Os estresses abióticos impostos à planta no campo, tais como o estresse salino, toxidez por alumínio, frio, seca, entre outros, afetam seu crescimento, desenvolvimento e produtividade. Dentre as gramíneas cultivadas, o arroz (Oryza sativa) é uma das culturas de maior importância no Brasil, cuja oscilação na produção acarreta prejuízos consideráveis à economia brasileira. Dessa forma, o estudo das interações entre os estresses abióticos e as respostas dos vegetais frente a esses estímulos ambientais é fundamental para um conhecimento detalhado desses mecanismos. O arroz é um dos cereais mais tolerantes ao alumínio (Al), sendo um ótimo modelo para o estudo de mecanismos de tolerância. Neste trabalho, foi estudado o papel das proteínas ASR na tolerância ao Al. A expressão dos genes ASR (ABA, Stress and Ripening) é induzida por ABA, estresses e amadurecimento do fruto. Essas proteínas foram caracterizadas como chaperonas e fatores de transcrição. Os genes da família OsASR também têm o nível de transcritos aumentado em resposta ao Al, sendo OsASR1 e OsASR5 os genes com expressão mais abundante em arroz. No entanto, as regiões promotoras responsivas ao Al e os fatores de transcrição reguladores da expressão desses genes ainda não foram descritos. Assim, este trabalho tem como objetivo geral a identificação dos fatores que regulam a transcrição dos genes OsASR1 e OsASR5. Para tanto, construções contendo fragmentos dos promotores desses genes dirigindo a expressão do gene repórter GUS foram obtidas, visando ensaio de expressão transiente em protoplastos de arroz submetidos ao tratamento com Al. Os fragmentos das regiões promotoras de ambos os genes OsASR1 e OsASR5 foram todos responsivos ao Al. Também foi realizado ensaio de transativação em protoplastos de Arabidopsis thaliana para verificar a existência de auto-regulação nesses genes. Tanto ASR1 quanto ASR5 foram capazes de transativar seus próprios promotores. Além disso, foi realizada uma triagem de biblioteca de cDNAs por mono-híbrido em levedura com fragmento da região promotora de OsASR5. Com essa abordagem, foram identificados sete genes candidatos a codificadores de fatores capazes de interação DNA-proteína. / The abiotic stress that plants in the field are subjected to, such as salt, aluminum, cold, drought, among others, affect their growth, development and productivity. Among cultivated grasses, rice (Oryza sativa) is one of the most important crops in Brazil, whose oscillation in production causes considerable costs to the Brazilian economy. Thus, the study of interactions between abiotic stresses and plant responses to these environmental stimuli is essential to a detailed knowledge of these mechanisms. Rice is one of the most Al-tolerant crops, being a great model for studying Al-tolerance mechanisms. In this work, the role of ASR proteins in Al tolerance was studied. The expression of ASR (ABA, Stress and Ripening) genes is induced by ABA, stresses and fruit ripening. These proteins were characterized as chaperones and transcription factors. The OsASR genes also have increased transcript accumulation in response to Al, and OsASR1 and OsASR5 have the most abundant expression in rice. However, the Al-responsive promoter regions and the transcription factors that regulate these genes have not yet been described. Therefore, the goal of this work is to identify regulating factors of OsASR1 and OsASR5 gene transcription. For this, vectors containing promoter fragments of these genes driving the expression of the GUS gene were constructed and transient expression assays were performed in rice protoplasts subjected to Al treatment. All of the promoter fragments were Al-responsive for both OsASR1 and OsASR5 genes. Transactivation assays in Arabidopsis thaliana protoplasts were also conducted in order to verify the existence of auto-regulation in these genes. Both ASR1 and ASR5 were able to transactivate its own promoters. Furthermore, a library screening was performed by yeast one-hybrid using a promoter fragment from OsASR5. With this approach, seven candidate genes encoding transcription factors capable of DNA-protein interactions were identified.

