High temperature stress (HTS) adversely affects reproduction in most plant species studied to date. HTS during flowering may result in an almost total inhibition of seed production in crop plants. Increasing our knowledge of the effects of HTS on seed production will aid the breeding of more thermotolerant crop plants and improve our understanding of the effects of stress on plants. An investigation of the effects of both drought and high temperature stress on the yields of barley, canola, flax, durum and spring wheat in five locations in Saskatchewan over a 25-year period was performed using multivariate analysis. Higher temperatures during June and July, when the plants were flowering, were correlated with reductions in yields of all the crops studied (except barley in June). A positive correlation between yields and precipitation during May and the winter preceding the growing season was observed.<p>In growth chambers, <i>Brassica napus</i> silique and seed production were inhibited during a ramping HTS treatment. This was due to a decrease in pollen germinability rather than a reduction in the number of flowers produced. HTS also caused reductions in megagametophyte fertility and disrupted embryo and/or seed development.<p>Transgenic plants were developed to overcome the effects of HTS on seed production. Two DNA constructs, one with the <i>Arabidopsis thaliana LEAFY</i> (<i>AtLFY</i>) promoter controlling <i>A. thaliana HEAT SHOCK PROTEIN 101</i> (<i>AtHSP101</i>) ORF expression and another with the <i>AtHSP101</i> promoter controlling <i>AtLFY</i> ORF expression, were inserted into <i>B. napus</i>. Other DNA constructs were made, using the constitutively expressed Cauliflower Mosaic Virus <i>35S</i> or the synthetic <i>EntCup4</i> promoters to control expression of the <i>AtHSP101</i> or <i>A. thaliana HEAT SHOCK TRANSCRIPTION FACTOR 3</i> (<i>AtHSF3</i>) ORFs. These constructs were inserted into both <i>B. napus</i> and <i>A. thaliana</i>. Transgenic plants were tested using a ramping temperature regime but were found not to have increased flower thermotolerance. During the manufacture of the DNA constructs it was determined that, in <i>A. thaliana</i>, 573 bp of <i>AtHSP101</i> had been copied between Terminal Inverted Repeats of a <i>Mu-Like Element</i> (<i>MULE</i>). This fragment was named <i>HSP101B</i>. In some transgenic <i>B. napus</i> and <i>A. thaliana</i> lines, containing 2046 bp of the <i>HSP101B</i> upstream regulatory region controlling <i>B</i>-glucuronidase (GUS) expression, cold-inducible GUS expression was observed. Methylation may have a role in control of endogenous <i>HSP101B</i> transcription.

Exposure of cells to environmental or chemical stressors will initiate the heat shock response, which is mediated by heat shock proteins. Heat shock proteins are molecular chaperones which are classified by size into six main families: HSP100, HSP90, HSP70, HSP60, HSP40 and the small heat shock proteins (sHsps). The sHsp family members bind to denatured proteins and maintain them in a folding competent state such that they may be refolded by other molecular chaperones. <br /><br /> The present study examined the expression and function of two amphibian sHsps, namely, <em>Rana catesbeiana</em> HSP30 and <em>Xenopus laevis</em> HSP27. Initially, an antisense riboprobe was produced to study the mRNA accumulation of <em>Rana hsp30</em> in cultured tongue fibroblast (FT) cells. Results showed that <em>Rana hsp30</em> mRNA was optimally induced when maintained at 35&deg;C for 2 h. An antibody to the recombinant <em>Rana</em> HSP30 protein was also produced in order to study HSP30 protein accumulation in <em>Rana</em> FT cells. Analysis showed that <em>Rana</em> HSP30 was heat-inducible and accumulated maximally at 4 h when maintained at 35&deg;C and then allowed to recover at 22&deg;C for 2 h. Immunocytochemical analysis indicated that <em>Rana</em> HSP30 protein was present primarily in the nucleus, with diffuse localization in the cytoplasm. Additional immunocytochemical analysis showed that <em>Rana</em> HSP30 remained in the nucleus following heat stress and extended periods of recovery. <br /><br /> The molecular chaperone function of <em>Rana</em> HSP30 was also studied. Recombinant <em>Rana</em> HSP30 was found to inhibit the heat induced aggregation of various target proteins including citrate synthase, luciferase and malate dehydrogenase. Also, no major difference was detected between <em>Rana</em> HSP30 and <em>Xenopus</em> HSP30C in the