Avaliação da atividade antimicrobiana das defensinas recombinantes de ervilha (drr230a) e café (cd1) produzidas em Pichia pastoris

The inefficiency of chemical pesticides to control phytopathogenic fungi in agriculture and the frequent incidence of human diseases caused by bacteria which are resistant to antibiotics lead to the search for alternative antimicrobial compounds. In this context, plant defensins are a promising tool for the control of both plant and human pathogenic agents. Plant defensins are cationic peptides of about 50 amino acid residues, rich in cysteine and whose tridimensional structure is considerably conserved among different plant species. These antimicrobial molecules represent an important innate component from plant defense response against pathogens and are expressed in various plant tissues, such as leaves, tubers, flowers, pods and seeds. The present work aimed at the evaluation of the antimicrobial activity of two plant defensins against different phytopathogenic fungi and pathogenic bacteria to humans. The defensin Drr230a, whose gene was isolated from pea (Pisum sativum), and the defensin CD1,whose gene was identified within coffee (Coffea arabica) transcriptome, were subcloned in yeast expression vector and expressed in Pichia pastoris. The gene cd1 was subcloned as two different recombinant forms: CD1tC, containing a six-histidine sequence (6xHis) at the peptide C-terminal region and CD1tN, containing 6xHis coding sequence at the N-terminal region. In the case of the defensin Drr230a, the 6xHis coding sequence was inserted only at the N-terminal region. Assays of the antimicrobial activity of the purified recombinant proteins rDrr230a and rCD1 against Phakopsora pachyrhizi, causal agent of soybean Asian rust, were performed to analyze the in vitro spore germination inhibition and disease severity caused by the fungus in planta. Both recombinant defensins were able to inhibit P. pachyrhizi uredospore germination, with no difference between the antimicrobial action of either CD1tC or CD1tN. Moreover, rDrr230a and rCD1 drastically reduced severity of soybean Asian rust, as demonstrated by in planta assays. In spite of the fact that rCD1 was not able to inhibit proliferation of the human pathogenic bacteria Staplylococcus aureus and Klebsiella pneumoniae, rCD1 was able to inhibit growth of the phytopathogenic fungus Fusarium tucumaniae, that causes soybean sudden death syndrome. The obtained results show that these plant defensins are useful candidates to be used in plant genetic engineering programs to control agriculture impacting fungal diseases.

Patogenicidade de isolados de espécies de fusarium em grão-de-bico e ervilha.

Este trabalho teve como objetivo avaliar a patogenicidade de isolados de espécies de fusarium em grão-de-bico e ervilha.

Diversificação de culturas de inverno na região tritícola do Sul do Brasil.

1984

Anais...

Estatuto Social. Programação. Resumos: Roadmapping da comercialização de inoculantes no Brasil: novas estratégias, articulações e demandas da ANPII. Evaluación de la sobrevida bacteriana en un pre-inoculante para soja. Utilização de um inoculante padrão como referência para a determinação de qualidade de produtos comerciais. Comparação e adequação de metodologias para controle de qualidade de inoculantes comerciais para leguminosas. Avaliação da produtividade da cultura do milho com diferentes doses de fósforo e inoculação com Penicillium bilaiae. Testes de eficiência agronômica da tecnologia de co-inoculação de rizóbios e azospirillum em soja e feijoeiro. Eficiência simbiótica de estirpes isoladas de áreas cultivadas com soja em Roraima. Interação entre cultivares de arroz irrigado com bactérias diazotrófricas associativas. A construção de uma rede de promoção do benefício da FBN através dos inoculantes: uma proposta metodológica em busca de uma Agricultura de Baixo Carbono. Efeitos da fertilização nas características de promoção de crescimento vegetal: direcionando a prospecção de inoculantes. Potencial de expansão da FBN para a produção de grãos no Brasil pela agricultura familiar. Promoção do crescimento inicial do milho estimulado por Bacillus sp. Validação e demandas de estirpes de rizóbio para a inoculação de espécies arbóreas, adubos verdes e forrageiras visando metas do programa ABC e novo código florestal. Tecnologia de bioprocessos aplicada ao desenvolvimento de inoculantes e novos insumos biológicos. A construção de uma rede de promoção do benefício da FBN através dos inoculantes: uma proposta metodológica em busca de uma Agricultura de Baixo Carbono. A pesquisa em Fixação Biológica do Nitrogênio na Embrapa Soja: passado, presente e perspectivas futuras. Embrapa Cerrados: 37 anos de contribuições para o avanço da FBN no Brasil. Especificidade de rizóbios em ervilha. Ata técnica da assembleia geral ordinária da XVI RELARE. Ata da eleição da diretoria da RELARE para o biênio 2012-2014. Relação de participantes da XVI RELARE.

