroxidation and chlorophyll autofluorescence of whole Ramalina farinacea thalli and its isolated microalgae (Trebouxia TR1 and T. TR9) was recorded. A genetic characterization of the microalgae present in the thalli used was also carried out in order to assess possible correlations among the relative abundance of each phycobiont, their individual physiological responses and that of the entire thallus. Unexpectedly, Pb decreased ROS and lipid peroxidation in thalli and its phycobionts, associated with a lower chlorophyll autofluorescence. Each phycobiont showed a particular pattern, but the oxidative response of the thallus paralleled the TR1's, agreeing with the genetic identification of this strain as the predominant phycobiont. We conclude that: (1) the lichen oxidative behaviour seems to be modulated by the predominant phycobiont and (2) Pb evokes in R. farinacea and its phycobionts strong mechanisms to neutralize its own oxidant effects along with those of rehydration

Melatonin as an antioxidant and its semi-lunar rhythm in green macroalga Ulva sp.

The presence and role of melatonin in plants are still under debate owing to difficulties of identification and quantification. Accordingly, although it has been frequently proposed that melatonin acts as an antioxidant in phototrophic organisms, experimental data on its physiological role are scarce. This study describes the use of a rapid and simple new method for quantification of melatonin in the marine macroalga Ulva sp., organisms routinely exposed to tide-related environmental stresses and known for their high tolerance to abiotic conditions. The method was used here to show that exposure to oxidative stress-inducing environmental conditions (elevated temperature and heavy metals) induced a rise in melatonin level in the algae. Addition of exogenous melatonin alleviated the algae from cadmium-induced stress. Interestingly, although the algae were taken from a culture growing free floating and kept under constant photoperiod and water level, they exhibited a semi-lunar rhythm of melatonin levels that correlated with predicted spring tides. The correlation can probably be interpreted as reflecting preparation for predicted low tides, when the algae are exposed to increasing temperature, desiccation, and salinity, all known to induce oxidative stress. Given the simplicity of the described method it can easily be adapted for the study of melatonin in many other phototrophic organisms. These results provide, for the first time, experimental data that support both an antioxidant role for melatonin and its semi-lunar rhythm in macroalgae

Multivariate modeling of chromium-induced oxidative stress and biochemical changes in plants of Pistia stratiotes L.

Biochemical changes in the plants of Pistia stratiotes L., a free floating macrophyte exposed to different concentrations of hexavalent chromium (0, 10, 40, 60, 80 and 160 μM) for 48, 96 and 144 h were studied. Chromium-induced oxidative stress in macrophyte was investigated using the multivariate modeling approaches. Cluster analysis rendered two fairly distinct clusters (roots and shoots) of similar characteristics in terms of their biochemical responses. Discriminant analysis identified ascorbate peroxidase (APX) as discriminating variable between the root and shoot tissues. Principal components analysis results suggested that malondialdehyde (MDA), superoxide dismutase (SOD), APX, non-protein thiols (NP-SH), cysteine, ascorbic acid, and Cr-accumulation are dominant in root tissues, whereas, protein and guaiacol peroxidase (GPX) in shoots of the plant. Discriminant partial least squares analysis results further confirmed that MDA, SOD, NP-SH, cysteine, GPX, APX, ascorbic acid and Cr-accumulation dominated in the root tissues, while protein in the shoot. Three-way analysis helped in visualizing simultaneous influence of metal concentration and exposure duration on biochemical variables in plant tissues. The multivariate approaches, thus, allowed for the interpretation of the induced biochemical changes in the plant tissues exposed to chromium, which otherwise using the conventional approaches is difficult

Transcriptome Analysis of H2O2-Treated Wheat Seedlings Reveals a H2O2-Responsive Fatty Acid Desaturase Gene Participating in Powdery Mildew Resistance

Hydrogen peroxide (H2O2) plays important roles in plant biotic and abiotic stress responses. However, the effect of H2O2 stress on the bread wheat transcriptome is still lacking. To investigate the cellular and metabolic responses triggered by H2O2, we performed an mRNA tag analysis of wheat seedlings under 10 mM H2O2 treatment for 6 hour in one powdery mildew (PM) resistant (PmA) and two susceptible (Cha and Han) lines. In total, 6,156, 6,875 and 3,276 transcripts were found to be differentially expressed in PmA, Han and Cha respectively. Among them, 260 genes exhibited consistent expression patterns in all three wheat lines and may represent a subset of basal H2O2 responsive genes that were associated with cell defense, signal transduction, photosynthesis, carbohydrate metabolism, lipid metabolism, redox homeostasis, and transport. Among genes specific to PmA, ‘transport’ activity was significantly enriched in Gene Ontology analysis. MapMan classification showed that, while both up- and down- regulations were observed for auxin, abscisic acid, and brassinolides signaling genes, the jasmonic acid and ethylene signaling pathway genes were all up-regulated, suggesting H2O2-enhanced JA/Et functions in PmA. To further study whether any of these genes were involved in wheat PM response, 19 H2O2-responsive putative defense related genes were assayed in wheat seedlings infected with Blumeria graminis f. sp. tritici (Bgt). Eight of these genes were found to be co-regulated by H2O2 and Bgt, among which a fatty acid desaturase gene TaFAD was then confirmed by virus induced gene silencing (VIGS) to be required for the PM resistance. Together, our data presents the first global picture of the wheat transcriptome under H2O2 stress and uncovers potential links between H2O2 and Bgt responses, hence providing important candidate genes for the PM resistance in wheat

