Made available in DSpace on 2014-06-11T19:28:37Z (GMT). No. of bitstreams: 0 Previous issue date: 2009-08-28Bitstream added on 2014-06-13T20:37:48Z : No. of bitstreams: 1 mituti_t_me_botfca.pdf: 570381 bytes, checksum: c452322034e0c18e9beeb7fcc73e8761 (MD5) / Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP) / O alho é propagado através de bulbilhos, prática que favorece a perpetuação de patógenos, especialmente os vírus. Pode ser infectado por vírus pertencentes aos gêneros Allexivirus, Carlavirus e Potyvirus. Garlic common latent virus (GarCLV) e Shallot latent virus (SLV) são os principais carlavirus encontrados em alho no mundo. No Brasil, existem poucas informações sobre as espécies de carlavirus ocorrendo em alho, de modo que os objetivos do trabalho foram realizar levantamento visando determinar a ocorrência de carlavirus em algumas regiões produtoras do País, caracterizar e identificar através de técnicas biológicas e moleculares os isolados obtidos. Novecentas e dezenove amostras foram coletadas nos estados de São Paulo, Paraná, Goiás e Minas Gerais. Através do teste de ELISA, 560 foram positivas somente para potyvirus, 53 verificou-se a infecção pelo complexo entre GarCLV e potyvirus, sete foram positivas para SLV e potyvirus, duas positivas para GarCLV e uma para SLV. Cento e oito amostras foram negativas para os testes realizados. Alguns isolados de GarCLV foram inoculados mecanicamente, utilizando extrato vegetal, em plantas de Chenopodium amaranticolor, Celosia argentea, C. murale, C. quinoa, Nicotiana occidentalis, Gomphrena globosa, Allium tuberosum (cebolinha japonesa), Allium porrum (alho-porro) e Allium fistulosum (cebolinha). A presença de anéis cloróticos e mosaico foram observados em plantas de Celosia argentea, pontos cloróticos em Nicotiana occidentalis e lesões locais em Chenopodium quinoa. 2 Os oligonucleotídeos universais e específicos para GarCLV foram eficientes para detecção de vírus. Quatorze isolados foram sequenciados, indicando se tratar de isolados de GarCLV. A identidade de nucleotídeos entre os isolados de GarCLV foram de 87% a 97% comparadas com sequências depositadas no GenBank. Pela primeira vez no Brasil, foi detectado... / Garlic is propagated by bulbs, practice that favors the transmission of pathogens, especially viruses. Garlic can be infected by viruses belonging to the genus Allexivirus, Carlavirus and Potyvirus. Garlic common latent virus (GarCLV) and Shallot latent virus (SLV) are the most important carlavirus species infecting garlic around the world. In Brazil there is no information about the species of carlavirus occurring in garlic, so the objective of this work was to evaluate the occurrence of carlavirus in some producing regions of Brazil and to identify and characterize the isolates by molecular and biological techniques. Nine hundred and nineteen samples were collected in the states of São Paulo, Paraná, Goiás and Minas Gerais. Through the ELISA test, 560 were positive for the presence of potyvirus, 53 were infected with GarCLV and potyvirus, seven were infected with SLV and potyvirus, two by GarCLV, and one for SLV. One hundred and eight samples were negative for the presence of viruses. 4 Some GarCLV isolates were inoculated by sap transmission on Chenopodium amaranticolor, Celosia argentea, C. murale, C. quinoa, Nicotiana occidentalis, Gomphrena globosa, Allium tuberosum Allium porrum and Allium fistulosum. The presence of rings spots and mosaic were observed on Celosia argentea, chlorotic spots on Nicotiana occidentalis and chlorotic local lesions on C. quinoa. The universal oligonucleotides for carlavirus and the specifics for GarCLV were efficiently used for the detection of the viruses. Fourteen isolates were sequenced, indicating the presence of GarCLV species. The identity of nucleotides between the GarCLV isolates was of 87% to 97% compared to the sequences deposited in GenBank. SLV was detected for the first time in Brazil on samples collected in São Manuel and Piraquara. The nucleotide identity of the complete CP sequence was of 90% to 99% compared to the sequences... (Complete abstract click electronic access below)

