Mémoire numérisé par la Direction des bibliothèques de l'Université de Montréal.

This study focuses on the soil- and water-borne plant pathogen Phytophthora cinnamomi Rands and the phenomenon of P. cinnamomi suppressive soil. In particular, this thesis reports on the outcome of field surveys and glasshouse assays undertaken to locate P. cinnamomi suppressive soils and to confirm the involvement of biological processes in suppression. The potential role of cellulase and laminarinase in suppression was investigated and a molecular technique known as length heterogeneity PCR (LH-PCR) was used to analyse the structure and diversity of bacterial and fungal communities in avocado orchard soils that were suppressive and conducive to P. cinnamomi. Four avocado orchards with P. cinnamomi suppressive soils were identified and soils were ã-irradiated to destroy their suppressive capacity, thus confirming biological suppression. Suppression was also partially transferred to ã-irradiated and conducive soils by mixing with 10% suppressive avocado soils. Cellulase and laminarinase activities measured in avocado orchard soils inoculated with P. cinnamomi were not associated with disease severity in lupin seedlings during glasshouse assays involving the same soil samples. Minor shifts in bacterial and fungal community structure were observed in response to mixing conducive and irradiated soils with suppressive soils. This was associated with decreased disease severity in avocado seedlings in these treatments. The shift in bacterial community structure was partially determined by the appearance and increased abundance of several bacterial 16S rDNA sequences, which were unique to the suppressive soils, in the mixed soil treatments. It is suggested that the bacteria and fungi from which these sequences originated may be involved in suppression and further work should be undertaken to determine their identity and confirm their potential role in the development and maintenance of P. cinnamomi suppressive soils. / Doctor of Philosophy (PhD)

Multiple sclerosis (MS) is an autoimmune disorder of the central nervous system (CNS). In the current thesis, we have preformed an immunological investigation of patients with MS and developed an immunosuppressive cell therapy that could be beneficial for these patients. MS has been considered to be driven by T helper type1 (Th1) lymphocytes but new data indicate the involvement of Th17 responses. T cells from patients with MS that were evaluated for immunological status secreted both interferon-γ and interleukin-17 upon stimulation. However, T cells from patients with MS in remission, in contrast to relapse, had poor proliferative capacity suggesting that they are controlled and kept in anergy. T regulatory cells (Tregs) are important to maintain self-tolerance and the role of CD4+CD25+FoxP3+ Tregs in autoimmunity has been extensively investigated. We analyzed Tregs from patients with MS in relapse and remission by multicolor flow cytometry for the expression of CD3, CD4, IL2R (CD25), FoxP3 and the IL7R (CD127). Patients in relapse exhibited higher levels of FoxP3-positive Tregs lacking CD25 compared to healthy controls, indicating that Tregs might attempt to restrain immune activity during relapse. In the murine experimental autoimmune encephalomyelitis (EAE) model of MS, therapy with suppressive cells such as Tregs or mesenchymal stromal cells (MSCs) has proven beneficial. However, systemic administration of such cells may immunologically compromise the recipient and promote infections due to general immunosuppression. We hypothesized that suppressive cells can be equipped with a CNS-targeting receptor and be delivered intra-nasally to avoid systemic exposure. CD4+ T cells were modified with a lentiviral vector system to express a myelin oligodendrocyte (MOG)-targeting receptor in trans with the FoxP3 gene that drives Treg differentiation. Genetically engineered Tregs demonstrated suppressive capacity in vitro and localized to the brain and suppressed ongoing encephalomyelitis in vivo. Cured mice were rechallenged with an EAE-inducing inoculum but remained healthy. MSCs are a heterogeneous population of stromal cells residing in most connective tissues and have the capacity to suppress effector cells of the immune system. MSCs were engineered to express MOG-targeting receptors using lentiviral vectors. Genetically engineered MSCs retained their suppressive capacity in vitro and successfully targeted the brain upon intranasal delivery. Engineered MSCs cured mice from disease symptoms and these mice were resistant to further EAE challenge. Encephalitic T cells isolated from cured mice displayed an anergic profile while peripheral T cells were still responsive to stimuli. In conclusion, MS patients have peripheral CNS-reactive T cells of both Th1 and Th17 type that, while in remission, are kept in anergy. Also, MS patients in relapse exhibit increased levels of CD25 negative Tregs indicating an attempt to restrain immune activity. Finally, immunosuppressive cells can be genetically engineered to target CNS and efficiently suppress encephalomyelitis in an active EAE model upon intranasal delivery.