Plant responses to abiotic stresses: heavy metal-induced oxidative stress and protection by mycorrhization

The aim of this review is to assess the mode of action and role of antioxidants as protection from heavy metal stress in roots, mycorrhizal fungi and mycorrhizae. Based on their chemical and physical properties three different molecular mechanisms of heavy metal toxicity can be distinguished: (a) production of reactive oxygen species by autoxidation and Fenton reaction; this reaction is typical for transition metals such as iron or copper, (b) blocking of essential functional groups in biomolecules, this reaction has mainly been reported for non-redox-reactive heavy metals such as cadmium and mercury, (c) displacement of essential metal ions from biomolecules; the latter reaction occurs with different kinds of heavy metals. Transition metals cause oxidative injury in plant tissue, but a literature survey did not provide evidence that this stress could be alleviated by increased levels of antioxidative systems. The reason may be that transition metals initiate hydroxyl radical production, which can not be controlled by antioxidants. Exposure of plants to non-redox reactive metals also resulted in oxidative stress as indicated by lipid peroxidation, H2O2 accumulation, and an oxidative burst. Cadmium and some other metals caused a transient depletion of GSH and an inhibition of antioxidative enzymes, especially of glutathione reductase. Assessment of antioxidative capacities by metabolic modelling suggested that the reported diminution of antioxidants was sufficient to cause H2O2 accumulation. The depletion of GSH is apparently a critical step in cadmium sensitivity since plants with improved capacities for GSH synthesis displayed higher Cd tolerance. Available data suggest that cadmium, when not detoxified rapidly enough, may trigger, via the disturbance of the redox control of the cell, a sequence of reactions leading to growth inhibition, stimulation of secondary metabolism, lignification, and finally cell death. This view is in contrast to the idea that cadmium results in unspecific necrosis. Plants in certain mycorrhizal associations are less sensitive to cadmium stress than non-mycorrhizal plants. Data about antioxidative systems in mycorrhizal fungi in pure culture and in symbiosis are scarce. The present results indicate that mycorrhization stimulated the phenolic defence system in the Paxillus-Pinus mycorrhizal symbiosis. Cadmium-induced changes in mycorrhizal roots were absent or smaller than those in non-mycorrhizal roots. These observations suggest that although changes in rhizospheric conditions were perceived by the root part of the symbiosis, the typical Cd-induced stress responses of phenolics were buffered. It is not known whether mycorrhization protected roots from Cd-induced injury by preventing access of cadmium to sensitive extra- or intracellular sites, or by excreted or intrinsic metal-chelators, or by other defence systems. It is possible that mycorrhizal fungi provide protection via GSH since higher concentrations of this thiol were found in pure cultures of the fungi than in bare roots. The development of stress-tolerant plant-mycorrhizal associations may be a promising new strategy for phytoremediation and soil amelioration measures

Physiological responses of a halophytic shub to salt stress by Na2SO4 and NaCl : oxidative damage and the role of polyphenols in antioxidant protection

Salt stress conditions lead to increased production of reactive oxygen species (ROS) in plant cells. Halophytes have the ability to reduce these toxic ROS by means of a powerful antioxidant system that includes enzymatic and non-enzymatic components. In this research, we used the halophytic shrub Prosopis strombulifera to investigate whether the ability of this species to grow under increasing salt concentrations and mixtures was related to the synthesis of polyphenolic compounds and to the maintenance of leaf pigment contents for an adequate photosynthetic activity. Seedlings of P. strombulifera were grown hydroponically in Hoagland’s solution, gradually adding Na2SO4 and NaCl separately or in mixtures until reaching final osmotic potentials of 21, 21.9 and 22.6 MPa. Control plants were allowed to develop in Hoagland’s solution without salt. Oxidative damage in tissues was determined by H2O2 and malondialdehyde content. Leaf pigment analysis was performed by high-performance liquid chromatography with ultraviolet, and total phenols, total flavonoids, total flavan-3-ols, condensed tannins, tartaric acid esters and flavonolswere spectrophotometrically assayed. Treatment with Na2SO4 increased H2O2 production and lipid peroxidation in tissues and induced a sharp increase in flavonoid compounds (mainly flavan-3-ols) and consequently in the antioxidant activity. Also, Na2SO4 treatment induced an increased carotenoid/chlorophyll ratio, which may represent a strategy to protect photosystems against photooxidation. NaCl treatment, however, did not affect H2O2 content, lipid peroxidation, pigments or polyphenols synthesis. The significant accumulation of flavonoids in tissues under Na2SO4 treatment and their powerful antioxidant activity indicates a role for these compounds in counteracting the oxidative damage induced by severe salt stress, particularly, ionic stress. We demonstrate that ionic interactions between different salts in salinized soils modify the biochemical and morpho-physiological responses of P. strombulifera plants to salinity