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

Submitted by Maria Beatriz Vieira (mbeatriz.vieira@gmail.com) on 2017-01-05T13:10:39Z No. of bitstreams: 2 license_rdf: 0 bytes, checksum: d41d8cd98f00b204e9800998ecf8427e (MD5) tese_fabio_schaun_harter.pdf: 3557391 bytes, checksum: 5b6d3757e05c02956957ee671553ba7f (MD5) / Approved for entry into archive by Aline Batista (alinehb.ufpel@gmail.com) on 2017-01-09T11:55:09Z (GMT) No. of bitstreams: 2 tese_fabio_schaun_harter.pdf: 3557391 bytes, checksum: 5b6d3757e05c02956957ee671553ba7f (MD5) license_rdf: 0 bytes, checksum: d41d8cd98f00b204e9800998ecf8427e (MD5) / Approved for entry into archive by Aline Batista (alinehb.ufpel@gmail.com) on 2017-01-09T11:56:59Z (GMT) No. of bitstreams: 2 tese_fabio_schaun_harter.pdf: 3557391 bytes, checksum: 5b6d3757e05c02956957ee671553ba7f (MD5) license_rdf: 0 bytes, checksum: d41d8cd98f00b204e9800998ecf8427e (MD5) / Made available in DSpace on 2017-01-09T11:57:10Z (GMT). No. of bitstreams: 2 tese_fabio_schaun_harter.pdf: 3557391 bytes, checksum: 5b6d3757e05c02956957ee671553ba7f (MD5) license_rdf: 0 bytes, checksum: d41d8cd98f00b204e9800998ecf8427e (MD5) Previous issue date: 2014-02-28 / Sem bolsa / O objetivo do trabalho foi avaliar a produção de sementes de arroz de diferentes cultivares, sob estresse salino. O experimento foi realizado em duas safras agrícolas; no primeiro ano foram avaliados cinco genótipos (IRGA 417, Avaxi CL, SCSBRS Tio Taka, Inov CL e IAS 12-9 - Formosa) submetidos a doses de 0, 12,5; 25; 37,5; 50; 75 e 100 mM. No segundo ano utilizou-se quatro genótipos (IRGA 417, Epagri 109, BRS Querência e IAS 12-9 - Formosa) nas doses 0, 4, 8, 12 e 16 mM. Avaliou-se a qualidade fisiológica das sementes sob estresse salino pelos testes de germinação, primeira contagem de germinação, teste frio, envelhecimento acelerado, índice de velocidade de emergência, emergência, área foliar, comprimento de parte aérea e sistema radicular, fitomassa seca parte aérea e sistema radicular. Os genótipos foram cultivados sob as mesmas condições de salinidade, avaliando-se no final do ciclo as características agronômicas e a qualidade da semente produzida da seguinte forma: número de panículas, peso total, peso de sementes cheias, peso de sementes chochas, porcentagem de sementes cheias e chochas, e peso de mil sementes; e qualidade fisiológica pelos teste de germinação, primeira contagem de germinação e teste de frio. Conclui-se que a qualidade fisiológica de sementes de arroz apresenta redução com o aumento dos níveis de estresse salino; cultivares de arroz mostram respostas diferenciadas sob condições de estresse salino; as cultivares de arroz IRGA 417, Epagri 109, BRS Querência e IAS 12-9 (Formosa) apresentam redução da produção de sementes sob condições de estresse salino; a qualidade fisiológica de sementes de arroz das cultivares IRGA 417, Epagri 109, BRS Querência e IAS 12-9 (Formosa), não são afetadas negativamente em concentrações salinas até 16 mM. / The objective of this work was to evaluate the production of rice seeds of different cultivars under salt stress. The experiment was conducted in two growing seasons, in the first year, the following five rice genotypes (IRGA 417, Avaxi CL, SCSBRS Tio Taka, Inov CL e IAS 12-9 - Formosa) were evaluated to salt tolerance and they were submitted to doses of 0; 12.5; 25; 37.5; 50; 75 and 100 mM. In the second year, four genotypes (IRGA 417, Epagri 109, BRS Querência e IAS 12-9 - Formosa) were used at dosages of 0; 4; 8; 12 and 16 mM. The following tests were performed to evaluate seed quality under stress: germination, first count, cold test, accelerated aging, emergence speed index, emergence, leaf area, shoot and root length, stem and root dry biomass. All genotypes were grown under the same salinity conditions and the agronomic characteristics and quality of seed produced were evaluated at the end of the cycle as follows: number of panicles, total weight, weight of full seeds, weight of empty seeds, percentage of full and empty seeds and thousand seed weight. Physiological seed quality was evaluated by tests of germination, first count and cold test. It can be concluded that physiological quality of rice seeds showed a decrease with the increase in salt stress levels. Rice cultivars show different responses under salt stress; salt stress reduce the production of seed of the cultivars IRGA 417, Epagri 109, BRS Querência and IAS 12-9 (Formosa) and the physiological quality of rice seeds of the cultivars IRGA 417, Epagri 109, BRS Querência and IAS 12-9 (Formosa), are not affected in salt concentrations up to 16 mM .

