Submitted by Gabriela Lopes (gmachadolopesufpel@gmail.com) on 2016-09-28T12:43:45Z No. of bitstreams: 2 license_rdf: 0 bytes, checksum: d41d8cd98f00b204e9800998ecf8427e (MD5) DISSERTAÇÃO CD 2.pdf: 3402780 bytes, checksum: 6767c83b34c1fb208e8cd7bc97b095d1 (MD5) / Approved for entry into archive by Aline Batista (alinehb.ufpel@gmail.com) on 2016-09-28T17:17:17Z (GMT) No. of bitstreams: 2 license_rdf: 0 bytes, checksum: d41d8cd98f00b204e9800998ecf8427e (MD5) DISSERTAÇÃO CD 2.pdf: 3402780 bytes, checksum: 6767c83b34c1fb208e8cd7bc97b095d1 (MD5) / Made available in DSpace on 2016-09-28T17:17:17Z (GMT). No. of bitstreams: 2 license_rdf: 0 bytes, checksum: d41d8cd98f00b204e9800998ecf8427e (MD5) DISSERTAÇÃO CD 2.pdf: 3402780 bytes, checksum: 6767c83b34c1fb208e8cd7bc97b095d1 (MD5) Previous issue date: 2014-02-28 / Conselho Nacional de Pesquisa e Desenvolvimento Científico e Tecnológico - CNPq / O crescimento populacional mundial tem contribuído para uma maior demanda de arroz, enquanto que alterações ambientais, causadas pelo homem, contribuem com o aumento da frequência de precipitações. Estas condições formam um ambiente de hipoxia, e restringem a produção do arroz. Frente a isso as plantas respondem com adaptações morfológicas e alterações em nível molecular. Portanto, este trabalho teve como objetivo avaliar os efeitos morfológicos e anatômicos em cultivares de arroz Oryza sativa L. submetidas a hipoxia e verificar o perfil de expressão dos genes de regulação complexa e simples de fatores de transcrição WRKY envolvidos na resposta ao estresse por hipoxia. Os resultados obtidos demonstraram que ocorre alterações morfológicas e anatômicas nas cultivares de arroz irrigado quando submetidas ao estresse por hipoxia, principalmente no sistema radicular. Ainda, o aerênquima é mais desenvolvido nas cultivares Epagri 108 e Nipponbare quando submetidas ao estresse por hipoxia. No estudo de expressão gênica através da técnica de qRT-PCR, foi possível observar que os genes WRKY responderam ao estresse por hipoxia, mas não foi possível, para este estresse, verificar uma diferença entre a resposta do perfil de expressão dos genes de regulação simples e complexa em plântulas de arroz sob estresse por hipoxia. / The growth of the world’s population has contributed for a greater demand of rice while environmental alterations caused by men contribute to an increase of rainfall. This conditions create a hypoxia environment and restrict the production of rice. Under these conditions, plants with morphological adaptations and alterations in molecular level. Thus, this work had as its objective to evaluate the morphological and anatomical effects in rice cultivars Oryza sativa L. submitted to hypoxia and to verify the expression profile of genes of complex and simple regulation of WRKY transcription factors involved in the response of stress by hypoxia. The results obtained demonstrated that morphological and anatomical alterations occur in cultivars of irrigated rice when submitted to stress by hypoxia, manly in their root system. Furthermore, the aerenchyma is more developed in the cultivars Epagri 108 and Nipponbare when submitted to stress by hypoxia. In the study of genic expression through the qRT-PCR technique, it was possible to observe that the WRKY genes responded to stress by hypoxia but it not possible, for this stress, to verify a difference between the response of the expression profile of genes of simple and complex regulation in seedlings of rice under stress by hypoxia.

