Several causes are pointed as responsible for the numbness in seeds of rice. The fenolic compounds have influence in the swinging between promoters and inhibitors of the germination of seeds, could represent an obstacle to the diffusion of gases. Several treatments are used with the purpose of overcoming the dormence in seeds and alternative methods appear to try to solve this problem, as the ultrasound radiations. That technique, for being little used in seeds, however, it presents optimization difficulty, as well as the lack of specific knowledge about the action of the radiation promoted by the ultrasonic waves in living tissues. The objective of the work was to evaluate the chemical effects produced by the ultrasound about the overcoming of the dormence and physiologic quality of seeds of rice. Seeds of irrigated rice cv. IRGA 417 and of drier cv. Spring, submitted to the exhibition of the ultrasonic waves by periods of 5, 10, 15 and 20 minutes and temperatures of 20, 30 and 40 oC. The variables analyzed after each treatment were: germination, first counting and index of germination speed. The results indicated that there was variation in the answers of the culture to the treatments, and the best results were found in the highest temperatures. It was concluded that the ultrasound radiations affect the quality of the seeds of rice, being a promising method for the overcoming of the dormence, still needing her standardization. / Inúmeras causas são apontadas como responsáveis pela dormência em sementes de arroz. Os compostos fenólicos destacam-se por interferirem no balanço entre promotores e inibidores da germinação de sementes, podendo representar um obstáculo à difusão de gases. Vários tratamentos são utilizados com a finalidade de superar a dormência em sementes e, portanto, surgem métodos alternativos para sua superação, como as radiações de ultra-som. Essa técnica, por ser pouco utilizada em sementes, no entanto, apresenta dificuldade de otimização, bem como a falta de conhecimento específico sobre o efeito da radiação promovida pelas ondas ultra-sônicas em tecidos vivos. O objetivo do trabalho foi avaliar os efeitos sonoquímicos produzidos pelo ultra-som sobre a superação da dormência e qualidade fisiológica de sementes de arroz. Utilizaram-se sementes de arroz irrigado cv. IRGA 417 e de sequeiro cv. Primavera, submetidas à exposição das ondas ultra-sônicas por períodos de 5, 10, 15 e 20 minutos e temperaturas de 20, 30 e 40 oC. As variáveis analisadas após cada tratamento foram: germinação, primeira contagem e índice de velocidade de germinação. Os resultados indicaram que houve variação nas respostas das cultivares aos tratamentos, sendo que os melhores resultados foram encontrados nas temperaturas mais elevadas. Concluiu-se que as radiações de ultra-som afetam a qualidade das sementes de arroz, sendo um método promissor para a superação da dormência, necessitando ainda sua padronização.

Made available in DSpace on 2016-01-26T18:56:22Z (GMT). No. of bitstreams: 1 Dissertacao.pdf: 249370 bytes, checksum: bb3ab2f95decedaeb7a114dc63aae796 (MD5) Previous issue date: 2011-01-25 / The production cost of new areas in the cerrado presents high values due to larger amount of farming operations when compared to conventional crops. The cerrado soils have low natural fertility and require large quantities of limestone for its correction significantly increasing the cost of production. The objective this study was to evaluate whether the Boro when applied in the cultivation of upland rice with low technological level, increases the productivity rice. The field work was conducted in the municipality of Nova Xavantina - MT in 2009/10 in two areas (first and second year of production). We used six treatments of boron in the source of boric acid (17 % of B), with doses of 0, 0.5, 1.0, 1.5, 2.0, and 2.5 kg B per ha. The plots were 10 m long and 3.4 m wide with 20-sowing rows, with spaced at 0.17 m. The fertilization of 160 kg of 08-28-16 fertilizer was formulated according to the results of