Um grande empecilho para a manutenção e estabilidade da produção nacional de soja reside na suscetibilidade dos diferentes genótipos aos estresses ambientais. Tendo em vista a importância social e econômica da leguminosa e os efeitos extremamente danosos dos estresses abióticos sobre a agricultura, faz-se necessário maior conhecimento acerca das interações entre os estímulos estressores e as respostas da planta. A seca é considerada o principal fator limitante na produtividade agrícola. Sendo assim, a identificação e caracterização de genes responsivos a essa condição é um passo inicial na compreensão das respostas adaptativas ao déficit hídrico. Os genes Asr (ABA, Stress and Ripening) são induzidos por estresse e ácido abscísico (ABA) e seus níveis de expressão são rapidamente aumentados em resposta à salinidade e seca. Nesse estudo os genes da família Asr de soja foram clonados. Estas proteínas são hidrofílicas e ricas nos aminoácidos Ala, His, Glu e Lis, apresentando homologia com ASRs de outras plantas, como atestado nas análises de múltiplos alinhamentos. O perfil de expressão foi avaliado através de RT-qPCR em tempo real e revelou que Asr1 tem um distinto padrão de indução no nível de transcritos em folha sob tratamento com ABA, sal e seca, enquanto Asr3 apresenta padrão distinto de indução na expressão em raiz, sob tratamento com ABA e seca. Além disto, foram construídos vetores para a superexpressão e localização subcelular das proteínas ASR1, ASR2 e ASR3 em plantas. Plantas de Arabidopsis thaliana foram submetidas a um protocolo de transformação genética mediada por Agrobacterium. / One of the major obstacles to maintain the stability of the national production of soybean (Glycine max) lies on the susceptibility of different genotypes to abiotic stress. In view of the social and economic importance of soybean and due to the extremely harmful effects of stress in agriculture, detailed knowledge of the interaction between these stresses and plant response to environmental stimuli is necessary. Drought is considered the main abiotic limitation factor for agricultural productivity. Identification and characterization of responsive genes to this condition is an initial step in understanding the adaptive responses to drought. The Asr (ABA, Stress and Ripening) genes are induced by stress and abscisic acid (ABA) in plants, and their expression levels are quickly increased in response to salinity and drought. In this study Asr genes from Glycine max were cloned. These proteins were found to be hydrophilic and rich in amino acids Ala, His, Glu and Lys, showing homology with those of other plant Asr genes via multiple alignment analysis. RT-qPCR analyses revealed that Asr1 had a distinct up-regulated transcript pattern in leaf under ABA, NaCl and drought treatments, while Asr3 had a distinct up-regulated transcript pattern in root under ABA and drought treatments. Besides, vectors for ASR1, ASR2 and ASR3 proteins overexpression and subcellular localization in plants were constructed. Arabidopsis thaliana plants were submitted to an Agrobacterium-mediated transformation procedure.

Um dos graves obstáculos para a manutenção e estabilidade da produção nacional de arroz (Oryza sativa) reside na susceptibilidade dos genótipos existentes a estresses abióticos. Tendo em vista a importância social e econômica do arroz e os efeitos extremamente danosos desses estresses sobre a agricultura, o conhecimento detalhado das interações entre os estresses abióticos e as respostas dos vegetais frente a esses estímulos ambientais faz-se necessário. O alumínio (Al) é considerado um dos principais fatores limitantes na produção agrícola, inibindo o crescimento das raízes e a absorção de minerais. A toxicidade do Al em plantas ocorre pela sua solubilização em solos com pH baixo ou solos ácidos. Os genes ASR (ABA, Stress and Ripening) são induzidos por estresse e ácido abscísico (ABA) em plantas, e seus níveis de expressão são rapidamente aumentados em resposta à salinidade e seca. Recentemente, foi demonstrado que o gene que codifica a proteína ASR5 é responsivo ao Al em raízes de arroz. Apesar do arroz ser considerado um dos cereais mais resistentes a Al, os mecanismos básicos de tolerância a este metal são pouco conhecidos no arroz em comparação a outros cereais. Por meio do presente trabalho objetivamos: i) a caracterização funcional dos membros da família gênica ASR de arroz em reposta ao Al; e ii) a construção de vetores binários de transformação de plantas visando o estudo da localização subcelular da proteína codificada pelo gene OsASR5, e o silenciamento gênico da família ASR de arroz. As análises dos transcritos por qRT-PCR mostraram que todos os genes da família ASR de arroz ssp Japonica respondem ao Al. Por outro lado, OsASR5 não sofre modulação de sua expressão em resposta ao Al em raízes de arroz ssp Indica. Essas diferenças de respostas dos genes OsASR5 em distintas variedades pode refletir diferenças no grau de tolerância ao Al de cada um desses genótipos. / One of the major obstacles to maintain the stability of the national production of rice (Oryza sativa) lies on the susceptibility of the different genotypes to abiotic stress. In view of the social and economic importance of rice and due to the extremely harmful effects of stress in agriculture, detailed knowledge of the interactions between these stresses and plant responses to environmental stimuli is necessary. Aluminum (Al) is considered one of the main limitation factors for agricultural productivity, inhibiting root growth and mineral absorption. Al toxicity occurs by its solubilization in soils with low pH or acid soils. The ASR (ABA, Stress and Ripening) genes are induced by stress and abscisic acid (ABA) in plants, and their expression levels are quickly increased in response to salinity and drought. Recently, it was demonstrated that the ASR5 gene is responsive to Al in rice roots. Despite the fact that rice is considered one of the most resistant crops to Al, the basic mechanisms of tolerance to this metal are poorly known when compared to other crops. This study aimed the functional characterization of the gene expression of rice ASR family members in response to Al, and the construction of binary vectors for the subcellular localization of the protein codified by the OsASR5 gene, and the construction of a gene silencing binary vector for the ASR family. Analyses of transcripts by qRT-PCR showed that in the ssp Japonica, all ASR genes responded to Al. In contrast, OsASR5 do not suffer expression modulation in response to Al in rice roots of ssp Indica. These differences in response of the OsASR5 gene in distinct varieties may reflect differences in the degree of Al tolerance in each genotype.