Pappelsorten (Populus) für den Anbau in Kurzumtriebsplantagen sollten neben einer hohen Biomasseproduktion und der Vermehrbarkeit über Steckhölzer auch Trockentoleranz aufweisen. Trockenheit stellt einen wichtigen abiotischen Stressor dar, der das Wachstum der Pflanze, ihre Vitalität und ihre Produktivität negativ beeinflussen kann. Polyploide Sorten von Getreidepflanzen (Triticum), krautigen Pflanzen (Lonicera, Spathiphyllum und Nicotiana) aber auch Baumarten (Betula) zeigen eine höhere Toleranz gegenüber Stress, wie z.B. Trockenstress, im Vergleich zu Sorten mit niedrigerem Ploidiegrad. Hybridaspen Populus tremula (L.) x P. tremuloides (Michx.) der Sektion Populus können auch auf Grenzertragsböden ökonomische Zuwächse verzeichnen, auf denen andere Baumarten ausfallen. Den Hybridaspen fehlt allerdings die Vermehrbarkeit über Steckhölzer. Anderen Pappelarten der Sektionen Tacamahaca und Aigeiros wie beispielsweise P. nigra (L.) and P. trichocarpa (Torr. & Gray) x P. deltoides (Bartram ex Marsh) zeigen diese entscheidende Eigenschaft. Da Kreuzungen zwischen den Sektionen Populus und Tacamahaca oder Aigeiros auf natürlichem Weg schwierig sind, wurde die Methode der somatischen Hybridisierung eingesetzt. Der Klon P. tremula x P. tremuloides ('Münden 2') wurden als ein Fusionspartner und einer der Klone P. x canescens (INRA clone No. 717 1‑B4), P. nigra oder P. trichocarpa x P. deltoides (B19) als zweiter Fusionspartner in Hybridisierungsexperimenten verwendet. Blattmorphologisch und durch Mikrosatellitenanalysen konnten die erhaltenen Fusionsprodukte dem Ausgangsklon P. tremula x P. tremuloides ('Münden 2') zugeordnet werden, wiesen dabei aber einen tetraploiden Chromosomensatz auf. In der vorliegenden Dissertation war das zentrale Ziel, Trockenstressreaktionen tetraploider Hybridaspenlinien (HAL) im Vergleich zur diploiden Ausgangslinie P. tremula x P. tremuloides ('Münden 2') zu untersuchen. Dazu wurden die folgenden Fragen aufgestellt. (i) Sind die tetraploiden HAL dem diploiden Ausgangsklon unter Trockenheit überlegen z.B. hinsichtlich des Wasserverbrauchs und des Verwelkungs- und Vertrocknungszeitpunktes der Blätter? (ii) Unterscheiden sich die tetraploiden HAL von dem diploiden Ausgangklon in morphologischen Ausprägungen wie beispielsweise der Stomatamorphologie, der Höhe, der Biomasse und der Gesamtblattfläche? (iii) Ist die höhere Trockentoleranz der tetraploiden HAL durch den erhöhten Ploidiegrad verursacht? (iv) Zeigen die tetraploiden HAL unter Trockenstress früh physiologische Veränderungen im Kohlenhydratgehalt, dem relativen Blattwassergehalt und der stomatären Leitfähigkeit? (v) Gibt es genetische Modifikationen abgesehen von der Tetraploidie wie z.B. Duplikationen und Deletionen, die eventuell Einfluss auf das Trockensressverhalten haben könnten? In einem ersten Experiment wurden vier tetraploide HAL untersucht. Zwei HAL zeigten einen geringeren Wasserverbrauch und drei HAL eine spätere Verwelkung als der diploide Ausgangsklon. Außerdem entwickelten die tetraploiden HAL eine geringere Stomatadichte, größere Stomata und eine vergleichbare oder geringere Höhe, Biomasse und Gesamtblattfläche im Vergleich zum diploiden Ausgangsklon. Das zweite Experiment wurde mit vier diploiden und 16 weiteren tetraploiden HAL durchgeführt. Auch die diploiden Linien stammten aus Protoplastenfusions-experimenten. Die tetraploiden HAL wiesen im Mittel eine höhere Überlebensrate als die diploiden HAL auf, entwickelten aber durchdschnittlich ein geringeres Höhenwachstum. Unter Berücksichtigung der Höhe verwelkten 11 tetraploide, aber keine diploide HAL später als der diploide Ausgangsklon. In einem dritten Experiment wurden zwei Linien aus dem