Biohemical and physiological characterization of three poplar clones (Populus spp.) during the copper, nickel and cadmium phytoextraction process

Predmet ovog istaživanja  bio je ispitivanje uticaja jona tri teška metala (Ni2+, Cu2+ i Cd2+) u dve toksične koncentracije u zemljištu na fiziološke i biohemijske karakteristike  odabranih klonova topola, M1, B229 i Pe 19/66. Ispitan je i potencijal odabranih klonova topola da vrše fitoekstrakciju-akumulaciju teških metala iz zemljišta u svoje nadzemne delove što dovodi  do dugoročnog uklanjanja ovih perzistentnih polutanata iz životne sredine. Takođe, ispitan je uticaj teških metala na antioksidantni potencijal, sposobnost  neutalizacije slobodnih radikala, aktivnosti antioksidantih enzima kao i  na sadržaj slobodnih i konjugovanih poliamina (Put, Spm, Spd),  određenih HPLC analizom,  i sadržaj biljnih hormona poput indol-3-sirćetne kiseline i  abscisinske  kiseline,  određenih GC/MS analizom, u listovima i korenovima klonova topola.  The aim of this study was to estimate and compare phytoextraction capacities of three poplar clones (M1, B229 and Pe 19/66) in soil. Furthermore, the goal was to assess  different biological responses among the poplar clones during exposure to different concentration of three heavy metal ions (Ni2+, Cu2+ i Cd2+). In order to track changes in  poplars’mineral, physiological, biochemical and antioxidant status during the  abiotic stress, quantification of physiological properties, free and conjugated polyamines, total phenolics  as well as quantification of phytohormones (indol-3-acetic and abscisic acid) was done. Furthermore, assessment of antioxidant potential by tracking radical scavenger capacities (RSC) against DPPH, ABTS, OH and NO radicals and by measuring enzymes activities (SOD, GSH-Px, GPx, GR) in vitro was performed in root and leaves of poplar clones

Drought Effects on Physiology and Biochemistry of Pedunculate Oak (Quercus robur L.) and Hornbeam (Carpinus betulus L.) Saplings Grown in Urban Area of Novi Sad, Serbia

Background and Purpose: Water stress is one of the major problems for urban trees. It affects a wide range of plant responses, from changes at the cellular level to the reduction in growth rates. Irrigation of trees in urban areas may provide numerous benefits important for increasing tree vitality to withstand other stresses that might occur. The aim of this study was to compare drought effects on some physiological and biochemical performances of Pedunculate oak (Quercus robur L.) and Hornbeam (Carpinus betulus L.) saplings grown in the urban area. Materials and Methods: The study was conducted during August 2012 at the Boulevard of Europe (Novi Sad, Republic of Serbia). Measurements were carried out on saplings grown in the part of the boulevard with drip irrigation system installed (Site 1) and on the saplings cultivated in the part without any irrigation system (Site 2). Results: Soil moisture content was significantly higher at Site 1 with approximately 57.2%, compared to 18.7% at Site 2. The results showed that irrigated saplings were characterized by significantly higher stomatal conductance in Q. robur and C. betulus. Similarly, the content of free proline, FRAP units and the amount of malonyldialdehyde showed increased values in trees subjected to soil water deficit. In contrast, net photosynthesis, chlorophyll and carotenoid contents did not differ notably in irrigated and non-irrigated Q. robur and C. betulus trees. Conclusions: Water stress significantly affected stomatal conductance and some biochemical properties of Q. robur and C. betulus saplings cultivated at the non-irrigated site. The results showed that the implementation of drip irrigation system in urban landscape is an important tool in the prevention of drought stress effects on the physiological processes of plants

Variability of Physiological Parameters of European Beech Provenances in International Provenance Trials in Serbia

In this study, the variability of physiological parameters of five provenances of Europeanbeech (Fagus sylvatica), which were planted at two locations with different ecological conditions atFruška Gora and Debeli Lug, was estimated. Provenance trials were established in the framework ofCOST Action E52: "Evaluation of Beech Genetic Resources for Sustainable Forestry". 2-3 years oldseedlings originating from Croatia, Germany, Bosnia, Austria and Serbia were planted in blocks offifty plants with a spacing of 2 x 1 m. Physiological parameters such as net photosynthesis, rate oftranspiration and stomatal conductance were measured with a portable gas analysis system. Generally,provenances from Fruška Gora Mountain showed higher intensity of all physiological parameters thanprovenances located at site Debeli Lug. High correlations among rates of net photosynthesis andtranspiration, on one side, and stomatal conductance, on the other side, were found. ANOVA testindicates that variability of net photosynthesis, transpiration and stomatal conductance of investigatedprovenances, at the two locations, was influenced both by environmental conditions of sites andgenetic constitution of provenances