soil analysis and used the cultivar BRS Sertaneja in amount of 115 kg of seeds per ha. Among the variables analyzed the productivity in the area of the first year was influenced by boron fertilization, whre the treatment that used a dose of 2.5 kg B hit productivity of 2.677 kg/ha-1. The cost analysis positive for both areas, where in the area of the first year, the cost benefit of boron fertilization was R$169,44 (with a dose of 1 kg B/ha). As for the second year the area of greatest cost benefit was obtained with a dose of 2.5 kg B, which resulted in na increase in productivity of 11.80 bags (60 kg) and therefore a cost benefit of R$333.83 per hectare. The use of boron into new areas of cerrado compensated the low technological level that was used in the formation of the crops, providing positive results in the cost benefit of the crop without significant increases inproduction costs. / O custo de produção de áreas novas no cerrado apresenta valores elevados devido á maior quantidade de operações agrícolas, quando comparado a lavouras convencionais. Os solos do cerrado apresentam baixa fertilidade natural e exigem grandes quantidades de calcário para sua correção, aumentando significamente o custo de produção. O objetivo deste trabalho foi avaliar se o Boro quando aplicado na cultura do arroz de terras altas com baixo nível tecnológico, proporcionaria um ganho de produtividade na cultura do arroz. O trabalho foi conduzido a campo no município de Nova Xavantina MT na safra 2009/10 em duas áreas (primeiro e segundo ano de produção). Foram utilizados 6 tratamentos de boro na fonte de ácido bórico (17% de B), sendo as doses de: 0; 0,5; 1,0; 1,5; 2,0; e 2,5 kg de B por ha. As parcelas tiveram 10 m de comprimento por 3,4 m de largura com 20 linhas de plantio no espaçamento de 0,17 m. A adubação de 160 kg do adubo formulado 08-28-16 foi em função dos resultados da análise do solo e utilizou-se a cultivar BRS Sertaneja na quantidade de 115 kg de sementes por ha. Dentre as variáveis avaliadas a produtividade da área de primeiro ano foi influenciada pela adubação boratada, onde o tratamento que utilizou a dose de 2,5 kg de B atingiu a produtividade de 2.677 kg/ha-1. A análise de custo foi positiva para as duas áreas, onde na área de primeiro ano, o custo beneficio da adubação boratada foi de R$169,44 (com a dose de 1 kg B/ha). Já para a área de segundo ano o maior custo beneficio foi obtido com a dose de 2,5 kg de B, que proporcionou um aumento na produtividade de 11,80 sacas (60 kg) e conseqüentemente um custo beneficio de R$333,83 por hectare. O uso do boro em áreas novas do cerrado compensou o baixo nível tecnológico que foi empregado na formação das lavouras, proporcionando resultados positivos no custo beneficio da lavoura sem acréscimos significativos no custo de produção.

Made available in DSpace on 2016-07-18T17:51:06Z (GMT). No. of bitstreams: 1 Dissertacao.pdf: 249370 bytes, checksum: bb3ab2f95decedaeb7a114dc63aae796 (MD5) Previous issue date: 2011-01-25 / The production cost of new areas in the cerrado presents high values due to larger amount of farming operations when compared to conventional crops. The cerrado soils have low natural fertility and require large quantities of limestone for its correction significantly increasing the cost of production. The objective this study was to evaluate whether the Boro when applied in the cultivation of upland rice with low technological level, increases the productivity rice. The field work was conducted in the municipality of Nova Xavantina - MT in 2009/10 in two areas (first and second year of production). We used six treatments of boron in the source of boric acid (17 % of B), with doses of 0, 0.5, 1.0, 1.5, 2.0, and 2.5 kg B per ha. The plots were 10 m long and 3.4 m wide with 20-sowing rows, with spaced at 0.17 m. The fertilization of 160 kg of 08-28-16 fertilizer was formulated according to the results of soil analysis and used the cultivar BRS Sertaneja in amount of 115 kg of seeds per ha. Among the variables analyzed the productivity