zweiten Experiment ausgewählt, die vergleichbares Höhenwachstum zeigten, aber später verwelkten als der diploide Ausgangklon. Es wurden physiologische Merkmale wie der Wasserverbrauch und der Verwelkungszeitpunkt der Blätter beobachtet und der relative Blattwassergehalt, die stomatäre Leitfähigkeit und der Kohlenhydratgehalt in einer Zeitreihe untersucht. Geringerer Wasserverbrauch im Vergleich zum diploiden Ausgangsklon konnte in beiden tetraploiden HAL gezeigt werden. Diese Beobachtung deckt sich mit dem höheren relativen Bodenwassergehalt der tetraploiden HAL im Vergleich zum diploiden Ausgangsklon. Insgesamt spricht das für geringeren Trockenstress in den tetraploiden HAL. Auf der anderen Seite sollte ein geringerer Wasserverbrauch auch eine geringere stomatäre Leitfähigkeit nach sich ziehen. Dies war in den tetraploiden HAL nicht durchgehend der Fall. Relativ zur Kontrolle konnten die tetraploiden Linien aber eine höhere stomatäre Leitfähigkeit bei Bodenwassergehalten aufrechterhalten, bei denen die diploide Ausgangslinie bereits ihre Stomata geschlossen hatte. Dies weist ebenfalls auf eine höhere Trockentoleranz der tetraploiden HAL hin. Genetische Modifikationen wie beispielsweise Duplikationen und Deletionen wurden mit Hilfe von “copy number variation”(“Kopienzahlvariation”)-Analysen an drei tetraploiden HAL und der diploiden Ausgangslinie durchgeführt. Die drei tetraploiden Linien zeigten vergleichbares Höhenwachstum, aber spätere Verwelkung als die diploide Ausgangslinie. Bei dieser Analyse konnten nur drei Segmente detektiert werden, die bei allen drei tetraploiden HAL im Vergleich zum diploiden Ausgangsklon verändert vorlagen. Die putativen Gene der detektierten Segmente wiesen keine Annotation auf. Eine der drei tetraploiden HAL zeigte eine hohe Anzahl von 90 Segmenten, die hier aber weder in dem diploiden Ausgangsklon noch in den beiden anderen tetraploiden Linien erhöht vorlagen. Diese Linie zeigte zwar eine höhere Trockentoleranz als der diploide Ausgangsklon, aber nicht gegenüber der anderen untersuchten tetraploiden HAL (Experiment 3). Beide tetraploiden HAL waren bezüglich des Wasserverbrauchs, des Verwelkungszeitpunktes und der stomatären Leitfähigkeit ähnlich. Dies lässt vermuten, dass die Trockentoleranz durch die Tetraploidie und nicht durch weitere genetische Modifikationen bedingt ist. Trotzdem ist eine weiterführende funktionelle Untersuchung der drei Segmente, die in allen tetraploiden HAL verändert auftraten, sinnvoll, da auch hier ein trockenrelevantes Gen verändert sein könnte. Um eine Relevanz der putativen Gene bezüglich Trockenheit zu testen, könnte zukünftig das Verhalten von Knock-out-Mutanten, denen die entsprechende Sequenz fehlt, unter Trockenstress untersucht werden. Abschließend ist zu bemerken, dass neben den hier untersuchten HAL mehrere hundert Fusionsprodukte in einem Verbundprojekt genetisch charakterisiert wurden. Die Heterofusionsfrequenz war äußerst gering und die Hybriden waren bereits in der in vitro Phase nicht überlebensfähig. Geringe Heterofusionsfrequenz wurde beteits in anderen Arten beobachtet (Cyclamen, Brassicaceae, Fabaceae, Poaceae, Solanaceae). Da in dieser Dissertation nur Homofusionslinien verwendet wurden, wurde die Vermehrbarkeit über Steckhölzer nicht getestet, sondern der Fokus auf das Trockenstressverhalten der tetraploiden Homofusionslinien gelegt. Zusammenfassend zeigen die Ergebnisse, dass veränderte morphologische Ausprägungen und eine höhere Trockentoleranz in den HAL mit erhöhten Ploidiegrad auftraten. Dies prädestiniert die tetraploiden Linien für Standorte, an denen Wasser als limitierender Faktor auftritt und hohe Ausfallraten durch Dürre erwartet werden.

