Expression profile of oxidative and antioxidative stress enzymes based on ESTs approach of citrus

Plants not only evolve but also reduce oxygen in photosynthesis. An inevitable consequence of this normal process is the production of reactive oxygen species (ROS). Plants are adequately protected by the presence of multiple antioxidative enzymes in the cytosol and also in the different cell organelles such as chloroplasts, mitochondria, and peroxisomes. Traditionally, ROS were considered to be only a toxic byproduct of aerobic metabolism. However, recently it has become apparent that plants actively produce these molecules which may control many different physiological processes such as abiotic and biotic stress response, pathogen defense and systemic signaling. The search results using the Citrus Genome Program in Brazil (CitEST) for oxidative stress and the antioxidant enzyme system in Citrus Sinensis variety ?Pera IAC? indicated that the multiple ROS-scavenging enzymes were expressed throughout all citrus tissues. The analyses demonstrated the ubiquitous expression of metallothioneins, probably indicating a constitutive expression pattern. Oxalate oxidase has been identified as the most abundant expressed gene in developing fruits, which suggests a specific function in the ripening of citrus fruit. Moreover, infected leaves with Xylella fastidiosa and Leprosis citri showed a massive change in their ROS gene expression profile which may indicate that the suppression of ROS detoxifying mechanisms may be involved in the induction of the diseases.87288

In vitro expression and antiserum production against the movement protein of Citrus leprosis virus C (CiLV-C)

Citrus leprosis, caused by Citrus leprosis virus C (CiLV-C), is currently considered the most important viral disease in the Brazilian citrus industry due to the high costs required for the chemical control of its vector, the mite Brevipalpus phoenicis. The pathogen induces a non-systemic infection and the disease is characterized by the appearance of localized lesions on citrus leaves, stems and fruits, premature fruit and leaf drop and dieback of stems. Attempts were made to promote in vitro expression of the putative cell-to-cell movement protein of CiLV-C in Escherichia coli and to produce a specific polyclonal antibody against this protein as a tool to investigate the virus-plant-vector relationship. The antibody reacted strongly with the homologous protein expressed in vitro by ELISA, but poorly with the native protein present in leaf lesion extracts from sweet orange caused by CiLV-C. Reactions from old lesions were more intense than those from young lesions. Western blot and in situ immunolocalization assays failed to detect the native protein. These results suggest low expression of the movement protein (MP) in host tissues. Moreover, it is possible that the conformation of the protein expressed in vitro and used to produce the antibody differs from that of the native MP, hindering a full recognition of the latter.Fundacsao de Apoio a Pesquisa do Estado de Sao Paulo - FAPESP [04/11799-0, 2008/52691-9]Fundacsao de Apoio a Pesquisa do Estado de Sao Paulo FAPES

In vitro expression and antiserum production against the movement protein of Citrus leprosis virus C (CiLV-C)

Citrus leprosis, caused by Citrus leprosis virus C (CiLV-C), is currently considered the most important viral disease in the Brazilian citrus industry due to the high costs required for the chemical control of its vector, the mite Brevipalpus phoenicis. The pathogen induces a non-systemic infection and the disease is characterized by the appearance of localized lesions on citrus leaves, stems and fruits, premature fruit and leaf drop and dieback of stems. Attempts were made to promote in vitro expression of the putative cell-to-cell movement protein of CiLV-C in Escherichia coli and to produce a specific polyclonal antibody against this protein as a tool to investigate the virus-plant-vector relationship. The antibody reacted strongly with the homologous protein expressed in vitro by ELISA, but poorly with the native protein present in leaf lesion extracts from sweet orange caused by CiLV-C. Reactions from old lesions were more intense than those from young lesions. Western blot and in situ immunolocalization assays failed to detect the native protein. These results suggest low expression of the movement protein (MP) in host tissues. Moreover, it is possible that the conformation of the protein expressed in vitro and used to produce the antibody differs from that of the native MP, hindering a full recognition of the latter.Fundacsao de Apoio a Pesquisa do Estado de Sao Paulo - FAPESP [04/11799-0, 2008/52691-9]Fundacsao de Apoio a Pesquisa do Estado de Sao Paulo FAPES