La diapause embryonnaire se manifeste par un arrêt réversible du développement embryonnaire durant la période de préimplantation et induit un retard de l’implantation. Chez le vison américain, une diapause embryonnaire obligatoire caractérise chaque gestation. Si les mécanismes de contrôle de la diapause embryonnaire obligatoire chez cette espèce sont bien connus, le rôle utérin impliqué dans la réactivation de l’embryon demeure, quant à lui, encore inconnu. Le sujet de ce doctorat a consisté dans un premier temps à explorer l’environnement utérin à la sortie de la diapause embryonnaire afin de caractériser, dans un deuxième temps, les principaux acteurs utérins qui provoquent la réactivation de l’embryon. Nous avons effectué une analyse du transcriptome utérin à l’émergence de la diapause embryonnaire ce qui a permis de construire une librairie de 123 séquences d’ADNc utérines différentiellement exprimées à la réactivation de l’embryon et homologues à des séquences de gènes connues chez d’autres espèces. Ces gènes sont impliqués dans la régulation du métabolisme (25 %), de l’expression génique (21 %), de la transduction de signal (15 %), du cycle cellulaire (15 %), du transport (10 %) et de la structure cellulaire (9 %), reflétant ainsi d’importantes modifications utérines à la réactivation embryonnaire. Nous avons validé l’expression différentielle de dix gènes ainsi identifiés : GDF3 (growth and differentiation 3), ALCAM (activated leukocyte cell adhesion molecule), ADIPOR1 (adiponectin receptor 1), HMGN1 (high mobility group N1), TXNL1 (thioredoxin like 1), TGM2 (tissue transglutaminase 2), SPARC (secreted protein acidic rich in cystein), et trois gènes codant pour AZIN1 (antizyme inhibitor 1), ODC1 (ornithine decarboxylase 1) et SAT1 (spermidine/spermine N1-acetyltransferase), des enzymes impliquées dans la biosynthèse des polyamines. Le patron de l’expression spatio-temporel de SPARC et d’HMGN1 illustrent spécifiquement un remodelage tissulaire et de la chromatine au niveau utérin à la sortie de la diapause embryonnaire. Ayant mesuré une augmentation des concentrations utérines en polyamines à la reprise du développement embryonnaire, nous avons émis l’hypothèse que les polyamines seraient impliquées dans les événements menant à la sortie de la diapause. L’inhibition de la biosynthèse des polyamines par un traitement à l’ α-difluoromethylornithine (DFMO) a provoqué une diminution significative de la proliferation cellulaire dans les embryons à la réactivation, un retard du moment de l’implantation, mais n’a pas affecté le succès de la reproduction. De manière similaire, nous avons induit un état de dormance dans les cellules de trophoblaste de vison en présence DFMO dans le milieu de culture, et constaté que cet état était réversible. En conclusion, cette étude a non seulement ouvert de nouveaux horizons quant à la compréhension du rôle utérin dans les événements menant à la sortie de la diapause embryonnaire, mais a démontré pour la première fois, l’existence de facteurs utérins indispensables à la réactivation de l’embryon: les polyamines. / Embryonic diapause is characterized by a reversible arrest of blastocyst development prior to implantation and delay in implantation. In the American mink, embryonic diapause is a characteristic of each gestation. Although the mechanisms which control obligate embryonic diapause of this species are well known, the role of the uterus involved in blastocyst reactivation remains elusive. The subject of this doctoral research consisted first in exploring the uterine environment at the emergence of embryonic diapause in order to subsequently determine, the main factors in the uterus that provoke reactivation of the embryo. We have undertaken an analysis of the uterine transcriptome at the emergence