Bioakumulacija Cu i Zu u nekim akvatičnim makrofitama na području ribnjaka Bardača

The paper deals with Cu and Zn content analysis in water, sediment and tissue of Phragmites communis Trin., Salvinia natans L. and Utricularia vulgaris L. in the area of Bardača fish pond (Necik and Sinjak- active fish ponds). Based on the measured value of Cu in water, and according to the Regulation on Classification of Water and Water Streams, the researched localities belong to category II waters, and based on the Zn content they belong to category III/ IV waters. The content of copper and zinc in sediments did not exceed the maximum permitted concentration values according to the RS Official Gazette (23/1994). The obtained results indicate that Cu and Zn content in plant tissue significantly varied in relation to species, locality and sampling period. In the tissue of Salvinia natans during the researched period, the lowest Zn concentration was measured in September on both localities (32 mg/kg), while the biggest concentration exceeded the maximum permitted values (163,55 mg/kg) which indicates a certain degree of contamination of researched localities. Zn concentration in the tissue of Utricularia vulgaris on both localities ranged from 39,14 to 55,20 mg/kg, whereby the obtained values were by 35 % lower in relation to Salvinia natans on Necik locality, while on Sinjak locality the Zn concentration at both species was similar (44,55 mg/kg). The most significant difference in Zn accumulation was determined for Phragmites communis where 3-6 times lower Zn content was obtained in relation to Salvinia natans and Utricularia vulgaris. The Cu content in tissue of Utricularia vulgaris during the research period was in the range from 1,7-10,18 mg/kg, while in the tissue of Salvinia natans insignificantly lower values were measured as compared to Utriculara vulgaris. The Cu concentration in the tissue of Phragmites communis during the researched period on both localities was below the detection limit (<0,023 mg/kg). Species Salvinia natans has shown to be the best Zn bioaccumulator, while Utricularia vulgaris accumulated copper the best, which indicates the existence of different trend in terms of accumulation of certain heavy metals during the season in different organs and tissues. The role of different types of aquatic macrophytes in heavy metal accumulation during the season was emphasized here and their potential of application in phytoremediation technique.Rad se bavi analizom sadržaja Cu i Zn u vodi, sedimentu i tkivu Phragmites communis Trin., Salvinia natans L. All. i Utricularia vulgaris L. na području ribnjaka Bardača (Necik i Sinjak-aktivni ribnjački bazeni). Na osnovu izmjerenih vrijednost Cu u vodi, a prema Uredbi o klasifikaciji voda i kategorizaciji vodotoka, istraživani lokaliteti pripadaju vodama II kategorije, a na osnovu sadržaja Zn vodama III/IV kategorije. Sadržaj bakra i cinka u sedimentu nije prelazio vrijednosti maksimalno dozvoljene koncentracije prema Službenom glasniku RS (23/1994). Dobijeni rezultati ukazuju da sadržaj Cu i Zn u biljnom tkivu znatno varira u odnosu na vrstu, lokalitet i period uzimanja uzoraka. U tkivu Salvinia natans tokom istraživanog perioda najniža koncentracija Zn izmjerena je tokom septembra na oba lokaliteta (32 mg/kg), dok je najveća koncentracija prelazila maksimalno dozvoljene vrijednosti (163,55 mg/kg) što ukazuje na izvjestan stepen zagađenosti istraživanog lokaliteta. Koncentracija Zn u tkivu Utricularia vulgaris na oba lokaliteta bila je u opsegu 39,14-55,20 mg/kg, pri čemu su dobijene vrijednosti za 35 % bile niže u odnosu na Salvinia natans na lokalitetu Necik, dok su na lokalitetu Sinjak koncentracije Zn kod obje vrste bile slične (44,55 mg/kg). Najznačajnija razlika u akumulaciji Zn utvrđena je za Phragmites communis kod koje je dobijen 3-6 puta niži sadržaj Zn u odnosu na Salvinia natans i Utricularia vulgaris. Sadržaj Cu u tkivu Utricularia vulgaris tokom perioda istraživanja je bio u opsegu 1,7-10,18 mg/kg, dok su u tkivu Salvinia natans izmjerene neznatno niže vrijednosti u odnosu na Utriculara vulgaris. Koncentracija Cu u tkivu Phragmites communis tokom istraživanog perioda na oba lokaliteta bila je ispod detekcionog limita (<0,023 mg/kg). Kao najbolji biokumulator Zn pokazala se vrsta Salvinia natans dok je Utricularia vulgaris bolje akumulirala bakar, što ukazuje na postojanje različitog trenda u pogledu akumulacije određenih teških metala tokom sezone u različitim organima i tkivima. U ovom radu je naglašena uloga različitih tipova vodenih makrofita u akumulaciji teških metala tokom sezone i njihov potencijal primjene u tehnici fitoremedijacije