Made available in DSpace on 2014-06-11T19:35:00Z (GMT). No. of bitstreams: 0 Previous issue date: 2011-05-13Bitstream added on 2014-06-13T18:45:54Z : No. of bitstreams: 1 silvajunior_taf_dr_botfca.pdf: 1093180 bytes, checksum: 562817be3d67ef9a5b0d63fa5e506cb7 (MD5) / Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES) / Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP) / A murcha-de-curtobacterium, causada por Curtobacterium flaccumfaciens pv. flaccumfaciens (Cff), é uma das principais doenças bacterianas da cultura do feijoeiro, acarretando grandes perdas na produção dessa cultura. Até o momento existem poucas informações sobre os diferentes nichos de sobrevivência desta bactéria e de sua gama de hospedeiras. Em vista disso, este trabalho teve por objetivos principais verificar a capacidade de sobrevivência saprofítica de Cff em restos de cultura de feijoeiro mantidos na superfície do solo e enterrados à 20 cm de profundidade; a influência da temperatura, umidade e do tipo de solo no período de sobrevivência da bactéria em solo; determinar a gama de hospedeiras de Cff inoculadas artificialmente, tanto por ferimento no caule, como por aspersão de suspensão bacteriana na parte aérea das plantas; a capacidade de colonização de Cff do rizoplano de plantas de aveia e trigo; e o efeito do pré-plantio de aveia e trigo na ocorrência da murcha-de-curtobacterium. Quanto à capacidade de sobrevivência de Cff em restos de cultura de feijoeiro, foi demonstrado que a bactéria possui menor capacidade de sobrevivência quando os restos vegetais são incorporados ao solo e também em épocas com maiores índices de precipitação e temperaturas mais altas. O período de sobrevivência do patógeno nos restos culturais de feijoeiro mantidos na superfície do solo variou entre 165 e 240 dias e nos restos vegetais enterrados a 20 cm de profundidade, o período de sobrevivência foi inferior a 30 dias. Quanto à sobrevivência de Cff na forma de células livres no solo, foi verificado que a temperatura, a umidade e o tipo do solo têm influencia na capacidade de sobrevivência da bactéria. O tempo de sobrevivência de Cff variou entre dois e quinze dias. Das 30 espécies botânicas inoculadas artificialmente com Cff, a bactéria causou lesões na parte... / Bacterial wilt caused by Curtobacterium flaccumfaciens pv. flaccumfaciens (Cff) is one of the main bacterial diseases affecting bean culture, leading to great losses in its production. So far there is scarce information about the different survival niches and host range of this bacterium. Thus, the present study had as major aims to verify Cff capability of saprophytically surviving in bean debris kept on the soil surface and buried at 20 cm depth; to assess the influence of temperature, humidity and soil type on the survival period of this bacterium in soil; to determine the host range for artificially inoculated Cff, either through stem injury or through bacterial sprinkling onto the shoot of plants; to verify Cff capability of colonizing the rhizoplane of oat and wheat plants; and to assess the effect of oat and wheat pre-planting on the occurrence of bean bacterial wilt. Cff had decreased capability of surviving in bean debris when the latter were incorporated into the soil and during periods of higher rainfall rates and temperatures. The pathogen survival period in bean culture remnants kept on the soil surface ranged from 165 and 240 days, while in plant debris buried at 20 cm depth the survival period was inferior to 45 days. The survival capability of Cff as free cells in soil was influenced by temperature, humidity and soil type. Cff survival time varied between two and fifteen days. Of 30 plant species artificially inoculated with Cff, bean and soy shoot had lesions caused by the bacterium which endophytically colonized wheat leaves and soy and wheat stem and leaves. Cff was also shown to have no capability of colonizing oat and wheat rhizoplane while the pre-planting of these grass plants, before bean culture establishment, had no effect on the occurrence of bean bacterial wilt

