The Gram-negative bacterium Pseudomonas aeruginosa is a life-threatening nosocomial pathogen due to its generally low susceptibility toward antibiotics. Furthermore, many strains have acquired resistance mechanisms requiring new antimicrobials with novel mechanisms to enhance treatment options. Proline-rich antimicrobial peptides, such as the apidaecin analog Api137, are highly efficient against various Enterobacteriaceae infections in mice, but less active against P. aeruginosa in vitro. Here, we extended our recent work by optimizing lead peptides Api755 (gu-OIORPVYOPRPRPPHPRL-OH; gu = N,N,N′,N′-tetramethylguanidino, O = L-ornithine) and Api760 (gu-OWORPVYOPRPRPPHPRL-OH) by incorporation of Ile-Orn- and Trp-Orn-motifs, respectively. Api795 (gu-O(IO)2RPVYOPRPRPPHPRL-OH) and Api794 (gu-O(WO)3RPVYOPRPRPPHPRL-OH) were highly active against P. aeruginosa with minimal inhibitory concentrations of 8–16 and 8–32 μg/mL against Escherichia coli and Klebsiella pneumoniae. Assessed using a quartz crystal microbalance, these peptides inserted into a membrane layer and the surface activity increased gradually from Api137, over Api795, to Api794. This mode of action was confirmed by transmission electron microscopy indicating some membrane damage only at the high peptide concentrations. Api794 and Api795 were highly stable against serum proteases (half-life times >5 h) and non-hemolytic to human erythrocytes at peptide concentrations of 0.6 g/L. At this concentration, Api795 reduced the cell viability of HeLa cells only slightly, whereas the IC50 of Api794 was 0.23 ± 0.09 g/L. Confocal fluorescence microscopy revealed no colocalization of 5(6)-carboxyfluorescein-labeled Api794 or Api795 with the mitochondria, excluding interactions with the mitochondrial membrane. Interestingly, Api795 was localized in endosomes, whereas Api794 was present in endosomes and the cytosol. This was verified using flow cytometry showing a 50% higher uptake of Api794 in HeLa cells compared with Api795. The uptake was reduced for both peptides by 50 and 80%, respectively, after inhibiting endocytotic uptake with dynasore. In summary, Api794 and Api795 were highly active against P. aeruginosa in vitro. Both peptides passed across the bacterial membrane efficiently, most likely then disturbing the ribosome assembly, and resulting in further intracellular damage. Api795 with its IOIO-motif, which was particularly active and only slightly toxic in vitro, appears to represent a promising third generation lead compound for the development of novel antibiotics against P. aeruginosa.

The Lon protease is a mitochondrial matrix quality-control protease belonging to the family of AAA+ proteins (ATPases associated with many cellular activities). We had previously found Lon to be upregulated in lung tumours with a non-angiogenic phenotype in a microarray study comparing these to conventional angiogenic tumours. In this project I set out to investigate whether Lon had any role in modulating the hypoxic response of tumour cells. Using a novel monoclonal antibody against Lon, I found that upregulation of Lon was present in breast and lung tumours and that higher levels of Lon are correlated with shorter overall survival in breast cancer patients. Targeting Lon with siRNA and shRNA in tumour cell lines reduced the normoxic and hypoxic stabilisation of HIF-α subunits. This is mediated through a mechanism independent of the activity of HIF-prolyl hydroxylases and independent of any changes in mitochondrial transcription. I found that the pre-imported form of Lon could bind and chaperone VHL in the cytoplasm potentially modulating VHL activity. In cell lines and human tumours, I observed that the proline-hydroxylated form of HIF-1α is induced by hypoxia and the hydroxylated form of HIF-1α is associated with shorter overall survival in breast cancer patients. This observation supports the notion that higher levels of Lon is associated with poor survival by downregulating VHL leading to higher levels of hydroxylated HIF. Finally I show that targeting Lon in cell lines is able to inhibit growth in a cell-line dependent fashion and partially reverses the Warburg effect, increasing oxygen consumption and reducing lactate production. In conclusion, I have demonstrated the broad therapeutic potential of targeting the Lon protease in tumours and highlighted a mechanism of post-hydroxylation HIF-regulation that has not been previously recognised in VHL competent tumours.