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

The effect of low-dose spermidine supplementation on polyamine content and antioxidative defence mechanisms in honey bees

Summary. The honey bee, a widespread pollinator, contributes to the conservation of biodiversity. In recent decades, a trend of declining colony numbers has emerged. The unsustainable exploitation of the environment may be the cause of this phenomenon. One protective strategy of organisms is to strengthen their antioxidative capacity. A class of positively charged molecules, polyamines, plays important roles in various cellular processes. They exert a regulatory effect on gene expression, have antioxidative properties, and promote longevity in model organisms. The three main representatives are putrescine, spermidine and spermine. The aim of this study was to determine whether supplementation of bees with low-dose spermidine leads to an increased level of the mentioned polyamines and whether this could strengthen the antioxidative defence system. Two experimental groups were established: C group (control), fed with a 50% (w/v) sucrose solution, and S0.01 group, whose diet was supplemented with 0.01 mM spermidine. The experiment lasted for 10 and 17 days. A significant increase in putrescine, spermidine and spermine content was noted in the supplemented group after 17 days, compared to its control. These results show a positive impact of spermidine supplementation on maintaining polyamine levels throughout aging. FRAP and MDA biochemical assays were used for the assessment of oxidative status. FRAP assay showed increased antioxidative capacity in the S0.01 group. These results are in accordance with the results obtained from the MDA assay, which showed a decreased level of lipid peroxidation in the supplemented group, in comparison to the control. The potential practical outcome of this study could be the use of spermidine in beekeeping practice to promote overall honey bee health

Heavy Metals Content in Foliar Litter and Branches of Quercus petraea (Matt.) Liebl. and Quercus robur L. Observed at Two ICP Forests Monitoring Plots

Background and Purpose: Due to the ability to uptake and accumulate heavy metals (HM) in their aboveground tissues, trees may be used for phytoremediation purposes, but also as bioindicators of environmental pollution. The aims of the present study were: a) to investigate the content and temporal variation of the studied HMs in different plant organs during the period of intensive leaf falling (September-October), in two species from genus Quercus; b) to evaluate the observed HMs content relative to plausible ranges of element concentration in foliar litter, as recommended by ICP Forests Foliar Co-ordinating Centre (IFFCC). Material and Methods: The contents of zinc (Zn), manganese (Mn), copper (Cu) and iron (Fe) were assessed in the litterfall (i.e. leaves and branches) of Quercus petraea (Matt.) Liebl. and Quercus robur L. grown at two ICP Forests Level II monitoring plots (Fruška Gora and Odžaci, Serbia). Plant material was sampled during September-October 2018 and analyzed by atomic absorption spectrophotometry. Results: Comparing the content of investigated HMs in foliar litter and branches of Q. robur and Q. petraea, notably higher concentrations of Mn and Fe have been observed in the foliage of both species, whereas Zn and Cu concentrations were higher in collected branch material of both of the above-mentioned tree species. The results further showed that, when compared to plausible ranges of element concentrations in foliar litter, given by ICP Forests Foliar Co-ordinating Centre (IFFCC), average concentrations of Fe and Zn in the leaves were within suggested limits, whereas certain concentrations of Mn and Cu exceeded the proposed ranges. Furthermore, the content of the studied HMs in plant material of both Quercus species significantly varied during sampling period. Conclusions: Although it was observed that concentrations of certain HMs were beyond plausible limits recommended by IFFCC, our results were in accordance with the findings of other authors who studied HMs content in Quercus species grown in natural forests. For that reason, these limits should be considered optionally, i.e. in the cases when the results significantly deviate from the suggested values. Moreover, we believe that the observed variation in the HMs content during sampling period is related to the sampling procedure commonly applied at ICP Forests monitoring plots, which is indirectly associated with the capacity of trees to accumulate HMs and their phenological properties

Jesu li testovi praćenja oksidacijskog stresa dovoljni za genetički monitoring hrasta lužnjaka u malom razmjeru?