Submitted by Maria Beatriz Vieira (mbeatriz.vieira@gmail.com) on 2017-03-17T12:23:54Z No. of bitstreams: 2 license_rdf: 0 bytes, checksum: d41d8cd98f00b204e9800998ecf8427e (MD5) dissertacao_paulo_eduardo_rocha_eberhardt.pdf: 1985819 bytes, checksum: bed6d72ca6a80b32c05abe648c7fd3d5 (MD5) / Approved for entry into archive by Aline Batista (alinehb.ufpel@gmail.com) on 2017-03-17T22:21:54Z (GMT) No. of bitstreams: 2 license_rdf: 0 bytes, checksum: d41d8cd98f00b204e9800998ecf8427e (MD5) dissertacao_paulo_eduardo_rocha_eberhardt.pdf: 1985819 bytes, checksum: bed6d72ca6a80b32c05abe648c7fd3d5 (MD5) / Approved for entry into archive by Aline Batista (alinehb.ufpel@gmail.com) on 2017-03-17T22:22:51Z (GMT) No. of bitstreams: 2 license_rdf: 0 bytes, checksum: d41d8cd98f00b204e9800998ecf8427e (MD5) dissertacao_paulo_eduardo_rocha_eberhardt.pdf: 1985819 bytes, checksum: bed6d72ca6a80b32c05abe648c7fd3d5 (MD5) / Made available in DSpace on 2017-03-17T22:23:00Z (GMT). No. of bitstreams: 2 license_rdf: 0 bytes, checksum: d41d8cd98f00b204e9800998ecf8427e (MD5) dissertacao_paulo_eduardo_rocha_eberhardt.pdf: 1985819 bytes, checksum: bed6d72ca6a80b32c05abe648c7fd3d5 (MD5) Previous issue date: 2015-02-26 / Sem bolsa / O arroz (Oryza sativa L.) é um dos cereais mais cultivados, especialmente na Ásia onde concentra 90% da produção e consumo mundial de arroz. A qualidade das sementes é determinada por fatores físicos, genéticos, fisiológicos e sanitários. O objetivo foi avaliar o comportamento de sementes de arroz durante o armazenamento e o uso de pós de rocha no recobrimento de sementes. O trabalho foi conduzido na Universidade Federal de Pelotas no ano agrícola 2013/2014. As sementes foram recobertas por dois pós de rocha sendo um granodiorito e um basalto nas doses de zero, 7,5%, 15% e 30% de pó de rocha e armazenadas por doze meses. A cada dois meses foram realizados os testes de germinação, primeira contagem de germinação, envelhecimento acelerado, teste de frio, comprimento de raiz, comprimento de parte aérea e teor de água. No mês de outubro, período de semeadura da cultura foi realizado os testes de emergência em areia, índice de velocidade de emergência e matéria seca de plântulas. Pode-se observar que o recobrimento de sementes com pós de rocha proporcionaram maior germinação, melhor desempenho no teste de frio, maior comprimento de parte aérea e redução no teor de umidade de sementes. O recobrimento proporcionou também maiores índices de velocidade de emergência e matéria seca de plântulas em relação as sementes não tratadas. / Rice (Oryza sativa L.) is one of the most cultivated cereals, especially in Asia where it concentrates 90% of world production and consumption of rice. The seed quality is determined by physical, genetic, physiological and health. The objective was to evaluate the rice seed storage behavior and the use of rock in the post seed coating. The work was conducted at the Federal University of Pelotas in the agricultural year 2013/2014. The seeds were coated with two rock powders being one granodiorite rock and basalt in the untreated seeds and the doses, 7.5%, 15% and 30% of rock dust, and stored for twelve months. Every two months were performed by standard germination, first count of germination, accelerated aging, cold test, root length, shoot length and water content. In October, crop sowing period was evaluated emergence test in sand, emergency speed index and seedling dry. It can be observed that the coating of seeds with rock powders showed higher germination, improved performance in the cold test, increased shoot length and reduction in moisture content of the seeds. The coating also provided higher speed of emergence index and seedling dry over the untreated seeds.

X-ray crystallography is one of the most powerful tools for the elucidation of the structure of biological macromolecules such as proteins and viruses. Crystallographic techniques are extensively used for investigations on protein structure, ligand-binding, mechanisms of enzyme catalyzed reactions, protein-protein interactions, role of metal ions in protein structure and function, structure of multi-enzyme complexes and viruses, protein dynamics and for a myriad other problems in structural biology. Crystallographic studies are essential for understanding the intricate details of the mechanism of action of enzymes at molecular level. Understanding the subtle differences between the pathogenic enzymes and host enzymes is necessary for the design of inhibitor molecules that specifically inhibit parasite enzymes. The current thesis deals with the application of biochemical and crystallographic techniques for understanding the structure and function of proteins from two pathogenic organisms – a plant virus Physalis Mottle Virus (PhMV), and a pathogenic bacterium, Salmonella typhimurium and also stress induced proteins from Oryza sativa. The thesis has been divided into seven chapters, with the first four chapters describing the work carried out on PhMV, while the rest of the chapters deal with the studies on stress response proteins from Oryza sativa and Salmonella typhimurium. The first part of the thesis deals with studies on viral capsids. Viruses are obligate parasites that have proteinaceous capsids enclosing the genetic material, which, in the case of small plant viruses, is invariably ss-RNA. X-ray diffraction studies on single crystals of viruses enable visualization of the structures of intact virus particles at near-atomic resolution. These studies provide detailed information regarding the coat protein folding, molecular interactions between protein subunits, flexibility of the N-and C-terminal segments and their probable importance in viral assembly, role of RNA in capsid assembly, nucleic acid (RNA)-protein interactions, the capsid structure and mechanism of assembly and disassembly. The present thesis deals with the capsid structure and analysis of the coat protein (CP) recombinant mutants of PhMV. Virus assembly, one of the important steps in the life cycle of a virus, involves specific interactions between the structural protein and cognate viral genome. This is a complex process that requires precise protein-protein and protein nucleic acid interactions. In fact, most of the biological functional units such as ribosomes and proteosomes also require highly co-ordinated macromolecular interactions for their functional expression. Viruses being simple in their architecture, serve as excellent model systems to understand mechanism of macromolecular assembly and provide necessary information for the development of antiviral therapeutics, especially in animal viruses. PhMV is a plant virus infecting several members of Solanaceae family. It belongs to the tymoviridae group of single stranded RNA viruses. Its genome is encapsidated in a shell comprising of 180 (architecture based on T = 3 icosahedral lattice) chemically identical coat protein (CP) subunits (~ 20,000Da) arranged with icosahedral symmetry. In an earlier phase of work, PhMV purified from infected plant leaves was crystallized in the space group R3 (a = 294.56 Å,  = 59.86). X-ray diffraction data to 3.8 Å resolution were recorded on films by screenless oscillation photography. Using this data of severely limited quality and poor completion (40%), the structure PhMV was determined by molecular replacement using the related turnip yellow mosaic virus (TYMV) structure as the phasing model. There