4-Alkyl-L-proline derivatives (APD) are specialized precursors involved in the biosynthesis of at least three groups of different natural compounds: some pyrrolo-1,4-benzodiazepines with antitumor activity, bacterial hormone hormaomycin and clinically used lincosamide antibiotic lincomycin. These compounds share a biosynthetic pathway encoded by 5 or 6 homologous genes present in the biosynthetic gene clusters of the producing organisms. Similarities in biosynthesis and differences between APD structures of these compounds could be used to prepare a hybrid producing strain of biologically more effective lincomycin derivative. Unusual amino acid 4-propyl-L-proline (PPL) is the APD precursor of lincomycin. The originally proposed scheme of the PPL pathway does not comply with our current knowledge. Therefore, it was necessary to revise this scheme according to new results. The first two steps of the PPL pathway are functionally proved. Probing the next step was the main aim of this work. The protein LmbW was overproduced and its methyltransferase activity was confirmed in vitro. LmbW is able to directly methylate intermediate of second step of the pathway while the originally scheme proposed methylation at a later stage of biosynthesis. LmbW is also able to attach a longer alkyl chain to its substrate. This...

The aims of this work were 1) to assess whether sorghum (Sorghum bicolor (L.) Moench) genotypes originating from the India can be grown and analyzed in the climatic conditions of central Europe and 2) to find out the utilization potential of selected non- destructive and destructive methods based mostly on the chlorophyll a fluorescence measurements and the determination of photosynthetic pigments' content for the differentiation of sorghum genotypes based on their presumed drought tolerance. Field experiments made during 2 years compared 15 genotypes of this species (2 stay-green parental lines, 2 senescent parental lines and 11 introgression lines with stay-green loci), 2 of these genotypes were further analyzed in greenhouse conditions where the water deficit was induced by a cessation of watering for 12 days. The field-grown plants showed some differences between individual genotypes in all measured parameters; however, for the majority of the genotypes these differences were not statistically significant. The stay-green parental genotype B35 differred the most from the other ones in both field seasons, but the other stay-green genotypes usually did not differ from the senescent genotypes. No significant differences between both greenhouse-tested genotypes (presumably contrasting in their...

A cultura de soja (Glycine max (L.) Merrill) é muito importante mundialmente em função de seu uso na alimentação animal, como principal fonte de proteína e óleo, além de constituir a matéria prima básica para diversos setores da indústria. Sabe-se que a produtividade potencial de uma cultura é determinada por fatores genéticos e pelos seguintes atributos do ambiente de produção: temperatura, radiação solar, dióxido de carbono e fotoperíodo, sem restrição de água, nutrientes, plantas daninhas, pragas e doenças. A disponibilidade hídrica afeta o crescimento e desenvolvimento da cultura de soja, especialmente durante o período reprodutivo, fase de elevada atividade fisiológica. A deficiência hídrica é o principal fator limitante da produção mundial da cultura, que em sua maioria é cultivada em sistema de sequeiro em áreas com consideráveis riscos de ocorrência de deficit hídrico ao longo do ciclo da cultura. As plantas, ao longo do tempo, desenvolveram mecanismos para tolerar e/ou evitar os efeitos negativos desse estresse. O presente estudo teve como objetivo caracterizar esses mecanismos de tolerância associando-os ao potencial matricial do solo em processo de secagem. Foi avaliado a umidade gravimétrica e potencial matricial do solo, conteúdo relativo de água na folha, potencial hídrico foliar, prolina e caracteres relacionados à produção de plantas de soja, durante o período de enchimento de grãos, submetidas a três, seis, nove e doze dias sem irrigação, comparados com o controle irrigado diariamente. De acordo com o observado, os mecanismos de tolerância da soja à deficiência hídrica foram