Um grande empecilho para a manutenção e estabilidade da produção nacional de soja reside na suscetibilidade dos diferentes genótipos aos estresses ambientais. Tendo em vista a importância social e econômica da leguminosa e os efeitos extremamente danosos dos estresses abióticos sobre a agricultura, faz-se necessário maior conhecimento acerca das interações entre os estímulos estressores e as respostas da planta. A seca é considerada o principal fator limitante na produtividade agrícola. Sendo assim, a identificação e caracterização de genes responsivos a essa condição é um passo inicial na compreensão das respostas adaptativas ao déficit hídrico. Os genes Asr (ABA, Stress and Ripening) são induzidos por estresse e ácido abscísico (ABA) e seus níveis de expressão são rapidamente aumentados em resposta à salinidade e seca. Nesse estudo os genes da família Asr de soja foram clonados. Estas proteínas são hidrofílicas e ricas nos aminoácidos Ala, His, Glu e Lis, apresentando homologia com ASRs de outras plantas, como atestado nas análises de múltiplos alinhamentos. O perfil de expressão foi avaliado através de RT-qPCR em tempo real e revelou que Asr1 tem um distinto padrão de indução no nível de transcritos em folha sob tratamento com ABA, sal e seca, enquanto Asr3 apresenta padrão distinto de indução na expressão em raiz, sob tratamento com ABA e seca. Além disto, foram construídos vetores para a superexpressão e localização subcelular das proteínas ASR1, ASR2 e ASR3 em plantas. Plantas de Arabidopsis thaliana foram submetidas a um protocolo de transformação genética mediada por Agrobacterium. / One of the major obstacles to maintain the stability of the national production of soybean (Glycine max) lies on the susceptibility of different genotypes to abiotic stress. In view of the social and economic importance of soybean and due to the extremely harmful effects of stress in agriculture, detailed knowledge of the interaction between these stresses and plant response to environmental stimuli is necessary. Drought is considered the main abiotic limitation factor for agricultural productivity. Identification and characterization of responsive genes to this condition is an initial step in understanding the adaptive responses to drought. The Asr (ABA, Stress and Ripening) genes are induced by stress and abscisic acid (ABA) in plants, and their expression levels are quickly increased in response to salinity and drought. In this study Asr genes from Glycine max were cloned. These proteins were found to be hydrophilic and rich in amino acids Ala, His, Glu and Lys, showing homology with those of other plant Asr genes via multiple alignment analysis. RT-qPCR analyses revealed that Asr1 had a distinct up-regulated transcript pattern in leaf under ABA, NaCl and drought treatments, while Asr3 had a distinct up-regulated transcript pattern in root under ABA and drought treatments. Besides, vectors for ASR1, ASR2 and ASR3 proteins overexpression and subcellular localization in plants were constructed. Arabidopsis thaliana plants were submitted to an Agrobacterium-mediated transformation procedure.