Resumo: O alho é propagado através de bulbilhos, prática que favorece a perpetuação de patógenos, especialmente os vírus. Pode ser infectado por vírus pertencentes aos gêneros Allexivirus, Carlavirus e Potyvirus. Garlic common latent virus (GarCLV) e Shallot latent virus (SLV) são os principais carlavirus encontrados em alho no mundo. No Brasil, existem poucas informações sobre as espécies de carlavirus ocorrendo em alho, de modo que os objetivos do trabalho foram realizar levantamento visando determinar a ocorrência de carlavirus em algumas regiões produtoras do País, caracterizar e identificar através de técnicas biológicas e moleculares os isolados obtidos. Novecentas e dezenove amostras foram coletadas nos estados de São Paulo, Paraná, Goiás e Minas Gerais. Através do teste de ELISA, 560 foram positivas somente para potyvirus, 53 verificou-se a infecção pelo complexo entre GarCLV e potyvirus, sete foram positivas para SLV e potyvirus, duas positivas para GarCLV e uma para SLV. Cento e oito amostras foram negativas para os testes realizados. Alguns isolados de GarCLV foram inoculados mecanicamente, utilizando extrato vegetal, em plantas de Chenopodium amaranticolor, Celosia argentea, C. murale, C. quinoa, Nicotiana occidentalis, Gomphrena globosa, Allium tuberosum (cebolinha japonesa), Allium porrum (alho-porro) e Allium fistulosum (cebolinha). A presença de anéis cloróticos e mosaico foram observados em plantas de Celosia argentea, pontos cloróticos em Nicotiana occidentalis e lesões locais em Chenopodium quinoa. 2 Os oligonucleotídeos universais e específicos para GarCLV foram eficientes para detecção de vírus. Quatorze isolados foram sequenciados, indicando se tratar de isolados de GarCLV. A identidade de nucleotídeos entre os isolados de GarCLV foram de 87% a 97% comparadas com sequências depositadas no GenBank. Pela primeira vez no Brasil, foi detectado... (Resumo completo, clicar acesso eletrônico abaixo) / Abstract: Garlic is propagated by bulbs, practice that favors the transmission of pathogens, especially viruses. Garlic can be infected by viruses belonging to the genus Allexivirus, Carlavirus and Potyvirus. Garlic common latent virus (GarCLV) and Shallot latent virus (SLV) are the most important carlavirus species infecting garlic around the world. In Brazil there is no information about the species of carlavirus occurring in garlic, so the objective of this work was to evaluate the occurrence of carlavirus in some producing regions of Brazil and to identify and characterize the isolates by molecular and biological techniques. Nine hundred and nineteen samples were collected in the states of São Paulo, Paraná, Goiás and Minas Gerais. Through the ELISA test, 560 were positive for the presence of potyvirus, 53 were infected with GarCLV and potyvirus, seven were infected with SLV and potyvirus, two by GarCLV, and one for SLV. One hundred and eight samples were negative for the presence of viruses. 4 Some GarCLV isolates were inoculated by sap transmission on Chenopodium amaranticolor, Celosia argentea, C. murale, C. quinoa, Nicotiana occidentalis, Gomphrena globosa, Allium tuberosum Allium porrum and Allium fistulosum. The presence of rings spots and mosaic were observed on Celosia argentea, chlorotic spots on Nicotiana occidentalis and chlorotic local lesions on C. quinoa. The universal oligonucleotides for carlavirus and the specifics for GarCLV were efficiently used for the detection of the viruses. Fourteen isolates were sequenced, indicating the presence of GarCLV species. The identity of nucleotides between the GarCLV isolates was of 87% to 97% compared to the sequences deposited in GenBank. SLV was detected for the first time in Brazil on samples collected in São Manuel and Piraquara. The nucleotide identity of the complete CP sequence was of 90% to 99% compared to the sequences... (Complete abstract click electronic access below) / Orientador: Marcelo Agenor Pavan / Coorientador: Renate Krause Sakate / Banca: Antonio Carlos Maringoni / Banca: Valdir Atsushi Yuki / Mestre

The presented thesis deals with interspecific competition and host range of entomopathogenic nematodes and assess the mechanisms enabling coexistence of multiple EPN species. The thesis further studied the scavenging ability of EPNs and their defense mechanisms against invertebrate scavengers. The results revealed that coexisting EPN species share the same niche and their interaction are complex and may be asymmetric. EPNs were found to be able to colonise and multiply in cadavers of different insects and scavenging seems to be an important alternative to normal infection. The defence of EPNs against scavengers seems to be an adaptation of the nematode-bacteria complex.