in the area of the first year was influenced by boron fertilization, whre the treatment that used a dose of 2.5 kg B hit productivity of 2.677 kg/ha-1. The cost analysis positive for both areas, where in the area of the first year, the cost benefit of boron fertilization was R$169,44 (with a dose of 1 kg B/ha). As for the second year the area of greatest cost benefit was obtained with a dose of 2.5 kg B, which resulted in na increase in productivity of 11.80 bags (60 kg) and therefore a cost benefit of R$333.83 per hectare. The use of boron into new areas of cerrado compensated the low technological level that was used in the formation of the crops, providing positive results in the cost benefit of the crop without significant increases inproduction costs. / O custo de produção de áreas novas no cerrado apresenta valores elevados devido á maior quantidade de operações agrícolas, quando comparado a lavouras convencionais. Os solos do cerrado apresentam baixa fertilidade natural e exigem grandes quantidades de calcário para sua correção, aumentando significamente o custo de produção. O objetivo deste trabalho foi avaliar se o Boro quando aplicado na cultura do arroz de terras altas com baixo nível tecnológico, proporcionaria um ganho de produtividade na cultura do arroz. O trabalho foi conduzido a campo no município de Nova Xavantina MT na safra 2009/10 em duas áreas (primeiro e segundo ano de produção). Foram utilizados 6 tratamentos de boro na fonte de ácido bórico (17% de B), sendo as doses de: 0; 0,5; 1,0; 1,5; 2,0; e 2,5 kg de B por ha. As parcelas tiveram 10 m de comprimento por 3,4 m de largura com 20 linhas de plantio no espaçamento de 0,17 m. A adubação de 160 kg do adubo formulado 08-28-16 foi em função dos resultados da análise do solo e utilizou-se a cultivar BRS Sertaneja na quantidade de 115 kg de sementes por ha. Dentre as variáveis avaliadas a produtividade da área de primeiro ano foi influenciada pela adubação boratada, onde o tratamento que utilizou a dose de 2,5 kg de B atingiu a produtividade de 2.677 kg/ha-1. A análise de custo foi positiva para as duas áreas, onde na área de primeiro ano, o custo beneficio da adubação boratada foi de R$169,44 (com a dose de 1 kg B/ha). Já para a área de segundo ano o maior custo beneficio foi obtido com a dose de 2,5 kg de B, que proporcionou um aumento na produtividade de 11,80 sacas (60 kg) e conseqüentemente um custo beneficio de R$333,83 por hectare. O uso do boro em áreas novas do cerrado compensou o baixo nível tecnológico que foi empregado na formação das lavouras, proporcionando resultados positivos no custo beneficio da lavoura sem acréscimos significativos no custo de produção.

Les éléments transposables (ET) sont des éléments génétiques ubiquitaires et potentiellement mobiles dans les génomes eucaryotes. Les génomes hôtes ont développé des mécanismes épigénétiques pour contrôler et prévenir la prolifération des ET. Néanmoins, certains ET semblent capables de s’activer en réponses à des stress ou à des facteurs développementaux. Les méthodes disponibles pour détecter l’activité transpositionnelle d’un ET sont souvent limitées au stade transcriptionnel ou sont adaptées à des génomes de petite taille. Relativement peu d’ET sont actuellement connus pour être actifs et les mécanismes spécifiques qui les contrôlent ne sont pas clairement identifiés.Durant mes travaux de thèse, nous avons développé une stratégie de séquençage à haut débit qui permet la détection d’ADN extrachromosomique circulaire (ADNecc) témoignant notamment de l’activité des ET et de la stabilité d’un génome. Ainsi nous avons pu caractériser chez plusieurs espèces le mobilome, défini comme l’ensemble des ADNecc présents dans un tissu.La technique du mobilome-seq s’est avérée être un outil puissant pour la détection des ET actifs notamment chez le riz asiatique Oryza sativa. Notre analyse du mobilome a permis l’identification d’un rétrotransposon PopRice actif dans l’albumen (tissu nourricier du grain) chez différentes variétés de riz. Pour la première fois