inhibition of heat-induced aggregation of target proteins. <br /><br /> This study also examined the expression and function of <em>Xenopus laevis</em> HSP27. Analysis of the putative amino acid sequence of the <em>Xenopus hsp27</em> cDNA revealed that it had an identity of 71% with chicken, 65% with zebrafish, 63% with human and 53% with topminnow. Most of the identity was located within the &alpha;-crystallin domain of the protein. Interestingly, <em>Xenopus</em> HSP27 shared only a 19% identity with 2 other <em>Xenopus</em> sHsps, HSP30C and HSP30D. <br /><br /> Western blot analysis using an anti-<em>Xenopus</em> HSP27 antibody revealed that HSP27 was not detectable in cultured kidney epithelial cells. However, examination of early <em>Xenopus</em> embryos revealed that HSP27 was first detected in tadpole embryos (stage 44). Heat-inducible HSP27 was also first detected at this stage. The accumulation pattern of <em>Xenopus</em> HSP27 protein was distinct from <em>Xenopus</em> HSP30, which was heat-inducible at midtailbud stage 26, approximately two and a half days earlier in development. <br /><br /> Analysis of recombinant HSP27 by native pore exclusion limit electrophoresis showed that it formed high molecular weight, multimeric complexes. The molecular chaperone function of HSP27 was assessed by means of thermal aggregation assays employing citrate synthase, luciferase and malate dehydrogenase. <em>Xenopus</em> HSP27 inhibited the heat-induced aggregation of all of these target proteins. This study has revealed that <em>Xenopus</em> HSP27 is a member of the HSP27 subfamily of small heat shock proteins in <em>Xenopus</em> and distinct from the HSP30 family. The accumulation of HSP27 under constitutive and stress-inducible conditions is developmentally regulated. Finally, this sHsp appears to function as a molecular chaperone.

Mycoplasma hyopneumoniae causes swine enzootic pneumonia (SEP) and leads to economic loss worldwide. The mechanism of pathogenesis is still not clear. Since this pathogen remains extracellulary after infection, the surface proteins on M. hyopneumoniae should play very important roles in adhering and affecting tracheal mucosal cells. Therefore, the potential of using the surface proteins as the basic to develop molecular vaccine is currently being investigated. The recombinant clone expressing the 42 kDa protein was isolated from the

Submitted by Amanda Silva (amanda.osilva2@ufpe.br) on 2015-03-11T13:42:17Z No. of bitstreams: 2 TESE Adriana da Silva Andrade Pereira.pdf: 6828087 bytes, checksum: 9d66e9700be0a70e927b955ee06cb850 (MD5) license_rdf: 1232 bytes, checksum: 66e71c371cc565284e70f40736c94386 (MD5) / Made available in DSpace on 2015-03-11T13:42:17Z (GMT). No. of bitstreams: 2 TESE Adriana da Silva Andrade Pereira.pdf: 6828087 bytes, checksum: 9d66e9700be0a70e927b955ee06cb850 (MD5) license_rdf: 1232 bytes, checksum: 66e71c371cc565284e70f40736c94386 (MD5) Previous issue date: 2014-02-17 / CNPq; CAPES / Schistosoma mansoni, é um dos mais importantes parasitas que infectam seres humanos. No nordeste do Brasil, a esquistossomose é historicamente endêmica e considerada como um problema de saúde pública. No estado de Pernambuco, particularmente no litoral e zona da Mata, predomina a cepa São Lourenço da Mata (SLM). Estudos ultraestruturais desta cepa até então ainda não foram realizados, embora análises morfométricas e morfológicas de outras cepas existentes no Brasil já tenham sido publicadas na literatura. Os vermes são bem adaptados ao hospedeiro, tendo a sua longevidade vista como uma conseqüência do eficaz escape do sistema imune. Na circulação sanguínea os vermes devem obter lipídios a partir do hospedeiro, já que eles não sintetizam tais compostos, sendo as lipoproteínas plasmáticas a possível fonte de tais lipídios, além de poderem atuar mascarando o reconhecimento do verme pelo sistema imune. Neste trabalho, microscopia eletrônica de varredura (SEM) foi utilizada para a análise morfológica e morfométrica de cercárias e vermes adultos de S. mansoni cepa SLM, bem como para estudar a interação da lipoproteína de baixa densidade (LDL) com o tegumento do parasito. Também foram usadas diferentes técnicas para a localização e identificação das proteínas envolvidas no processo de absorção e/ou transporte de lipoproteínas no verme adulto de S. mansoni. As cercárias foram obtidas de caramujos Biomphalaria glabrata e os vermes adultos de camundongos machos Mus musculus e Swiss, ambos infectados pela cepa SLM de S. mansoni. Os resultados