Antonio Scopa

Ascorbate Peroxidase and Catalase Activities and Their Genetic Regulation in Plants Subjected to Drought and Salinity Stresses

Hydrogen peroxide (H2O2), an important relatively stable non-radical reactive oxygen species (ROS) is produced by normal aerobic metabolism in plants. At low concentrations, H2O2 acts as a signal molecule involved in the regulation of specific biological/physiological processes (photosynthetic functions, cell cycle, growth and development, plant responses to biotic and abiotic stresses). Oxidative stress and eventual cell death in plants can be caused by excess H2O2 accumulation. Since stress factors provoke enhanced production of H2O2 in plants, severe damage to biomolecules can be possible due to elevated and non-metabolized cellular H2O2. Plants are endowed with H2O2-metabolizing enzymes such as catalases (CAT), ascorbate peroxidases (APX), some peroxiredoxins, glutathione/thioredoxin peroxidases, and glutathione sulfo-transferases. However, the most notably distinguished enzymes are CAT and APX since the former mainly occurs in peroxisomes and does not require a reductant for catalyzing a dismutation reaction. In particular, APX has a higher affinity for H2O2 and reduces it to H2O in chloroplasts, cytosol, mitochondria and peroxisomes, as well as in the apoplastic space, utilizing ascorbate as specific electron donor. Based on recent reports, this review highlights the role of H2O2 in plants experiencing water deficit and salinity and synthesizes major outcomes of studies on CAT and APX activity and genetic regulation in drought- and salt-stressed plants.

Lipids and proteins—major targets of oxidative modifications in abiotic stressed plants

Stress factors provoke enhanced production of reactive oxygen species (ROS) in plants. ROS that escape antioxidant-mediated scavenging/detoxification react with biomolecules such as cellular lipids and proteins and cause irreversible damage to the structure of these molecules, initiate their oxidation, and subsequently inactivate key cellular functions. The lipid- and protein-oxidation products are considered as the significant oxidative stress biomarkers in stressed plants. Also, there exists an abundance of information on the abiotic stress-mediated elevations in the generation of ROS, and the modulation of lipid and protein oxidation in abiotic stressed plants. However, the available literature reflects a wide information gap on the mechanisms underlying lipid- and protein-oxidation processes, major techniques for the determination of lipid- and protein-oxidation products, and on critical cross-talks among these aspects. Based on recent reports, this article (a) introduces ROS and highlights their relationship with abiotic stress-caused consequences in crop plants, (b) examines critically the various physiological/biochemical aspects of oxidative damage to lipids (membrane lipids) and proteins in stressed crop plants, (c) summarizes the principles of current technologies used to evaluate the extent of lipid and protein oxidation, (d) synthesizes major outcomes of studies on lipid and protein oxidation in plants under abiotic stress, and finally, (e) considers a brief cross-talk on the ROS-accrued lipid and protein oxidation, pointing to the aspects unexplored so far.

The Importance of Pathogenic Organisms in Sewage and Sewage Sludge

ABSTRACT Deficient sanitation poses a serious threat to human and animal health, involving complex relationships between environments, animals, refuse, food, pathogens, parasites, and man. However, by sanitizing and stabilizing the organic matter of sewage sludge, agriculture can utilize it to maintain soil, water, and air quality. As ingredients in soil amendments, such bioresidues are a source of nutrients for plants. Stabilization and sanitation of sewage sludge safely couple its recycling and disposal. This coupling becomes increasingly important as economic and environmental constraints make strategies for waste disposal more difficult to apply. The occurrence of viruses, bacteria, yeasts, fungi, and zooparasites in sewage sludge is reviewed in this article, and consequential epidemiologic concerns that arise from sewage sludge recycling is also addressed.

Nutraceutical properties and polyphenolic profile of berry skin and wine of Vitis vinifera L. (cv. Aglianico)

Red grapes are rich in phenolics, flavonoids, anthocyanins and resveratrol, all substances which have been suggested as having nutraceutical and health benefits. The berry skin and wine of grape cultivar Vitis vinifera L. (cv. Aglianico), grown in Basilicata (Southern Italy) were examined to determinate the presence of the above mentioned compounds as well as to establish the inorganic cation profile. HPLC analysis coupled with LC-ESI/MS/MS detected high contents of total flavonols and anthocyanins in berry skin and wine. The wine made with the same grape used for berry skin assays showed a notable presence of quercetin-3-O-glucoside (39.4% of total flavonols), and malvidin and petunidin derivatives (63.9% and 10.8% of total anthocyanins, respectively). The strong antioxidant ROS-scavenging activity, determined by both DPPH and FRAP assays, and the high resveratrol content confer high sensory characteristics resulted to be associated with positive nutraceutical properties of these grapes and wine. The level of cis-resveratrol was lower than trans-resveratrol in both berry skin and wine reaching 44.1mg/kg and 0.3mg/l, respectively. The cation profile presents low levels of Ca, Cu, K, Fe, Zn and Cd compared to numerous, important red wines, such as Monastrell and Tempranillo.