of embryonic diapause which has enabled us to set up a library of 123 cDNA uterine sequences differentially expressed at blastocyst reactivation, and homologue gene sequences known in other species. Twenty-five percent of these genes are implicated in genetic expression, 15 % in cell signal transduction, 15 % in cell cycle, 10 % in transport and 9 % in cell structure. All of them reflect significant uterine modifications at blastocyst reactivation. We have validated differential expression of ten genes, identified as: GDF3 (growth and differentiation 3), ALCAM (activated leukocyte cell adhesion molecule), ADIPOR1 (adiponectin receptor 1), HMGN1 (high mobility group N1), TXNL1 (thioredoxine like 1), TGM2 (tissue transglutaminase 2), SPARC (secreted protein acidic rich in cystein), and three genes encoding for AZIN1 (antizyme inhibitor 1), ODC1 (ornithine decarboxylase 1) and SAT1 (spermidine/spermine N1-acetyltransferase), which are enzymes implicated in polyamine biosynthesis. The spatio-temporal expression patterns of SPARC and HMGN1 illustrate tissue and chromatin remodelling in the uterus at the termination of embryonic diapause. Having measured an increase in concentration of polyamines in the uterus at the resumption of blastocyst development, we have hypothetized that polyamines are implicated in the emergence of blastocysts from diapause. We inhibited polyamine biosynthesis in pregnant mink females during early blastocyst reactivation. The inhibition of polyamine biosynthesis through treatment with α-difluoromehtylornithine (DFMO) provoked a major reduction in cell proliferation in blastocysts at reactivation and a delay in the timing of implantation, but did not affect the success of reproduction. Similarly, we induced a reversible dormant state in cultured mink trophoblast cells traited with DFMO. To conclude, not only are results of this study a breakthrough in the understanding of the role of the uterus in stimulating at the emergence of blastocysts from embryonic diapause, but also, for the very first time, they indicate the existence of uterine factors, the polyamines, that are responsible for blastocysts reactivation.

Notre travail qui sinscrit dans le cadre général de létude du développement embryonnaire de Phaseolus a pour objectif principal disoler et de caractériser des gènes différemment exprimés chez les embryons en voie davortement, et donc nécessaires au développment normal des embryons. Des embryons en cours de dégénérescence issus des hybridations interspécifiques et de la mutagenèse induite ont été analysés. Des ADNc différemment exprimés chez ces embryons ont été identifiés par les techniques de lHybridation Soustractive Suppressive (HSS) et de la dot blot. Les hybridations interspécifiques ont été réalisées entre lespèce P. vulgaris L. utilisée comme parent mâle et les espèces P. coccineus L. et P. polyanthus Greenm. utilisées comme parents femelles (formes sauvages et cultivées). La mutagenèse induite à lEthyl Méthyl Sulfonate (EMS) a été appliquée sur le génotype BAT93 de P. vulgaris, une variété améliorée du CIAT. Dans les croisements P. coccineus x P. vulgaris, 938 hybridations ont été effectuées et le taux de gousses avortées au-delà de 8 JAP est denviron 12%. Quatre gousses supposées hybrides ont été obtenues. Pour les croisements P. polyanthus x P. vulgaris, 733 hybridations ont été réalisées. Le taux de gousses avortées au-delà de 8 JAP est denviron 18% et une seule gousse supposée hybride a été produite. Les caractères hybrides dune plante de chacune des deux combinaisons interspécifiques ont été mis en évidence au moyen de caractères morphologiques des fleurs et des graines, mais aussi grâce à lutilisation dun marqueur moléculaire, le microsatellite BM160. La mise en évidence et la caractérisation des