Exopolysaccharides produced by plant pathogenic bacteria affect ascorbate metabolism in Nicotiana tabacum

The role of the exopolysaccharides (EPSs) produced by plant pathogenic bacteria has not completely clarified, they are considered either molecules able to avoid or delay the activation of plant defences, or acting as signal in the plant-pathogen cross-talk. In order to understand whether EPSs are recognized by infected plant cells and are able to induce the activation of plant defence responses, their capability to induce metabolic alteration in tobacco cells has been analysed. The results indicate that several EPSs, even if not chemically related, induce increases in phenylalanine ammonia-lyase, a marker enzyme of defence responses of plants against stress; but others are completely ineffective. The EPSs affecting phenylalanine ammonia-lyase also induce an increase in hydrogen peroxide production. Moreover, they alter the metabolism of ascorbate, another parameter indicative of the presence of stress conditions and the involvement of which in the hypersensitive reaction has been recently reported. The possibility that specific EPSs could act as signals in the plant-pathogenic bacteria interaction is discussed.     

Induction of chromosome aberrations in vitro by phenolphthalein: mechanistic studies

Phenolphthalein induces tumors in rodents but because it is negative in assays for mutation in Salmonella and in mammalian cells, for DNA adducts and for DNA strand breaks, its primary mechanism does not seem to be DNA damage. Chromosome aberration (Ab) induction by phenolphthalein in vitro is associated with marked cytotoxicity. At very high doses, phenolphthalein induces weak increases in micronuclei (MN) in mouse bone marrow; a larger response is seen with chronic treatment. All this suggests genotoxicity is a secondary effect that may not occur at lower doses. In heterozygous TSG-p53((R)) mice, phenolphthalein induces lymphomas and also MN, many with kinetochores (K), implying chromosome loss. Induction of aneuploidy would be compatible with the loss of the normal p53 gene seen in the lymphomas.Here we address some of the postulated mechanisms of genotoxicity in vitro, including metabolic activation, inhibition of thymidylate synthetase, cytotoxicity, oxidative stress, DNA damage and aneuploidy. We show clearly that phenolphthalein does not require metabolic activation by S9 to induce Abs. Inhibition of thymidylate synthetase is an unlikely mechanism, since thymidine did not prevent Ab induction by phenolphthalein. Phenolphthalein dramatically inhibited DNA synthesis, in common with many non-DNA reactive chemicals that induce Abs at cytotoxic doses. Phenolphthalein strongly enhances levels of intracellular oxygen radicals (ROS). The radical scavenger DMSO suppresses phenolphthalein-induced toxicity and Abs whereas H(2)O(2) potentiates them, suggesting a role for peroxidative activation. Phenolphthalein did not produce DNA strand breaks in rat hepatocytes or DNA adducts in Chinese hamster ovary (CHO) cells. All the evidence points to an indirect mechanism for Abs that is unlikely to operate at low doses of phenolphthalein. We also found that phenolphthalein induces mitotic abnormalities and MN with kinetochores in vitro. These are also enhanced by H(2)O(2) and suppressed by DMSO. Our findings suggest that induction of Abs in vitro is a high-dose effect in oxidatively stressed cells and may thus have a threshold. There may be more than one mechanism operating in vitro and in vivo, possibly indirect genotoxicity at high doses and also chromosome loss, both of which would likely have a threshold.     

Redox regulation in plant programmed cell death

Programmed cell death (PCD) is a genetically controlled process described both in eukaryotic and prokaryotic organisms. Even if it is clear that PCD occurs in plants, in response to various developmental and environmental stimuli, the signalling pathways involved in the triggering of this cell suicide remain to be characterized. In this review, the main similarities and differences in the players involved in plant and animal PCD are outlined. Particular attention is paid to the role of reactive oxygen species (ROS) as key inducers of PCD in plants. The involvement of different kinds of ROS, different sites of ROS production, as well as their interaction with other molecules, is crucial in activating PCD in response to specific stimuli. Moreover, the importance is stressed on the balance between ROS production and scavenging, in various cell compartments, for the activation of specific steps in the signalling pathways triggering this cell suicide process. The review focuses on the complexity of the interplay between ROS and antioxidant molecules and enzymes in determining the most suitable redox environment required for the occurrence of different forms of PCD.     

Global Phosphorus Flows and Environmental Impacts from a Consumption Perspective

Human activities have significantly intensified natural phosphorus cycles, which has resulted in some serious environmental problems that modern societies face today. This article attempts to quantify the global phosphorus flows associated with present day mining, farming, animal feeding, and household consumption. Various physical characteristics of the related phosphorus fluxes as well as their environmental impacts in different economies, including the United States, European countries, and China, are examined. Particular attention is given to the global phosphorus budget in cropland and the movement and transformation of phosphorus in soil, because these phosphorus flows, in association with the farming sector, constitute major fluxes that dominate the anthropogenic phosphorus cycle. The results show that the global input of phosphorus to cropland, in both inorganic and organic forms from various sources, cannot compensate for the removal in harvests and in the losses by erosion and runoff. A net loss of phosphorus from the world's cropland is estimated at about 10.5 million metric tons (MMT) phosphorus each year, nearly one half of the phosphorus extracted yearly.     

