UTJECAJ ONEČIŠĆENOSTI TLA TEŠKIM METALIMA NA KLONOVE TOPOLA IZ SRBIJE

Oxidative stress is known as disturbed balance between antioxidative protection mechanism and production of reactive oxygen species, which can negatively influence on normal biological and metabolical processes in living organisms, such as poplar species. Phytoremediation is promising biotechnical method of cleaning of polluted soils by various pollutants: heavy metals, organic contaminants, pesticides, oil etc. Until today, poplars showed potential for regenerating polluted soils during phytoremediation process. This study represents results of oxidative stress profiles of three poplar clones (M1, B229 and PE 19/66) shoots from Institute of Lowland Forestry and Environment, University of Novi Sad, Serbia, while being treated by different concentration of three heavy metals in soil: Ni3+, Cu2+ and Cd2+. Biochemical parameters of oxidative stress which have been analyzed were: content of soluble proteins, intensity of lipid peroxidation, antioxidative capacity by ferric reducing antioxidative power assay and activity of superoxid dismutase. Results showed that the most acceptable phytoremediation response to heavy metal pollution in soil showed clone M1. Great differences between B229 and PE 19/66 clones were in response on soil heavy metal contamination, directly suggesting of not being suitable for possible phytoremediation application.Oksidacijski stres je poznat kao narušena ravnoteža između antioksidacijskog mehanizma zaštite i proizvodnje reaktivnih kisikovih vrsta, što može negativno utjecati na normalne biološke i metaboličke procese u živim organizmima, kao na primjer kod topola. U cilju opstanka, aerobni organizmi su stekli mehanizme antioksidacijske zaštite, gdje su od enzimskih najistraženije aktivnosti antioksidativnih enzima kao što su katalaze, peroksidaze, glutation peroksidaze i superoksid dismutaza. Fitoremedijacija je obećavajuća biotehnička metoda čišćenja zagađenih tala raznim onečišćujućim tvarima poput: teških kovina, organskih kontaminanata, pesticida i dr. Do danas, topole su pokazale potencijal za regeneraciju onečišćenih tala tijekom fitoremedijacijskog procesa.Biokemijsko profiliranje statusa oksidacijskog stresa u drvenastim biljkama nije često istraživana u Srbiji i stoga je cilj ovoga pokusa bio ispitati utjecaj različitih koncentracija tri jona teških kovina, Ni3+, Cu2+ i Cd2+ na razinu oksidacijskog stresa tri klona, dvije različite vrste topole (Populus euramericana-M1; PE 19/66 i B-229 oba Populus deltoides vrsta). Biokemijski parametri za određivanje