Johnsongrass mosaic virus (JGMV) é uma espécie do gênero Potyvirus. A sua distribuição geográfica, até o início da década de 1990, estava limitada à Austrália e aos Estados Unidos, onde causa doença em sorgo, milho e várias gramíneas. Em 2001, o JGMV foi detectado pela primeira vez no Brasil em amostras de híbridos e variedades de milho provenientes da região de Ribeirão Preto, SP mediante análise sorológica (DAS-ELISA), e em 2013 foi detectado mediante RT-PCR em amostras de Pennisetum purpureum provenientes do Estado da Bahia. Em Fevereiro de 2012 a Clínica Fitopatológica da ESALQ/USP recebeu amostras de Panicum maximum cv. Mombaça, com sintomas de mosaico, de São Luiz do Paraitinga, SP. Exames preliminares de contrastação negativa em microscópio eletrônico de transmissão indicaram a presença de partículas virais características de potyvirus. Diante disso, o principal objetivo deste trabalho foi caracterizar o agente etiológico associado às plantas doentes de capim Mombaça mediante testes biológicos, sorológicos e moleculares. Extratos foliares de plantas sintomáticas de capim Mombaça foram inoculados mecanicamente em 69 genótipos da família Poaceae. As avaliações foram feitas com base nos sintomas e por PTA-ELISA usando antissoro policlonal contra a proteína capsidial do potyvirus produzido nesse trabalho, após purificação do isolado viral. As espécies susceptíveis foram Brachiaria brizantha, B. decumbens, B. plantaginea, Cenchrus echinatus, Echinochloa colona, E. crus-galli, E. cruspavonis, Melinis minutiflora, Panicum maximum cv. Colonião, Pennisetum setosum, Rhynchelytrum repens, Rottboellia exaltata, Sorghum bicolor BRS 332, S. bicolor BRS 509, S. bicolor x S. sudanense BRS 802 e S. verticilliflorum. Espécies cultivadas como arroz, aveia, cana-de-açúcar, centeio, milho e trigo não foram infectadas com esse isolado. O peso molecular da proteína capsidial deste potyvirus foi estimado em cerca de 33 kDa por meio de Western blot. Sequência de nucleotídeos do genoma completo (9.885 nt) obtida neste estudo revelou identidade de 82,03% com a única sequência completa do genoma de um isolado do JGMV da Austrália, depositada no GenBank. A partir dessa sequência foram obtidos oligonucleotídeos iniciadores específicos para a detecção do isolado de SP do JGMV mediante RT-PCR. / Johnsongrass mosaic virus (JGMV) is a species of the genus Potyvirus. The geographical distribution, until the early 1990s, was limited to Australia and the United States, where it causes disease in sorghum, corn and various grasses. In 2001, JGMV was first detected in Brazil in samples of hybrids and varieties of corn from the region of Ribeirão Preto, São Paulo State by serological analysis (DASELISA), and in 2013 it was detected by RT-PCR in samples of Pennisetum purpureum from the State of Bahia. In February 2012, the Disease Diagnostic Clinic ESALQ/USP received samples of Panicum maximum cv. Mombaça, exhibiting mosaic symptoms, from the region of São Luiz do Paraitinga, SP. Preliminary examination of negatively stained sap in a transmission electron microscope indicated the presence of potyvirus-like particles. Therefore, the main objective of this study was to characterize the etiologic agent associated with P. maximum cv. Mombaça diseased plants by biological, serological and molecular tests. Leaf extract from Mombaça infected plants was mechanically inoculated in 69 genotypes of the Poaceae family. Evaluations were done based on symptoms expression and PTAELISA using polyclonal antiserum against the capsid protein of the potyvirus produced in the preset work virus purification. Susceptible species were Brachiaria brizantha, B. decumbens, B. plantaginea, Cenchrus echinatus, Echinochloa colona, E. crus-galli, E. crus-pavonis, Melinis minutiflora, Panicum maximum cv. Colonião, Pennisetum setosum, Rhynchelytrum repens, Rottboellia exaltata, Sorghum bicolor BRS 332, S. bicolor BRS 509, S. bicolor x S. sudanense BRS 802 and S. verticilliflorum. Cultivated species such as rice, oats, sugarcane, rye, corn and wheat were not infected with this isolate. The molecular weight of the coat protein of this potyvirus was estimated at about 33 kDa by Western blot. The nucleotide sequence of the complete genome (9885 nt) obtained in this study showed 82.03% identity with an unique sequence for the complete genome of an isolate of JGMV from Australia, deposited in GenBank. From this nucleotide sequence, specific pair of primers was designed for the detection of the São Paulo isolate of JGMV by RT-PCR.