chez les plantes, nous avons également détecté des insertions somatiques d’ET par re-séquençage de génome entier. À partir de nos résultats, nous avons combiné nos données mobilomiques avec une analyse GWAS pour proposer des pistes afin d’identifier de nouveaux mécanismes de régulation de cet élément.En parallèle, nous avons appliqué la technique du mobilome-seq à différents organismes animaux et végétaux révélant ainsi des spécificités de mobilome propre à chaque espèce. Nos travaux en collaboration avec d’autres équipes ont notamment contribué à préciser le rôle de l’ARN polymérase II dans le contrôle des ET chez O. sativa et à mettre en évidence le lien entre la présence d’ADNecc viral et la réponse immunitaire chez Drosophila melanogaster.Mes travaux de thèse ouvrent des perspectives pour l’étude du mobilome, ce répertoire génomique encore largement inexploré et qui se révèle être à la fois une source d’information au niveau des mouvements des ET mais aussi de la stabilité des génomes. L’étude future des mobilomes promet d’apporter des réponses sur notre compréhension de la dynamique des génomes. / Transposable elements (TEs) are mobile genetic elements that constitute a major part of eukaryotic genomes. Host genomes have developed epigenetic mechanisms to control and prevent their proliferation. While efficiently silenced by the epigenetic machinery, they can be reactivated upon stress or at precise developmental stages. However, available methods to detect TE activity are often limited to transcriptional level or more adapted to small genomes. Today, only few TEs are known to be active and specific mechanisms controlling TEs are not well defined. 
To address this question during my phD, we developed a strategy of high throughput sequencing that detects extrachromosomal circular DNA (eccDNA) forms which reflect TE activity and genome stability. We characterised mobilomes from different organisms defined as all eccDNA in a cell. 
Our mobilome-seq technique successfully identified active TEs especially in asian rice Oryza sativa. We identified an active retrotransposon PopRice in endosperm tissue from different rice varieties. Interestingly and for the first time in plants, we detected somatic insertions from genome- wide resequencing. We combined our mobilome-seq results with a GWAS analysis to propose new PopRice regulation mechanisms. 
 In a second step, we applied our mobilome seq technique to different animal and plant organisms showing mobilome specificities from each species. Our work in collaboration with different labs help contributed to define role of RNA polymerase II in the control of TEs in O. sativa and have revealed a link between presence of eccDNA from virus and immune response in Drosophila melanogaster. 
Altogether, our mobilome-sequencing method opens the possibility to explore unexplored genomic compartment. Future mobilome analysis represents new possibilities to improve our understanding of dynamics of genomes.

Submitted by Nayara Fernanda Siviero Garcia null (nayaragarcia.agro@gmail.com) on 2017-03-27T15:53:43Z No. of bitstreams: 1 Dissertação_Nayara_F_S_Garcia.docx: 5783562 bytes, checksum: 96991caf24ea2205403970813df47675 (MD5) / Rejected by Luiz Galeffi (luizgaleffi@gmail.com), reason: Solicitamos que realize uma nova submissão seguindo as orientações abaixo: A versão final da dissertação/tese deve ser submetida no formato PDF (Portable Document Format). O arquivo PDF não deve estar protegido e a dissertação/tese deve estar em um único arquivo, inclusive os apêndices e anexos, se houver. Por favor, corrija o formato do arquivo e realize uma nova submissão. Agradecemos a compreensão. on 2017-03-29T19:01:33Z (GMT) / Submitted by Nayara Fernanda Siviero Garcia null (nayaragarcia.agro@gmail.com) on 2017-03-29T21:55:00Z No. of bitstreams: 1 Dissertação_Nayara_F_S_Garcia.pdf: 3991651 bytes, checksum: a711929970ccc2436829f10c78ae5af5 (MD5) / Approved for entry into archive by Luiz Galeffi (luizgaleffi@gmail.com) on 2017-03-30T16:37:43Z (GMT) No. of bitstreams: 1 