Effects of hemazin SC 500 (terbuthylazine) on antioxidative enzymes in human erythrocytes in vitro

The aim of this work was to investigate the effect of the commercial formulation hemazin SC 500, an herbicide containing terbuthylazine as the active compound, on the isoenzyme patterns and activities of Cu-Zn superoxide dismutase (SOD1) and catalase (CAT), as well as on the glutathione S-transferase (GST) activity, in human erythrocytes in vitro. The human erythrocytes were treated with hemazin SC 500 over a broad range of terbuthylazine concentrations (37 nmol L-1– –37 mol L-1) for 1 and 3 h at a temperature of 37°C. Native electrophoresis of the control and treated samples revealed two SOD1 and one CAT isoform. Treatment did not affect the SOD1 and CAT isoenzyme profile, but induced a change in their activities. Terbuthylazine at lower concentration induced a significant increase of the total SOD1 activity and decreased the GST activity in samples incubated for 1 and 3 h. On the other hand, the highest increase in the CAT activity was observed for the sample treated for 1 h with a higher concentration of terbuthylazine. Hemazin SC 500 containing terbuthylazine induces changes in the erythrocyte antioxidative system whereby the response of individual enzymatic antioxidants depends on the concentration of the pesticide and the incubation time

Cell wall-bound cationic and anionic class III isoperoxidases of pea root: biochemical characterization and function in root growth

Cell wall isolated from pea roots was used to separate and characterize two fractions possessing class III peroxidase activity: (i) ionically bound proteins and (ii) covalently bound proteins. Modified SDS-PAGE separated peroxidase isoforms by their apparent molecular weights: four bands of 56, 46, 44, and 41 kDa were found in the ionically bound fraction (iPOD) and one band (70 kDa) was resolved after treatment of the cell wall with cellulase and pectinase (cPOD). Isoelectric focusing (IEF) patterns for iPODs and cPODs were significantly different: five iPODs with highly cationic pI (9.5-9.2) were detected, whereas the nine cPODs were anionic with pI values between pH 3.7 and 5. iPODs and cPODs showed rather specific substrate affinity and different sensitivity to inhibitors, heat, and deglycosylation treatments. Peroxidase and oxidase activities and their IEF patterns for both fractions were determined in different zones along the root and in roots of different ages. New iPODs with pI 9.34 and 9.5 were induced with root growth, while the activity of cPODs was more related to the formation of the cell wall in non-elongating tissue. Treatment with auxin that inhibits root growth led to suppression of iPOD and induction of cPOD. A similar effect was obtained with the widely used elicitor, chitosan, which also induced cPODs with pI 5.3 and 5.7, which may be specifically related to pathogen defence. The differences reported here between biochemical properties of cPOD and iPOD and their differential induction during development and under specific treatments implicate that they are involved in specific and different physiological processes

Induction of peroxidase isoforms in the roots of two Verbascum thapsus L. populations is involved in adaptive responses to excess Zn2+ and Cu2+