embryons en voie davortement ont été effectuées à partir de matériels issus des hybridations interspécifiques et de la mutagenèse à lEthyl Méthyl Sulfonate (EMS). Les observations, faites sur des embryons extraits et sur des coupes histologiques dovules, révèlent des malformations au niveau du suspenseur et des cotylédons et des retards de croissance. Les plantes issues de la mutagenèse et produisant des graines avortant avant la maturité ont été croisées avec des plantes normales. Lanalyse de la F2 effectuée sur 96 plantes révèle une proportion mendélienne 3:1 de plantes avec des graines normales et de plantes avec graines qui avortent. Ce résultat suggère un contrôle du caractère « avortement des graines » par une paire dallèles récessifs. La technique de lHSS a permis disoler des fragments dADNs complémentaires différemment exprimés dans les graines en voie davortement. Lanalyse des séquences de ces ADNs complémentaires montre quils codent pour plusieurs protéines intervenant dans les développements cellulaire et embryonnaire. Les principales protéines sont le cytochrome P450, la myo-inositol 1-phosphate synthase, la peroxydase cationique, le voltage-dependent anion channel et la sucrose synthase. A lexception du cytochrome P450, les niveaux dexpression des autres gènes sont plus faibles dans les graines en voie davortement issues de la mutagenèse par rapport aux graines normales. The main objective of this study was to isolate and to characterize cDNAs differentially expressed in Phaseolus degenerating embryos. Aborting embryos from interspecific hybridizations and induced mutation were analysed. cDNAs differentially expressed in these embryos were isolated using the Suppressive Subtractive Hybridization (SSH) and the dot blot techniques. The interspecific hybridizations were performed between P. vulgaris L. used as male parent and P. coccineus L. and P. polyanthus Greenm. used as female parents (wild and cultivated forms). The induced mutation was performed whith Ethyl Methyl Sulfonate (EMS) applied on the genotype BAT93 of P. vulgaris, a breeding line from CIAT. A total number of 938 crosses P. coccineus x P. vulgaris and 733 crosses P. polyanthus x P. vulgaris were carried out. In the crosses P. coccineus x P. vulgaris, the rate of pod abortion after 8 days after pollination (DAP) is 12%. Four putative hybrid pods were obtained. The rate of pod abortion after 8 DAP in the crosses P. polyanthus x P. vulgaris is 18% and one putative hybrid pod was produced. The hybrid nature of one plant from each interspecific combination was confirmed using morphological characters of flowers and seeds and molecular marker (microsatellite BM160). The isolation and the characterization of degenerating embryos were realised with materials from interspecific hybridizations and from chemical mutagenesis with EMS. The observations of these two materials revealed abnormalities mainly in suspensor and cotyledons; and the embryos failed to grow normally. Plants from mutagenesis which produce degenerating seeds were crossed with normal plants. Genetic analysis on 96 F2 plants revealed a 3:1 Mendel ratio of plants with normal seeds and plants with degenerating seeds. This result suggests the control of the seed abortion trait by a single recessive gene. The SSH technique was used to isolate cDNAs fragments differentially expressed in aborting seeds. Analysis of the cDNAs sequences revealed that these cDNAs encode for proteins involved in cellular and embryonic development. The main proteins are cytochrome P450, myo-inositol 1-phosphate synthase, cationic peroxidase, voltage-dependent anion channel and sucrose synthase. All the genes showed a reduction of their expression in developing seeds of the mutagenized plants, compared to those observed in wild-type plants.