Distribution of cytosolic ascorbate peroxidase in Angiosperms

Abstract

Over 80 Angiosperms have been screened for cytosolic ascorbate peroxidase by means of native-polyacrylamide gel electrophoresis techniques. The results of our analysis show that the presence of a single cytosolic ascorbate peroxidase form is the most common case in the Angiosperms investigated when seedlings or young tissues are analized. This is a conserved character in orders of the Dicots. Two electrophoretic distinct AApx forms have been identified in Magnoliales, the ancestor group from which both Monocots and Dicots were originated and in few other orders. A notable increase in the isoenzyme number is observed in the advanced Monocots (Poales), thus suggesting the existence of an evolutionary trend leading in Monocots to an apparently progressive rise in the ascorbate-dependent enzymatic scavenging of hydrogen peroxide.     

Reduced expression of top1beta gene induces programmed cell death and alters ascorbate metabolism in Daucus carota cultured cells

Topoisomerase I (topo I) is a nuclear enzyme which plays a fundamental role in several pathways involving changes in DNA topology. The topo I-mediated reaction is accomplished by the transient covalent binding of the enzyme to DNA (topo I-DNA complex). Stabilization of the topo I-DNA complex, leading to irreversible double-strand breaks, has been reported to occur in animal cells under oxidative stress conditions and during apoptosis. In order to study the existence of a putative link between the topo I-mediated DNA damage and ascorbate (ASC) metabolism, also involved in the responses against oxidative stress and in the apoptotic process in plants, Daucus carota cells showing reduced expression of the top1beta gene encoding the topo Ibeta isoform were produced, using an antisense RNA strategy. Two independent transgenic lines (AT1-beta/22 and beta/36), characterized by a slow growth phenotype, resistance to camptothecin, a specific inhibitor of topo I, but sensitivity to etoposide, an inhibitor of topo II, were investigated in this study. In the absence of external stimuli, AT1-beta/22 and beta/36 cells underwent programmed cell death (PCD) in a precocious phase of the growth curve. ASC metabolism showed remarkable differences in AT1-beta/22 and beta/36 cells, compared with control, and the observed alterations were similar to those occurring in tobacco Yellow Bright-2 cells induced to enter PCD by exogenous stimuli. However, differently from other studied examples of PCD, overproduction of reactive oxygen species was not detected in AT1-beta/22 and beta/36 cells. The relevance of these findings in relation to the signalling pathways leading to PCD is discussed.     

Production of reactive species and modulation of antioxidant network in response to heat shock: a critical balance for cell fate

Exposure to adverse temperature conditions is a common stress factor for plants. In order to cope with heat stress, plants activate several defence mechanisms responsible for the control of reactive oxygen species (ROS) and redox homeostasis. Specific heat shocks (HSs) are also able to activate programmed cell death (PCD). In this paper, the alteration of several oxidative markers and ROS scavenging enzymes were studied after subjecting cells to two different HSs. Our results suggest that, under moderate HS, the redox homeostasis is mainly guaranteed by an increase in glutathione (GSH) content and in the ascorbate peroxidase (APX) and catalase (CAT) activities. These two enzymes undergo different regulatory mechanisms. On the other hand, the HS-induced PCD determines an increase in the activity of the enzymes recycling the ascorbate- and GSH-oxidized forms and a reduction of APX; whereas, CAT decreases only after a transient rise of its activity, which occurs in spite of the decrease of its gene expression. These results suggest that the enzyme-dependent ROS scavenging is enhanced under moderate HS and suppressed under HS-induced PCD. Moreover, the APX suppression occurring very early during PCD, could represent a hallmark of cells that have activated a suicide programme.     

Increase in ascorbate-glutathione metabolism as local and precocious systemic responses induced by cadmium in durum wheat plants

Durum wheat plants (Triticum durum cv Creso) were grown in thepresence of cadmium (0–40 µM) and analysed after3 and 7 d for their growth, oxidative stress markers, phytochelatins,and enzymes and metabolites of the ascorbate (ASC)–glutathione(GSH) cycle. Cd exposure produced a dose-dependent inhibitionof growth in both roots and leaves. Lipid peroxidation, proteinoxidation and the decrease in the ascorbate redox state indicatethe presence of oxidative stress in the roots, where H₂O₂ overproductionand phytochelatin synthesis also occurred. The activity of theASC–GSH cycle enzymes significantly increased in roots.Consistently, a dose-dependent accumulation of Cd was evidentin these organs. On the other hand, no oxidative stress symptomsor phytochelatin synthesis occurred in the leaves; where, atleast during the time of our analysis, the levels of Cd remainedirrelevant. In spite of this, enzymes of the ASC–GSH cyclesignificantly increased their activity in the leaves. When ASCbiosynthesis was enhanced, by feeding plants with its last precursor,L-galactono--lactone (GL), Cd uptake was not affected. On theother hand, the oxidative stress induced in the roots by theheavy metal was alleviated. GL treatment also inhibited theCd-dependent phytochelatin biosynthesis. These results suggestthat different strategies can successfully cope with heavy metaltoxicity. The changes that occurred in the ASC–GSH cycleenzymes of the leaves also suggest that the whole plant improvedits antioxidant defense, even in those parts which had not yetbeen reached by Cd. This precocious increase in the enzymesof the ASC–GSH cycle further highlight the tight regulationand the relevance of this cycle in the defense against heavymetals.