SummaryAlthough being present much widely in terms of its geographical distribution, pedunculate oak (Quercus robur L.) represents one of the crucial species in forest ecosystems in South Eastern Europe. We wanted to contribute to existing efforts in designing genetic monitoring methodology for this noble broadleaved species, through the screening of genotypes with different health status and belonging to two phenological varieties [early (var. praecox) vs late (var. tardissima)] using seven oxidative stress parameters (total protein content, total phenolic compounds content, total flavonoids content, DPPH, ABTS, NO radical scavenger activities and Ferric reducing antioxidant power essay - FRAP). Results of this study provided significant biological message, contributing to better understanding of existing biodiversity of pedunculate oak in Serbia. Four out of seven oxidative stress screening tests (total phenolic compounds content, total flavonoids content, RSC NO and FRAP) clearly discriminated late vs early individuals sampled from selected pedunculate oak gene pool. Although presented results did not show significancy in using mentioned parameters to distinct damaged from vital individuals, nevertheless, knowledge gained through discussion and comparison with previous studies contributed to precise determination of biochemical tests which might be used in future efforts of genetic monitoring of this species.SažetakPrema klimatskim scenarijima RCP-a, hrast lužnjak (Q. robur L.) označen je kao najteže pogođena vrsta drveća zbog stalnih, intenzivnih klimatskih promjena. Propadanje hrastovih šuma može uzrokovati velike ekonomske gubitke, budući da se hrastovi smatraju vrlo vrijednim primjercima drveća zbog duge povijesti iskorištavanja za proizvodnju drva i gradnju (Mikac i sur., 2018.). Uzimajući u obzir da uobičajeni biokemijski parametri mogu doprinijeti razumijevanju temeljnih obrambenih mehanizama biljaka protiv stresa od suše i stoga se koristiti kao pouzdani deskriptori prilagodljivosti u kontekstu klimatskih promjena, cilj ove studije bio je utvrditi postoji li neki od 7 predloženih testova praćenja oksidacijskog stresa (ukupni sadržaji bjelančevina, fenola, flavonoida i testovi hvatači slobodnih radikala – ABTS, NO, FRAP i DPPH) koji može poslužiti za dugoročni genetski monitoring dviju sorti hrasta lužnjaka (rani vs kasni). Fenološki varijeteti [rani (var. praecox) vs kasni (var. tardissima)] i fiziološka skupina (vitalne ili oštećene) bile su promatrane varijable u testiranih jedinki zbirnog radnog hrasta vitalnog (PEOV), zbirnog ranog hrasta oštećenog (PEOD), zbirnog kasnog vitalnog (PLOV) i zbirnog ranog oštećenog (PLOD). Svaka objedinjena (zbirna) skupina sadržavala je ukupno 10 ili 11 uzoraka, mjerenih u tri primjerka kako bi se postigla optimalna statistička značajnost za svaku skupinu (za više, pogledajte pododjeljak Statistika). Sakupljeno lišće odmah je označeno i stavljeno u aluminijski folij, konzervirano na suhom ledu i transportirano u Laboratorij za kemijske i biokemijske analize Instituta za nizinsko šumarstvo i okoliš (ILFE) na daljnju obradu. Dva grama hrastovog lišća homogenizirano je u ohlađenom mužaru s tučkom s 10 ml fosfatnog pufera (0,1 M KH2PO4, pH=7), centrifugirano 10 min na 11000 okretaja u minuti i supernatant je korišten za daljnji test praćenja oksidativnog stresa (Tablica 1), prema uputama autora. Sve analize određene su spektrofotometrijski u mikropločicama s čitačem Multiplate MultiScan GO (Thermo Scientific, Njemačka). Dobiveni rezultati biokemijskih parametara pokazali su da je lišće kasnog hrasta imalo veći sadržaj bjelančevina i fenolnih spojeva od ranog, bez obzira na fiziološku skupinu (vitalno ili oštećeno) (Tablica 2). Štoviše, radi boljeg prikaza rezultata iz Tablice 2. prikazali smo ih grafički (Graf 1. i 2.). Najveći udio ukupnih flavonoida imali su listovi PLOD i PLOV, a najmanji listovi PEOV i PEOD (Tablica 2). Što se tiče ispitivanja kapaciteta hvatanja radikala, svi testirani listovi imali su nizak kapacitet hvatanja. ABTS- i NO- testovi hvatači radikala pokazali su manje od 50% hvatanja za sve uzorke, dok je DPPH-test pokazao neznatno više od 50% hvatanja za PEOD, PEOV, PLOV listove (56,15%, 57,28% odnosno 51,58%) i 41,69 % za listove PLOD. Kasno lišće hrasta ima veću antioksidacijsku moć reduciranja željeza (203,5 odnosno 180,4 mg TE/g FW) nego rano (93,9 odnosno 85,5 mg TE/g FW), bez obzira na fiziološku skupinu (Tablica 2). Razlike između ranih i kasnih genotipova hrasta bile su značajnije u pogledu ukupnog sadržaja fenola i flavonoida, kao i antioksidativne moći reduciranja željeza (FRAP). Ovo zapažanje također je potvrđeno PCA analizom, gdje su sva tri parametra grupirana zajedno s malom razlikom u najdominantnijoj komponenti PC1 (Graf 3). Dvosmjerna ANOVA u našem slučaju pokazala je da fiziološko grupiranje nije imalo utjecaja na ukupni sadržaj fenola i naknadno na aktivnost hvatača radikala FRAP, ABTS i NO. Dobiveni rezultati mogu pomoći da se objasne razlike u biokemijskim odgovorima na abiotičke stresove stabala hrasta lužnjaka na različitim mikrolokalitetima i na razini varijeteta unutar jednog šumskog kompleksa te nam mogu pružiti vrijedne informacije o varijabilnosti unutar vrste u otpornosti na sušu (rano naspram kasno). Stoga ovo može biti važan alat za poboljšanje strategija uzgoja i planova umjetne regeneracije hrasta lužnjaka u sušnijim nizinskim regijama. S obzirom na rezultate, potrebno je dugoročno mjerenje parametara oksidacijskog stresa stabala hrasta na individualnoj i populacijskoj razini, kako bi se dobila jasnija slika o tome kakve se promjene događaju na molekularnoj razini. Ova mjerenja također mogu biti korisna u genetskom praćenju stabala hrasta postavljenih u polukontroliranim i kontroliranim uvjetima. Na taj način bi se mogao dobiti uvid u točan genetski kapacitet vrsta drveća u pogledu njihove otpornosti na abiotski stres na pojedinim mikrolokalitetima unutar krajolika