A protocol for cauliflower micro-propagule production was developed and optimised for both micropropagation and artificial seed production techniques using meristematic tissues from cauliflower curd. All steps in the protocol were empirically optimised including: blending, sieving, culture methods, liquid culture media composition and plant growth regulator combinations and concentrations. The cost of the micro-propagules could be reduced by as much as 50% on the initial costings reported previously since treatments doubled the number of microshoots produced per culture unit. The research confirmed the suitability of cauliflower microshoots to be encapsulated as artificial seeds and an effective protocol for microshoot encapsulation was designed through the optimization of 1) the production of cauliflower microshoots suitable for encapsulation, 2) encapsulation procedures, 3) artificial seed artificial endosperm structure, 4) conversion materials. The possibility of culturing cauliflower artificial seeds in commercial substrates such as perlite, sand, vermiculite and compost was confirmed. The use of plant preservative mixture (PPM) for the control of contamination in cauliflower culture media and artificial seeds was optimised and the effect of this material on the development of plant material was assessed. It was confirmed that cauliflower artificial seed could be stored in a domestic refrigerator for up to 6 months which could have a great impact in cauliflower breeding programmes. The huge number of cauliflower microshoots that could be produced using this protocol and the homogeneity of the culture system, provided a tool for the molecular analysis of cauliflower microshoots (and artificial seed) abiotic stress tolerance analysis. Various treatments were conducted to improve microshoot cold tolerance and the up-regulation of the CBF/DREB1 transcription factor including low temperature acclimation, mannitol, ABA (abscisic acid) and Mo (molybdenum). Microshoots were confirmed to acclimate successfully using low temperature. Mo was shown to improve the cold tolerance of cauliflower microshoots and to up-regulate CBF/DREB1 in the absence of low temperature acclimation. Acclimation did not increase the accumulation of dehydrin proteins and it is concluded that dehydrins do not play a significant role in the cold tolerance of cauliflower microshoots. Since cauliflower breeding and seed multiplication protocols make extensive use of micropropagation, the studies reported in this research could make a significant impact by decreasing the cost of micropropagation and increasing its reliability. It also opens new perspectives for further research for cauliflower artificial seed production and the possibility of sowing these seeds directly in the field. Furthermore, this research helps to facilitate cauliflower breeding programmes by improving the understanding of abiotic stress tolerance mechanisms and the relationship between different types of abiotic stresses such as cold and drought.

Plant microRNAs play important roles in plant growth and development. Here we investigated the roles of miRNAs in the plant abiotic stress, development and viral infection. MicroRNA membrane array analysis using five different abiotic stress treatments resulted in the identification of 8 novel stress inducible miRNA-families. Functional studies on novel stress inducible miR168 revealed its functional relation with abiotic stress. Over expression of miR168 in Arabidopsis showed upregulation of four stress related miRNAs (miR163, miR167, miR398 and miR408). Analysis of 9 independent transgenic lines showed induction of miR398, an oxidative stress responsive miRNA with a corresponding down regulation of its target genes. Heavy metal oxidative stress tolerance bioassays confirmed the susceptibility of transgenics compared to the wild types indicating the fact that the miR168 is indirectly involved in plant abiotic stress by inducing other stress responsive miRNAs. MicroRNAs are highly conserved across the plant kingdom. A miRNA atlas was drafted for different tomato organs and fruit stages using the known miRNA sequences from different plants species. A large variation in both number and level of miRNA expression pattern was observed among different organs as well as among fruit stages. In the present investigation, we have found a window of expression for different miRNAs during the fruit development. A gradual decrease in the expression levels of miR160h, miR167a and miR399d and a gradual increase in miR164a have been noticed towards the fruit maturation while miR398b showed dual peaks during fruit development indicating a potential role of various miRNAs in fruit development and maturation. Sonchus yellow net virus (SYNV) infected Nicotinana benthamiana leaves showed severe disease symptoms at two weeks post infection (WPI) and gradually recovered from the SYNV infection after 4-5 WPI correlating with the overall miRNA levels. The miRNA array and northern analysis showed an overall reduction of miRNA biogenesis during 2WPI followed by restoration to normal levels supporting the idea that the SYNV indeed interfered with the host miRNA levels which caused the symptoms and recovery phenotypes. Overall studies on plant abiotic stress, development and viral infection showed important roles of miRNAs in different aspects of plant life.