Método E Kit De Detecção E Diagnóstico Do Esporo Phakopsora Pachyrhizi

MÉTODO E KIT DE DETECÇÃO E DIAGNÓSTICO DO ESPORO PHAKOPSORA PACHYRHIZI. A presente invenção está relacionada à imobilização física das moléculas químicas e/ou biológicas à placa de poliestireno ou policarbonato com a manutenção da sua estrutura conformacional intacta. Esta acessibilidade permite o diagnóstico de moléculas quimicas e/ou biológicas que apresentem um tamanho variável entre 1 a 50 gm. Este dispositivo pode ser utilizado em uma variabilidade de ensaios incluindo diagnóstico de detecção e ou métodos de quantificação de moléculas. Este dispositivo de diagnóstico está relacionado a um método rápido e eficiente do ensaio de ELISA (enzymelinked immunosorbent assay) para detecção específica do esporo do fungo Phakopsora pachyrhizi. O método com resultados reproduzíveis é perfeitamente adequado para sua utilização no campo, é extremamente simples, rápido e econômico. Este procedimento inventado é extremamente útil para diagnósticos clínicos, biologia molecular, agricultura, tecnologia de alimentos e etc. O método apresenta potencial de automação.BRPI0500452 (A)G01N33/549G01N33/549C12R1/645BR2005PI00452G01N33/549G01N33/549C12R1/64

Biossensor Para Detecção Precoce Da Ferrugem Asiática Na Soja

A presente invenção refere-se a um biossensor baseado na interação antígeno-anticorpo pare a detecção precoce (antes do aparecimento dos sintomas da doença) da ferrugem asiática em lavouras de soja. Este biossensor irá contribuir na redução dos prejuízos causados pela dispersão do fungo Phakopsora pachyrhizi (patógeno causador da doença) na sojicultura nacional e internacional. O monitoramento da doença e sua identificação nos estágios iniciais são essenciais para a eficiência do método utilizado pare manter a saúde da lavoura. Os biossensores que utilizam anticorpo como elemento de reconhecimento biológico do antígeno (imunossensores) apresentam características únicas como análises rápidas, sensíveis e com alto grau de seletividade, que os tornam atrativos para a determinação de compostos alvos em matrizes complexas (coma as plantas), além de possibilitar medidas diretas e em tempo real. O principal atrativo do dispositivo proposto é a obtenção do diagnóstico da doença precocemente e de maneira simples, rápida e segura, sem a necessidade de pré-tratamento da amostra e de testes confirmatórios.BRPI0805316 (A2)G01N33/569BR2008PI05316G01N33/56

Thymic Alterations Induced By Plasmodium Berghei: Expression Of Matrix Metalloproteinases And Their Tissue Inhibitors.

The thymus plays a crucial role in the generation of T-cells, and so our laboratory has been interested in the study of the intrathymic events that occur during infection diseases and may cause disruption in its functions. Previously, we showed that thymus from experimentally Plasmodium berghei-infected mice present histological alterations with high levels of apoptosis, changes in cell migration-related molecules, and premature egress of immature thymocytes to periphery. In addition, parasites were found inside the thymus. In this work we investigated alterations in the expression pattern and activity of matrix metalloproteinases MMP-2 and -9, and their tissue inhibitors, TIMP-1 and TIMP-2. Our results show enhanced expression and widespread distribution of these molecules in thymus from infected animals. Also, the presence of active MMP-2 was detected. These data are suggestive of MMPs and TIMPs importance in the earlier observed changes in the extracellular matrix during thymic alterations after plasmodium infection.27953-

Thymic alterations induced by Plasmodium berghei: Expression of matrix metalloproteinases and their tissue inhibitors

The thymus plays a crucial role in the generation of T-cells, and so our laboratory has been interested in the study of the intrathymic events that occur during infection diseases and may cause disruption in its functions. Previously, we showed that thymus from experimentally Plasmodium berghei-infected mice present histological alterations with high levels of apoptosis, changes in cell migration-related molecules, and premature egress of immature thymocytes to periphery. In addition, parasites were found inside the thymus. In this work we investigated alterations in the expression pattern and activity of matrix metalloproteinases MMP-2 and -9, and their tissue inhibitors, TIMP-1 and TIMP-2. Our results show enhanced expression and widespread distribution of these molecules in thymus from infected animals. Also, the presence of active MMP-2 was detected. These data are suggestive of MMPs and TIMPs importance in the earlier observed changes in the extracellular matrix during thymic alterations after plasmodium infection27915359COORDENAÇÃO DE APERFEIÇOAMENTO DE PESSOAL DE NÍVEL SUPERIOR - CAPESFUNDAÇÃO DE AMPARO À PESQUISA DO ESTADO DE SÃO PAULO - FAPESPsem informação08/58604-0