da SEM mostraram que os corpos da cercárias são cobertos por espinhos, com uma ventosa ventral, uma ventosa oral com receptores sensoriais, e um par de glândulas de penetração na cabeça. Seu comprimento total variou de 174 a 290μm. O comprimento dos vermes adultos machos foi de 4mm e das fêmeas 5mm. O comprimento da região anterior do macho foi de 470μm e 271μm para o sexo feminino. Todos os parâmetros foram realizados em dez amostras. Os valores encontrados na morfometria da cepa SLM foram menores que em outras cepas de S. mansoni já descritas. Através da microscopia eletrônica de transmissão (TEM), os experimentos de imunomarcação mostraram partículas de ouro localizadas dentro do tegumento, na região muscular e espículas dos machos e em torno das células vitelínicas das fêmeas. A utilização de técnicas tais como imunoblotting e “liquid chromatography–mass spectrometry” (LC-MS) proporcionaram a identificação no S. mansoni de duas formas diferentes da chaperona Hsp 70, proteínas envolvidas na interação com a apo-B e o seu transporte para o retículo endoplasmático, sendo responsável também pela ruptura das interações clatrina-clatrina na endocitose humana. Com a descoberta destas proteínas, novos estudos serão necessários para esclarecer o papel funcional das mesmas no S. mansoni.

Equine spermadhesin HSP-7 is a 14 kDa protein isolated from stallion seminal plasma and present on the surface of spermatozoa. HSP-7 displays carbohydrate and zona-pellucida binding properties, but the physiological role in equine fertilization is not well defined. HSP-7 has 98% amino acid sequence homology with the well-studied boar spermadhesin, AWN. Currently, these two proteins are considered to have the same reproductive function. Immunofluorescence studies presented here show that the stallion and boar spermadhesins are localized to different segments on spermatozoa. The variation in molecular compartmentalization of spermadhesin molecules in different species suggests that these structurally related proteins could be involved in independent events of fertilization. While the variation in HSP-7 abundance was not statistically significant between fertile and subfertile stallions, capacitated spermatozoa displayed a marked increase in HSP-7 when compared to neat sperm (P < 0.05). These results indicate that rather than aiding in capacitation, HSP-7 is exposed with capacitation and may have a more significant role in the acrosome reaction and sperm-oocyte recognition than previously documented.

O objetivo deste trabalho foi contribuir para o melhor entendimento dos mecanismos intracelulares envolvidos na regeneração muscular esquelética, através do estudo da influência das proteínas de choque térmico (HSPs) e do mTORC1 (mammalian target of rapamycin complex 1) no processo regenerativo muscular. O tratamento com radicicol (indutor de HSPs) em músculos lesados induziu aumento da área de secção transversal das fibras musculares em 10 e 21 dias após lesão e aumento do número de células satélites e de fibras musculares em diferenciação em 1 e 10 dias após lesão, respectivamente, quando comparado aos seus respectivos controles apenas lesados. O tratamento com rapamicina (inibidor de mTORC1) em músculos lesados induziu uma diminuição maior da área de secção transversal das fibras musculares em 10 e 21 dias após lesão e menor síntese protéica muscular em 10 dias após lesão quando comparado aos músculos somente lesados. Nossos resultados sugerem que as HSPs e o mTORC1 são importantes para o processo de regeneração muscular esquelética. / The goal of this work was to contribute to a better understanding about the intracellular mechanisms involved in skeletal muscle regeneration by studying the influence of heat shock proteins (HSPs) and mTORC1 (mammalian target of rapamycin complex 1) in the muscle regeneration process. The treatment with radicicol (a HSP inductor) in injured muscles induced increase of myofiber cross section area at 10 and 21 days post lesion and increased number of satellite cells and differentiating myofibers at 1 and 10 days post lesion, respectively, when compared to their respective injured controls. The treatment with rapamycin (a mTORC1 inhibitor) in injured muscles induced a more accentuated decrease in myofiber cross section area at 10 and 21 days post lesion and decreased muscle protein synthesis at 10 days post lesion when compared to only-injured muscles. Our results suggest that HSPs and mTORC1 are important to the process of skeletal muscle regeneration.