Increases in temperature damage plant cells, and plants react to heat stress by inducing of a number of protective mechanisms. In this study, it has been shown that heat damages cells both directly while the plants are being heated, and indirectly through heat induced oxidative stress during recovery from heating. Different stress response pathways are induced in each case. Evidence implicates salicylic acid, abscisic acid, ACC (a precursor of ethylene), calcium ions and active oxygen species in pathways resulting in thermotolerance (i.e. increased survival at high temperature). Addition of these potential second messengers results in increased survival and decreased oxidative damage after heating (as measured using the TBARS assay), while silencing the pathways through use of mutants, inhibitors or transgenes results in decreased thermotolerance. In vivo calcium measurements show cytosolic calcium transients only at initiation of cooling after heat stress. Calcium chloride also induces thermotolerance when added after heating, as does ACC. These substances can induce early increases in ascorbate peroxidase activity after heating, and induce expression of antioxidant genes. Thus they may play a role in heat induced oxidative stress response pathways in recovery. Two different pathways induced during heating appear to result in the expression of genes for heat shock proteins. The predominant pathway induced at 30oC involves ABA, while that at 35-40oC involves an oxidative burst generated through the NADPH oxidase, atrbohB. Addition of SA can induce HSP (heat shock protein) expression, but there is little evidence that this occurs endogenously in plant cells, although nahG plants unable to signal via SA are thermosensitive. Results in this study indicate that there are several signalling pathways associated with heat shock. The pathway induced during recovery from heating involves calcium ions and ethylene, and results in increased antioxidant capacity. During heating two pathways induce expression of HSPs: one involving abscisic acid and possibly salicylic acid and one involving an oxidative burst. At least one further pathway is believed to exist, which involves protein kinases and phosphatases as heat shock causes up-regulation of expression of certain genes for these signalling components.

Doctor of Philosophy / Department of Horticulture, Forestry, and Recreation Resources / Sunghun Park / Plants are continuously exposed to numerous abiotic stresses, which adversely affect plant growth, development, and yield. Plants have developed different signaling pathways to cope with abiotic stresses, and some of the pathways converge to help plants tolerate simultaneous stresses. Here, we report ectopic expression of an Arabidopsis glutaredoxin AtGRXS17 that confers tolerance to multiple abiotic stresses in tomato plants. In yeast assays, AtGRXS17 co-localized with yeast ScGrx3 in the nucleus and suppressed the sensitivity of yeast grx3grx4 double mutants to oxidative stress and heat shock. In plants, GFP-AtGRXS17 fusion proteins initially localized in the cytoplasm but migrated to the nucleus during heat stress. Ectopic expression of AtGRXS17 in tomato plants minimized photo-oxidation of chlorophyll and reduced oxidative damage of cell membrane systems under heat stress. Furthermore, expression of the heat shock transcription factor (HSF) and heat shock protein (HSP) genes was up-regulated in AtGRXS17-expressing tomato plants during heat stress when compared to wild-type controls. Under cold, drought, and oxidative stress conditions, AtGRXS17-expressing tomato plants also displayed more vigorous growth and less physiological damage than those of the wild-type control plants. Quantitative real-time PCR (qRT-PCR) analysis indicated that expression of AtGRXS17 alters multiple stress defense signaling pathways, including the Abscisic Acid (ABA) and C-Repeat Binding Factors (CBF) pathways. The results revealed a conserved function for a glutaredoxin protein in abiotic stress adaptation, and manipulation of AtGRXS17 may be a useful approach to improve crop stress tolerance and understand plant signaling under abiotic stress conditions. Deregulated expression of an Arabidopsis H[superscript]+/Ca[superscript]2[superscript]+ antiporter (sCAX1) in agricultural crops increases total calcium (Ca[superscript]2[superscript]+) but may result in yield loses due to calcium-deficiency like symptoms. Here we demonstrate that co-expression of a maize calreticulin (CRT, a Ca[superscript]2[superscript]+ binding protein located at endoplasmic reticulum) in sCAX1-expressing plants mitigated these adverse effects while maintaining enhanced Ca[superscript]2[superscript]+ content. Co-expression of CRT and sCAX1 could alleviate the hypersensitivity to ion imbalance in tobacco plants. Furthermore, blossom-end rot (BER) in tomato may be linked to changes in CAX activity and enhanced CRT expression mitigated BER in sCAX1 expressing lines. These findings suggest that co-expressing Ca[superscript]2[superscript]+ transporters and binding protein at different intracellular compartments can alter the content and distribution of calcium within the plant matrix.