Para alimentar a população mundial crescente é necessário um aumento sustentável na produtividade agrícola. Nesse sentido, Rizobactérias Promotoras de Crescimento de Plantas (RPCPs) têm sido continuamente buscadas para formulações inoculantes por sua capacidade de incremento na produção vegetal aliado ao seu potencial de redução e/ou substituição do uso de fertilizantes minerais, insumos que causam grandes impactos ambientais, na saúde humana e econômicos. A RPCP Bacillus sp. RZ2MS9, um representante da biodiversidade amazônica brasileira, é uma forte candidata a bionoculante por seu efeito benéfico, previamente descrito, em uma ampla gama de culturas, incluindo milho e soja. Essas duas culturas representam mais de 80% da área cultivada com grãos no Brasil, de forma que incrementos relativamente modestos de crescimento e produtividade poderiam gerar ganhos significativos. Membros do gênero Bacillus apresentam vantagem em formulações inoculantes, principalmente devido a sua capacidade de formação de esporos resistentes ao calor e dissecação. Seus modos de ação são diversos, tornando o entendimento da sua interação com plantas bastante desafiador. Bacillus sp. RZ2MS9 apresentou, dentre os mecanismos envolvidos na promoção de crescimento vegetal, a produção de Ácido Indol Acético (AIA) e sideróforos, solubilização de fosfato e fixação biológica de nitrogênio, in vitro. No presente trabalho, foi buscado um entendimento detalhado dos mecanismos de ação dessa rizobactéria, explorando desde seu genoma até seu desempenho em condições de campo. O draft genômico (genoma parcial) bacteriano foi obtido utilizando a tecnologia de sequenciamento Illumina, o qual possibilitou a detecção de genes envolvidos nos mecanismos potencialmente relacionados ao efeito benéfico dessa bactéria, que vão desde sua formação de esporos, atração por exsudatos radiculares, motilidade e competição na rizosfera até mecanismos de solubilização de fosfato, produção de sideróforos, entre outros. As informações obtidas permitem uma exploração genética desses mecanismos, fornecendo uma oportunidade de maximizar essa interação e, futuramente, favorecer os benefícios em campo. Adicionalmente, foi demonstrado o potencial de quimiotaxia (atração) de RZ2MS9 em direção a raízes de milho. Um estudo filogenético dessa RPCP, utilizando um método de tipagem com o gene pycA (piruvato carboxilase), mostrou que o Bacillus sp. RZ2MS9 apresentou-se distante do clado altamente monomórfico de B. anthracis, patógeno humano, e se afiliou a um grupo composto por linhagens de B. thuringiensis (Bt) comercializadas como produtos biopesticidas há mais de 60 anos, o que sugere a potencial possibilidade de seu uso seguro no campo. Sabe-se que a maioria, se não todas, atividades fisiológicas das plantas é regulada por fitormônios como a auxina AIA, os quais podem ser sintetizados também por RPCPs. Com mais detalhamento, os genes envolvidos nas vias biossintéticas desse fitormônio foram detectados no draft genômico de RZ2MS9, indicando que sua produção ocorre através da via IPA (Indol-3-Piruvato). Além disso, plantas de tomate anão Micro-Tom (MT) e seu mutante Δdgt, defectivo na sensibilidade a auxinas, foram utilizadas para caracterizar especificamente o efeito do AIA produzido por Bacillus sp. RZ2MS9 na promoção de crescimento vegetal. A aplicação de RZ2MS9 causou inibição no crescimento de raízes primárias, aumento no comprimento de raízes laterais e na área superficial total de raízes de plantas MT, efeitos característicos daqueles proporcionados por auxinas. Esse incremento radicular refletiu, ainda, em aumento da biomassa da parte aérea de plantas MT. Os mesmos efeitos não foram observados em plantas Δdgt, insensíveis a auxinas, indicando que a elicitação de promoção de crescimento em MT por RZ2MS9 ocorre por meio desses fitormônios. Finalmente, foi demonstrado o efeito sobre o desenvolvimento e produtividade de milho e soja da aplicação de Bacillus sp. RZ2MS9 em condições de campo, sendo comparado com o desempenho de bioinoculantes comerciais. No milho, o efeito da inoculação bacteriana foi, ainda, associado à adubação nitrogenada para verificar a possibilidade de redução desses insumos. Bacillus sp. RZ2MS9 apresentou efeitos significativos sobre o desenvolvimento tanto da soja (comparáveis aos efeitos de rizóbios) quanto do milho, os quais, porém, não refletiram em aumento significativo de produtividade em ambas as culturas. No entanto, o potencial dessa rizobactéria é bastante claro pois, com um custo de produção inferior a R$1,00 por hectare, sua inoculação causou incremento de 16 sacas de milho por hectare com redução de 30% na adubação nitrogenada, assim como um incremento de 11 sacas de soja por hectare, ambos comparados ao controle não inoculado. Os