garcia_nfs_me_ilha.pdf: 3991651 bytes, checksum: a711929970ccc2436829f10c78ae5af5 (MD5) / Made available in DSpace on 2017-03-30T16:37:43Z (GMT). No. of bitstreams: 1 garcia_nfs_me_ilha.pdf: 3991651 bytes, checksum: a711929970ccc2436829f10c78ae5af5 (MD5) Previous issue date: 2017-02-03 / Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES) / Dentre os cereais de importância no cenário mundial merecem destaque o arroz e feijão fazendo parte da alimentação básica dos brasileiros. A integração de métodos como a diversificação de culturas e a utilização da biotecnologia que aumentem a produtividade de grãos do arroz e feijão pode contribuir ambientalmente e economicamente na adoção de sistemas de produção agrícolas mais sustentáveis. Assim, o objetivo do trabalho foi avaliar o efeito de diferentes culturas antecessoras (milho e/ou Urochloa ruziziensis) e inoculação de Azospirillum brasilense sobre o desenvolvimento e a produtividade do arroz de terras altas e do feijão cultivado em sucessão, além da inoculação de Rhizobium tropici na cultura do feijão de inverno. O estudo foi realizado no município de Selvíria-MS, em um LATOSSOLO VERMELHO Distrófico em área de plantio direto há mais de 10 anos. O delineamento experimental utilizado foi o de blocos casualisados, disposto em esquema de faixas, com quatro repetições. Os tratamentos foram constituídos pelo arranjo de oito combinações de milho e Urochloa ruziziensis e ausência ou presença de inoculação com Azospirillum brasilense. Durante a condução do experimento foram realizadas as seguintes avaliações: características agronômicas, componentes de produção e produtividade das culturas do arroz e feijão. O cultivo antecessor de Urochloa ruziziensis propiciou plantas mais altas e com maior teor de nitrogênio e plantas de feijão com maior número de vagens. As diferentes culturas antecessoras utilizadas não influenciaram na produtividade de grãos do arroz de terras altas e do feijão de inverno. A inoculação de sementes de arroz com Azospirillum brasilense promoveu incremento de 19% na produtividade de grãos de arroz de terras altas e, consequentemente, seu efeito resultou em incremento de aproximadamente 25% na produtividade do feijoeiro de inverno inoculado com Rhizobium tropici. / Among the grains of importance in the world scenario, rice and common bean should be highlighted as part of the basic diet of Brazilians. The integration of methods such as crop diversification and the use of biotechnology that increase grain yield of rice and common bean can contribute environmentally and economically to the adoption of more sustainable agricultural production systems. The objective of this work was to evaluate the effect of different predecessor crops (corn e/ou Urochloa ruziziensis) and inoculation of Azospirillum brasilense on the development and productivity of upland rice and common bean grown in succession, as well as the inoculation of Rhizobium tropici on winter common bean crop. The study was carried out in Selvíria-MS country, in a Red Latosol Distrofic in no-tillage area for more than 10 years. The experimental design was randomized blocks, arranged in strips scheme, with four replications. The treatments were constituted by the arrangement of eight combinations of the corn and Urochloa ruziziensis and absence or presence of inoculation with Azospirillum brasilense. During the conduction of the experiment the following evaluations were carried out: agronomic characteristics, production components and yield of rice and bean crops. The predecessor crop of Urochloa ruziziensis provided higher plants with higher nitrogen content and bean plants with higher number of pods. The different predecessor crops used did not influence grain yield of upland rice and winter common bean. The inoculation of rice seeds with Azospirillum brasilense promoted a 19% increase in the productivity of upland rice grains and, consequently, its effect resulted in an increase of approximately 25% in the yield of winter common bean inoculated with Rhizobium tropici.