Des systèmes injectables de formation in situ ont été utilisés dans les dernières années pour l'obtention de formulations de préparation facile et permettant la libération prolongée de principes actifs. Ces systèmes utilisant des solvants biocompatibles et des polymères biodégradables sont des liquides (solutions ou émulsions) qui une fois injectés dans l'organisme donnent lieu à des implants (ISI) ou à des microparticules (ISM) solides. La formation de ces systèmes est induite par la précipitation du polymère à partir des solutions polymériques qu'ils contiennent lors du contact avec les fluides corporaux aqueux. Dans ce travail, des ISI et des ISM, réalisés à partir des polymères de l'acide lactique et/ou glycolique (PLA et PLGA) et des différents solvants biocompatibles, pour la libération prolongée d?ivermectine (IVM), un principe actif antiparasitaire faiblement biodisponible par la voie orale, ont été développés. Les profils de libération du principe actif in vitro et in vivo à partir de ces systèmes, ont été comparés avec ceux obtenus à partir de microparticules réalisées par la méthode classique dite d'émulsion simple - évaporation de solvant ; il s'agit d'une technique aux multiples étapes, à coût élevé et dont l'utilisation de solvants toxiques la font difficilement industrialisable. La libération du principe actif à partir des microparticules obtenues par émulsion simple/évaporation du solvant a été influencée par la forte interaction du principe actif avec les polymères mais aussi par la porosité. Dans le cas des systèmes in situ, la vitesse de libération d'IVM a été conditionnée par la solubilité dans l'eau du solvant biocompatible sélectionné et par les interactions solvant/polymère. Pour les ISM, des paramètres tels que la nature de la phase externe, aqueuse (ISM-O/W) ou huileuse (ISM-O/O), la solubilité dans l'eau du solvant de la phase interne, l'affinité entre les phases et l'affinité de l'IVM pour chacune des phases, ont déterminé la vitesse de libération du principe actif. La bonne stabilité ainsi que les profils de libération plus prolongés et présentant une faible libération initiale du principe actif in vivo et in vitro, ont montré que les ISI et les ISM réalisés à partir de solvants biocompatibles de faible solubilité dans l'eau tels que la triacetine sont les plus indiqués pour l'encapsulation d'IVM par rapport à ceux plus solubles dans l'eau comme la N-methyl-2-pyrrolidone et la 2-pyrrolidone. Ces systèmes représentent donc une alternative intéressante par rapport aux formulations conventionnelles d'IVM / In situ forming injectable systems have been used in the past years to obtain sustained drug release formulations which are easy to prepare. These systems using biocompatible solvents and biodegradable polymers are liquids (solutions or emulsions) that upon injection on the body lead to solid implants (ISI) or microparticles (ISM). These systems are formed in contact with water body fluids by polymer precipitation from the polymeric solution. In this work, ISI and ISM made from lactide and/or glycolide polymers (PLA and PLGA) and different biocompatible solvents were performed to obtain sustained release of ivermectin (IVM), an antiparasitic drug with a low oral bioavailability. In vitro and in vivo drug release profiles from these systems were compared with those from microparticles obtained by the classical simple emulsion/solvent evaporation method, which is difficult to propose in industry because of its multiple steps, high cost and the solvent toxicity. Drug release from simple emulsion/solvent evaporation microparticles was affected by the strong polymer/drug interactions and porosity. Concerning to in situ forming systems, the rate of IVM release was dependent on solvent water solubility and solvent/polymer interactions. The nature of the external phase, water (ISM-O/W) or oil (ISM-O/O), the water solubility of the solvent in the internal phase, phase affinity and IVM/phase affinity determined drug release from ISM. The good stability, the in vitro and in vivo sustained release and the low burst effect of IVM, indicated that ISI and ISM formulated from low hydrosoluble biocompatible solvents such as triacetin are more appropriated to IVM formulation instead of those based on more hydrophilic solvent (N-methyl-2-pyrrolidone and 2-pyrrolidone). These systems are an interesting alternative to conventional IVM formulations

