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Besides anthropogenic factors, climate change causes altered precipitation patterns that indirectly affect the increase of heavy metals in soils due to hydrological effects and enhanced leaching (i.e., Cd and Ni), especially in the vicinity of mines and smelters. Phytoextraction is a well-known, powerful “green” technique for environmental clean-up that uses plants to extract, sequester, and/or detoxify heavy metals, and it makes significant contributions to the removal of persistent inorganic pollutants from soils. Poplar species, due to their growth features, high transpiration rate, large biomass, and feasible reproduction represent great candidates for phytoextraction technology. However, the consequences of concomitant oxidative stress upon plant metabolism and the mechanism of the poplar’s tolerance to heavy metal-induced stress are still not completely understood. In this study, cuttings of poplar species (Populus deltoides W. Bartram ex Marshall) were separately exposed to two heavy metals (Cd2+ and Ni2+) that were triple the maximum allowed amount (MAA) (according to national legislation). The aim of the study was to estimate the effects of heavy metals on: (I) the accumulation of free and conjugated polyamines, (II) plant hormones (including abscisic acid-ABA and indole-3-acetic acid-IAA), and (III) the activities of different antioxidant enzymes at root and leaf levels. By using the selected ion monitoring (SIM) mode of gas chromatography with mass spectrometry (GC/MS) coupled with the isotopically labeled technique, amounts of ABA and IAA were quantified, while polyamine amounts were determined by using high-performance liquid chromatography (HPLC) with fluorometric detection after derivatization. The results showed that P. deltoides responded to elevated concentrations of heavy metals in soils by exhibiting metal- and organ-specific tolerance. Knowledge about tolerance mechanisms is of great importance for the development of phytoremediation technology

In light of climate change, pedunculate oak (Q. robur L.) was marked as the most threatened European tree species. Pedunculate oak is particularly jeopardized by powdery mildew disease caused by Erysiphe alphitoides. We hypothesized that priming of this tree species with ectomycorrhizal fungi could mitigate biotic stress and produce bioprotective properties against the disease. In this study, we have compared oaks’ foliar physiological and biochemical responses upon infection with E. alphitoides in the presence and absence of ectomycorrhizal fungi (ECM). The main aim of this study was to inspect how ECM modulate an oak’s biochemical response to infection with E. alphitoides, particularly at the level of the accumulation of the main polyamines (putrescine, spermidine, and spermine), soluble osmolytes (proline and glycine betaine), and phenolics (total phenolic content, flavonoids, and condensed tannins). A polyamine quantification was performed after derivatization by using high-performance liquid chromatography (HLPC) coupled with fluorescent detection. Oak seedlings inoculated with ECM fungi exhibited significantly higher levels of putrescine, spermine, and proline compared to non-inoculated seedlings, indicating the priming properties of the ECM. E. alphitoides caused an increase in individual and total polyamine content and lipid peroxidation in oak leaves regardless of the effect of ECM, while causing a decrease in physiological and antioxidative parameters and water use efficiency (WUE). Common biochemical parameters may contribute to understanding the underpinning plant defense mechanisms in three-way interactions among plants and pathogenic and ectomycorrhizal fungi and can be used as reliable adaptability descriptors in the context of climate change.

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 spektrofoto

Although 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., Prema 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 spektrofotometrijs

The influence of different groundwater table depths on pre-dawn water potential (PWP), soil moisture content and antioxidant activity (FRAP, RSC against DPPH·, NO and ABTS radicals) of adult pedunculate oak (Quercus robur L.) trees was assessed in three managed stands (localities 1-3) and one unmanaged (locality 4 - forest reserve). The study sites were located within single forest complex at different distances from the Sava riverbed. The measurements were performed during July 2015, when the first evidence of drought was evidenced. We hypothesised that the trees at locations farther from the river would be more drought-stressed due to lower groundwater depth and will show higher antioxidant capacity. Indeed, trees at localities 2 and 3 were more drought-stressed as compared to trees at locations 1 and 4, as indicated by the PWP results. Since antioxidant capacity can be directly correlated to drought tolerance, the present results indicate a higher antioxidant capacity in leaf extracts from locality 3, but also good oxidative adaptation in leaf extracts from locality 4, where the obtained values for almost all examined antioxidant tests were without statistically significant differences compared to those on locality 3. Obtained results may help to explain differences in biochemical responses to water deficit stress of pedunculate oak trees from different microsites within the single forest complex and provide us valuable information about intra-species drought resistance. Therefore, this can be an important tool for the improvement of breeding strategies and artificial regeneration plans for pedunculate oak in dry lowland regions., U ovom radu, procenjen je uticaj različitih dubina podzemnih voda na vodni potencijal lista (PWP), sadržaj vlage u zemljištu i antioksidativnu aktivnost (FRAP, inhibicija DPPH,·NO i ABTS radikala) odraslih stabala hrasta lužnjaka (Quercus robur L.) u tri gazdovne sastojine (lokaliteti 1-3) i u jednoj neuređenoj (lokalitet 4 – šumski rezervat). Lokacije istraživanja su se nalazile u okviru jedinstvenog šumskog kompleksa na različitim udaljenostima od korita Save. Merenja su obavljena tokom jula 2015. godine, kada su zabeleženi prvi dokazi o suši. Pretpostavili smo da će drveće na lokacijama udaljenim od reke biti pod većim sušnim stresom zbog manje dubine podzemne vode i da će pokazati veći antioksidantni kapacitet. Zaista, drveće na lokalitetima 2 i 3 bilo je pod većim stresom od suše u poređenju sa drvećem na lokacijama 1 i 4, kao što pokazuju rezultati PWP. Obzirom da antikosidativni kapacitet može biti direktno povezan sa tolerancijom na sušu, sadašnji rezultati ukazuju na veći antioksidativni kapacitet u ekstraktima listova sa lokaliteta 3 ali i na dobru oksidativnu adaptaciju listova sa lokaliteta 4, gde su dobijene vrednosti za skoro sve ispitivane antioksidativne testove bile su bez statistički značajnih razlika u odnosu na vrednosti dobijene na lokalitetu 3. Dobijeni rezultati mogu pomoći da se objasne razlike u biohmeijskim odgovorima na stres vodnog deficita kod stabala hrasta lužnjaka sa različitih mikrolokacija u okviru jedinstvenog šumskog kompleksa i daju nam dragocene informacije o intra-specijskoj otpornosti na sušu. Stoga, ovo može biti važna alatka za unapređivanje strategije oplemenjivanja i planova veštačke regeneracije hrasta lužnjaka u suvim ravničarskim regionima.