No description available.

As reações luminosas da fotossíntese em plantas envolvem quatro complexos proteicos multi-unidades na membrana dos tilacóides incluindo o fotossistema II (PSII), o complexo citocromo b6f, o fotossistema I (PSI) e o complexo ATP sintase. Uma atividade apropriada desse processo exige um mecanismo de controle de qualidade mediado por chaperonas, DnaJs e proteases, como o complexo FtsH. Esse conjunto de proteínas garantem um dobramento correto de proteínas, as montagens devidas dos complexos e a degradação de algumas subunidades danificadas quando necessário. Neste trabalho nós mostramos o envolvimento de FIP, uma proteína com um domínio dedo-de-zinco localizada nos tilacóides de cloroplastos de A. thaliana, no mecanismo de resposta à estresses abióticos. Plantas mutantes fip foram, fenotipicamente, mais tolerantes à estresses abióticos de alta luminosidade, elevado potencial osmótico e excesso de sal. Também mostramos que a expressão de FIP é diminuída em resposta às diferentes condições de estresse, assim como o acúmulo de transcritos de genes relacionados à estresse foi menor nas plantas mutantes fip. Análises por immunoblot mostraram que os mutantes fip acumulam menos proteínas PsaA e PsaB do fotossistema I e plastocianina (PC) do que as plantas selvagens, no entanto não são afetados quanto ao acúmulo de proteínas do fotossistema II e do Complexo do Citocromo b6f sob condições controle. Esses mutantes também acumulam menos FtsH5 nos tilacóides, sem afetar a eficiência dos fotossistemas I e II. Foi testado também o potencial redutase do domínio dedo-de-zinco da proteína recombinante FIP (6xHis-FIP) em ensaios in vitro de redução de insulina. Vimos que FIP apresenta atividade redutase, significantemente, maior que o controle negativo nas condições testadas. Considerando todos os resultados obtidos até o momento, acreditamos que FIP possa estar agindo como uma redutase na membrana dos tilacóides, tendo como alvos não somente FtsH5, mas também outras proteínas com resíduos de cisteína nas suas estruturas, e que sua atividade tem influência no acúmulo de proteínas dependentes de redução para a maturação como PsaA, PsaB e PC. Uma investigação mais aprofundada da atividade de FIP nos cloroplastos ainda é necessária para o completo entendimento da sua função. / The light-driven photosynthetic reactions in plants take place within four multi- subunit protein complexes in the thylakoid membranes, including photosystem II (PSII), the cytochrome b6f complex, photosystem I (PSI) and the ATP synthase complex. Regulation of all these molecular machineries requires a fine-tuning control mechanism mediated by specific proteins, including chaperones, DnaJs, and proteases, such as the FtsH complex. These set of proteins guarantee the proper folding, assembly and degradation of the photosynthetic complexes\' subunits. In this work we showed the involvement of FIP, a zinc-finger protein localized in the thylakoid membranes in A. thaliana, in the abiotic stress response mechanism. Mutants fip knockdown plants were phenotypically more tolerant to abiotic stresses like high light, increased osmotic potential and salt excess. We also showed that FIP is down-regulated by different abiotic stresses, with lower levels of stress-related gene transcripts accumulation in mutant fip plants. Analysis of accumulation of photosynthetic proteins by immunoblot under control conditions showed that mutants fip displayed lower levels of PsaA, PsaB (PSI) and Plastocyanin (PC) proteins than wild-type plants, however are not affected for PSII and Cyt b6f proteins accumulation under the same growth conditions. In addition, the mutants accumulated slightly less FtsH5 proteins in thylakoid membranes, without affecting PSII and PSI efficiency. We tested the putative reductase activity probably mediated by FIP zinc-finger domain, using the recombinant form of the protein 6xHis-FIP in in vitro insulin reduction assays. FIP presented a reductase activity higher than the negative control under the same assay conditions. Taking all together, these results suggest that FIP may be acting as a reductase in the thylakoid membranes, having as targets not only FtsH5 but other targets with available cysteine residues, depending on the reduction step for proper accumulation such as PsaA, PsaB and PC. Further investigations regarding the role of FIP in chloroplasts are still necessary to completely understand its function.