Lors d'un stress, les cellules activent un mécanisme de défense appelé “la réponse au stress”. Ce mécanisme empêche notamment l'accumulation de protéines mal enroulées grâce à la synthèse des protéines du choc thermique, les HSPs (Heat Shock Proteins),via l'activation du facteur de transcription HSF1 (Heat Shock Factor 1). Les HSPs empêchent l'agrégation des protéines et aident au repliement protéique. Deux partenaires associés à HSF1 ont récemment été identifiés : le chaperon moléculaire VCP (Valosin Containing Protein) et l'histone déacétylase 6 (HDAC6). Notre projet était d'étudier les rôles d'HDAC6 et de VCP dans la réponse au stress thermique. Nous avons mis en évidence un rôle prépondérant du domaine de fixation à l'ubiquitine d'HDAC6 dans la régulation de la durée d'activation d'HSF1 suite à un choc thermique. Lorsque HDAC6 ne peut pas se fixer à l'ubiquitine, VCP favorise une inactivation rapide de HSF1 empêchant la transcription du chaperon HSP25. Ces travaux montrent également que la réponse activée suite à un stress dépend de la nature de celui-ci. En effet, nous avons montré que les mécanismes activés dans la réponse au stress suite à un choc thermique sont différents de ceux activés suite à une inhibition du protéasome. La myopathie à corps d'inclusion associée à la maladie osseuse de Paget et à une démence fronto-temporale, appelée IBMPFD, Inclusion Body Myopathy associated with Paget disease of the bone and Frontotemporal Dementia, est une maladie autosomale dominante rare. La myopathie est la caractéristique clinique la plus commune parmi celles qui sont causées par des mutations de VCP. La perturbation de la fonction de VCP entraîne l'accumulation de protéines poly-ubiquitinées et la formation de corps d'inclusion en partie responsables de la pathogenèse de l'IBMPFD. Nous avons montré que l'activation de la réponse au stress via un choc thermique dans des cellules murines déficientes en VCP mimant le phénotype de l'IBMPFD, entraîne une diminution du taux de cellules ayant des agrégats de protéines poly-ubiquitinées. Nos résultats préliminaires montrent que HSF1 ne serait pas à l'origine de cette diminution des agrégats au contraire de HSP90 et HDAC6 qui interviendraient suite à l'activation de la réponse au stress. Cette stratégie est actuellement en cours de test sur des modèles murins. / Under stress, cells activate a defense mechanism named “cellular stress response”. This mechanism prevents especially unfolded proteins accumulation thanks to Heat Shock Proteins (HSPs) synthesis through the activation of Heat Shock Factor 1 (HSF1) transcription factor. HSPs prevent aggregation and help protein refolding. Two partners associated to HSF1, have recently been identified: the molecular chaperone, VCP (Valosin Containing Protein) and the Histone DeACetylase 6 (HDAC6). Our project was to characterize the roles of HDAC6 and VCP in the Heat Shock Response (HSR). We have highlighted a preponderant role for the HDAC6 ubiquitin binding domain in the HSF1 activation time regulation after a heat shock. When HDAC6 can't bind ubiquitin, VCP promotes a rapid HSF1 inactivation preventing HSP25 chaperone transcription. This work also emphasizes a stimulus-dependent stress response. Indeed, we showed that mechanisms activated during the stress response following a heat shock differ from those activated after a proteasome inhibition. Inclusion Body Myopathy, Paget disease of the bone, and Frontotemporal Dementia (IBMPFD) is a rare autosomal dominant disorder. Myopathy is the most common clinical feature of IBMPFD. It is caused by mutations of VCP. Alteration of VCP function leads to the accumulation of poly-ubiquitinated proteins and to the formation of inclusion bodies thought to be responsible, at least in part, for the pathogenesis of IBMPFD. We have shown that activation of the heat shock stress response in mouse cells VCP deficient mimicking IBMPFD phenotype, results in the decrease of cells with ubiquitinated protein aggregates. Our preliminary results show that HSF1 is not responsible for this decrease unlike HSP90 and HDAC6 that seems to intervene following the stress response activation. This strategy is currently tested on mouse models.