La diapause embryonnaire se manifeste par un arrêt réversible du développement embryonnaire durant la période de préimplantation et induit un retard de l’implantation. Chez le vison américain, une diapause embryonnaire obligatoire caractérise chaque gestation. Si les mécanismes de contrôle de la diapause embryonnaire obligatoire chez cette espèce sont bien connus, le rôle utérin impliqué dans la réactivation de l’embryon demeure, quant à lui, encore inconnu. Le sujet de ce doctorat a consisté dans un premier temps à explorer l’environnement utérin à la sortie de la diapause embryonnaire afin de caractériser, dans un deuxième temps, les principaux acteurs utérins qui provoquent la réactivation de l’embryon. Nous avons effectué une analyse du transcriptome utérin à l’émergence de la diapause embryonnaire ce qui a permis de construire une librairie de 123 séquences d’ADNc utérines différentiellement exprimées à la réactivation de l’embryon et homologues à des séquences de gènes connues chez d’autres espèces. Ces gènes sont impliqués dans la régulation du métabolisme (25 %), de l’expression génique (21 %), de la transduction de signal (15 %), du cycle cellulaire (15 %), du transport (10 %) et de la structure cellulaire (9 %), reflétant ainsi d’importantes modifications utérines à la réactivation embryonnaire. Nous avons validé l’expression différentielle de dix gènes ainsi identifiés : GDF3 (growth and differentiation 3), ALCAM (activated leukocyte cell adhesion molecule), ADIPOR1 (adiponectin receptor 1), HMGN1 (high mobility group N1), TXNL1 (thioredoxin like 1), TGM2 (tissue transglutaminase 2), SPARC (secreted protein acidic rich in cystein), et trois gènes codant pour AZIN1 (antizyme inhibitor 1), ODC1 (ornithine decarboxylase 1) et SAT1 (spermidine/spermine N1-acetyltransferase), des enzymes impliquées dans la biosynthèse des polyamines. Le patron de l’expression spatio-temporel de SPARC et d’HMGN1 illustrent spécifiquement un remodelage tissulaire et de la chromatine au niveau utérin à la sortie de la diapause embryonnaire. Ayant mesuré une augmentation des concentrations utérines en polyamines à la reprise du développement embryonnaire, nous avons émis l’hypothèse que les polyamines seraient impliquées dans les événements menant à la sortie de la diapause. L’inhibition de la biosynthèse des polyamines par un traitement à l’ α-difluoromethylornithine (DFMO) a provoqué une diminution significative de la proliferation cellulaire dans les embryons à la réactivation, un retard du moment de l’implantation, mais n’a pas affecté le succès de la reproduction. De manière similaire, nous avons induit un état de dormance dans les cellules de trophoblaste de vison en présence DFMO dans le milieu de culture, et constaté que cet état était réversible. En conclusion, cette étude a non seulement ouvert de nouveaux horizons quant à la compréhension du rôle utérin dans les événements menant à la sortie de la diapause embryonnaire, mais a démontré pour la première fois, l’existence de facteurs utérins indispensables à la réactivation de l’embryon: les polyamines. / Embryonic diapause is characterized by a reversible arrest of blastocyst development prior to implantation and delay in implantation. In the American mink, embryonic diapause is a characteristic of each gestation. Although the mechanisms which control obligate embryonic diapause of this species are well known, the role of the uterus involved in blastocyst reactivation remains elusive. The subject of this doctoral research consisted first in exploring the uterine environment at the emergence of embryonic diapause in order to subsequently determine, the main factors in the uterus that provoke reactivation of the embryo. We have undertaken an analysis of the uterine transcriptome at the emergence of embryonic diapause which has enabled us to set up a library of 123 cDNA uterine sequences differentially expressed at blastocyst reactivation, and homologue gene sequences known in other species. Twenty-five percent of these genes are implicated in genetic expression, 15 % in cell signal transduction, 15 % in cell cycle, 10 % in transport and 9 % in cell structure. All of them reflect significant uterine modifications at blastocyst reactivation. We have validated differential expression of ten genes, identified as: GDF3 (growth and differentiation 3), ALCAM (activated leukocyte cell adhesion molecule), ADIPOR1 (adiponectin receptor 1), HMGN1 (high mobility group N1), TXNL1 (thioredoxine like 1), TGM2 (tissue transglutaminase 2), SPARC (secreted protein acidic rich in cystein), and three genes encoding for AZIN1 (antizyme inhibitor 1), ODC1 (ornithine decarboxylase 1) and SAT1 (spermidine/spermine N1-acetyltransferase), which are enzymes implicated in polyamine biosynthesis. The spatio-temporal expression patterns of SPARC and HMGN1 illustrate tissue and chromatin remodelling in the uterus at the termination of embryonic diapause. Having measured an increase in concentration of polyamines in the uterus at the resumption of blastocyst development, we have hypothetized that polyamines are implicated in the emergence of blastocysts from diapause. We inhibited polyamine biosynthesis in pregnant mink females during early blastocyst reactivation. The inhibition of polyamine biosynthesis through treatment with α-difluoromehtylornithine (DFMO) provoked a major reduction in cell proliferation in blastocysts at reactivation and a delay in the timing of implantation, but did not affect the success of reproduction. Similarly, we induced a reversible dormant state in cultured mink trophoblast cells traited with DFMO. To conclude, not only are results of this study a breakthrough in the understanding of the role of the uterus in stimulating at the emergence of blastocysts from embryonic diapause, but also, for the very first time, they indicate the existence of uterine factors, the polyamines, that are responsible for blastocysts reactivation.