razine oksidacijskog stresa su: lipidna peroksi­dacija (LPx), test redukcijske snage željeza (FRAP), superoksid dismutaze (SOD) aktivnost i sadržina topivih bjelančevina. Ovi rezultati mogli bi dati smjernice u odabiru klonova iz kolekcije Instituta za nizijsko šumarstvo i životnu sredinu Univerziteta u Novom Sadu za buduće fitoremedijacijske uporabe ovih genotipova topola. Pješčano fluvisol tlo u pokusnim posudama je zagađeno različitim tretmanima teških kovina, koji su prikazani u tablici 1. Biljni ekstrakti izrađeni su od 2 g biljnog materijala (izbojci) i homogenizirani s kvarcnim pijeskom i suspenzirani u 10 cm3 0,1 mol/dm3 K2HPO4 pH 7,0. Homogenati su centrifugirani za 10 min na 4000 g (Quy Hai sur., 1975). Sva očitavanja apsorbancija za sadržaj topivih proteina, SOD i FRAP odrađeno pomoću Janway UV / VIS spektrofotometra 6505 i za čitanje količine nastalog MDA korišten je višefrekvencijski Spectrum Termo Corporation.Sve analize izvedene su u tri ponavljanja. Statistička usporedba između uzoraka izvedena je u programu Statistica 9, koristeći Duncan test, s statističkom značajnosti p <0,05. Dobiveni rezultati prikazani su grafički preko histograma (slike 1. do 4.) i iznad njih su slova koja označuju statistički značajne razlike između rezultata i kontrola (od a do e).Usporedni postotak je izračunat prema formuli: Δ (%) = (100 * uzorak / kontrola) –100, gdje vrijednosti mogu rezultirati pozitivno (+) ako je došlo do povećanja u odnosu na kontrolu i rezultiralo negativno (–), ako je došlo do smanjenja u odnosu na kontrolu.Trudić i sur. (2012) analizirali su kroz iste metode okdisacijskog stresa ekstrakte lišća i korijena spomenutih klonova topola. Uspoređujući ove rezultate, B229 klon je kroz lišće i korijenje pokazao više prihvatljiv antioksidacijski odgovor, dok je, u ovom istraživanju, M1 klon pokazao najprihvatljivije antioksidacijske odgovore kroz ekstrakte izbojaka. Parametri iz Pilipović i sur. (2012), Trudić i sur. (2012) i naše studije, mogu predstavljati udružen fiziološki i antioksidacijski marker sustav za buduće različite fitoremedijacijske primjene ovih klonova na tlima onečišćenim raznim tvarima.Rezultati ovog istraživanja mogu ukazati na genotipske specifičnosti svih ispitivanih biokemijskih parametara i obilježava klonove topola, kao što je M1 kao primarni, a B229 klon kao potencijalno sljedeći u primjeni fitoremedijacije tala onečišćenih teškim kovinama. Ipak, za stjecanje novih, dubljih rezultata u vezi s oksidacijskim stresom izazvanim onečišćavanjem tala teškim kovinama, potrebne su dalje in vitro antioksidacijske analize