The general aim of this work was to compare the leaf-level responses of different protective components to water deficit and high temperatures in Quercus cerris L. and Quercus robur L. Several biochemical components of the osmotic adjustment and antioxidant system were investigated together with changes in hormones. Q. cerris and Q. robur seedlings responded to water deficit and high temperatures by: (1) activating a different pattern of osmoregulation and antioxidant mechanisms depending on the species and on the nature of the stress; (2) upregulating the synthesis of a newly-explored osmoprotectant, dimethylsulphoniopropionate (DMSP); (3) trading-off between metabolites; and (4) modulating hormone levels. Under water deficit, Q. cerris had a higher antioxidant capacity compared to Q. robur, which showed a lower investment in the antioxidant system. In both species, exposure to high temperatures induced a strong osmoregulation capacity that appeared largely conferred by DMSP in Q. cerris and by glycine betaine in Q. robur. Collectively, the more stress-responsive compounds in each species were those present at a significant basal level in non-stress conditions. Our results were discussed in terms of pre-adaptation and stress-induced metabolic patterns as related to species-specific stress tolerance features.

U okviru ove doktorske disertacije vršeno je istraživanje zajednica makrogljiva u okviru 5 šumskih staništa na Vidliču, Kopaoniku i Tari. Ispitivan je mikodiverzitet sa morfološkog, funkcionalnog i genetskog stanovišta. U istraživanju morološkog i funkcionalnog diverziteta, korišćene su različite klasične metode čiji rezultati su omogućili procenu stanja posmatranih mikocenoza, kao i samih šumskih staništa. Za analizu sastava vrsta u okviru mikocenoza, kao i procenu uticaja abiotičkih faktora na brojnost i sastav vrsta u okviru različitih funkcionalnih grupa, korišćeno je nekoliko statističkih metoda (PCA, PLS, CA i CCA). Osam vrsta, koje su pripadale najrasprostranjenijim i najzastupljenijim vrstama su odabrane za molekularne analize, koje su podrazumevale sekvenciranje ITS regiona rDNK, analizu njihovih polimorfizama kao i filogenetske analize u okviru vrste/roda. U cilju procene zagađenja staništa, u plodnim telima makrogljiva i njihovom supstratu je određen sadržaj metala (atomskom apsorpcionom spektrofotometrijom) i radionuklida (gamaspektrometrijom). Dobijeni rezultati ukazuju na to da diverzitet makrogljiva oslikava stanje samog staništa i da dugoročnim monitoringom mogu ukazati na promene u njemu., Within the framework of this doctoral dissertation, monitoring of macrofungal communities, within 5 forest habitats on Vidlič, Kopaonik and Tara, was done. Mycodiversity was investigated from the morphological, functional and genetic point of view. Various classical methods used, enabled the assessment of the condition of macrofungal communities, as well as the observed forest habitats. Several statistical methods (PCA, PLS, CA and CCA) were used to analyze the composition of species within the mycocenosis, as well as the assessment of the effects of abiotic factors on the species richness and species composition within different functional groups.Some of the most represented species have been selected for molecular analyzes, which includedsequencing of the ITS region, the analysis of polymorphisms, as well as phylogenetic analyzes within the species/genus. In order to assess the pollution of habitats, the content of metals (atomic absorption spectrophotometry) and radionuclides (gamma spectrometry) was determined in the sporocarps of macrofungi and their substrate. The obtained results indicate that diversity of macrofungi reflects the state of the habitat itself and that long-term monitoring can indicate changes in it.