satisfatórios para a manutenção do conteúdo relativo de água e potencial hídrico foliar em níveis adequados até o valor de potencial matricial de água no solo de -0,8 atm, tendo a prolina papel importante nesse mecanismo. As avaliações hídricas, conteúdo relativo de água e potencial foliar se mostraram relacionados com a manutenção da produtividade de soja sob situação de deficiência hídrica, mostrando-se adequados para o estudo de tolerância de cultivares de soja à restrição hídrica. / Soybean crop (Glycine max (L.) Merrill) is very important worldwide because it is used in animal feeding, a source of protein and oil, and also raw material for many manufactured goods. It is known that a crop potential yield is determined by genetic factors and by the following production environment characteristics: temperature, solar radiation, carbon dioxide, photoperiod, water availability, nutrients, weeds, pests and diseases. Water availability affects soybean growth and development especially during the reproductive phase which has high physiological activity. Water deficit is a key factor on world`s soybean production which is generally grown in dry land systems with considerable high risks of water deficit along soybean crop season. Plants have developed mechanisms over time to tolerate and/or avoid negative effects of this kind of stress. The present study had as the main goal to characterize these mechanisms of tolerance associating them with soil matric potential of a soil in a drying process. It was assessed the gravimetrical humidity and soil matric potential, leaf relative water content, leaf water potential, proline and yield component of plants under three, six, nine and twelve days without irrigation during grain filling phase. In accordance with the observed, soybean water deficit tolerance mechanisms were enough to maintain leaf relative water content and leaf water potential at acceptable levels until soil matric potential were around -0.8 atm, which proline was a key factor in this mechanism. Leaf relative water content and leaf water potential showed to be related in maintaining soybean yield under water deficit and they may be used in studies of soybean tolerant cultivars to water restriction.

Na África subsaariana, as serpentes do gênero Bitis são de extrema importância, pois suas vítimas apresentam sintomas como dano local, hemorragia e uma severa hipotensão. Este trabalho identificou moléculas capazes de inibir a atividade da enzima conversora de angiotensina I (ECA) presentes no veneno de quatro serpentes do gênero Bitis. Para isto, as porções de baixa massa molecular desses 4 venenos foram fracionadas em RP-HPLC e as frações com boa inibição sobre a atividade da ECA foram analisadas por espectrometria de massas. Foram identificados 34 oligopeptídeos ricos em prolina (PRO), sendo 8 sintetizados e suas constantes de inibição (Ki) determinadas. Em testes com substratos naturais da ECA, angiotensina I e bradicinina, foi constatada a maior inibição da hidrólise da angiotensina I por quatro PROs. Todos os PROs in vivo reduziram a pressão arterial, e seis deles aumentaram a frequência cardíaca em ratos Wistar. Com isto, conclui-se que existem toxinas no veneno de serpentes do gênero Bitis responsáveis pela hipotensão. / In the sub-saharian Africa, snakes from the Bitis genus are of extreme medical importance, since its victims show symptoms as local tissue damage, hemorrhage and a severe hypotension. This work identified molecules that inhibit the angiotensin I converting enzyme (ACE) in the venom of 4 snakes from the Bitis genus. The low molecular portions of the venom of these snakes were fractionated in RP-HPLC and the fractions that efficiently inhibited the ACE activity were analyzed by mass spectrometry. 34 proline-rich oligopeptides were identified, 8 of them synthesized and had their inhibition constants (Ki) determined. In tests using natural substrates of ACE, angiotensin I and bradykinin, the angiotensin I hydrolysis were better inhibited by four PROs. In vivo tests results showed that all PROs decreased the mean arterial pressure and six of them increased the heart rate. Therefore, we can conclude that there are toxins present in the venom of Bitis capable of cause hypotension.