In this study poplar trees have been examined under different stress conditions. Apart from the detailed descriptions above two main conclusions might be drawn: i) A small plant like Arabidopsis thaliana is highly susceptible to stress situations that might become life-threatening compared to a tree that has extremely more biomass at its disposal. Such an organism might be able to compensate severe stress much longer than a smaller one. It seems therefore reasonable that a crop like Arabidopsis reacts earlier and faster to a massive threat. ii) In poplar both tested stress responses seemed to be regulated by hormones. The reactions to abiotic salt stress are mainly controlled by ABA, which also has a strong impact upon cold and drought stress situations. The term commonly used for ABA is “stress hormone” and is at least applicable to all abiotic stresses. In case of herbivory (biotic stress), jasmonic acid appears to be the key-player that coordinates the defence mechanism underlying extrafloral nectary and nectar production. Thus the presented work has gained a few more insights into the complex network of general stress induced processes of poplar trees. Future studies will help to understand the particular role of the intriguing indirect defence system of the extrafloral nectaries in more detail. / In dieser Arbeit wurden Pappelbäume unter verschiedenen Stressbedingungen untersucht. Zusammenfassend und zusätzlich zu den obigen Beschreibungen lassen sich zwei Schlussfolgerungen ziehen: i.) Eine kleine Pflanze wie Arabidopsis ist viel empfindlicher für Stresssituationen, die möglicherweise lebensbedrohlich werden könnten, im Gegensatz zu einem Baum mit wesentlich grösserer Biomasse . Solch ein Organismus kann schwerwiegendem Stress viel länger kompensieren als ein kleinerer Organismus. Es erscheint daher sinnvoll, dass eine Pflanze wie Arabidopsis viel früher und schneller auf eine massive Bedrohung reagiert. ii.) In Pappeln scheinen beide untersuchten Arten von Stressreaktion durch Hormone reguliert zu werden. Die Reaktionen auf abiotischen Salzstress werden hauptsächlich durch ABA kontrolliert, welches auch einen starken Einfluss auf Kälte- und Trockenstressszenarien hat. Üblicherweise wird für ABA der Ausdruck "Stress-Hormon" verwendet, was zumindest für abiotischen Stress zutreffend ist. Im Fall von Herbivorie (biotischer Stress) scheint Jasmonsäure die Schlüsselrolle zu spielen, die die Abwehrmechanismen koordiniert, die den extrafloralen Nektarien und der Nektarproduktion zu Grunde liegt. Demzufolge hat die vorliegende Arbeit ein paar neue Einsichten in das komplexe Netzwerk der Stress-induzierten Prozesse der Pappel ermöglicht. Zukünftige Studien werden dazu beitragen die besondere Rolle des faszinierendem indirektem Abwehrmechanismus der extrafloralen Nektarien en detail zu verstehen.