Aim of study: In the present paper the nature and level of co-dependence between leaf functional traits and intrinsic water-use efficiency (WUEi) were studied in one-year-old Quercus robur L. seedlings, grown in a common garden experiment under moderate drought conditions. The study was established to identify those traits that might potentially be utilized to improve leaf-level WUEi, and therefore be used in breeding programmes to enhance drought adaptation of Q. robur.Area of study: The study was carried out at the common garden site within the UNESCO Biosphere Reserve Mura-Drava-Danube.Material and methods: The study involved one-year-old seedlings of eight half-sib lines of Q. robur. Eighteen leaf parameters were analyzed; i.e. physiological, biochemical, morphological and anatomical. The data were processed using multivariate statistical methods: a) principal component analysis, b) stepwise regression analysis, and c) path coefficient analysis.Main results: The results showed that leaf stomata traits, particularly stomatal density (SD), and leaf dry mass per unit leaf area (LMA) were the most important traits, closely associated with WUEi. Stomatal density achieved the highest score on PC1 (0.825), in which WUEi had the highest loading (0.920), as well. SD was also included first in stepwise regression model.

Aim of study: In the present paper the nature and level of co-dependence between leaf functional traits and intrinsic water-use efficiency (WUEi) were studied in one-year-old Quercus robur L. seedlings, grown in a common garden experiment under moderate drought conditions. The study was established to identify those traits that might potentially be utilized to improve leaf-level WUEi, and therefore be used in breeding programmes to enhance drought adaptation of Q. robur. Area of study: The study was carried out at the common garden site within the UNESCO Biosphere Reserve Mura-Drava-Danube. Material and methods:The study involved one-year-old seedlings of eight half-sib lines of Q. robur. Eighteen leaf parameters were analyzed; i.e. physiological, biochemical, morphological and anatomical. The data were processed using multivariate statistical methods: a) principal component analysis, b) stepwise regression analysis, and c) path coefficient analysis. Main results: The results showed that leaf stomata traits, particularly stomatal density (SD), and leaf dry mass per unit leaf area (LMA) were the most important traits, closely associated with WUEi. Stomatal density achieved the highest score on PC1 (0.825), in which WUEi had the highest loading (0.920), as well. SD was also included first in stepwise regression model. Research highlights: These results demonstrate that under moderate water stress WUEi in Q. robur half-sib lines were mainly the result of the plants' structural acclimation to surrounding environmental conditions.

There is insufficient available information on structural changes within wood-pastures including their relationship to abiotic influences such as livestock grazing, flooding and available soil nutrients. In this paper, we address the links between important environmental variables and different stages of the wood-pasture cycle, with the aim of understanding fluctuations in this relationship and processes that follow changes in wood-pasture condition. We used satellite and aerial image interpretation to identify structural vegetation shifts over 44 years under significantly declining livestock numbers. We used ground truthing of 24 plots to assess the current field scenario and employed canonical correspondence analysis (CCA) to evaluate the relationship between plant communities and environmental influences. Three dominant structural vegetation types grassland, transitional vegetation with thorny shrubs and woody encroachment were surveyed and the following set of variables was chosen: grazing intensity, inundation frequency, elevation, soil total nitrogen, soil available phosphorus, soil potassium, soil magnesium, soil calcium, soil pH and soil carbon to nitrogen ratio. Interpretation of satellite images revealed dominance of wood-pasture in the past, which alternated structurally between more open and more closed physiognomies. CCA with ground truthing data and forward selection revealed grazing intensity as the predominant ecological driver modifying vegetation structure, as well as transitioning vegetation patterns between open herbaceous and closed woody cover. Each structural vegetation type demonstrated a collective distribution pattern and a close relationship to certain abiotic drivers, indicating strong interactions between soil parameters, grazing pressure and vegetation composition.

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.

Background and Purpose: This paper analyses the lead tolerance and accumulation in white poplar genotypes in vitro, in order to optimize genotype evaluation and other procedures in their implementation in phytoremediation projects and landscaping in areas endangered by lead accumulation. Material and Methods: The lead tolerance and accumulation of five white poplar genotypes after 35 days in vitro cultivation on media supplemented with lead was examined. The following Pb(NO3)2 concentrations were used: 0, 10-6, 10-5, 10-4 and 10-3 M. Tolerance analysis (described by tolerance indices) was based on morphological parameters, biomass accumulation and the content of photosynthetic pigments, while lead accumulation was described by shoot lead accumulation and shoot lead content. Results and Conclusions: The chosen lead concentrations appeared not to be lethal. Moreover, the obtained results showed that the tested lead concentrations had a positive effect on: number of formed roots, shoot moisture content and shoot height. The best differentiation among the examined genotypes was gained by the tolerance index based on the shoot height on 10-4 M Pb(NO3)2. The shoot lead accumulation and shoot lead content significantly increased on 10-4 and 10-3 M Pb(NO3)2 media. Thus, the concentration of 10-4 M Pb(NO3)2 is recommended for further research. Two examined genotypes of horticultural value (LCM and LBM) achieved a significantly higher lead shoot content compared to the wide spread genotype “Villafranca” (almost 200% and 125% higher, respectively).