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No. of bitstreams: 2 license_rdf: 0 bytes, checksum: d41d8cd98f00b204e9800998ecf8427e (MD5) tese_isabel_lopes_vighi.pdf: 2167536 bytes, checksum: ac573c14d5ef3a4795a4bf594d60bd85 (MD5) Previous issue date: 2016-03-10 / Coordenação de Aperfeiçoamento de Pessoal de Nível Superior - CAPES / O arroz (Oryza sativa L.) é o segundo cereal mais cultivado no mundo, sendo o Brasil o 9º maior produtor. No entanto sua produção é severamente influenciada por condições ambientais adversas, dentre as quais se destaca a salinidade e baixas temperaturas. O objetivo deste estudo foi avaliar respostas, bioquímicas e moleculares desencadeadas pela salinidade e por baixas temperaturas em plantas de arroz, com respostas contrastantes. Para atingir este objetivo foram realizados três estudos: No primeiro, foram quantificados os teores do ânion Superóxido (O2•−), Peróxido de hidrogênio (H2O2), Peroxidação lipídica (MDA), atividade enzimática e expressão gênica diferencial das isoformas das enzimas Superóxido dismutase (SOD), Catalase (CAT), Ascorbato peroxidase APX e Glutationa reduzida (GR) em genótipos com tolerância contrastante ao sal. Para o genótipo sensível (BRS Pampa), ocorreu aumento na atividade enzimática da SOD, CAT, APX e diminuição nos níveis de MDA. No genótipo tolerante (BRS Bojuru), também foi observado aumento da atividade das enzimas SOD e CAT, porém diminuição nos níveis de MDA. A regulação positiva das diferentes isoformas da SOD, CAT e APX demonstrou a sua contribuição para o aumento na atividade da enzima. No segundo estudo, foram quantificados os teores de H2O2, MDA, atividade enzimática e expressão gênica da CAT, bem como análise do promotor e cis elementos, nos mesmos dois genótipos contrastantes frente ao estresse por salinidade e baixa temperatura. O genótipo BRS Pampa (sensível) foi o que mostrou maior proteção contra danos oxidativos frente ao estresse por frio, bem como aumento na atividade da CAT e do número de transcritos das isoformas OSCATA e OSCATB. No terceiro estudo, avaliou-se o metabolismo da prolina em plantas de arroz, através do padrão de expressão dos genes envolvidos na biossíntese e catabolismo desse aminoácido e verificou-se a correlação com o conteúdo de prolina produzido em condições de estresse por salinidade e baixa temperatura. O conteúdo de prolina foi maior no genótipo BRS Bojuru (tolerante) frente às duas condições de estresse testadas. Sob estresse salino, os níveis de transcritos dos genes de biossíntese estão mais correlacionados com o conteúdo de prolina no genótipo tolerante, enquanto que em condições de baixa temperatura a correlação é maior no genótipo sensível. Os resultados obtidos permitem concluir que, sob estresse salino e por baixa temperatura, as plantas de arroz apresentam respostas diferenciais a nível bioquímico e molecular, sendo estas respostas dependentes do tempo, intensidade do estresse e composição genética dos genótipos. / Rice (Oryza sativa L.) is the second most cultivated cereal in the world, and Brazil is the 9th largest producer. However production is severely affected by adverse environmental conditions, among which the mains ones are the salinity and low temperatures. The aim of this study was to evaluate responses, biochemical and molecular triggered by salinity and low temperatures in rice plants with contrasting response. To achieve this goal there were three studies: In the first one, the anion content superoxide (O2•-) were quantified, hydrogen peroxide (H2O2), lipid peroxidation (MDA), enzyme activity and differential gene expression of isoforms of the enzyme superoxide dismutase (SOD), catalase (CAT), ascorbate peroxidase APX and reduced glutathione (GR) in genotypes with contrasting tolerance to salt. For the sensitive genotype (BRS Pampa), there was an increase in the enzymatic activity of SOD, CAT, APX and a decrease in MDA levels. In tolerant genotype (BRS Bojuru) was also observed an increased activity of enzymes SOD and CAT, but a decrease in MDA levels. The upregulation of the different isoforms of SOD, CAT and APX has demonstrated its contribution to the increase in the enzyme activity. In the second study, H2O2 levels were quantified, MDA, enzyme activity and gene expression of CAT, and promoter analysis and cis-elements, the same two contrasting genotypes against salinity stress and low temperature. The BRS Pampa genotype (sensitive) was the one that showed greater protection against oxidative damage compared to the stress by cold, as well as an increase in CAT activity and the number of transcripts OSCATA and OSCATB isoforms. In the third study evaluated the metabolism of proline in rice plants through the expression pattern of genes involved in biosynthesis and catabolism of amino-acid and was found to correlate with the proline content produced under stress by salinity and low temperature. The proline content was higher in BRS Bojuru genotype (tolerant) against the two tested stress conditions. Under salt stress, the biosynthesis gene transcript levels are more correlated with the proline content in tolerant genotype, whereas in low temperature conditions the correlation is highest in sensitive genotype. The results showed that under salt stress and low temperature, the rice plants have differential responses to biochemical and molecular level, which are dependent responses of time, stress intensity and genetic makeup of the genotypes.