O objetivo deste trabalho foi contribuir para o melhor entendimento dos mecanismos intracelulares envolvidos na regeneração muscular esquelética, através do estudo da influência das proteínas de choque térmico (HSPs) e do mTORC1 (mammalian target of rapamycin complex 1) no processo regenerativo muscular. O tratamento com radicicol (indutor de HSPs) em músculos lesados induziu aumento da área de secção transversal das fibras musculares em 10 e 21 dias após lesão e aumento do número de células satélites e de fibras musculares em diferenciação em 1 e 10 dias após lesão, respectivamente, quando comparado aos seus respectivos controles apenas lesados. O tratamento com rapamicina (inibidor de mTORC1) em músculos lesados induziu uma diminuição maior da área de secção transversal das fibras musculares em 10 e 21 dias após lesão e menor síntese protéica muscular em 10 dias após lesão quando comparado aos músculos somente lesados. Nossos resultados sugerem que as HSPs e o mTORC1 são importantes para o processo de regeneração muscular esquelética. / The goal of this work was to contribute to a better understanding about the intracellular mechanisms involved in skeletal muscle regeneration by studying the influence of heat shock proteins (HSPs) and mTORC1 (mammalian target of rapamycin complex 1) in the muscle regeneration process. The treatment with radicicol (a HSP inductor) in injured muscles induced increase of myofiber cross section area at 10 and 21 days post lesion and increased number of satellite cells and differentiating myofibers at 1 and 10 days post lesion, respectively, when compared to their respective injured controls. The treatment with rapamycin (a mTORC1 inhibitor) in injured muscles induced a more accentuated decrease in myofiber cross section area at 10 and 21 days post lesion and decreased muscle protein synthesis at 10 days post lesion when compared to only-injured muscles. Our results suggest that HSPs and mTORC1 are important to the process of skeletal muscle regeneration.

Made available in DSpace on 2014-08-20T13:59:14Z (GMT). No. of bitstreams: 1 dissertacao_cristina_moll_huther.pdf: 1943888 bytes, checksum: 19ce036e997295b4526288308a80fe3c (MD5) Previous issue date: 2011-08-10 / The abiotic stress often cause metabolic disorders which cause a decrease in plant growth. In response to stress, an increase in the synthesis of certain proteins called ―heat shock proteins‖ (HSPs), might occur. The tomato (Lycopersicon esculentum) cv. Micro-Tom is regarded as a model for experimental studies, and that is because it has features that make it appropriate, such as small size, short development time and easy modification. This study aimed to evaluate the stress effects by flooding and by high and low temperatures in both modified and not modified ‗Micro-Tom tomato plants which have different mitochondrial expressions (MT- sHSP22). The first experiment was conducted in growth chambers under controlled conditions. When the plants were 50 days, they were submitted to stress by flooding during 14 days, and after 7 days, half the plants were removed from the flooding. In the second trial, plants were grown as in the first experiment, and when they presented themselves in a complete vegetative stage, they were submitted to thermal stress for a period of 24 hours of 10 ºC or 37 °C, soon after that, the plants were transferred to the initial conditions for 24 hours. After this phase they were again subjected to a new cycle of stress and recovery. On both experiments were evaluated parameters of the kinetic emission of chlorophyll fluorescence and of gas exchange, taking into account that in the first was also evaluated the content of chlorophyll, and, at the end of the test, the leaf area and the dry mass of the upper area and root. The flooding interfered on the the chlorophyll content of all genotypes, and on the genotype with a high expression of MT-sHSP22, a decrease in the leaf area was not identified and neither was in the total dry mass. The kinetics analysis of the transient fluorescence of chlorophyll showed differential effects of stress by flooding in the different parts of the photosynthetic machinery, and