UTJECAJ ONEČIŠĆENOSTI TLA TEŠKIM METALIMA NA KLONOVE TOPOLA IZ SRBIJE

Oxidative stress is known as disturbed balance between antioxidative protection mechanism and production of reactive oxygen species, which can negatively influence on normal biological and metabolical processes in living organisms, such as poplar species. Phytoremediation is promising biotechnical method of cleaning of polluted soils by various pollutants: heavy metals, organic contaminants, pesticides, oil etc. Until today, poplars showed potential for regenerating polluted soils during phytoremediation process. This study represents results of oxidative stress profiles of three poplar clones (M1, B229 and PE 19/66) shoots from Institute of Lowland Forestry and Environment, University of Novi Sad, Serbia, while being treated by different concentration of three heavy metals in soil: Ni3+, Cu2+ and Cd2+. Biochemical parameters of oxidative stress which have been analyzed were: content of soluble proteins, intensity of lipid peroxidation, antioxidative capacity by ferric reducing antioxidative power assay and activity of superoxid dismutase. Results showed that the most acceptable phytoremediation response to heavy metal pollution in soil showed clone M1. Great differences between B229 and PE 19/66 clones were in response on soil heavy metal contamination, directly suggesting of not being suitable for possible phytoremediation application.Oksidacijski stres je poznat kao narušena ravnoteža između antioksidacijskog mehanizma zaštite i proizvodnje reaktivnih kisikovih vrsta, što može negativno utjecati na normalne biološke i metaboličke procese u živim organizmima, kao na primjer kod topola. U cilju opstanka, aerobni organizmi su stekli mehanizme antioksidacijske zaštite, gdje su od enzimskih najistraženije aktivnosti antioksidativnih enzima kao što su katalaze, peroksidaze, glutation peroksidaze i superoksid dismutaza. Fitoremedijacija je obećavajuća biotehnička metoda čišćenja zagađenih tala raznim onečišćujućim tvarima poput: teških kovina, organskih kontaminanata, pesticida i dr. Do danas, topole su pokazale potencijal za regeneraciju onečišćenih tala tijekom fitoremedijacijskog procesa.Biokemijsko profiliranje statusa oksidacijskog stresa u drvenastim biljkama nije često istraživana u Srbiji i stoga je cilj ovoga pokusa bio ispitati utjecaj različitih koncentracija tri jona teških kovina, Ni3+, Cu2+ i Cd2+ na razinu oksidacijskog stresa tri klona, dvije različite vrste topole (Populus euramericana-M1; PE 19/66 i B-229 oba Populus deltoides vrsta). Biokemijski parametri za određivanje razine oksidacijskog stresa su: lipidna peroksi­dacija (LPx), test redukcijske snage željeza (FRAP), superoksid dismutaze (SOD) aktivnost i sadržina topivih bjelančevina. Ovi rezultati mogli bi dati smjernice u odabiru klonova iz kolekcije Instituta za nizijsko šumarstvo i životnu sredinu Univerziteta u Novom Sadu za buduće fitoremedijacijske uporabe ovih genotipova topola. Pješčano fluvisol tlo u pokusnim posudama je zagađeno različitim tretmanima teških kovina, koji su prikazani u tablici 1. Biljni ekstrakti izrađeni su od 2 g biljnog materijala (izbojci) i homogenizirani s kvarcnim pijeskom i suspenzirani u 10 cm3 0,1 mol/dm3 K2HPO4 pH 7,0. Homogenati su centrifugirani za 10 min na 4000 g (Quy Hai sur., 1975). Sva očitavanja apsorbancija za sadržaj topivih proteina, SOD i FRAP odrađeno pomoću Janway UV / VIS spektrofotometra 6505 i za čitanje količine nastalog MDA korišten je višefrekvencijski Spectrum Termo Corporation.Sve analize izvedene su u tri ponavljanja. Statistička usporedba između uzoraka izvedena je u programu Statistica 9, koristeći Duncan test, s statističkom značajnosti p <0,05. Dobiveni rezultati prikazani su grafički preko histograma (slike 1. do 4.) i iznad njih su slova koja označuju statistički značajne razlike između rezultata i kontrola (od a do e).Usporedni postotak je izračunat prema formuli: Δ (%) = (100 * uzorak / kontrola) –100, gdje vrijednosti mogu rezultirati pozitivno (+) ako je došlo do povećanja u odnosu na kontrolu i rezultiralo negativno (–), ako je došlo do smanjenja u odnosu na kontrolu.Trudić i sur. (2012) analizirali su kroz iste metode okdisacijskog stresa ekstrakte lišća i korijena spomenutih klonova topola. Uspoređujući ove rezultate, B229 klon je kroz lišće i korijenje pokazao više prihvatljiv antioksidacijski odgovor, dok je, u ovom istraživanju, M1 klon pokazao najprihvatljivije antioksidacijske odgovore kroz ekstrakte izbojaka. Parametri iz Pilipović i sur. (2012), Trudić i sur. (2012) i naše studije, mogu predstavljati udružen fiziološki i antioksidacijski marker sustav za buduće različite fitoremedijacijske primjene ovih klonova na tlima onečišćenim raznim tvarima.Rezultati ovog istraživanja mogu ukazati na genotipske specifičnosti svih ispitivanih biokemijskih parametara i obilježava klonove topola, kao što je M1 kao primarni, a B229 klon kao potencijalno sljedeći u primjeni fitoremedijacije tala onečišćenih teškim kovinama. Ipak, za stjecanje novih, dubljih rezultata u vezi s oksidacijskim stresom izazvanim onečišćavanjem tala teškim kovinama, potrebne su dalje in vitro antioksidacijske analize