was therefore a need to re-determine and improve the structure, which could be useful for understanding the earlier detailed studies on its biophysical properties. As a continuation of these studies, the present investigations were conceived with the goal of determining the natural top and bottom component capsid structures of PhMV. Investigations were also carried out to examine the possibility of enhancing the diffraction quality of PhMV crystals. The thesis begins with a review of the current literature on the available crystal structures of viruses and their implications for capsid assembly (chapter I). All experimental and computational methods used during the course of investigations are described in chapter II, as most of these are applicable to all the structure determinations and analyses. The experimental procedures described include cloning, overexpression, purification, crystallization and intensity data collection. Computational methods covered include details of various programs used during data processing, structure solution, refinement, model building, validation and analysis. Chapter III describes structural studies on top and bottom components of PhMV. Purified tymoviruses including PhMV are found to contain two classes of particles that sediment at different velocities through sucrose gradients and are called the top (sedimentation coefficient 54 Svedberg units(S)) and the bottom (115S) components. The top component particles are either devoid of RNA or contain only a small subgenomic RNA (5%) while the bottom component particles contain the full length genomic RNA. Only the bottom component is infectious. The top and bottom components were separately crystallized in P1 and R3 space groups, respectively. It is of interest to note that crystals of the bottom component obtained earlier belonged to R3 space group while recombinant capsids that lack of full length RNA as in natural top component crystallized in the P1 space group. A polyalanine model of the homologous TYMV was used as the phasing model to determine the structures of these particles by molecular replacement using the program AMoRe. The refinement of top and bottom component capsid structures were carried out using CNS version 1.1 and the polypeptide models were built into the final electron-density map using the interactive graphics program O. The quality of the map was sufficient for building the model and unambiguous positioning of the side chains. There is a significant difference in the radius of the top and bottom component capsids, the top component being 5 Å larger in radius. Thus, RNA makes the capsid more compact, even though RNA is not a pre-requisite for capsid assembly. Partially ordered RNA was observed in the bottom component. The refined models could form the basis for understanding the architecture, protein-protein interactions, protein-nucleic acid interactions, stability and assembly of PhMV. Chapter IV provides a detailed description of the mutations carried out on PhMV coat protein towards enhancing the diffraction quality of crystals. The gene coding for PhMV coat protein (PhMVCP) and several of its deletion and substitution mutants were originally cloned in pRSETC and pET-21 vectors by Mira Sastri and Uma Shankar in Prof. Savithri’s laboratory at the Department of Biochemistry. It was observed that the recombinant intact coat protein and several mutants lacking up to 30 amino acids from the N-terminal end could assemble into empty shells resembling the natural top component. None of these deletion mutants crystallized in forms that diffracted to high resolution. Based on the intersubunit contacts observed, three more site-specific mutants were designed. These three mutants were expressed in BL21 (DE3), purified and crystallized. Even these mutant crystals did not diffract to high resolution. The polypeptide fold of PhMV coat protein therefore was carefully examined for probable reasons. It was found that PhMV subunit has three major cavities. Three cavities are likely to increase the flexibility of protein subunits, which in turn may result in crystals of poor quality. Mutations V52W, S158Q and A160L were shown to fill up these cavities and with the view of obtaining better crystals. These site specific mutations were carried out the mutant proteins were purified. It was shown that the recombinant capsids are stable and possess T=3 architecture. Two mutants were crystallized and a data set for V52W extending to 6.0 Å resolution could be collected. Due to the limited resolution, further work was not pursued. It is plausible that the triple mutant will diffract to higher resolution. The second part of the thesis deals with stress response proteins from Oryza sativa and Salmonella typhimurium. It is known that viral infection and abiotic and biotic stresses induce a network of proteins in plants. Chapter V presents a review of the current literature on stress proteins, focusing mainly on Oryza sativa and S. typhimurium stress response proteins. Chapter VI describes the over expression of stress proteins SAP1 and SAP2 from rice. These stress related proteins confer tolerance to cold, dehydration and salt stress in rice. These proteins have been cloned in the expression vector pEt-28(a) and expressed in E. coli strain BL21 CodonPlus(DE3)RIL. The proteins were purified and crystallization trials were made. However, there were no hits. In an attempt to get crystals, nine deletion constructs of SAP1 were designed eliminating potentially disordered and unfolded regions based on a bioinformatics analysis. Crystallization trails are being carried out on three of the constructs. Structural studies on a universal stress protein from Salmonella typhimurium, which shares homology with the rice universal stress proteins, was initiated. Apart from this, several other stress related proteins of Salmonella typhimurium have also been selected for structural and functional studies. These include YdaA, YbdQ, Yic, Ynaf, Yec, Spy and Usb. All these were cloned and expressed in E. coli. Out of seven proteins, Ynaf, YdaA and YbdQ were found in the soluble fraction and were expressed in quantities suitable for structural studies. I could crystallize YdaA and Ynaf. X-ray diffraction data to resolutions of 3.6 Å and 2.3 Å were collected on crystals of YdaA and YnaF, respectively. A tentative structure of YnaF has been obtained. Further attempts to determine these structures are in progress. Biophysical, Biochemical functional characterization of YdaA and YnaF proteins are described. Structural studies on mannose-6-phosphate isomerase, an enzyme related to stress regulatory proteins from S. typhimurium are dealt with in Chapter VII. Mannose 6-phosphate isomerase (MPI) catalyzes the interconversion of mannose 6-phosphate and fructose 6-phosphate. The structure could be solved in its apo and holo forms (with two different metal atoms, Y3+ and Zn2+), and complexed with the cyclic form of the substrate fructose 6-phosphate (F6P) and Zn2+. Isomerization involves acid/base catalysis with proton transfer between C1 and C2 atoms of the substrate. Lys 132, His 131, His 99 and Asp 270 are close to the substrate and are likely to be the residues involved in proton transfer. Interactions observed at the active site suggest that the ring opening step is catalyzed by His 99 and Asp 270. An active site loop consisting of residues 130-133 undergoes conformational changes upon substrate binding. The metal ion is not close to the substrate atoms involved in proton transfer. Binding of the metal induces structural order in the loop consisting of residues 50-54. Hence, the metal atom does not appear to play a direct role in catalysis, but is probably important for maintaining the architecture of the active site. Based on these structures and earlier biochemical work, a probable isomerization mechanism has been proposed. The thesis concludes with a brief discussion on the future prospects of the work. The following manuscripts have been published or will be communicated for publication based on the results presented in the thesis:

Resumo: O cultivo de arroz de terras altas sob condições de sequeiro é considerado um cultivo de risco, sendo a água o fator ambiental mais ligado à baixa produtividade. A identificação e a compreensão dos mecanismos de tolerância à seca, bem como o desenvolvimento de práticas culturais que aumentem essa tolerância são fundamentais para alcançar maior estabilidade na produção. A adubação silicatada tem aumentado a tolerância das plantas aos estresses biótico e abiótico. O trabalho teve como objetivo avaliar o desenvolvimento de cultivares de arroz de terras altas sob condição de déficit hídrico e adubação silicatada e a marcha de absorção e acúmulo de silício na parte aérea. Foram instalados dois experimentos em casa de vegetação, o primeiro foi delineado em blocos ao acaso em esquema fatorial 2 x 2 x 2 com quatro repetições. Os tratamentos foram constituídos por duas cultivares (Caiapó - tradicional e Maravilha - moderno), em duas tensões de água no solo (-0,025 MPa e -0,050 MPa) e duas doses de silício ( 0 e 350 kg ha-1Si). O segundo experimento foi conduzido de maneira inteiramente casualizado em esquema fatorial 2 x 2 x 7 com quatro repetições, sendo duas cultivares (Caiapó e Maravilha) dois corretivo de solo (carbonato de cálcio e silicato de cálcio) e sete estádios de desenvolvimento (início do perfilhamento, perfilhamento máximo, diferenciação do primórdio da panícula, emborrachamento, antese, grão leitoso e maturação fisiológica.) O estresse hídrico reduziu o 2 número de panícula por metro quadrado, o número total de espiguetas por panícula, a fertilidade das espiguetas e, consequentemente, a produtividade de grãos. Sob condições de estresse hídrico, o silício reduz o teor de prolina na fase vegetativa e reprodutiva e aumenta a atividade da peroxidase na fase reprodutiva, podendo ser um indicativo de tolerância a esse estresse. As cultivares de arroz acumulam silício em quantidades. / Abstract: The highland rice cultivate under upland condition is considered a risk cultivate, being the water enviroment factor most linked to low productivity. The identification and the understanding of the dry tolerance mechanisms as well as the development of culture practical that increase this tolerance are fundamental to achieve more stabitity, productivity on highland rice cultivate specially on upland system. The silicon fertilization have been increased the plants tolerance under biotic and abiotic stress. This silicon fertilization have been increased the plants tolerance to biotic and abiotic stress. This research had the objective to evaluate the development of the upland rice cultivar under water stress condition and silicon fertilization and uptake dynamic of silicon in the aerial part of the plant. The were installed two experiment in greenhouse, the first experiment was outlined at random blocks design in a factorial draft 2x2x2 with four replication. The treatments were 4 two cultivars ( Caiapó- Traditional and Maravilha Modern) for two tension of water on the ground ( -0,025 MPa e -0,0050 MPa) and for two silicon rates ( 0 and 350 kg ha-1Si). The second experiment was carried out as entirely randomized design factorial draft 2x2x7 with four replication, being two cultivares (Caiapó and Maravilha) two soil amendments (limestone and calcium silicate) and seven estage of development. The water stress decreased the number of panicle per square meter, the total number of spikets per panicle, spiketel fertility and consequently the grains productivity. Under water stress, the silicon decrease the proline level at vegetation and reproductive stage and increase the peroxide activity at reproductive stage, could be a tolerance indicative to this stress. The rice cultivares accumulate silicon in different quantity. The silicon uptake dynamic and silicon accumulate is gradual at the Caiapó cultivar, since the tillering to the booting, while. / Orientador: Hélio Grassi Filho / Coorientador: Carlos Alexandre Costa Crusciol / Banca: Orivaldo Arf / Banca: Sonia Regina de Souza / Banca: Gaspar Henrique Korndorfer / Banca: José Salvador Simonetti Foloni / Doutor

Rice (Oryza sativa L.) is an important crop cultivated worldwide, and abiotic stresses limit its productivity. Different approaches were carried out to understand the mechanisms of rice defense responses against abiotic stresses, mainly drought. The NADPH-generating enzymes engaged in response to dehydration and salt stresses during the seedling stage of Nipponbare cultivar were analyzed. Enzyme activities of 6-phosphogluconate dehydrogenase (6PGDH), NADP-dependent aldehyde dehydrogenase, NADP-malic enzyme (NADP-ME) in leaves, and 6PGDH in roots were significantly increased in response to dehydration stress. NADP-ME and NADP-glutamate dehydrogenase activities in roots increased significantly in response to salt stress. These results suggest the involvement of NADPH-generating enzymes in plant responses to dehydration and salinity stresses, and the increased demands of NADPH in plants under abiotic stress can be furnished by enhanced activities of NADPH-producing enzymes. Also, a dehydration-induced protein was detected and identified as serine-hydroxymethyltransferase. This result indicates that serine-hydroxymethyltransferase can play a key role in regulating dehydration response in rice. Moreover, comparative proteomic analyses of CL163 (drought-tolerant), Cheniere (drought-sensitive), and Rex (moderately-drought-sensitive) rice varieties were performed. Drought-responsive proteome changes were profiled in leaves and roots at the seedling stage in response to drought stress imposed by polyethylene glycol (PEG-6000). Eighteen significantly differentially expressed proteins were identified by mass spectrometry. Elongate factor1 alpha and 17.9-kDa classI heat shock protein appear to have different expression patterns between CL163 and Cheniere, which may be attributable to the difference in drought response of the two rice varieties. Furthermore, a compendium of 103 drought resistance genes in rice was compiled to construct and analyze networks formed by associations between genes/proteins and to identify the most significant genes, biological processes/pathways. Genes were classified based on gene ontology and protein class into 26 groups. Forty-two genes were classified as transcription factors. Proteins encoded by the genes were localized in 8 subcellular locations and classified into three classes. Two pathways from KEGG whose genes were overrepresented in the compendium were identified. Gene expression, network presenting pairwise interactions between genes/proteins, and co-expression network were constructed. This study provides a systematic view of the crucial genes that can be contributing collectively to drought tolerance.