To investigate metal specific responses of root class III peroxidases (POD, EC 1.11.1.7), two populations of Verbascum thapsus L. were exposed to excess Zn2+ or Cu2+ for three weeks in hydroponic culture. One population originating from an uncontaminated area (NMET) and one from an industrial disposal area for jarosite residues from zinc refining (MET) were chosen to test the capacity of V. thapsus to adapt to excess metal in the soil. Exposure to 60 μM Zn2+ led to increased levels of protein carbonyl groups only in the roots of NMET, which was accompanied by higher increase of POD activity and NADH-oxidase activity compared with MET plants. New anionic and cationic POD isoforms were induced in the roots of both populations in response to Zn2+ treatment, while IAA-oxidase activity decreased. On the other hand, root growth was more affected by Cu2+ than Zn2+ in both populations, which was correlated with increased auxin-oxidase (IAA-oxidase) activity. Cu2+ induced an increased activity of anionic POD isoforms in the roots of both populations, yet the ratio of NADH-oxidative to peroxidative POD activity remained higher in NMET than in MET plants. Overall results show differential effects of Zn2+ and Cu2+ on POD activity in the roots of V. thapsus L. In addition, higher tolerance to Zn2+ in MET plants than in NMET indicated that these plants have developed an adaptive mechanism to cope with Zn2+ excess

Peroxidase, phenolics, and antioxidative capacity of common mullein (verbascum thapsus l.) Grown in a zinc excess

Common mullein (Verbascum thapsus L.) is the dominant plant species at a disposal site polluted with metal from the hydrometallurgical jarosite zinc production process. Seeds collected at the site were germinated and plants were grown hydroponically under controlled conditions in a excess of Zn. Induction of total soluble POD activity in the root occurred at 1, 5, and 10 mM Zn, indicating Zn accumulation within the root. Accumulation of Zn in leaves was not accompanied by changes in POD activity, but resulted in gradual increase of total antioxidative capacity, which could be partly attributed to accumulation of soluble phenolics. The role of the phenolics/POD system in defense of V thapsus against zinc is discussed

Lipid composition of pea (Pisum sativum L.) and maize (Zea mays L.) root plasma membrane and membrane-bound peroxidase and superoxide dismutase

Plasma membrane was isolated from roots of pea and maize plants and used to analyze POD and SOD isoforms, as well as lipid composition. Among lipids, phospholipids were the main lipid class, with phosphatidylcholine being the most abundant individual component in both pea and maize plasma membranes. Significant differences between the two plant species were found in the contents of cerebrosides, free sterols, and steryl glycosides. Most maize POD isoforms were with neutral and anionic pI values, but the opposite was observed in pea. While both anionic and cationic SOD isoforms were isolated from maize, only two anionic SOD isoforms were detected in pea

Effects of mixed saline and alkaline stress on the morphology and anatomy of pisum sativum l.: the role of peroxidase and ascorbate oxidase in growth regulation

The effects of hyperalkaline, thermo-mineral water from Slatina on the morphology and anatomy of pea plants (Pisum sativum L.), were examined after eleven days of treatment with a mixture of tap water and Slatina water in 3:1 (T1) and 1:1 ratios (T2). Complete growth arrest of seedlings was observed in the Slatina water (T3). The alkalinity of external media was recovered to pH 8 within four days only in T1 and T2. Analysis of morphological parameters (the length of the main root, root application zone, number of lateral roots) indicated that the thermo-mineral water either promoted (T1) or inhibited (T2) the formation of lateral roots and plant growth. Comparative histological and anatomical analyses showed that inhibition of lateral roots was accompanied by an increase in the xylem and phloem. These changes in root morphology were accompanied by an increase in the activity of superoxide dismutase (SOD: E.C. 1.15.1.1) and peroxidase (POD: E.C 1.1.1.17) in the soluble fraction, whereas the activities of ascorbate oxidase (AAO: E.C. 1.10.3.3) bound to the cell wall and ionic POD decreased. The lower ratio of Slatina water in the hydroponic solution contributed to a more developed mesophyll with significantly higher AAO activity in the leaves and the induction of ionic POD isoforms. Besides alkalinity and excess NaCl, we suggest that a specific combination of metals (e.g. Ca and Mg) might be responsible for subtle changes in the cell area and xylem development, leading to dramatic changes in root anatomy