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

Mycorrhizal Fungal Community of Poplars Growing on Pyrite Tailings Contaminated Site near the River Timok

Background and Purpose: Mycorrhizal fungi are of high importance for functioning of forest ecosystems and they could be used as indicators of environmental stress. The aim of this research was to analyze ectomycorrhizal community structure and to determine root colonization rate with ectomycorrhizal, arbuscular mycorrhizal and endophytic fungi of poplars growing on pyrite tailings contaminated site near the river Timok (Eastern Serbia). Materials and Methods: Identification of ectomycorrhizal types was performed by combining morphological and anatomical characterization of ectomycorrhizae with molecular identification approach, based on sequencing of the nuclear ITS rRNA region. Also, colonization of poplar roots with ectomycorrhizal, arbuscular mycorrhizal and dark septated endophytic fungi were analysed with intersection method. Results and Conclusions: Physico-chemical analyses of soil from studied site showed unfavourable water properties of soil, relatively low pH and high content of heavy metals (copper and zinc). In investigated samples only four different ectomycorrhizal fungi were found. To the species level were identified Thelephora terrestris and Tomentella ellisi, while two types remained unidentified. Type Thelephora terrestris made up 89% of all ectomycorrhizal roots on studied site. Consequently total values of Species richness index and Shannon-Weaver diversity index were 0.80 and 0.43, respectively. No structures of arbuscular mycorrhizal fungi were recorded. Unfavourable environmental conditions prevailing on investigated site caused decrease of ectomycorrhizal types diversity. Our findings point out that mycorrhyzal fungal community could be used as an appropriate indicator of environmental changes

Response of Salix alba L. to heavy metals and diesel fuel contamination

Pot experiment was set in a greenhouse in order to determine the physiological response of Salix alba grown on soil co-contaminated with individual and combination of Cd, Ni, Pb-ethylenediaminetetraacetic acid (EDTA) and diesel fuel treatments. EDTA enhanced the uptake of Cd and Ni, whereas the antagonism between Cd, Ni and Pb led to reduced accumulation of Pb. Presence of 5 g/kg of diesel fuel in soil significantly increased toxic influence of applied heavy metals by further reducing plant growth, photosynthetic rate, transpiration rate and water use efficiency. This concentration of diesel fuel also reduced the uptake and accumulation of Cd (soil concentration of Cd was 4.36 mg/kg), while Ni and Pb accumulation (soil concentrations of 118.7 and 186.7 mg/kg) remained at the same level, but with significant reduction of plant growth, thus reducing S. alba phytoextracion potential. At lower applied contaminant concentrations (3 g of diesel fuel/kg and Cd 2.15, Ni 70.8 and Pb 116.1 mg/kg), growth disturbances were low and diesel fuel presence reduced the uptake of Pb only in roots and old leaves, whereas accumulation capacity of Cd and Ni remained unaffected, indicating that S. alba plants have potential for remediation and re-development of co-contaminated sites with moderate levels of polluters.Key words: Cd, Ni, Pb, phytoremediation, phytoextraction, willow, Salix alba