A cultura de soja (Glycine max (L.) Merrill) é muito importante mundialmente em função de seu uso na alimentação animal, como principal fonte de proteína e óleo, além de constituir a matéria prima básica para diversos setores da indústria. Sabe-se que a produtividade potencial de uma cultura é determinada por fatores genéticos e pelos seguintes atributos do ambiente de produção: temperatura, radiação solar, dióxido de carbono e fotoperíodo, sem restrição de água, nutrientes, plantas daninhas, pragas e doenças. A disponibilidade hídrica afeta o crescimento e desenvolvimento da cultura de soja, especialmente durante o período reprodutivo, fase de elevada atividade fisiológica. A deficiência hídrica é o principal fator limitante da produção mundial da cultura, que em sua maioria é cultivada em sistema de sequeiro em áreas com consideráveis riscos de ocorrência de deficit hídrico ao longo do ciclo da cultura. As plantas, ao longo do tempo, desenvolveram mecanismos para tolerar e/ou evitar os efeitos negativos desse estresse. O presente estudo teve como objetivo caracterizar esses mecanismos de tolerância associando-os ao potencial matricial do solo em processo de secagem. Foi avaliado a umidade gravimétrica e potencial matricial do solo, conteúdo relativo de água na folha, potencial hídrico foliar, prolina e caracteres relacionados à produção de plantas de soja, durante o período de enchimento de grãos, submetidas a três, seis, nove e doze dias sem irrigação, comparados com o controle irrigado diariamente. De acordo com o observado, os mecanismos de tolerância da soja à deficiência hídrica foram satisfatórios para a manutenção do conteúdo relativo de água e potencial hídrico foliar em níveis adequados até o valor de potencial matricial de água no solo de -0,8 atm, tendo a prolina papel importante nesse mecanismo. As avaliações hídricas, conteúdo relativo de água e potencial foliar se mostraram relacionados com a manutenção da produtividade de soja sob situação de deficiência hídrica, mostrando-se adequados para o estudo de tolerância de cultivares de soja à restrição hídrica. / Soybean crop (Glycine max (L.) Merrill) is very important worldwide because it is used in animal feeding, a source of protein and oil, and also raw material for many manufactured goods. It is known that a crop potential yield is determined by genetic factors and by the following production environment characteristics: temperature, solar radiation, carbon dioxide, photoperiod, water availability, nutrients, weeds, pests and diseases. Water availability affects soybean growth and development especially during the reproductive phase which has high physiological activity. Water deficit is a key factor on world`s soybean production which is generally grown in dry land systems with considerable high risks of water deficit along soybean crop season. Plants have developed mechanisms over time to tolerate and/or avoid negative effects of this kind of stress. The present study had as the main goal to characterize these mechanisms of tolerance associating them with soil matric potential of a soil in a drying process. It was assessed the gravimetrical humidity and soil matric potential, leaf relative water content, leaf water potential, proline and yield component of plants under three, six, nine and twelve days without irrigation during grain filling phase. In accordance with the observed, soybean water deficit tolerance mechanisms were enough to maintain leaf relative water content and leaf water potential at acceptable levels until soil matric potential were around -0.8 atm, which proline was a key factor in this mechanism. Leaf relative water content and leaf water potential showed to be related in maintaining soybean yield under water deficit and they may be used in studies of soybean tolerant cultivars to water restriction.