Made available in DSpace on 2014-07-29T14:52:04Z (GMT). No. of bitstreams: 1 TESE LUICE gomes.pdf: 3188724 bytes, checksum: c4156187d61efe808b3d021846f674dd (MD5) Previous issue date: 2010-08-31 / The plant genetic resources stored ex situ are considered as a genetic repository, and are raw material for the development of the world agriculture. In rice, despite its high genetic variability, the lack of information of accessions to compose a databank prevents its use to help the choice of genitors for the breeding programs. The Embrapa Rice Core Collection (ERiCC) was developed from 10,000 accessions from Embrapa GeneBank, and it was set up by 550 accessions, divided in three subsets: 1) 94 lines and cultivars from Brazil (LCB); 2) 148 lines and cultivars from abroad (LCI); and 3) 308 traditional varieties (VT), obtained from germplasm collection expeditions in Brazil. This work aimed: 1) to evaluate the extension of genetic variability of 550 accessions from ERiCC by means of agronomic traits characterization using mixed models and multivariate statistics; 2) to perform a comparative analysis of the genetic divergence considering the agronomical and SSR markers characterizations; and 3) to identify the genotypes with higher genetic diversity and with the best agronomic performances, aiming to promote the most efficient use of such germplasm in breeding programs. The agronomic characterization of 550 accessions was performed in nine field experiments, evaluating 18 phenological-agronomic traits. The data were analyzed using the mixed linear and AMMI models. There was wide variation range of genotypical values for most evaluated traits. In different environments, it was observed VT accessions among the high-yielding materials, demonstrating the potential of this group of germplasm, particularly important due to its high genetic variability, to contribute to the development of cultivars regionally adapted. The AMMI approach allowed a good discrimination of ERiCC rice genotypes in relation to the adaptive performance, identifying the accessions CA880078, CA990001, CA870071 (subset VT), and CNA0009113 (LCI) as having good yield and broad adaptation to distinct environments. The comparative analysis of genetic diversity between agronomic and molecular data was performed using the 242 lines and cultivars accessions from ERiCC, which were characterized by 86 fluorescent SSR markers, and five agronomic traits with genotypic values predicted (values without from the effects of interaction genotypes x environment, from a joint analysis of nine experiments. The genetic divergence among accessions was estimated by the average Euclidian distance for phenotypical data, and by the Rogers modified by Wright (RW) genetic distance. The datasets were jointly analyzed by descriptive and multivariate statistics, using correlation analyses from hierarchical grouping of Ward and UPGMA methods. The phenotypical and molecular data showed a broad distribution of dissimilarity indexes, despite they showed different patterns of variation between them. Low molecular distances were associated to low phenotypical distances, however to high molecular distances, occurred a high broad range of phenotypical variation. The correlation between genetical and phenotypical dissimilarities was significant for both lowland and upland accessions, despite with different values (r=0.156 and r=0.409, respectively). Due to the low relation between phenotypical and molecular data, the analysis of genotypes to be used in breeding programs must include both evaluations to a better accession characterization. Considering the high yielding accessions, the higher molecular distances were identified among the accessions from lowland system of cultivation, among which BR IRGA 413 and CNA0005014, BR IRGA 413 and CNA0005853, and CNA0004552 and CNA0005014. Considering the upland accessions, maximum genetic distances were identified in CNA0000482 and CNA0006422, CNA0001006 and CNA0006422, and CNA0001006 and CNA0003490. The molecular analysis was able to identify accessions with reduced genetic relationship, that if used as genitors, will result in a progeny with a high probability to find new allelic combinations. On the other hand, the phenotypical characterization is important to identify accessions not just genetically divergent, but with superior agronomic trait performances for breeding programs. The results of this work will permit to increase the activities related to the characterization of accessions from rice Genebank, giving support of breeding programs to choose the best accessions to obtain new cultivars, with favorable traits, and broad genetic basis. In addition, a continuous program of phenotypical and molecular characterization of germplasm will be able to identify accessions to increase the genetic variability of ERiCC. / Os recursos genéticos vegetais armazenados ex situ são considerados reservatórios de genes e funcionam como matéria-prima para o desenvolvimento da agricultura mundial. Na cultura do arroz, apesar da extensa variabilidade genética existente, a deficiência de informações que integrem dados que possam efetivamente auxiliar na escolha de genótipos importantes para os programas de melhoramento constitui o principal fator que limita a utilização mais ampla dos acessos armazenados nos bancos de germoplasma. A Coleção Nuclear de Arroz da Embrapa (CNAE) representa a variabilidade genética de mais de 10 mil acessos constituintes do Banco Ativo de Germoplasma (BAG) da Embrapa Arroz e Feijão, e é composta por 550 acessos subdivididos em três estratos: 1) 94 Linhagens e Cultivares Brasileiras (LCB), provenientes de programas de melhoramento de instituições brasileiras; 2) 148 Linhagens e Cultivares Introduzidas (LCI), provenientes de programas de melhoramento de outros países; e 3) 308 Variedades Tradicionais (VT), que reúne acessos obtidos por expedições de coleta de germoplasma realizadas em vários estados do Brasil. Este trabalho teve como principais objetivos: 1) avaliar a extensão da variabilidade genética dos 550 acessos pertencentes à CNAE por meio da caracterização agronômica via metodologias de modelos mistos e estatísticas multivariadas; 2) realizar a análise comparativa da divergência genética entre acessos, determinada pela avaliação de caracteres agronômicos e marcadores moleculares SSR; e 3) identificar os genótipos com maior diversidade genética e com melhores atributos agronômicos, a fim de indicar uma melhor utilização destes recursos genéticos em programas de melhoramento. Na caracterização agronômica foram avaliados 550 acessos em experimentos conduzidos em nove locais no Brasil, envolvendo um total de 18 caracteres fenológico-agronômicos. Os dados foram analisados empregando-se a abordagem de modelos lineares mistos e modelo AMMI