Salt-induced changes in the antioxidant system and viability of oilseed rape

This work was aimed to investigate the effect of salt (NaCl) stress on three oilseed rape (Brassica napus L.) cultivars: 'Banacanka', 'Jasna' and 'Kata'. The following parameters were determined: antioxidant enzymes superoxide dismutase (SOD) and guaiacol peroxidase (GPX), reduced glutathione (GSH) quantity, lipid peroxidation (LP) intensity, radical-scavenging capacity (DPPH-RSC), hydroxyl radical scavenging ((OH)-O-center dot radicals) and ferric reducing antioxidant power (FRAP). Seed viability was determined using the standard laboratory test for germination as well as the seed accelerated aging test measuring the germination, shoot and root length. The plants were subjected to three salt treatments: 0, 100, 150 and 200 mmol dm(-3) NaCl for 7 days. All three cultivars responded with an increase of lipid peroxidation in seedlings and with a decrease of GPX activity. 'Banacanka' was found to be the most vulnerable cultivar, with an increase of 237% of LP intensity in comparison to the control at the highest NaCl concentration. The induction of the SOD activity was observed for all three cultivars by different NaCl concentrations. The antioxidant capacity measured by FRAP, DPPH-RSC and (OH)-O-center dot radicals was also induced, especially FRAP at the highest concentration in the cultivar 'Jasna' (by 282%). Germination, shoot and root length decreased for all investigated cultivars. Our results showed that cultivar 'Jasna' is the most adapted to salt stress because of the highest SOD activity and accumulation of antioxidants. Furthermore, this research suggests that some of the antioxidant parameters can be useful biochemical markers for the selection of oilseed rape tolerant cultivars under the conditions of salinity

Enhancement of Antioxidant and Isoflavones Concentration in Gamma Irradiated Soybean

Serbian soybean genotype Ana was gamma irradiated at doses of 1, 2, 4, and 10 kGy in order to evaluate the influence of gamma irradiation on isoflavone (genistein, daidzein, and their glycosides genistin and daidzin) contents and hydroxyl radical scavenging effect (HRSE). The increase in genistin and daidzin contents as well as antioxidant activities was observed especially at doses of 4 and 10 kGy. Results were also compared with our previous results relating to total phenol content (TPC), DPPH radical scavenger capacity (DPPH RSC), and ferric reducing antioxidant power (FRAP). Our results indicated that doses up to 10 kGy improve the antioxidant activities of soybean and also nutritional quality with respect to isoflavone content. All results were analyzed by multivariate techniques (correlation matrix calculation and autoscaling transformation of data). Significant positive correlations were observed between genistin, daidzin, DPPH RSC, and HRSE

Antioxidative response of poplar tissue culture exposed to peg 6000

The aim of this work was to investigate the influence of the water deficit in poplar tissue culture (M-1 genotype) through proline content, level of lipid peroxidation and activities of antioxidative enzymes. Plants were exposed to PEG 6000 (100 and 200 mOsm) in in vitro conditions for 6 days. Malondialdehyde (MDA) content showed an increase only at 200 mOsm stress. Under the 100 mOsm stress, catalase and guaiacol peroxidase activities were induces indicating their important role in elimination of reactive oxygen species (ROS). Under the 200 mOsm stress, superoxide dismutase and guaiacol peroxidase activities were induced

Comparison between Organic and Conventional Spelt and Wheatgrass juice

This work is aimed to compare antioxidant and scavenger properties of conventionally and organically produced speltgrass. Moreover, the antioxidant status of spelt was compared with wheatgrass. Three spelt varieties were analyzed:Nirvana, Ebners–Rotkorn variety and Eco-10. Antioxidant enzymes, superoxide-dismutase and guiacol-peroxidase, were determined as well as ferric reducing antioxidant power and DPPH-antiradical power. Total soluble proteins and total phenols were determined and also lipid peroxidation as a parameter of oxidative stress. The organically produced spelt was exposed to the higher levels of oxidative stress than conventionally produced spelt, and also DPPH antiradical power was induced by conventional production, which is also demonstrated using the relative antioxidant capacity index. The best antioxidant properties, better than wheat, showed organically produced spelt genotype Ebners–Rotkorn which is determined by the highest phenol content, superoxide-dismutase activity, DPPH antiradical power and ferric reduced antioxidant power and consequently with the lowest lipid peroxidation intensity