Os estresses abióticos impostos à planta no campo, tais como o estresse salino, toxidez por alumínio, frio, seca, entre outros, afetam seu crescimento, desenvolvimento e produtividade. Dentre as gramíneas cultivadas, o arroz (Oryza sativa) é uma das culturas de maior importância no Brasil, cuja oscilação na produção acarreta prejuízos consideráveis à economia brasileira. Dessa forma, o estudo das interações entre os estresses abióticos e as respostas dos vegetais frente a esses estímulos ambientais é fundamental para um conhecimento detalhado desses mecanismos. O arroz é um dos cereais mais tolerantes ao alumínio (Al), sendo um ótimo modelo para o estudo de mecanismos de tolerância. Neste trabalho, foi estudado o papel das proteínas ASR na tolerância ao Al. A expressão dos genes ASR (ABA, Stress and Ripening) é induzida por ABA, estresses e amadurecimento do fruto. Essas proteínas foram caracterizadas como chaperonas e fatores de transcrição. Os genes da família OsASR também têm o nível de transcritos aumentado em resposta ao Al, sendo OsASR1 e OsASR5 os genes com expressão mais abundante em arroz. No entanto, as regiões promotoras responsivas ao Al e os fatores de transcrição reguladores da expressão desses genes ainda não foram descritos. Assim, este trabalho tem como objetivo geral a identificação dos fatores que regulam a transcrição dos genes OsASR1 e OsASR5. Para tanto, construções contendo fragmentos dos promotores desses genes dirigindo a expressão do gene repórter GUS foram obtidas, visando ensaio de expressão transiente em protoplastos de arroz submetidos ao tratamento com Al. Os fragmentos das regiões promotoras de ambos os genes OsASR1 e OsASR5 foram todos responsivos ao Al. Também foi realizado ensaio de transativação em protoplastos de Arabidopsis thaliana para verificar a existência de auto-regulação nesses genes. Tanto ASR1 quanto ASR5 foram capazes de transativar seus próprios promotores. Além disso, foi realizada uma triagem de biblioteca de cDNAs por mono-híbrido em levedura com fragmento da região promotora de OsASR5. Com essa abordagem, foram identificados sete genes candidatos a codificadores de fatores capazes de interação DNA-proteína. / The abiotic stress that plants in the field are subjected to, such as salt, aluminum, cold, drought, among others, affect their growth, development and productivity. Among cultivated grasses, rice (Oryza sativa) is one of the most important crops in Brazil, whose oscillation in production causes considerable costs to the Brazilian economy. Thus, the study of interactions between abiotic stresses and plant responses to these environmental stimuli is essential to a detailed knowledge of these mechanisms. Rice is one of the most Al-tolerant crops, being a great model for studying Al-tolerance mechanisms. In this work, the role of ASR proteins in Al tolerance was studied. The expression of ASR (ABA, Stress and Ripening) genes is induced by ABA, stresses and fruit ripening. These proteins were characterized as chaperones and transcription factors. The OsASR genes also have increased transcript accumulation in response to Al, and OsASR1 and OsASR5 have the most abundant expression in rice. However, the Al-responsive promoter regions and the transcription factors that regulate these genes have not yet been described. Therefore, the goal of this work is to identify regulating factors of OsASR1 and OsASR5 gene transcription. For this, vectors containing promoter fragments of these genes driving the expression of the GUS gene were constructed and transient expression assays were performed in rice protoplasts subjected to Al treatment. All of the promoter fragments were Al-responsive for both OsASR1 and OsASR5 genes. Transactivation assays in Arabidopsis thaliana protoplasts were also conducted in order to verify the existence of auto-regulation in these genes. Both ASR1 and ASR5 were able to transactivate its own promoters. Furthermore, a library screening was performed by yeast one-hybrid using a promoter fragment from OsASR5. With this approach, seven candidate genes encoding transcription factors capable of DNA-protein interactions were identified.