resultados apresentados no presente trabalho vão, portanto, de encontro à grande expectativa na obtenção de linhagens microbianas promissoras visando sistemas agrícolas mais sustentáveis. / To feed the growing global population, a sustainable increase of agricultural production and crop yield is required. In this sense, Plant Growth Promoting Rhizobacteria (PGPR) have been continuously sought to inoculant formulation due to their capacity to increase plant yield along with their potential to reduce and/or replace the use of mineral fertilizers, inputs that cause serious impacts on environment, human health and economy. The PGPR Bacillus sp. RZ2MS9, a representative of the Brazilian Amazonian biodiversity, is a great candidate to bioinoculant because of its beneficial effect on a broad range of crops, including maize and soybean. These two crops represent more than 80% of the area planted with grains in Brazil, so relatively modest growth and yield increases could generate significant gains. Bacillus spp. have advantage in inoculant formulations, mainly due to their ability to form heat- and dissecation-resistant spores. Their modes of action are diverse, making the understanding of its interaction with plants quite challenging. Bacillus sp. RZ2MS9 displays, between the mechanisms involved in plant growth, Indole Acetic Acid (IAA) and siderophore production, phosphate solubilization and biological nitrogen fixation, in vitro. In the present work, we seek a detailed understanding of this rhizobacterium mechanisms of action, exploring from its genome to its performance in field conditions. The bacterial draft genome was obtained using Illumina sequencing technology, making possible the detection of genes involved in mechanisms potentially related to the beneficial effect of this bacterium, and range from its spore formation, attraction by root exudates, motility and competition in the rhizosphere to mechanisms of phosphate solubilization, siderophore production, among others. The information obtained allow a genetic exploration of these mechanisms, providing an opportunity to maximize this interaction and, in the future, favor benefits in field. Additionally, it was demonstrated the chemotaxis (attraction) potential of RZ2MS9 towards maize roots. A phylogenetic study of this PGPR, using a typing method with the pycA (pyruvate carboxylase) gene, showed that Bacillus sp. RZ2MS9 was distant from the highly monomorphic clade of B. anthracis, a human pathogen, and affiliated with B. thuringiensis (Bt) strains marketed as biopesticides for more than 60 years, suggesting the potential possibility of its safe use in the field. It is known that most, if not all, physiological activities of plants are regulated by phytormones such as the auxin IAA, which can also be synthesized by PGPRs. With more detail, genes involved in biosynthetic pathways of this phytormone were detected in the RZ2MS9 draft genome, indicating that its production occurs via the IPA (indole-3-pyruvate) pathway. In addition, plants of the dwarf tomato Micro-Tom (MT) and its mutant Δdgt, impaired in auxin sensibility, were used to specifically characterize the effects of IAA produced by Bacillus sp. RZ2MS9 in the plant growth promotion. The inoculation of RZ2MS9 caused inhibition in the primary roots growth, increase in lateral roots length and in roots total surface area of MT plants, characteristic effects of those provided by auxins. This root growth also reflected in an increase of MT plants shoot biomass. The same effects were not observed in Δdgt plants, insensitive to auxins, suggesting that the elicitation of growth promotion in MT by RZ2MS9 occurs through these phytormones. Finally, we demonstrated the effect of inoculation with Bacillus sp. RZ2MS9 on maize and soybean development and productivity under field conditions, being compared with the performance of commercial bioinoculants. In maize, the effect of bacterial inoculation was also associated with nitrogen fertilization to verify the possibility of reducing these inputs. Bacillus sp. RZ2MS9 showed significant effects on the development of both soybean (comparable to the effects of rhizobia) and maize, which, however, did not reflect a significant increase in productivity in both crops. However, the potential of this rhizobacterium is very clear because, with a cost of production of less than R$1.00 per hectare, its inoculation caused an increase of 16 sacks of maize per hectare with a 30% reduction in nitrogen fertilization, as well as an increase of 11 sacks of soybean per hectare, both compared to uninoculated control. The results presented in this study meet the great expectation of obtaining promising microbial strains aiming at more sustainable agricultural systems.