de análise. Verificou-se grande amplitude de variação dos valores genotípicos para a maioria dos caracteres avaliados. Nos diferentes ambientes, houve ocorrência de genótipos do estrato VT entre os mais produtivos, o que demonstra o potencial deste grupo de germoplasma, particularmente importante por sua grande variabilidade genética, em contribuir para o desenvolvimento de cultivares regionalmente adaptadas. A abordagem AMMI permitiu uma boa discriminação dos genótipos de arroz da CNAE quanto ao seu comportamento adaptativo, identificando os acessos CA880078, CA990001, CA870071 (do estrato VT), e CNA0009113 (LCI) com estabilidade, produtividade satisfatória e ampla adaptação à diferentes ambientes. Para a análise comparativa da diversidade genética entre dados agronômicos e moleculares foram considerados 242 acessos da CNAE, os quais foram caracterizados utilizando-se 86 marcadores SSR fluorescentes, sendo que para os dados agronômicos, foram realizadas análises conjuntas dos experimentos e considerados os valores genotípicos preditos de cinco caracteres (valores livres dos efeitos de interação genótipos x ambientes). A divergência genética entre os acessos foi estimada pelo procedimento de distância Euclidiana média para os dados fenotípicos, e por meio da distância de Rogers modificada por Wright (RW) para os dados moleculares, analisando-se os conjuntos de dados por meio de estatísticas descritivas e multivariadas, empregando-se análises de correlação entre matrizes de dissimilaridade e análises de agrupamento hierárquico de Ward e UPGMA. Os dados fenotípicos e moleculares apresentaram uma ampla distribuição dos índices de dissimilaridade, embora tenham apresentado diferentes padrões dessa variação. Baixas distâncias moleculares estiveram associadas a baixas distâncias baseada nos valores genotípicos, no entanto para elevadas distâncias moleculares houve ocorrência de ampla escala de variação fenotípica. A correlação entre as dissimilaridades genéticas e valores genotípicos foi significativa tanto no conjunto de acessos irrigados quanto no de sequeiro, porém, com diferentes magnitudes (r=0,156 e r=0,409, respectivamente). Devido esta baixa relação entre os dados fenotípicos e moleculares, o estudo de genótipos para fins de uso no melhoramento genético deve incluir ambas avaliações para a melhor caracterização dos acessos. Entre os materiais mais produtivos, as maiores distâncias moleculares foram identificadas entre os genótipos do sistema de cultivo irrigado, dentre eles BR IRGA 413 e CNA0005014, BR IRGA 413 e CNA0005853, e CNA0004552 e CNA0005014. Entre os materiais de sequeiro, máximas distâncias genéticas foram identificadas entre os acessos CNA0000482 e CNA0006422, CNA0001006 e CNA0006422, e CNA0001006 e CNA0003490. A análise molecular permitiu que fossem identificados genótipos de vínculo genético reduzido, que quando utilizados como parentais em cruzamentos, possibilitarão que as progênies obtidas apresentem maiores chances de combinações alélicas inéditas. Por sua vez, a caracterização fenotípica tem papel fundamental na identificação de materiais que além de divergentes, apresentem desempenho agronômico superior para os programas de melhoramento. Os resultados deste trabalho permitirão aumentar eficazmente as atividades relacionadas à caracterização de acessos do Banco Ativo de Germoplasma de arroz, subsidiando os programas de melhoramento na escolha de genótipos a serem utilizados para a obtenção de novas cultivares, com características favoráveis, de ampla base genética. Em adição, um programa contínuo de caracterização fenotípica e molecular de germoplasma permitirá ainda a escolha de acessos para a ampliação da variabilidade genética da CNAE.

Made available in DSpace on 2014-07-29T16:24:18Z (GMT). No. of bitstreams: 1 Dissertacao Rogerio Gomes Pereira.pdf: 1311913 bytes, checksum: 594d4803b27bb1b817ac16d754681cbb (MD5) Previous issue date: 2011-02-14 / The average national rice yield was 11,3 million tones, with a harvested area of about 2,7 million hectares (IBGE 2010). Part of this comes from upland production systems in savannas that have high acidity, low fertility, with low phosphorus and potassium levels, low cation exchange capacity and high aluminum saturation. Upland rice in the Brazilian savannas (Cerrados) experiences multiple abiotic stresses and is characterized by high levels of uncertainty caused by rainfall variability. The increasing consumption of plant biomass to generate energy has produced products and by-products with high potential for agricultural use. Among these products is charcoal, that, due to its physical-chemical and molecular properties, have beneficial effects for the physical, chemical and biological soil characteristics. The objective of this work was to investigate the effect of charcoal on soil chemical properties and on the development of upland rice, and on the transpiration rate response of upland rice as an alternative to decrease the effect of drought on its growth and development. Two pot experiments were carried out using a random split-plot experimental design. The main treatments (plots) were water stress (WS) and no-water stress (NWS) and the sub treatments (subplots) were different biochar doses, 0% or control (T1), 6% (T2), 12% (T3) and 24% (T4) in growing medium (sand). The plastic pots were randomized within the sub plots with six replications. The total transpirable soil water (TTSW), the p factor, defined as the average fraction of TTSW that can be depleted from the root zone before reduction in transpiration occurs and the normalized transpiration rate (NTR) were determined. Charcoal significantly increased dry matter accumulation and leaf area of rice plants in both experiments and positively affected the chemical and physical soil properties such as water retention and nitrate availability to the plants. Biochar addition increased TTSW, p factor and reduced NTR. Consequently, biochar addition was able to change the moisture threshold (p factor) of the growing medium, up to a maximum concentration of 12%, hence delaying the point where transpiration declines and affects yield. / A safra nacional de arroz foi de 11,3 milhões de toneladas, com uma área colhida de aproximadamente 2,7 milhões hectares (IBGE 2010). Parte desta produção é cultivada em sistema de sequeiro sob solos de cerrados que é caracterizado por apresentar elevada acidez e baixa fertilidade natural, com baixos teores de fósforo, potássio, capacidade de troca catiônica e elevada saturação por alumínio. O arroz de terras altas cultivado no Brasil Central (Cerrados) experimenta múltiplos estresses abióticos e sua produtividade é caracterizada pela incerteza devido à variabilidade na precipitação. O crescente consumo de biomassa vegetal para a geração de energia tem disponibilizado