Os estresses abióticos impostos à planta no campo, tais como o estresse salino, toxidez por alumínio, frio, seca, entre outros, afetam seu crescimento, desenvolvimento e produtividade. Dentre as gramíneas cultivadas, o arroz (Oryza sativa) é uma das culturas de maior importância no Brasil, cuja oscilação na produção acarreta prejuízos consideráveis à economia brasileira. Dessa forma, o estudo das interações entre os estresses abióticos e as respostas dos vegetais frente a esses estímulos ambientais é fundamental para um conhecimento detalhado desses mecanismos. O arroz é um dos cereais mais tolerantes ao alumínio (Al), sendo um ótimo modelo para o estudo de mecanismos de tolerância. Neste trabalho, foi estudado o papel das proteínas ASR na tolerância ao Al. A expressão dos genes ASR (ABA, Stress and Ripening) é induzida por ABA, estresses e amadurecimento do fruto. Essas proteínas foram caracterizadas como chaperonas e fatores de transcrição. Os genes da família OsASR também têm o nível de transcritos aumentado em resposta ao Al, sendo OsASR1 e OsASR5 os genes com expressão mais abundante em arroz. No entanto, as regiões promotoras responsivas ao Al e os fatores de transcrição reguladores da expressão desses genes ainda não foram descritos. Assim, este trabalho tem como objetivo geral a identificação dos fatores que regulam a transcrição dos genes OsASR1 e OsASR5. Para tanto, construções contendo fragmentos dos promotores desses genes dirigindo a expressão do gene repórter GUS foram obtidas, visando ensaio de expressão transiente em protoplastos de arroz submetidos ao tratamento com Al. Os fragmentos das regiões promotoras de ambos os genes OsASR1 e OsASR5 foram todos responsivos ao Al. Também foi realizado ensaio de transativação em protoplastos de Arabidopsis thaliana para verificar a existência de auto-regulação nesses genes. Tanto ASR1 quanto ASR5 foram capazes de transativar seus próprios promotores. Além disso, foi realizada uma triagem de biblioteca de cDNAs por mono-híbrido em levedura com fragmento da região promotora de OsASR5. Com essa abordagem, foram identificados sete genes candidatos a codificadores de fatores capazes de interação DNA-proteína. / The abiotic stress that plants in the field are subjected to, such as salt, aluminum, cold, drought, among others, affect their growth, development and productivity. Among cultivated grasses, rice (Oryza sativa) is one of the most important crops in Brazil, whose oscillation in production causes considerable costs to the Brazilian economy. Thus, the study of interactions between abiotic stresses and plant responses to these environmental stimuli is essential to a detailed knowledge of these mechanisms. Rice is one of the most Al-tolerant crops, being a great model for studying Al-tolerance mechanisms. In this work, the role of ASR proteins in Al tolerance was studied. The expression of ASR (ABA, Stress and Ripening) genes is induced by ABA, stresses and fruit ripening. These proteins were characterized as chaperones and transcription factors. The OsASR genes also have increased transcript accumulation in response to Al, and OsASR1 and OsASR5 have the most abundant expression in rice. However, the Al-responsive promoter regions and the transcription factors that regulate these genes have not yet been described. Therefore, the goal of this work is to identify regulating factors of OsASR1 and OsASR5 gene transcription. For this, vectors containing promoter fragments of these genes driving the expression of the GUS gene were constructed and transient expression assays were performed in rice protoplasts subjected to Al treatment. All of the promoter fragments were Al-responsive for both OsASR1 and OsASR5 genes. Transactivation assays in Arabidopsis thaliana protoplasts were also conducted in order to verify the existence of auto-regulation in these genes. Both ASR1 and ASR5 were able to transactivate its own promoters. Furthermore, a library screening was performed by yeast one-hybrid using a promoter fragment from OsASR5. With this approach, seven candidate genes encoding transcription factors capable of DNA-protein interactions were identified.