Orientador: Orivaldo Arf / Resumo: Dentre os cereais de importância no cenário mundial merecem destaque o arroz e feijão fazendo parte da alimentação básica dos brasileiros. A integração de métodos como a diversificação de culturas e a utilização da biotecnologia que aumentem a produtividade de grãos do arroz e feijão pode contribuir ambientalmente e economicamente na adoção de sistemas de produção agrícolas mais sustentáveis. Assim, o objetivo do trabalho foi avaliar o efeito de diferentes culturas antecessoras (milho e/ou Urochloa ruziziensis) e inoculação de Azospirillum brasilense sobre o desenvolvimento e a produtividade do arroz de terras altas e do feijão cultivado em sucessão, além da inoculação de Rhizobium tropici na cultura do feijão de inverno. O estudo foi realizado no município de Selvíria-MS, em um LATOSSOLO VERMELHO Distrófico em área de plantio direto há mais de 10 anos. O delineamento experimental utilizado foi o de blocos casualisados, disposto em esquema de faixas, com quatro repetições. Os tratamentos foram constituídos pelo arranjo de oito combinações de milho e Urochloa ruziziensis e ausência ou presença de inoculação com Azospirillum brasilense. Durante a condução do experimento foram realizadas as seguintes avaliações: características agronômicas, componentes de produção e produtividade das culturas do arroz e feijão. O cultivo antecessor de Urochloa ruziziensis propiciou plantas mais altas e com maior teor de nitrogênio e plantas de feijão com maior número de vagens. As diferentes cul... (Resumo completo, clicar acesso eletrônico abaixo) / Abstract: Among the grains of importance in the world scenario, rice and common bean should be highlighted as part of the basic diet of Brazilians. The integration of methods such as crop diversification and the use of biotechnology that increase grain yield of rice and common bean can contribute environmentally and economically to the adoption of more sustainable agricultural production systems. The objective of this work was to evaluate the effect of different predecessor crops (corn e/ou Urochloa ruziziensis) and inoculation of Azospirillum brasilense on the development and productivity of upland rice and common bean grown in succession, as well as the inoculation of Rhizobium tropici on winter common bean crop. The study was carried out in Selvíria-MS country, in a Red Latosol Distrofic in no-tillage area for more than 10 years. The experimental design was randomized blocks, arranged in strips scheme, with four replications. The treatments were constituted by the arrangement of eight combinations of the corn and Urochloa ruziziensis and absence or presence of inoculation with Azospirillum brasilense. During the conduction of the experiment the following evaluations were carried out: agronomic characteristics, production components and yield of rice and bean crops. The predecessor crop of Urochloa ruziziensis provided higher plants with higher nitrogen content and bean plants with higher number of pods. The different predecessor crops used did not influence grain yield of upland ric... (Complete abstract click electronic access below) / Mestre

O ferro é um micronutiente essencial em plantas, como também para praticamente todos os demais organismos. Porém, as formas livres de ferro intracelular podem ser extremamente danosas. A proteína ferritina tem papel crucial neste contexto, com a função de acumular ferro de uma forma segura e biodisponível. Cada proteína pode acumular aproximadamente 4500 átomos de ferro em sua cavidade interna. Em plantas, existe um número variado de cópias gênicas para ferritina e estas cópias têm expressão modulada por fatores bióticos e abióticos. No genoma do arroz foram caracterizadas duas cópias para o gene da ferritina. Como existem poucos estudos funcionais para ferritina em arroz, este trabalho teve como objetivos: (a) silenciar as duas cópias da ferritina da subespécie japonica variedade Nipponbare; (b) caracterizar, por PCR, mutantes para ferritina por inserção do retroelemento TOS17; (c) caracterizar a expressão da ferritina da subespécie de arroz japonica, variedade Nipponbare, em plantas cultivadas em meio hidropônico sob excesso de ferro. Utilizando o sistema Gateway (Invitrogen) nós desenvolvemos uma construção que expressa um RNA em grampo projetado para silenciar ambas as cópias dos genes da ferritina de arroz. Baseando-se em um protocolo bem estabelecido de regeneração de plantas transgênicas de arroz, nós regeneramos plantas transgênicas silenciadas para os genes da ferritina. Foi obtido 75% de sucesso na geração das plantas silenciadas, o que está de acordo com a literatura. Os transformates primários (T0) não apresentaram anormalidades morfológicas evidentes. É possível que uma rota compensatória para armazenar ferro de forma segura seja ativada quando os níveis de ferritina