Spelt (Triticum spelta L.) Genotypes from the Western Balkan Countries

The yield components participate in the formation of the overall yield and vary within the genotype and environmental factors. The aim of this research was to evaluate yield components of spelt genotypes originating from different Western Balkan countries. Spelt genotypes were examined during two growing seasons 2015/16 and 2016/17 in the agro-ecological conditions of Banja Luka. Nine spelt genotypes were used, i.e. landrace Sitnica from Manjača mountain (the Republic of Srpska, B&H), seven Montenegrin landraces KP34LJ5-1/34 (SP1), KR16LJ5-1/16 (SP2), KR29LJ5-1/29 (SP3), KR12LJ5-1/12 (SP4), KR15LJ5-1/15 (SP5), LJ5-1/9 (SP6), KR20LJ5-1/20 (SP7) and cultivar Nirvana from Serbia. Standard production practices were applied for these spelt genotypes. Five traits were analyzed: the number of spikes m-2, plant height, spike length, the number of grains spike-1 and spike index. A two-factorial analysis of variance was performed and significant differences between treatment means were tested by the Fisher's LSD test at P≤0.05 and P≤0.01 significance level. Genotypes SP4 (140.38 cm), SP5 (138.82 cm), Sitnica (137.66 cm) and SP6 (135.65 cm) showed tendency of forming a relatively higher plants. Sitnica had the longest spikes (14.07 cm), while Montenegrin genotypes had relatively shorter spikes of 6.79 cm on average. Genotypes SP2, SP4, SP5, SP6 and SP7 obtained the highest number of grains per spike while the highest average spike index was obtained in SP7 (0.81)

WHEAT STRAW CONVERSION BY ENZYMATIC SYSTEM OF GANODERMA LUCIDUM

The purpose of this study was to resolve the question of whether various nitrogen sources and concentrations affect characteristics of selected G. lucidum ligninolytic enzymes participating in wheat straw fermentation. This is the first study reporting the presence of versatile peroxidase activity in crude extract of G. lucidum culture, as well as isoforms profile of Mn-oxidizing peroxidases. NH4NO3 was the optimum nitrogen source for laccase and Mn-dependent peroxidase activity, while peptone was the optimum one for versatile peroxidase activity. Four bands with laccase activity were obtained by native PAGE and IEF separations from medium enriched with inorganic nitrogen source, and only two bands from medium containing organic source. Medium composition was not shown to affect isoenzyme patterns of Mn-oxidizing peroxidases. Four isoforms of Mn-dependent peroxidase and three of versatile peroxidase were obtained on native PAGE. By IEF separation, five isoforms of Mn-dependent peroxidase and only two of versatile peroxidase were observed. The results demonstrated that G. lucidum has potential for mineralization and transformation of various agricultural residues and should take more significant participation in large-scale biotechnological processes

Senescence-related changes in the antioxidant status of ginkgo and birch leaves during autumn yellowing

The antioxidant status of birch and ginkgo leaves during autumnal senescence was characterized by the activities of catalase (CAT), peroxidase (POD), ascorbate peroxidase (APX) and superoxide dismutase (SOD). The contents of leaf H2O2 and ascorbate were used as indicators of oxidative stress. Degradation of chlorophyll (chl) during natural senescence was not accompanied either by an increase of H2O2 or by a decrease of reduced ascorbate. A transient decrease of reduced ascorbate in ginkgo and birch leaves in early senescence was accompanied by CAT inactivation. The activity of ionically-bound PODs was stimulated in late senescence in both species, when more than 30% of chl was degraded. Induction of MnSOD in both species and new isoforms of CuZnSOD in birch in late senescence was accompanied by the disappearance of other CuZnSOD isoforms in birch and FeSOD in ginkgo. The role of antioxidative enzymes in keeping ascorbate reduced and endogenous H2O2 at low levels in senescent leaves of deciduous trees was discussed