Aquifer storage and recovery (ASR) is a strategy in which water is injected into an aquifer when it is plentiful and pumped from the aquifer when water is scarce. An impediment to ASR in Florida is leaching of naturally-occurring arsenic from limestone of the Upper Floridan Aquifer System (UFAS) into stored water. The concentration of arsenic in surface water, which serves as the recharge water for many ASR systems, and native groundwater is usually much less than 3.0 µ/L. However, data from ASR wells in Florida show that arsenic in recovered water frequently exceeded the 10 µg/L maximum contaminant level (MCL) established by the Environmental Protection Agency and were as high as 130.0 µg/L. The cause of elevated arsenic concentrations is displacement of reduced native groundwater with oxygenated surface water that dissolves arsenic-bearing pyrite in limestone. Although arsenic can be removed from recovered water during final treatment, mobilization of arsenic in the aquifer at levels that exceed the MCL is problematic under federal regulations. This dissertation investigated a number of aspects of the ASR/arsenic problem to provide additional insights into the mechanisms of arsenic mobilization and measures that could be taken to avoid or reduce the release of arsenic during ASR operations. Chapter 2, involved development of a geochemical model to simulate an ASR system’s injection of oxygenated surface water into reduced groundwater to determine whether aquifer redox conditions could be altered to the degree of pyrite instability. Increasing amounts of injection water were added to the storage-zone in a series of steps and resulting reaction paths were plotted on pyrite stability diagrams. Unmixed storage-zone water in wells plotted within the pyrite stability field indicating that redox conditions were sufficiently reducing to allow for pyrite stability. Thus arsenic is immobilized in pyrite and its concentration in groundwater should be low. During simulation, as the injection/storage-zone water ratio increased, redox conditions became less reducing and pyrite became unstable. The result would be release of arsenic from limestone into storage-zone water. Chapter 3 examined the importance of maintaining a substantial volume of stored water around an ASR well to prevent recovery of reduced native groundwater to the vicinity of the well. Depleting the stored water and recovering reduced native groundwater would result in dissolution of arsenic-bearing hydrous ferric oxide (HFO) and release of arsenic into water recovered from the ASR well. Injection/recovery volumes for each cycle for each well were tracked to determine if a substantial volume of stored water was maintained for each cycle or if it was depleted so that reduced native groundwater was brought back to the well. Each well was assigned to either the “storage zone maintained group” where a zone of stored water was established in early cycles and largely maintained through the period of investigation, or the “storage-zone depleted group” where a zone of stored water was either established in later cycles and/or was depleted during the period of investigation. Graphical and statistical analyses verified that maximum arsenic concentrations for storage-zone maintained wells were nearly always lower in each cycle and declined below the MCL after fewer cycles than those of storage-zone depleted wells. Chapter 4 was a mineralogical investigation of cores located at 20 m (ASR core 1), 152 m (ASR core 2), and 452 m (ASR core 3) from operating ASR wells to determine where mobilized arsenic in limestone is precipitated during ASR. If arsenic is precipitated distally, reduced concentrations of elements in pyrite, (iron, sulfur, arsenic, etc.) would be expected in ASR core 1 relative to more distant cores and there would be noticeable changes in appearance of pyrite crystals due to enhanced oxidation. The results showed that mean concentrations of the elements were lowest in ASR core 2, which did not support distal precipitation. However, scanning electron microscopy identified well-defined pyrite framboids only in core 3 while framboids in ASR cores 1 and 2 were less clear and distinct, indicating pyrite oxidation in cores closest to ASR wells. Statistical comparison of concentrations of iron, sulfur, and arsenic between the three ASR cores and 19 control cores not subject to ASR, showed that mean concentrations in ASR cores 1 and 2 were statistically similar to concentrations in control cores. This indicated that concentrations in ASR cores 1 and 2 had not been significantly reduced by ASR. The concentrations of elements were higher in ASR core 3 than in ASR cores 1 and 2 and control cores and statistically dissimilar to all but one control core. This indicated natural heterogeneity in core 3 rather than diminution of elements in ASR cores 1 and 2 due to ASR. The statistical analysis supported local precipitation. Once arsenic is mobilized from dissolved pyrite, it is rapidly complexed with precipitated HFO near the well. As long as all of the stored water is not removed during recovery so that reduced native groundwater is brought back to the well, HFO remains stable and complexed with arsenic. The concentration of elements would not have been lowest in ASR core 1 for this reason and because calculations showed that the mass of arsenic removed during recovery events prior to coring was minor compared to the total in limestone surrounding the well. The implications of this are that while large quantities of arsenic are present near the ASR well, only a small percentage may be available for dissolution. Most arsenic occurs with pyrite in limestone, which may insulate it from exposure to oxidized injection water. Water recovered from ASR wells may continue to have low concentrations of arsenic indefinitely because as limestone is dissolved, more pyrite becomes exposed and available for dissolution. The primary contribution of this dissertation to understanding and overcoming the arsenic problem in ASR systems is the empirical data developed to support or challenge important ASR/arsenic hypotheses. These data were used to 1) establish that background concentrations of arsenic in groundwater of the Suwannee Limestone were less than 1µg/L, 2) demonstrate that redox conditions necessary for pyrite in limestone to become unstable and dissolve occur when oxygenated surface water is injected into the aquifer, 3) demonstrate that the concentration of pyrite in the Suwannee Limestone is spatially variable to a high degree, 4) support the hypothesis that following injection of oxygenated surface water, pyrite in limestone dissolves and releases arsenic into solution and HFO forms and complexes with the arsenic near the ASR well, 5) propose that only a small percentage of pyrite near an ASR well may be available for dissolution during each cycle because most occurs in the limestone matrix and is isolated from injection water, 6) propose that as a result of the previous conclusion, water recovered from ASR systems may continue to have low concentrations of arsenic indefinitely because as limestone that contains pyrite is dissolved with each cycle, additional pyrite is exposed and is available for dissolution, and 7) support the effectiveness of maintaining a zone of stored water in an ASR well as an effective means of minimizing arsenic in recovered water during ASR.