Made available in DSpace on 2017-07-10T17:37:10Z (GMT). No. of bitstreams: 1 Sarah_Romano_Ortolan.pdf: 975526 bytes, checksum: d72ab61fc7bdc88b6ddbab0eb3a86e42 (MD5) Previous issue date: 2015-07-28 / Coordenação de Aperfeiçoamento de Pessoal de Nível Superior / Wheat is considered the main cereal diet of the world population, but in recent years has achieved some gain in productivity of this culture despite having increased the use of nitrogen fertilizer. The use of plant growth promoting bacteria such as Herbaspirillum seropedicae SmR1 among others has been studied to obtain development of plants with less use of nitrogen fertilizers. However there is little information relating the effects of this interaction in plant development and grain yield. Objective of this study was to evaluate the carbon and nitrogen metabolism by certain enzymes, metabolites and indices related to the response to infectious stress on the wheat cultivars CD 104 and CD 120 in association with Herbaspirillum seropedicae bacteria. Two experiments were conducted. The experimental design was completely randomized with four replications in a 4x2. The first factor relates to the conditions inoculation with bacteria and/or nitrogen source in coverage are: control without inoculation with bacteria or added nitrogen fertilizer (C); application of nitrogen fertilizer (50 kg.ha-1) 30 days after sowing (N); inoculation with 106 cells of the bacterium H. seropedicae/seed at planting (Hs) and inoculation with bacteria combined with the application of nitrogen fertilizer (Hs + N) and the second factor refers to the phenological stages (tillering and booting). The results indicated that inoculation with H. seropedicae in wheat seeds of cv.s CD 104 and CD 120 in the two growth stages answered in relation to the indices related to stress with the involvement of enzymes of carbon and nitrogen metabolism. However prominent effect was not noticed to promote plant growth of wheat in late development, nor a deleterious effect of the bacterium for inoculation cv. CD 104 under the experimental conditions. For cv. CD 120 the differential effects indicate lower levels of stress and some level of association to positive effect on productivity when combined inoculation of bacteria to nitrogen fertilization. It was concluded that as well as pathogenic and stressors, H. seropedicae able to beneficially associate with wheat also provides similar interference pattern of carbon and nitrogen metabolism and stress levels / O trigo é considerado o principal cereal da dieta da população mundial, entretanto nos últimos anos tem se obtido pouco ganho de produtividade desta cultura apesar de se ter aumentado o uso de fertilizante nitrogenado. O uso de bactérias promotoras do crescimento vegetal, como Herbaspirillum seropedicae SmR1 entre outras, tem sido estudado para se obter desenvolvimento de plantas com menor uso de fertilizantes nitrogenados. Entretanto existem poucas informações que relacionam os efeitos desta interação no desenvolvimento da planta e de produtividade de grãos. Objetivo deste trabalho foi avaliar o metabolismo de carbono e nitrogênio através de algumas enzimas, metabólitos e índices relacionados à resposta ao estresse infeccioso em trigo das cultivares CD 104 e CD 120 em associação com a bactéria Herbaspirillum seropedicae em dois estádios fenológicos. Foram realizados dois experimentos. O delineamento experimental utilizado foi o inteiramente ao acaso, com 4 repetições, em esquema fatorial 4x2. O primeiro fator refere-se às condições de inoculação com bactéria e/ou fertilização nitrogenada em cobertura, sendo: controle, sem inoculação com bactéria ou adição de fertilizante nitrogenado (C); aplicação de fertilizante nitrogenado (50 kg.ha-1) após 30 dias da semeadura (N); inoculação de 106 células da bactéria H. seropedicae/semente na semeadura (Hs) e inoculação com a bactéria combinada com a aplicação de fertilizante nitrogenado (Hs + N) e o segundo