subprodutos com alto potencial para o uso agrícola. Dentre esses subprodutos, destaca-se o carvão vegetal, que devido as suas características físico-químicas e moleculares, tem-se mostrado benéfico para as propriedades físicas, químicas e biológicas do solo. O objetivo deste estudo foi analisar os efeitos de diferentes doses de carvão vegetal de eucalipto na produção de matéria seca na cultura do arroz de terras altas e nas características físico-químicas e hídricas em substrato areia, e na resposta da taxa de transpiração do arroz de terras altas como uma alternativa para minimizar os efeitos da seca no seu desenvolvimento e crescimento. Dois experimentos em vasos foram realizados por meio do delineamento em parcela sub-dividida. A parcela principal foi constituída pelo tratamento deficiência hídrica (WS) e não deficiência hídrica (NWS) e as sub-parcelas pelas diferentes doses de carvão; 0% (controle, T1), 6% (T2), 12% (T3) e 24% (T4) adicionadas no substrato (areia). Os vasos foram inteiramente casualizados nas sub-parcelas com seis repetições por dose de carvão. A quantidade de água transpirável no solo (TTSW), o fator de depleção (p), definido como a fração de água disponível no solo que pode ser retirada da zona radicular antes de ocorrer a redução da taxa de transpiração, e a taxa de transpiração normalizada (NTR) foram determinadas. A adição de carvão ao substrato proporcionou um aumento significativo na produção de matéria seca e área foliar das plantas de arroz em ambos os experimentos. Nos tratamentos T2, T3 e T4, a adição de carvão vegetal alterou significativamente as características químicas e físico-hídricas do solo, aumentando a retenção de água e a disponibilidade de nitrato para as plantas. A adição de carvão aumentou a TTSW, o fator p e reduziu a NTR. Conseqüentemente, o carvão foi responsável por modificar o fator p, até uma concentração máxima de 12% e retardar o ponto no qual o declínio da transpiração afeta o rendimento.

Molecular chaperones are proteins that interact with and aid in stabilization and activation of other proteins. Chaperones help proteins attain their three dimensional conformation, without forming a part of the final structure. Many of the chaperones are stress proteins known as Heat shock proteins (Hsps). Their expression is upregulated in response to various kinds of stress such as heat stress, oxidative stress etc., which threaten the protein homeostasis, by structurally destabilizing cellular proteins, and increasing the concentration of aggregation-prone folding intermediates. The Hsps are classified according to their molecular weight into Hsp40, Hsp60, Hsp70, Hsp90, Hsp100, and the small Hsp families. Some of them are constitutively expressed and play a fundamental role in de novo protein folding. They further aid in proteome maintenance by assisting in oligomeric assembly, protein trafficking, refolding of stress denatured protein, preventing protein aggregation and protein degradation. Heat shock protein 90 (Hsp90) are one of the important representatives of this class of proteins. Hsp90 are highly conserved class of molecular chaperones. They are found in bacteria, eukaryotes, but not in archaea. In contrast to the eukaryotes which require a functional cytoplasmic Hsp90 for viability, the bacterial counterpart (HtpG) is typically nonessential. Hsp90 is an ATP dependent chaperone. Hsp90 form dimers, with each protomer consisting of three functional domains: N- terminal, ATP binding domain, Middle domain and C-terminal domain. Hsp90 is a dynamic protein, and undergoes an elaborate conformational cycle during its ATPase cycle, which is essential for its chaperoning activity. The Hsp90 chaperone cycle is regulated by interaction with diverse cochaperones. Hsp90 interacts with specific set of substrate proteins. Many of these substrate proteins function at the heart of several cellular processes like signalling, cell cycle, apoptosis. Studies from protozoans like Leishmania, Plasmodium, Trypanosoma etc. have also implicated the role of Hsp90 in their growth and stage transitions. Thus, selective inhibition of Hsp90 has been explored as an intervention strategy against important human diseases such as cancer, malaria and other protozoan diseases. The ATP binding N-terminal domain (NTD), has been explored as the target domain for inhibition of Hsp90 using competitive inhibitors of ATP. Several chemical classes of Hsp90 inhibitors are known, including ansamycins, macrolides, purines, pyrazoles, and coumarin antibiotics. However, many inhibitors are observed to be toxic, less soluble and unstable. Hence, there is a requirement for new approach to design inhibitors which are more soluble and less toxic and serve as effective therapeutic drugs.inhibitors are observed to be toxic, less soluble and unstable. Hence, there is a requirement for new approach to design inhibitors which are more soluble and less toxic and serve as effective therapeutic drugs. The work presented in this thesis mainly concerns with the structural studies and biochemical and biophysical characterization of Hsp90 from two different sources viz. Dictyostelium discoideum, a cellular slime mould and a plant source Oryza sativa (rice). The structural analyses of these two proteins have been carried out by X-ray crystallography. Though yeast has been explored extensively as a model system to understand the different roles of Hsp90, it lacks the various signalling pathways essential for growth and development present in case of higher eukaryotes. D. discoideum has been employed as a model system to understand multicellular development, which occurs in response to starvation induced stress. D. discoideum has the advantages due to its ease of manipulation. The organism's genome also shows many signalling pathway for growth and differentiation that are conserved between D. discoideum and mammals. With this motivation, we have studied several structural aspects of the cytosolic isoform of Hsp90 from D. discoideum called HspD. HspD was also observed to play a role in the multicellular development of D. discoideum. It has been demonstrated that the treatment of D. discoideum with inhibitors like Geldanamycin or Radicicol causes an arrest in the multicellular development at the mound stage, and the few which escaped this arrest gave rise to abnormal fruiting bodies. A subset of the proteins involved in this mound arrest phenotype, were observed to have homologs in humans, which are clients of Hsp90. Therefore, a structural perspective of HspD can aid in better understanding of the role of this protein in the organism, as well as, elucidate any structural differences observed as compared to other species, which may have an impact on its activity. Studies on the physiological role of Hsp90 in plants began much later as