Toxic effects of four sulphonylureas herbicides on soil microbial biomass

The effect of four triazinyl-sulfonylurea herbicides (cinosulfuron, prosulfuron, thifensulfuron methyl, triasulfuron) on soil microbial biomass, soil respiration, metabolic activity, metabolic quotient, and some enzymatic activities (acid and alkaline phosphatase, β-glucosidase, arylsulphatase, and fluorescein diacetate hydrolysis) were monitored under controlled conditions over 30 days. The herbicides were applied at the normal field dose (FD) and at ten-fold (10 FD) the field dose, in order to mimic a long term toxic effect. The measured soil microbial parameters showed that the FD had slight effects on soil microflora, while at 10 FD the tested herbicides exerted a stronger detrimental effect on soil microbial biomass and its biochemical activities.

Correlation between hormonal homeostasis and morphogenic responses in Arabidopsis thaliana seedlings growing in a Cd/Cu/Zn multi-pollution context

To date, almost no information is available in roots and shoots of the model plant Arabidopsis thaliana about the hierarchic relationship between metal accumulation, phytohormone levels, and glutathione/phytochelatin content, and how this relation affects root development. For this purpose, specific concentrations of cadmium, copper and zinc, alone or in triple combination, were supplied for 12 days to in vitro growing seedlings. The accumulation of these metals was measured in roots and shoots, and a significant competition in metal uptake was observed. Microscopic analyses revealed that root morphology was affected by metal exposure, and that the levels of trans-zeatin riboside, dihydrozeatin riboside, indole-3-acetic acid and the auxin/cytokinin ratio varied accordingly. By contrast, under metal treatments, minor modifications in gibberellic acid and abscisic acid levels occurred. Real-time polymerase chain reaction analysis of some genes involved in auxin and cytokinin synthesis (e.g. AtNIT in roots and AtIPT in shoots) showed on average a metal up-regulated transcription. The production of thiol-peptides was induced by all the metals, alone or in combination, and the expression of the genes involved in thiol-peptide synthesis (AtGSH1, AtGSH2, AtPCS1 and AtPCS2) was not stimulated by the metals, suggesting a full post-transcriptional control. Results show that the Cd/Cu/Zn-induced changes in root morphology are caused by a hormonal unbalance, mainly governed by the auxin/cytokinin ratio.

Auxin and Cytokinin Metabolism and Root Morphological Modifications in Arabidopsis thaliana Seedlings Infected with Cucumber mosaic virus (CMV) or Exposed to Cadmium

Arabidopsis thaliana L. is a model plant but little information is available about morphological root changes as part of a phytohormonal common response against both biotic and abiotic stressors. For this purpose, two-week-old Arabidopsis seedlings were treated with 10 μM CdSO4 or infected with CMV. After 12 days the entire aerial parts and the root system were analyzed, and the presence of CMV or the accumulation of Cd were detected. Microscopic analysis revealed that both CMV and Cd influenced root morphology by a marked development in the length of root hairs and an intense root branching if compared to controls. Among the three treatments, Cd-treated seedlings showed a shorter root axis length and doubled their lateral root diameter, while the lateral roots of CMV-infected seedlings were the longest. The root growth patterns were accompanied by significant changes in the levels of indole-3-acetic acid, trans-zeatin riboside, dihydrozeatin riboside, as a probable consequence of the regulation of some genes involved in their biosynthesis/degradation. The opposite role on root development played by the phythormones studied is discussed in detail. The results obtained could provide insights into novel strategies for plant defense against pathogens and plant protection against pollutants.

Abscisic acid root and leaf concentration in relation to biomass partitioning in salinized tomato plants.