Phytochemical characteristics and antioxidant capacity of fruit extracts of different prunus species

Fruits are one of the major sources of polyphenol compounds in human diet. These compounds are known to have many health-promoting activities, especially anticancer, antiradical and antioxidant effects. In this work, seven different Prunus species traditionally grown in south Bačka region of Vojvodina were investigated: blackthorn (P. spinosa), plum (P. domestica), apricot (P. armeniaca), cherry plum (P. cerasifera), sweet cherry (P. avium), sour cherry (P. cerasus) and mahaleb cherry (P. mahaleb). Freeze-dried fruits were evaluated in terms of their phytochemical characteristics and bioactivity, determining total content of phenolics, flavonoids and anthocyanins, total antioxidant capacity and antiproliferative effect on human colon cancer cells (HT29). Blackthorn fruits are the richest in phenolic and flavonoid contents, while mahaleb cherry and sweet cherry had much higher content of total anthocyanins than other examined species. Apricot and cherry plum fruits had the lowest levels of polyphenol compounds, but very high antiproliferative effect, almost the same as blackthorn. This indicates that not only polyphenol compounds contribute to antiproliferative effects. Concerning total antioxidant activity, blackthorn, sweet cherry, sour cherry and mahaleb cherry showed the highest capacity of scavenging DPPH radical and ferric reducing activity power

Irradiation Effects on Phenolic Content, Lipid and Protein Oxidation and Scavenger Ability of Soybean Seeds

The effect of medium doses of γ-irradiation (1-10 kGy) on total phenolic andtannin contents, lipid peroxidation, protein oxidation intensity and soluble protein contentof soybean seeds, (genotype Ana), was investigated. Screening for antioxidant ability wasperformed using FRAP and DPPH methods. Total phenolic and tannin contents and DPPHscavenger activity were increased, while protein oxidation intensity was decreased byapplied doses of γ-irradiation. Gamma irradiation provoked insignificant changes in lipidperoxidation and soluble protein content, while protein oxidation intensity wassignificantly decreased when dose of 10 kGy was applied. Presented results implicated thatincreased antioxidant capacity and protein stability of soybean seeds were increased afterapplication of γ-irradiation

Comparative Study of Antioxidant Status in Androgenic Embryos of Aesculus hippocastanum and Aesculus flava

In vivo (leaves and seed embryos) and in vitro (androgenic embryos) antioxidant scavenging activity of Aesculus hippocastanum and Aesculus flava medical plants was examined. Here we report antioxidant enzyme activities of superoxide dismutase, catalase, guaiacol peroxidase and glutathione peroxidase, reduced glutathione quantity, flavonoids, soluble protein contents, quantities of malondialdehyde, and (OH)-O-center dot radical presence in the investigated plant samples. Total antioxidant capacity of all the samples of A. hippocastanum and A. flava was determined using FRAP, DPPH, and NO center dot radical scavenger capacity. The leaves of A. flava collected from the botanical garden exhibited stronger antioxidant activity (higher activities of SOD, and higher quantities of GSH, TSH, TPC, and scavenging abilities of DPPH and NO center dot, and higher FRAP values and lowest quantities of (OH)-O-center dot and MDA) than in vitro obtained cultures. However, the leaves of A. flava showed higher antioxidant activity than the leaves of A. hippocastanum, and therefore they have a stronger tolerance of oxidative stress. Androgenic embryos of both species had low amount of antioxidants due to controlled in vitro environmental conditions (T, photoperiod, humidity, nutritive factors, and pathogen-free). Our results confirmed that we found optimal in vitro conditions for producing androgenic embryos of both Aesculus species. Also, we assume that horse chestnut androgenic embryos can be used as an alternative source for large-scale aescin production.Ministry of Education, Science, and Technology Development of Serbia {[}III43002, 173015