são diminuídos. Além disso, as plantas produzidas neste trabalho são uma ferramenta potencial para estudar a relação ferro-planta. Baseando-se em análises in silico e por PCR, nós caracterizamos três linhagens mutantes contendo inserção do retroelemento TOS17 no gene OsFer2, entretanto, ainda não identificamos mutantes homozigotos. Em plantas de arroz crescidas em meio hidropônico, o aumento da concentração de ferro resultou em maiores níveis de expressão de ferritina, avaliados por RT-PCR semi-quantitativo, após 6 h e 12 h de exposição aos tratamentos de 50 e 500 ppm de FeSO4 do que na condição controle (5,6 ppm). / Iron is an essential micronutrient for plants, as for virtually all organisms. However, free intracelular iron forms can be extremely dangerous. The ferritin protein has a crucial role in this context, storing iron in a safe and bioavailable form. Each protein molecule can accumulate about 4500 iron atoms in its internal cavity. In plants, there is a variable number of ferritin gene copies and their expression is modulated by biotic and abiotic factors. There are two copies of the ferritin gene in the rice genome. As there are few functional studies for the ferritin genes in rice, this work had the objetives of: (a) to silence both copies of the ferritin genes in the japonica Nipponbare variety; (b) to identify and characterize TOS17 insertional mutants for the ferritin genes using in silico and PCR essays; (c) to characterize the expression of ferritin in the japonica Nipponbare variety under iron stress conditions. Using the Gateway system we generated a construct that expresses a hairpin RNA designed to silence both rice ferritin gene copies. Based on a well-established protocol to regenerate transgenic plants, we developed transgenic ferritin silenced lines. We obtained 75% success in generating rice silenced lines against ferritin. The primary transformants (T0) had no clear morphological abnormalities. It is possible that a compensatory pathway to store iron in a safe form can be induced when levels of ferritin are downregulated. Furthermore, the plants generated in this work are a potential tool to study iron-plant relations. Based on in silico and PCR essays we characterized three TOS17 insertional mutant lines for the OsFer2 gene, but until now we could not identify TOS17 homozygous mutants. In rice plants grown in hydroponic culture, increasing iron concentrations resulted in higher expression levels of ferritin, evaluated by semi-quantitative RT-PCR, after 6h and 12h exposure to 50 and 500 ppm of FeSO4, than in the control treatment (5,6 ppm).

The RCg2 promoter was identified in a search for root-specific genes to combat the rice water weevil (RWW) but expressed at low frequency (~10%). Spatial expression of RCg2 was investigated using two reporter constructs YXA (RCg2-gus-ocs) and YXB (RCg2-gus-RCg2) that included 1.6 kb of the RCg2 5' sequence fused to the ?-glucuronidase (gus) coding region. YXB plants were generated via Agrobacterium-mediated transformation but only 8 of 158 plants analyzed showed strong GUS activity despite the presence of an intact construct. Reactivation of RCg2 gene in rice was investigated by treatment of R0 and R1 of YXB transgenic plants with 5-azacytidine. Reactivation of RCg2-gus was observed in some transgenic plants indicating different mechanisms involved in the gene silencing of the YXB lines. DNA methylation analysis, northern blotting, RT-PCR and small RNA analysis supported the conclusion that PTGS and TGS are present in the silenced plants. Promoter analysis in silico and using promoter deletion assays predicted that the RCg2 promoter contains a complex region that includes miRNA homologs, MITEs and repetitive sequences. The high frequency of promoter-related silencing suggests functional interactions of these elements of the transgene and the homologous endogenous gene. To identify key elements contributing to the root-preferential expression of RCg2 and the high frequency of silencing observed in transgenic (YXB) lines, several RCg2 promoter deletion constructs were designed. These include 5' deletions MC1, MC2, MC4, MC7 and MC8 and internal deletions MC5, MC11, MC12 and MC13. The frequency with which silencing was encountered in populations of the deletion mutants was used to characterize the effects of various promoter elements. Deletion of the region from -406 to -208 (compared MC11 to YXB, and MC13 to MC1) revealed that region contains a negative element. Among 36 independent transformants, 33% with MC11 expressed GUS and 85% with MC13 showed GUS expression. Comparing MC7 transgenic plants to MC1 revealed that the region ?888 to ?729 is another negative regulatory element, and comparing MC11 to MC12, the proportion of expression of transgenic plants indicated the region ?729 to ?406 is a positive regulatory element.