Studies on arbuscular mycorrhiza (AM) in the Alentejo (Portugal) using pea mutants resistant to AM fungi as a control tool for field conditions

The utilization and management of arbuscular mycorrhiza (AM) symbiosis may improve production and sustainability of the cropping system. For this purpose, native AM fungi (AMF) were sought and tested for their efficiency to increase plant growth by enhanced P uptake and by alleviation of drought stress. Pot experiments with safflower (Carthamus tinctorius) and pea (Pisum sativum) in five soils (mostly sandy loamy Luvisols) and field experiments with peas were carried out during three years at four different sites. Host plants were grown in heated soils inoculated with AMF or the respective heat sterilized inoculum. In the case of peas, mutants resistant to AMF colonization were used as non-mycorrhizal controls. The mycorrhizal impact on yields and its components, transpiration, and P and N uptake was studied in several experiments, partly under varying P and N levels and water supply. Screening of native AMF by most probable number bioassays was not very meaningful. Soil monoliths were placed in the open to simulate field conditions. Inoculation with a native AMF mix improved grain yield, shoot and leaf growth variables as compared to control. Exposed to drought, higher soil water depletion of mycorrhizal plants resulted in a haying-off effect. The growth response to this inoculum could not be significantly reproduced in a subsequent open air pot experiment at two levels of irrigation and P fertilization, however, safflower grew better at higher P and water supply by multiples. The water use efficiency concerning biomass was improved by the AMF inoculum in the two experiments. Transpiration rates were not significantly affected by AM but as a tendency were higher in non-mycorrhizal safflower. A fundamental methodological problem in mycorrhiza field research is providing an appropriate (negative) control for the experimental factor arbuscular mycorrhiza. Soil sterilization or fungicide treatment have undesirable side effects in field and greenhouse settings. Furthermore, artificial rooting, temperature and light conditions in pot experiments may interfere with the interpretation of mycorrhiza effects. Therefore, the myc- pea mutant P2 was tested as a non-mycorrhizal control in a bioassay to evaluate AMF under field conditions in comparison to the symbiotic isogenetic wild type of var. FRISSON as a new integrative approach. However, mutant P2 is also of nod- phenotype and therefore unable to fix N2. A 3-factorial experiment was carried out in a climate chamber at high NPK fertilization to examine the two isolines under non-symbiotic and symbiotic conditions. P2 achieved the same (or higher) biomass as wild type both under good and poor water supply. However, inoculation with the AMF Glomus manihot did not improve plant growth. Differences of grain and straw yields in field trials were large (up to 80 per cent) between those isogenetic pea lines mainly due to higher P uptake under P and water limited conditions. The lacking N2 fixation in mutants was compensated for by high mineral N supply as indicated by the high N status of the pea mutant plants. This finding was corroborated by the results of a major field experiment at three sites with two levels of N fertilization. The higher N rate did not affect grain or straw yields of the non-fixing mutants. Very efficient AMF were detected in a Ferric Luvisol on pasture land as revealed by yield levels of the evaluation crop and by functional vital staining of highly colonized roots. Generally, levels of grain yield were low, at between 40 and 980 kg ha-1. An additional pot trial was carried out to elucidate the strong mycorrhizal effect in the Ferric Luvisol. A triplication of the plant equivalent field P fertilization was necessary to compensate for the mycorrhizal benefit which was with five times higher grain yield very similar to that found in the field experiment. However, the yield differences between the two isolines were not always plausible as the evaluation variable because they were also found in (small) field test trials with apparently sufficient P and N supply and in a soil of almost no AMF potential. This similarly occurred for pea lines of var. SPARKLE and its non-fixing mycorrhizal (E135) and non-symbiotic (R25) isomutants, which were tested in order to exclude experimentally undesirable benefits by N2 fixation. In contrast to var. FRISSON, SPARKLE was not a suitable variety for Mediterranean field conditions. This raises suspicion putative genetic defects other than symbiotic ones may be effective under field conditions, which would conflict with the concept of an appropriate control. It was concluded that AMF resistant plants may help to overcome fundamental problems of present research on arbuscular mycorrhiza, but may create new ones.