Salinization is one of the most important causes of crop productivity reduction in many areas of the world. Mechanisms that control leaf growth and shoot development under the osmotic phase of salinity are still obscure, and opinions differ regarding the Abscisic acid (ABA) role in regulation of biomass allocation under salt stress. ABA concentration in roots and leaves was analyzed in a genotype of processing tomato under two increasing levels of salinity stress for five weeks: 100 mM NaCl (S10) and 150 mM NaCl (S15), to study the effect of ABA changes on leaf gas exchange and dry matter partitioning of this crop under salinity conditions. In S15, salinization decreased dry matter by 78% and induced significant increases of Na(+) and Cl(-) in both leaves and roots. Dry matter allocated in different parts of plant was significantly different in salt-stressed treatments, as salinization increased root/shoot ratio 2-fold in S15 and 3-fold in S15 compared to the control. Total leaf water potential (Ψ(w)) decreased from an average value of approximately -1.0 MPa, measured on control plants and S10, to -1.17 MPa in S15. In S15, photosynthesis was reduced by 23% and stomatal conductance decreased by 61%. Moreover, salinity induced ABA accumulation both in tomato leaves and roots of the more stressed treatment (S15), where ABA level was higher in roots than in leaves (550 and 312 ng g(-1) fresh weight, respectively). Our results suggest that the dynamics of ABA and ion accumulation in tomato leaves significantly affected both growth and gas exchange-related parameters in tomato. In particular, ABA appeared to be involved in the tomato salinity response and could play an important role in dry matter partitioning between roots and shoots of tomato plants subjected to salt stress.

Berry morphology and composition in irrigated and non-irrigated grapevine (Vitis vinifera L.).

The present study was carried out in a 5-year-old vineyard (Vitis vinifera L., cv. Aglianico) located in Southern Italy. Half of the plants (IRR) were fully irrigated, whereas the other half were not irrigated (NIRR). In both of the treatments, plant water status, gas exchange, photosynthetic efficiency and productive performance were determined. The arid conditions resulted in significant decreases in stem water potential in NIRR (minimum values of -1.34 and -1.52 MPa in IRR and NIRR, respectively). The values of yield per plant, cluster weight and total berry weight were significantly higher in IRR. Grape berries were separated into four weight classes, and morphometric and microscopic analyses were carried out to measure and calculate berry skin characteristics. Irrigation determined a marked shift toward heavier (+23% in the class ≥ 1.25 g) and bigger (336.35 mm³ vs 299.15 mm³) berries, and induced significant changes in other morphometric berry parameters. No differences among berry weight classes and irrigation treatments were observed for berry skin thickness. In all of the berry weight classes, total anthocyanins extracted from berry skins were significantly higher in NIRR than in IRR (12301.53 and 9585.52 mg kg⁻¹ fresh berry skin, respectively), and appeared to be positively related to berry weight, whereas total flavonols were not significantly different between the two treatments. Qualitative changes in the levels of single anthocyanin and flavonol compounds were detected between IRR and NIRR. In addition, iron, copper and zinc, whose high concentration can negatively affect wine quality, were significantly higher in the IRR treatment. The results highlighted that the absence of irrigation did not determine decreases in grape quality. Such data can be of primary importance in environments where water availability is by far the most important limiting factor for plant growth.

Regulation of the Ascorbate–Glutathione Cycle in Plants Under Drought Stress

Acclimation of plants to drought is often associated with increased levels of reactive oxygen species (ROS), such as superoxide anion (O2 · −), hydrogen peroxide (H2O2), hydroxyl radical (HO·) and singlet oxygen (1O2), which are toxic for the cells. ROS are by-products of aerobic metabolism, and their production is enhanced during drought conditions through the disruption of electron transport system and oxidizing metabolic activities occurring in chloroplasts, mitochondria and microbodies. Under non-stressful conditions, ROS are efficiently eliminated by non-enzymatic and enzymatic antioxidants, whereas during drought conditions the production of ROS exceeds the capacity of the antioxidative systems to remove them, causing oxidative stress. The non-enzymatic antioxidant system includes ascorbate and glutathione, located both within the cell and in the apoplast. They are two constituents of the antioxidative ascorbate–glutathione cycle which detoxify H2O2 in the chloroplasts. Ascorbate (AsA) is a major primary antioxidant compound synthesized on the inner membrane of the mitochondria which reacts chemically with 1O2, O2·−, HO· and thiyl radical, and acts as the natural substrate of many plant peroxidases. Moreover, AsA is involved in other functions such as plant growth, gene regulation, modulation of some enzymes, and redox regulation of membrane-bound antioxidant compounds. Glutathione (GSH) is a tripeptide synthesized in the cytosol and chloroplasts which scavenges 1O2 and H2O2, and it is oxidized to glutathione disulfide (GSSG) when acts as an antioxidant and redox regulator. GSH is the substrate of glutathione S-transferases, which have a protective role in the detoxification of xenobiotics, and dehydroascorbate reductase (DHAR). Finally, GSH is a precursor of phytochelatins, which regulate cellular heavy metals levels, and is involved in gene expression. This review, based on the most significant studies published in the last decade, focuses on the changes of antioxidant enzyme activities (ascorbate peroxidase, APX; monodehydroascorbate reductase, MDHAR; dehydroascorbate reductase, DHAR; glutathione reductase, GR), and of the levels of some compounds involved in the ascorbate–glutathione cycle (ascorbate and glutathione pools, H2O2 and α-tocopherol) in plants grown under water shortage.