O crescente aumento da contaminação do solo por metais pesados, em especial o cádmio (Cd), associado à importância dos cereais, sendo estes nos países subdesenvolvidos e alguns em desenvolvimento a maior fonte protéica, são aspectos importantes que originaram a ideia deste estudo que procura analisar o metabolismo do aminoácido essencial lisina em milho (Zea mays), submetido à contaminação por Cd. Foram realizados vários ensaios a fim de se avaliar a atividade das principais enzimas do sistema antioxidante, a quantidade de aminoácidos solúveis totais e a atividade das enzimas da via do anabolismo e catabolismo da lisina. Os resultados foram apresentados na forma de dois capítulos. O capítulo 1 apresenta a caracterização bioquímica das plantas de milho (linhagem 161) submetidas ao estresse pelo Cd analisando-se a quantificação do metal nos diferentes tecidos vegetais, análise das principais enzimas do sistema antioxidante, quantificação de proteínas de reserva e aminoácidos solúveis totais, além da análise das principais enzimas da via da lisina. Os resultados da caracterização bioquímica mostraram que estes parâmetros avaliados não diferiram significativamente entre as plantas contaminadas e as do controle. No capítulo 2 foi realizada a análise das principais enzimas da via da lisina em um outro genótipo de milho, sendo uma variedade comercial (AL Avaré). De maneira geral, os resultados mostraram que o Cd induziu leves alterações nas atividades de algumas enzimas da via da lisina. Além disso, foram observadas diferenças entre os dois genótipos, tanto nas atividades específicas de enzimas como na quantificação de aminoácidos livres. Além dos dois capítulos, foram incluídos dois estudos complementares, que trouxeram análises adicionais ao estudo principal. O estudo complementar 1, apresenta os resultados de um estudo com plântulas de milho, de diferentes genótipos, sendo um com maiores teores de lisina (opaco2) e os outros para alta eficiência no uso de nitrogênio (EUN) e para baixa EUN. Tanto os resultados da peroxidação lipídica, quanto a quantidade de aminoácidos solúveis totais apresentaram diferenças entre os genótipos e doses de Cd utilizadas. O estudo complementar 2 apresenta alguns dados de enzimas antioxidantes, em PAGE não-desnaturante, de coleóptilos e radículas de embriões imaturos (sem a presença de endosperma) e germinados na presença de Cd. Estes dados mostraram uma nítida resposta antioxidante. Além disso, foram analisadas algumas células provenientes de radículas de sementes de milho germinada na presença de Cd, a fim de se avaliar os danos do metal sobre o DNA. Em resumo, os resultados obtidos neste estudo nos ajudaram a entender um pouco mais sobre o efeito do Cd nos processos fisiológicos e bioquímicos das plantas analisadas. Entretanto, estes dados podem ser considerados o ponto de partida para estudos mais detalhados. / The increase of soil contamination by heavy metals, especially cadmium (Cd), associated with the importance of cereals, which are the major source of protein in the underdeveloped and some developing countries, are important aspects that gave rise to the idea for this study which tries to analyze the metabolism of the essential amino acid lysine in maize (Zea mays), submitted to the contamination by Cd. Several tests were carried out to evaluate the activity of the main enzymes of the antioxidant system, the amount of total soluble amino acids and the activity of enzymes of lysine metabolism. The results were presented two chapters. Chapter 1 reports the biochemical characterization of the maize plants (a line with higher levels of lysine, L161) submitted to stress by Cd analyzing the metal quantification in different plant tissues, analysis of the main enzymes of the antioxidant system, quantification of reserve proteins and total soluble amino acids, in addition to the analysis of the main enzymes of lysine pathway The results showed that these parameters did not differ significantly between contaminated plants and those of the control. In Chapter 2 the main enzymes of lysine metabolism were analyzed in a commercial variety of maize (AL Avaré). Overall, the results showed that Cd induced slight changes in the activities of some enzymes of lysine pathway. In addition, differences were observed between the two genotypes, both in the specific activities of enzymes and on quantification of soluble amino acids. In addition to the two chapters, two complementary studies were included, which provided additional analyzes to the main study. Complementary study 1 presents the results of a study with maize seedlings of different genotypes, one with higher lysine (opaque2) and others for high efficiency in nitrogen use (EUN) and low EUN. Lipid peroxidation results and the amount of total soluble amino acids showed differences between the genotypes and the Cd concentrations used. Complementary study 2 presents some non-denaturing antioxidant enzyme data from coleoptiles and immature embryo rootlets (without the presence of endosperm) and germinated in the presence of Cd. These data showed a clear antioxidant response. In addition, some cells from germinated maize seedlings were analyzed in presence of Cd in order to evaluate the damage of the metal on the DNA. In summary, the results obtained in this study helped us to understand a little more about the effect of Cd on the physiological and biochemical processes of the plants analyzed. However, these data can be considered the starting point for more detailed studies.