Injúrias causadas por mosca-das-frutas sul-americana em cultivares de videira

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

Temperature during soybean seed storage and the amount of electrolytes of soaked seeds solution

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Teste de condutividade elétrica para a avaliação do vigor de sementes de abobrinha

O teste de condutividade elétrica da solução de embebição é recomendado para avaliar o vigor de sementes de ervilha e indicado para outras espécies, mas ainda há necessidade da continuidade de estudos sobre o assunto. A presente pesquisa foi desenvolvida com o objetivo de comparar procedimentos no teste de condutividade elétrica para avaliar o potencial fisiológico de sementes de abobrinha. Utilizaram-se sementes de abobrinha (Cucurbita pepo L.), cultivar Menina Brasileira, representadas por quatro lotes na primeira etapa e por outros cinco, na segunda. Foram conduzidos os teste de germinação, primeira contagem de germinação, emergência de plântulas, índice de velocidade de emergência e de condutividade elétrica. Para o teste de condutividade foram estudadas variações no volume de água (50 e 75mL), na temperatura (20, 25 e 30ºC) e no tempo de embebição (1, 2, 4, 8, 12, 16, 20 e 24 horas). O tempo de embebição para o teste de condutividade elétrica pode ser reduzido para 8 horas. Independentemente do volume de água, a temperatura de 30ºC foi a mais favorável para a ordenação consistente dos lotes quanto ao vigor. Assim, conclue-se que a condição mais adequada para o teste de condutividade elétrica em sementes de abobrinha é a utilização de 50 sementes em 75mL de água por oito horas, a 30ºC.

Electrical conductivity as an indicator of pea seed aging of stored at different temperatures

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Electrical conductivity and mineral composition of the imbibition solution of bean seeds during storage

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Alternative procedure for the cold test for soybean seeds

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

Utilização de marcador molecular SCAR na identificação de Fusarium subglutinans, agente causal da malformação da mangueira

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Desempenho de cultivares transgênicas de soja em sucessão a culturas de inverno em semeadura direta

The objective of this research was to evaluate crop yield and some characteristics and yield components of transgenic soybean cultivars sown after different winter cover crops in the first year under no tillage system. The experimental design was the completely randomized block with split plots and four replications. The main plots consisted of five winter cover crops, white oat (Avena sativa L.), forage turnip (Raphanus sativus L.), barley (Hordeum vulgare L.), wheat (Triticum aestivum L.) and ground pea (Pisum sativum L.) and an area under fallow (spontaneous vegetation). The subplots consisted of six soybean cultivars (BRS 243 RR, BRS 245 RR, BRS 247 RR, BRS 255 RR, BRS 256 RR and BRS 244 RR). Variance analysis for agronomic characteristics showed that soybean yield components were influenced by the interaction between winter crop and soybean cultivar. Thus, final population, number of nodes and pods per plant, nodes dry matter per plant, number of grains per pod and grain yield were affected significantly. When soybean nodulation was evaluated, the treatment with the area under fallow showed lower values. There was difference among winter crops for BRS 243 RR grain yield, white oat showed the highest values.

Caracterização de um isolado de Bidens mosaic virus proveniente de alface

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Inheritance of resistance to powdery mildew in pea and pathogenesis-related aspects

A herança da resistência ao oídio na cultivar de ervilha MK-10 e alguns aspectos histológicos da infecção foram estudados. Para o estudo da herança, as gerações F1, F2, retrocuzamentos e geração F3 de MK-10 com duas populações suscetíveis foram avaliadas. Nas avaliações histológicas observou-se a porcentagem de conídios germinados, porcentagem de conídios que formaram apressório, porcentagem de conídios que estabeleceram colônia e número de haustórios por colônia. Para comparar as razões de segregação obtidas no estudo da herança da resistência, adotou-se o teste do Qui-quadrado (X²) e para os dados das análises histológicas, utilizou-se o teste Tukey a 5% de probabilidade. Concluiu-se que a resistência de MK-10 ao oídio é devida a um par de alelos recessivos e que a resistência é expressa na fase de pré-penetração, completada por uma morte celular localizada pós-penetração, característica da presença do par de alelos recessivos er1er1.