compared to fungi and humans. In plants Hsp90 are involved in various abiotic stress responses. In addition, their roles have also been implicated in plant growth and development, innate immune response and buffering genetic variations. However, the molecular mechanisms of these various actions are not clearly understood. Also, the structural aspects of plant Hsp90 are yet to be explored. The structure of the NTD of Hsp90 from barley is the only one available from a plant source till now. We have initiated the studies on rice Hsp90 with the objective to understand the mechanism of Hsp90 in plants, which may aid in improving stress tolerance in plants. The thesis has been divided into five chapters. The first chapter introduces the various aspects of Hsp90 protein. The chapter starts with a general overview of concept of molecular chaperones and describes briefly the different classes of molecular chaperones. This is followed by a detailed description of different aspects of Hsp90 with main emphasis on the structure and its conformational flexibility. The chapter describes the association of Hsp90 with other accessory proteins like cochaperones and its interaction with its substrate proteins and explains the functional significance of Hsp90 as a drug target and the need for the development of new class of inhibitors, followed by the significance of the study of Hsp90 in the two model systems (D. discoideum and rice) chosen to be studied. The second chapter gives a brief overview of the principles behind the different experimental methods employed during the course of this research, which includes the tools of X-ray crystallography and other biochemical and biophysical techniques employed for the characterization of the protein. Chapter 3 describes the crystal structure of NTD of Hsp90 from D. discoideum. The structure of NTD was solved in two different native (ligand-free) forms viz. monoclinic and hexagonal. The two forms differed in local structural rearrangement of a segment of NTD known as the lid region. The lid region in the hexagonal form showed a shift in its position as compared to the other solved structures of NTD. The structure of NTD was also solved in complex with various ligands which include ADP, substrate analogs and an inhibitor molecule. A comparison of all the structures showed that the overall structure is well-conserved. One of the crystal structures of NTD showed a heptapeptide (part of the vector) bound at the active site. The peptide was observed to make several complementary interactions with the residues of the ATP binding pocket and retain several interactions which the nucleotide makes with the NTD. The NTD showed subtle conformational differences when compared with the NTD of Hsp90 from yeast. Chapter 4 details the structural and functional characteristics of full length Hsp90 protein from D. discoideum. Due to the large size and flexibility, the full length protein did not crystallize in spite of several attempts. Hence, HspD was studied using different solution studies like Small Angle X-ray Scattering (SAXS) and Dynamic Light Scattering (DLS). Both the studies showed the presence of higher oligomers. The SAXS data showed the presence of tetramers and hexamers while, the addition of the ligand shifts the protein from a dimer to a higher oligomer as observed from DLS studies. The chapter also describes the study of interaction of HspD with a cochaperone protein p23. The interactions were studied using ITC, which showed a strong binding. The ATPase activity was also evaluated in the presence of increasing concentrations of p23, which was observed to decline with increasing concentrations of p23. In chapter 5, we describe the biochemical characterization of Hsp90 from Oryza sativa (rice) and the crystallographic analysis of its NTD. Binding of the rice Hsp90 to ATP and an inhibitor were studied by fluorescence. The ATPase activity of rice Hsp90 was checked by radioactive assay and the protein was observed to be active. The NTD of rice Hsp90 crystallized as a monomer in complex with a substrate analog AMPPCP and the structure was determined.

The objective of this research was to investigate the occurrence of mycoflora in rice plants and rice seeds in South Africa and their negative impact. A total of six species of Fusarium were isolated from diseased rice plants and rice seeds and identified as F. anthophilum, F. chlamydosporum, F. compactum, F. equiseti, F. fujikuroi and F. semitectum. In the translation elongation factor data set, Fusarium equiseti isolates grouped together within the F. incarnatum - equiseti Species Complex (FIESC). The isolates from rice clustered together in a single clade with the F. equiseti and F. incarnatum isolates forming two separate sub-clades.The isolates of F. equiseti present a new phylogenetically distinct species in FIESC. In the pathogenicity tests, isolates of both F. anthophilum and F. fujikuroi caused bakanae disease to rice plants. Fifty four rice cultivars and lines were tested by the standardized test tube inoculation method for resistance and susceptibility against bakanae isolate of F. anthophilum and the bakanae isolate of F. fujikuroi. None of the rice cultivars and lines was found to be resistant to bakanae isolates of Fusarium spp. The fungicide, benomyl was found to be most effective as a seed treatment for controlling bakanae disease of rice due to isolates of both F. anthophilum and F. fujikuroi. Thiram was found to be the least effective fungicide for controlling bakanae disease of rice caused by isolates of both the Fusarium spp. Apart from Fusarium species, other fungi that were also isolated from diseased rice plants and rice seeds were identified as Alternaria alternata, Alternaria longipes, Cochliobolus miyabeanus, Nigrospora sphaerica, Phoma eupyrena, Phoma jolyana, Phoma sorghina and Pithomyces sp. In mycotoxin tests, the isolates of both F. anthophilum and F. fujikuroi produced moniliformin. None of the isolates of F. anthophilum and F. fujikuroi produced fumonisins. This research is important as it identifies many fungal species in rice plants and seeds in South Africa for the first time. Currently, there is very little literature that makes reference to such findings under South African conditions. In addition, this investigation unravels previously unknown information on the resistance of rice to bakanese disease. Finally, information is provided on the effectiveness of commonly used fungicides (benomyl and thiram) to control rice diseases. This knowledge is crucial information that is useful to plant pathologists, the farming community and the scientists that are involved in strategies of fighting or reducing rice diseases so as to help contribute to food security. / Environmental Sciences / D. Phil. (Environmental Science)