fator refere-se aos estádios fenológicos (perfilhamento e emborrachamento). Os resultados indicaram que a inoculação com H. seropedicae em sementes de trigo das cv.s CD 104 e CD 120 nos dois estádios fenológicos responderam em relação aos índices relacionados ao estresse com envolvimento das enzimas do metabolismo de carbono e nitrogênio. Entretanto não foi percebido efeito proeminente de promoção do crescimento vegetal no final do desenvolvimento do trigo, tampouco efeito deletério da inoculação de bactéria para a cv. CD 104, nas condições experimentais. Para a cv. CD 120 os efeitos diferenciais indicam menor nível de estresse e algum nível de associação para efeito positivo na produtividade quando combinada a inoculação da bactéria com a fertilização nitrogenada. Foi possível concluir que assim como para agentes patogênicos e estressantes, a H. seropedicae, capaz de associar beneficamente com trigo também apresenta padrão semelhante de interferência do metabolismo de carbono e nitrogênio e índices de estresse

<p>Cilj ove doktorske disertacije predstavljalo je ispitivanje uticaja vodnog stresa izazvanog&nbsp; sa polietilen glikolom (PEG)&nbsp; 6000 na biohemijske osobine klonova topole. Reznice tri klona topole (M-1, B-229 i PE19/66) su hidroponski gajene i izložene 100 mOsm i 200 mOsm PEG 6000&nbsp; tokom &scaron;est dana. Nakon tretmana u kontrolnim i stresiranim<br />biljkama su&nbsp; ispitane promene aktivnosti antioksidantnih enzima,&nbsp; aktivnost dva<br />enzimska markera polifenolnog metabolizma, fenolni profil, antioksidantna aktivnost, kao i sadržaj prolina i glicin betaina&nbsp; (GB). Takođe, ispitan je biohemijski odgovor kulture tkiva klona M-1 na vodni stres izazvan sa PEG 6000.</p><p>U oba eksperimenta uočeno je da je akumulacija prolina i GB u uslovima vodnog stresa izazvanog sa PEG 6000&nbsp;najvažnija strategija u otpornosti na stres i prevazilaženju vodnog deficita, tako da se&nbsp;ovi parametri mogu definisati kao najbolji indikatori otpornosti topole na su&scaron;u.</p><p>Rezultati ovog istraživanjaupotpunjavaju saznanja o povezanosti oksidativnog stresa uzrokovanog vodnim stresom sa antioksidantnim odgovorom, nivoom o&scaron;tećenja lipida i proteina, uključivanjem osmolita kao i promenama u polifenolnom profilu. Takođe, dobijeni rezultati su značajni za odabir klonova topola tolerantnih na vodni stres&nbsp; i&nbsp; za<br />po&scaron;umljavanje su&scaron;nih stani&scaron;ta, kojih će biti sve vi&scaron;e usled negativnog uticaja klimatskih promena.</p> / <p>The aim of presented doctoral thesis was investigation of the impact of water deficit caused by polyethylene glycol (PEG) 6000 on biochemical features of poplar clones.&nbsp; During six days, cuttings of three poplar clones (M-1, B-229 and PE 19/66) were grown in hydroponics and subjected to 100 and 200 mOsm PEG 6000. Changes in the activity of antioxidant enzymes, polyphenol characterization, two enzymatic markers of polyphenol metabolism, antioxidant capacity, as well as proline and glycine betaine content were investigated in stressed plants. Furthermore,&nbsp; assessment of&nbsp; biochemical response of poplar clone M-1 tissue culture to water stress induced by PEG 6000 was performed.</p><p>In both experiments, the most important strategy for stress resistance and overcoming water deficiency was accumulation of proline and glycine betaine so these parameters can be defined as the best indicators of poplar resistance to drought.</p><p>The results of this study complete&nbsp; the findings on the oxidative stress caused by water stress&nbsp; and their relationship with the antioxidant response, the level of lipid andprotein damage, accumulation of the osmolites and changes in the polyphenol profile. Also, the obtained results may be important for the selection of poplar clones resistant to water stress and for afforestation in arid sites, which appeared to be more numerous due to the effects of&nbsp; global climate change.</p>