showed different responses between genotypes. When plants were submitted to stress by low temperature, a reduction in the liquid CO2 rate was noticed immediately after the stress, but, after recovery, all genotypes returned close to the controls. For plants submitted to high temperatures of the wild genotype and with high expression of MT-sHSP22 increased the liquid assimilation rate in comparison to control. The chlorophyll fluorescence data indicated different effects on absorption and utilization of light energy between the two genotypes for two of the temperature stress. The interpretation of the JIP test parameters helped to identify that three genotypes exhibit different behavior, mainly on high-temperature stress, and the plants of high expression genotypes of HSP22 and not transformed showed an increase in the parameters related to the photosystem I activity. These results suggest that plants with high doses of mitochondrial sHSP22 may present mechanisms of tolerance against the applied stress. / Os estresses abióticos geralmente causam disfunções metabólicas que provocam diminuição no crescimento das plantas. Em respostas a estresses pode ocorrer o aumento na síntese de algumas proteínas chamadas proteínas de choque térmico (heat shock proteins, HSPs). O tomate (Lycopersicon esculentum), cv. Micro-Tom é considerado como um modelo para estudos experimentais, pois ele possui características que o tornam adequado, tais como o tamanho pequeno, tempo de geração curto, e facilidade de transformação. Este trabalho teve como objetivo avaliar os efeitos dos estresses por alagamento e por altas e baixas temperatura em plantas transformadas e não transformadas de tomate ‗Micro-Tom com diferentes expressões da sHSP22 mitocondrial (MT-sHSP22). O primeiro experimento foi conduzido em câmaras de crescimento, sob condições controladas. Quando as plantas apresentavam 50 dias foram submetidas a estresse por alagamento por um período de 14 dias, sendo que após 7 dias metade das plantas foram retiradas do alagamento. No segundo ensaio, as plantas foram cultivadas como no primeiro experimento, e quando se apresentavam em estádio vegetativo pleno foram submetidas a estresse térmico por período de 24h a 10ºC ou 37ºC, a seguir as plantas foram transferidas para as condições iniciais por 24h, após este período foram novamente submetidas a um novo ciclo de estresse e recuperação. Em ambos os experimentos foram avaliados os parâmetros da cinética de emissão da fluorescência da clorofila e trocas gasosas, sendo que no primeiro avaliou-se também o índice de clorofila, e ao final do ensaio a área foliar e massa seca da parte aérea e raiz. O alagamento interferiu nos teores de clorofila de todos os genótipos, sendo que no genótipo com elevada expressão da MT-sHSP22, não foi observada redução da área foliar e em relação a massa seca total. A análise da cinética da fluorescência transiente da clorofila a mostrou efeitos diferenciais do estresse por alagamento nos diferentes locais da maquinaria fotossintética, e apresentou variação de resposta entre os genótipos. Quando as plantas foram submetidas ao estresse por baixa temperatura, observou-se redução na taxa assimilatória liquida de CO2 imediatamente após o estresse, mas depois da recuperação todos os genótipos retornaram próximos aos controles. Para as plantas submetidas a altas temperaturas do genótipo selvagem e do com elevada expressão da MT-sHSP22 aumentaram a taxa assimilatória líquida em relação ao controle. Os dados de fluorescência das clorofilas indicaram efeitos diferentes na absorção e aproveitamento da energia luminosa entre os genótipos para os dois estresses de temperatura. A interpretação dos parâmetros do Teste JIP possibilitou identificar que os três genótipos apresentam comportamento distinto principalmente sobre estresse de alta temperatura, sendo que as plantas dos genótipos com alta expressão de sHSP22 e não transformado apresentaram elevação nos parâmetros relacionados a atividade do fotossistema I. Estes resultados sugerem que as plantas com elevada expressão das sHSP22 mitocondrial podem apresentar mecanismos de tolerância frente as estresses aplicados.