Safeners are the chemicals used to protect plants from detrimental effects of herbicides, but their mode of action at the molecular level is not well understood. As an initial step towards understanding the molecular mechanism of safener action in trees, homologous genes in hybrid poplar (Populus nigra x Populus maximowiczii) that were induced by a safener were identified. We here describe the identification of differentially expressed genes in Populus that are induced by Concep-III, a herbicide safener. Expressed sequence tags (ESTs) enriched for transcriptionally induced genes were isolated by suppressive subtractive hybridization (SSH). The SSH library cDNA inserts were used to construct a cDNA microarray for high-throughput validation of the up-regulated expression of safener-induced genes. Single-pass and partial sequences of 1,344 safener-induced ESTs were assembled into 418 single-tons and 328 clusters, but the putative functions of almost 53% of the ESTs are not known. Genes encoding proteins involved in all three different phases of safener action, viz., oxidation, conjugation, and sequestration, were found in the SSH library. Almost 75% of genes that showed greater than 2-fold expression upon safener treatment were redundant in the SSH library. The expression pattern for selected genes was validated by reverse transcription-polymerase chain reaction. A few safener-induced genes that were not previously reported to be induced by safeners, but which may have a role in herbicide metabolism, were identified. The newly identified genes could have potential for application in genetic engineering of plants for herbicide detoxification and tolerance.

Staphylococcus xylosus est une bactérie ubiquitaire, commensale de la peau et des muqueuses de l'homme et des animaux. C'est un des principaux ferments utilisé en salaison. Ses propriétés technologiques étaient bien connues mais peu de données génétiques sur cette espèce étaient disponibles. Nous avons établi la première carte physique et génétique de la souche modèle S. xylosus C2a. Cette carte nous a permis d'avoir une estimation de la taille (2,89 Mb) et de l'organisation globale d'un chromosome de S. xylosus. Au sein de cette espèce, nous avons montré qu'il existait une grande diversité, tant au niveau des caractères phénotypiques (métabolisme des sucres, formation de colonies géantes) que génotypiques (profils génomiques, taille du chromosome). Nous avons étudié, par hybridation soustractive, la diversité du contenu génétique de souches de diverses origines. Le génome de la souche C2a a été soustrait à ceux d'un ferment et de deux souches isolées d'infections opportunistes. Au total, 78 kb de séquences d'ADN ont été identifiés, la majorité correspondant à des gènes du métabolisme. La distribution de ces fragments souchesspécifiques au sein de l'espèce S. xylosus révèle deux groupes dont un composé des souches présentant un risque potentiel. Ces fragments pourraient être utilisés dans des études épidémiologiques. La région de l'oriC est une zone d'insertion privilégiée de ces fragments sur les chromosomes de S. xylosus. Elle permet l'acquisition de matériel génétique important pour son adaptation à différentes niches écologiques. Le séquençage de cette zone chez différentes souches permettra d'évaluer la plasticité des génomes de S. xylosus.

No description available.