The RCg2 promoter was identified in a search for root-specific genes to combat the rice water weevil (RWW) but expressed at low frequency (~10%). Spatial expression of RCg2 was investigated using two reporter constructs YXA (RCg2-gus-ocs) and YXB (RCg2-gus-RCg2) that included 1.6 kb of the RCg2 5' sequence fused to the ?-glucuronidase (gus) coding region. YXB plants were generated via Agrobacterium-mediated transformation but only 8 of 158 plants analyzed showed strong GUS activity despite the presence of an intact construct. Reactivation of RCg2 gene in rice was investigated by treatment of R0 and R1 of YXB transgenic plants with 5-azacytidine. Reactivation of RCg2-gus was observed in some transgenic plants indicating different mechanisms involved in the gene silencing of the YXB lines. DNA methylation analysis, northern blotting, RT-PCR and small RNA analysis supported the conclusion that PTGS and TGS are present in the silenced plants. Promoter analysis in silico and using promoter deletion assays predicted that the RCg2 promoter contains a complex region that includes miRNA homologs, MITEs and repetitive sequences. The high frequency of promoter-related silencing suggests functional interactions of these elements of the transgene and the homologous endogenous gene. To identify key elements contributing to the root-preferential expression of RCg2 and the high frequency of silencing observed in transgenic (YXB) lines, several RCg2 promoter deletion constructs were designed. These include 5' deletions MC1, MC2, MC4, MC7 and MC8 and internal deletions MC5, MC11, MC12 and MC13. The frequency with which silencing was encountered in populations of the deletion mutants was used to characterize the effects of various promoter elements. Deletion of the region from -406 to -208 (compared MC11 to YXB, and MC13 to MC1) revealed that region contains a negative element. Among 36 independent transformants, 33% with MC11 expressed GUS and 85% with MC13 showed GUS expression. Comparing MC7 transgenic plants to MC1 revealed that the region ?888 to ?729 is another negative regulatory element, and comparing MC11 to MC12, the proportion of expression of transgenic plants indicated the region ?729 to ?406 is a positive regulatory element.

The weed red rice is a major problem in rice producing areas world wide. All of the red rice in commercial rice fields in the United States has traditionally been considered to be the same species as commercial rice, Oryza sativa. However, using DNA markers it was found that most of the red rice with black hulls was sufficiently divergent to be considered a separate species. This includes TX4, a red rice ecotype that has been reported to have considerable natural tolerance to the herbicide glufosinate. TX4 is closely related to samples that have been classified as Oryza rufipogon. However, it was shown that both the TX4-like red rice from commercial fields and most of the Oryza rufipogon accessions in the US National Small Grains Collection are more accurately classified as Oryza nivara. This is significant since Oryza rufipogon is regulated under the Federal Noxious Weed Act, while Oryza nivara is not. Oryza nivara closely related to TX4 was found to be widely distributed across the rice production areas of Texas and was also found in Arkansas, Louisiana, and Mississippi. Of 240 samples from across Texas, 23 samples from six different counties were identical with TX4 with all 18 DNA markers tested. The reported glufosinate tolerance of TX4 is a potential problem since this same herbicide would be used in conjunction with genetically modified (GM) that is being developed as a method of red rice control. Thus, field, greenhouse and tissue culture studies were conducted to evaluate the degree of glufosinate tolerance in TX4. TX4 typically was severely damaged by glufosinate, but not efficiently controlled. Even with the maximum number of herbicide applications at the proposed maximum label rate, TX4 often re-sprouted and produced viable seed. Herbicide tolerance was found to be variable, but appears to be sufficient to present a problem with the use of the GM glufosinate resistant varieties currently under development, particularly when combined with variation in the response of ??sensitive?? varieties.