Les troubles mentaux sont une problématique considérable depuis plusieurs années. Afin de mieux répondre aux besoins des personnes atteintes de troubles mentaux, le Code des professions a redéfini les champs d’exercices de certaines professions touchées par cette problématique, dont les conseillers et conseillères d'orientation. Pour l’évaluation des personnes atteintes de troubles mentaux, les instruments psychométriques s’avèrent des outils d’évaluation fort utiles aux c.o. Le questionnaire Adult Self -Report (ASR), nouvellement traduit en français pour le Canada, est l’instrument de l’ASEBA permettant l’évaluation de l’adulte selon sa propre autoévaluation. Des analyses bivariées et multivariées ont été effectuées et ont donné lieu à des résultats appuyant la validité critériée de l’ASR. Les résultats suggèrent que l’ASR a une bonne validité critériée pour l’identification des personnes atteintes de problèmes psychiatriques nécessitant une hospitalisation.

People with motor function disorders that cause dysarthric speech find difficulty using state-of- the-art automatic speech recognition (ASR) systems. These systems are developed based on non- dysarthric speech models, which explains the poor performance when used by individuals with dysarthria. Thus, a solution is needed to compensate for the poor performance of these systems. This thesis examines the possibility of quantifying vowels of dysarthric and non-dysarthric speech into codewords regardless of inter-speaker variability and possible to be implemented on limited- processing-capability machines. I show that it is possible to model all possible vowels and vowel- like sounds that a North American speaker can produce if the frequencies of the first and second formants are used to encode these sounds. The proposed solution is aligned with the use of neural networks and hidden Markov models to build an acoustic model in conventional ASR systems. A secondary finding of this study includes the feasibility of reducing the set of ten most common vowels in North American English to eight vowels only. / Graduate / 2021-05-11

Research into Automatic Speech Recognition (ASR), the task of transforming speech into text, remains highly relevant due to its countless applications in industry and academia. State-of-the-art ASR models are able to produce nearly perfect, sometimes referred to as human-like transcriptions; however, accurate ASR models are most often available only in high-resource languages. Furthermore, the vast majority of ASR models are monolingual, that is, only able to handle one language at a time. In this thesis, we extensively evaluate the quality of existing monolingual ASR models for Swedish, Danish, and Norwegian. In addition, we search for parallels between monolingual ASR models and the cognition of foreign languages in native speakers of these languages. Lastly, we extend the Swedish monolingual model to handle all three languages. The research conducted in this thesis project is divided into two main sections, namely monolingual and multilingual models. In the former, we analyse and compare the performance of monolingual ASR models for Scandinavian languages in monolingual and cross-lingual settings. We compare these results against the levels of mutual intelligibility of Scandinavian languages in native speakers of Swedish, Danish, and Norwegian to see whether the monolingual models favour the same languages as native speakers. We also examine the performance of the monolingual models on the regional dialects of all three languages and perform qualitative analysis of the most common errors. As for multilingual models, we expand the most accurate monolingual ASR model to handle all three languages. To do so, we explore the most suitable settings via trial models. In addition, we propose an extension to the well-established Wav2Vec 2.0-CTC architecture by incorporating a language classification component. The extension enables the usage of language models, thus boosting the overall performance of the multilingual models. The results reported in this thesis suggest that in a cross-lingual setting, monolingual ASR models for Scandinavian languages perform better on the languages that are easier to comprehend for native speakers. Furthermore, the addition of a statistical language model boosts the performance of ASR models in monolingual,  cross-lingual, and multilingual settings. ASR models appear to favour certain regional dialects, though the gap narrows in a multilingual setting. Contrary to our expectations, our multilingual model performs comparably with the monolingual Swedish ASR models and outperforms the Danish and Norwegian models. The multilingual architecture proposed in this thesis project is fairly simple yet effective. With greater computational resources at hand, further extensions offered in the conclusions might improve the models further.