O Brasil é o segundo maior produtor de feijão, sendo a espécie mais cultivada o Phaseolus vulgaris L. Entre as três possíveis safras exploradas no Brasil, aquela que gera a maior produção é a da seca. Por outro lado, como a maioria das lavouras emprega pouca tecnologia, um dos problemas desta cultura é o estresse hídrico, que leva a uma redução na produtividade. Dessa forma, a identificação de genes que controlam os mecanismos de defesa e adaptação do feijoeiro à falta de água seria de grande utilidade. Nos últimos anos, muitas informações ômicas do feijoeiro foram geradas, criando uma visão integrada deste organismo e oferecendo uma complexa rede de interações entre genes e seus produtos. Este trabalho teve como objetivo central à identificação de genes diferencialmente expressos no sistema radicular de um genótipo de feijoeiro resistente ao estresse hídrico (BAT 477), quando submetido a uma interrupção de irrigação durante seu desenvolvimento. Foi construída uma biblioteca subtrativa de cDNA (SSH), que representou os genes diferencialmente expressos no genótipo resistente, utilizando-se como driver o genótipo Carioca 80SH (suscetível a seca). Foram obtidos 1572 reads válidos, sendo 931 destes singletons e 189 contigs com uma média de seis reads por cluster. A anotação das sequências foi conduzida via BLASTX, sendo consideradas para anotação somente os melhores resultados dos produtos gênicos similares com E- Value \'<OU=\' 10-5. A classificação funcional foi feita tendo-se como base modelos descritos para plantas (modelo CS e MIPS) e os resultados foram agrupados em seis classes funcionais distintas. As análises de bioinformática ajudaram na identificação de genes descritos como envolvidos na resposta da planta ao estresse hídrico. Entre eles: proteínas do grupo LEA; fatores de transcrição como DREB, NAC e proteínas ricas em leucina; enzimas sintetizadoras de carboidratos incluindo trehalose, sacarose e rhamnose; proteínas ricas em prolina; receptores de hormônios (ABA, etileno); aquaporinas; chaperonas; ubiquitinas; nodulinas; e proteínas associadas à fotossíntese e à respiração. A fim de se obter a validação das ESTs anotadas, foi conduzido um experimento de PCR em tempo real confrontando os padrões de expressão de 15 genes sob quatro tratamentos: ambos os genótipos sob estresse e respectivos controles. Três replicatas biológicas foram adotadas e dois genes de referência (act e skip2) foram escolhidos para normalização interna dos dados. Os padrões de expressão gênica obtidos confirmam a hipótese de que tais genes são mesmo mais expressos no genótipo resistente, embora não sejam exclusivos já que uma quantidade menor de tais transcritos também foi detectada no genótipo suscetível / Brazil is the second biggest producer of common bean, being Phaseolus vulgaris L. the most cultivated species. Among the three possible harvests exploited in Brazil, drought is the one which generates the greatest production. On the other hand, as the majority of the households employees low technology, one of the problems of this culture is drought stress that leads to a reduction in the productivity. So, the identification of gene that controls the mechanisms of defense and adaptation of common bean to the lack of water would be very useful. In the past years, many omics information of common bean have been generated, creating an integrated view of this organism and providing a complex network between genes and its products. The main goal of this work was the identification of differentially expressed genes in a genotype of common bean resistant to drought stress (BAT 477), when submitted to a interruption of irrigation during its development. It was build a cDNA suppression subtractive hybridization library (SSH), which represented the differentially expressed genes, on the resistant genotype, having as driver the genotype Carioca 80SH (susceptible to drought). It was obtained 1572 valid reads, being 931 singletons and 189 contigs, with the average of 6 reads per cluster. The sequences annotation was conducted via BLAST X, considering only the best similarity results with E value \'<OU=\' 10-5. The functional classification was done adopting models described for plants (CS and MIPS) and the results were grouped into six different functional classes. Bioinformatic analyses contribuited to the identification of genes described as involved on plants response to drought stress. Among them: LEA proteins; transcription factors like DREB, NAC and leucine-rich proteins; carbohydrates synthesizers enzymes like the ones for trehalose, sucrose and rhamnose; proline-rich proteins; hormone receptors (ABA and ethylene); aquaporins; chaperones; ubiquitins; nodulins; and proteins associated with photosynthesis and respiration. In order to validate the ESTs annotated, a RT-qPCR experiment was conducted comparing the expression patterns of 15 genes under four treatments: both genotypes under stress and their respective controls. Three technical replicates were used and two reference genes (act and skip2) were chosen for intern data normalization. The gene expression patterns obtained confirm the hypothesis that such genes are more expressed on the resistant genotype although they are not exclusive since a lower levels of these transcripts were also detected in the susceptible genotype