Several wild-type poxviruses have emerged as potential oncolytic viruses (OVs), including orf virus (OrfV), and vaccinia virus (VV). Oncolytic VVs have been modified to include attenuating mutations that enhance their tumour selective nature, but these mutations also reduce overall viral fitness in cancer cells. Previous studies have shown that a VV (Western Reserve) with its E3L gene replaced with the E3L homologue from, OrfV (designated VV-E3LOrfV), maintained its ability to infect cells in vitro, but was attenuated compared to its parental VV in vivo. Our goal was to determine the safety and oncolytic potential VV-E3LOrfV, compared to wild type VV and other attenuated recombinants. VV-E3LOrfV, was unable to replicate to the same titers and was sensitive to IFN compared to its parental virus and other attenuated VVs in normal human fibroblast cells. The virus was also less pathogenic when administered in vivo. Viral replication, spread and cell killing, as measures of oncolytic potential in vitro, along with in vivo efficacy, were also observed.. The Parapoxvirus, OrfV has been shown to have a unique immune-stimulation profile, inducing a number of pro-inflammatory cytokines, as well as potently recruiting and activating a number of immune cells. Despite this unique profile, OrfV is limited in its ability to replicate and spread in human cancer cells. Various strategies were employed to enhance the oncolytic efficacy of wild-type OrfV. A transient transfection/infection screen was created to determine if any of the VV host-range genes (C7L, K1L, E3L or K3L) would augment OrfV oncolysis. Combination therapy, including the use of microtubule targeting agents, Viral Sensitizer (VSe) compounds and the addition of soluble VV B18R gene product were employed to see if they also enhance OrfV efficacy. Unfortunately, none of the strategies mentioned were able to enhance OrfV.

Microorganisms interact frequently with each other and with higher organisms. This contact and communication takes place at the molecular level. Microbial interactions with eukaryotes can be pathogenic or mutualistic. One of the best-studied symbioses is the complex interaction between nitrogen-fixing soil bacteria, termed rhizobia, and legumes. This symbiosis culminates in the elaboration of a new organ, the root nodule. Many of the molecular signals exchanged between the host plant and the invading rhizobia have been deduced, but there is still much that remains to be discovered. The molecular determinant of host range at the genus level of the plant host has been determined to be lipochitooligomers called Nod factors. The molecular determinants of host range at the species and cultivar level are less well-defined. Part of my work has been to identify and characterize accessory plasmids that disrupt the normal progression of symbiosis between legumes of the genus Medicago and their rhizobial symbiont, Sinorhizobium meliloti. A cre--loxP-based system capable of making large, defined deletions was developed for the analysis of these plasmids. This system is also being employed to cure the laboratory strain, S. meliloti Rm1021 of its two megaplasmids-a loss of nearly half of its genome. I have also done work to determine whether locally-collected sinorhizobia are native, invasive, or native with symbiosis genes acquired horizontally from invasive sinorhizobia. Finally, I have studied Sinorhizobium meliloti as a host by identifying an outer membrane porin that several bacteriophages use to adsorb to the S. meliloti cell surface.

No description available.

Ailanthus altissima (Mill.) Swingle is a tree native to Asia that was intentionally introduced into the United States in the late eighteenth century. Ailanthus altissima has become an invasive species that has spread throughout most of North America. Lack of effective management tactics for suppression of A. altissima has lead to alternate control methods. Investigations into using biological control with the weevil Eucryptorrhynchus brandti were initiated in 2004. Studies were conducted to understand the general biology of E. brandti, rearing efficacy, and host specificity. Eucryptorrhynchus brandti is univoltine, has a life cycle similar to the closely related species Cryptorhynchus lapathi (L.), with 6 instars, and completes development in 126 ± 6.5 d at 25°C. Efficient egg to adult rearing was accomplished by caging 12 m and 12 f for 7 days on 23 – 92 cm long billets. Males and females can be differentiated by the structure of the metathoracic sternite and 1st abdominal segment. Host specificity experiments show that E. brandti preferentially feeds on North American A. altissima when tested against 29 species from 14 families. Larval development in the rare species Leitneria floridana Chapm. was apparent. Additional studies show that A. altissima does not occur across L. floridana distribution but may have the potential to invade L. floridana sites. / Ph. D.