Soybean (Glycine max. L.) and bacteroid glyoxylate cycle activities during nodular senescence

Soybean (Glycine max. L.) nodular senescence results in the dismantling of the peribacteroid membrane (PBM) and in an increase of soybean isocitrate lyase (ICL; EC 4.1.3.1) and malate synthase (MS; EC 4.1.3.2) mRNA and protein levels. This suggests that in senescing soybean nodular cells, the specific glyoxylate cycle enzyme activities might be induced to reallocate carbon obtained from the PBM degradation. In order to evaluate as well the carbon metabolism of the nitrogen-fixing Bradyrhizobium japonicum endosymbiotic bacteroids during nodular senescence, their glyoxylate cycle activities were also investigated. To this end, partial DNA sequences were isolated from their icl and ms genes, but the corresponding mRNAs were not detected in the microorganisms. It was also observed that the bacteroid ICL and MS activities were negligible during nodular senescence. This suggests that glyoxylate cycle activities are not reinitiated in the bacteroids under these physiological conditions. In case the microorganisms nevertheless feed on the PBM degradation products, this might occur via the citric acid cycle exclusively.

Metabolic and structural rearrangement during dark-induced autophagy in soybean (Glycine max L.) nodules: An electron microscopy and P-31 and C-13 nuclear magnetic resonance study

The effects of dark-induced stress on the evolution of the soluble metabolites present in senescent soybean (Glycine max L.) nodules were analysed in vitro using C-13- and P-31-NMR spectroscopy. Sucrose and trehalose were the predominant soluble storage carbons. During dark-induced stress, a decline in sugars and some key glycolytic metabolites was observed. Whereas 84% of the sucrose disappeared, only one-half of the trehalose was utilised. This decline coincides with the depletion of Gln, Asn, Ala and with an accumulation of ureides, which reflect a huge reduction of the N-2 fixation. Concomitantly, phosphodiesters and compounds like P-choline, a good marker of membrane phospholipids hydrolysis and cell autophagy, accumulated in the nodules. An autophagic process was confirmed by the decrease in cell fatty acid content. In addition, a slight increase in unsaturated fatty acids (oleic and linoleic acids) was observed, probably as a response to peroxidation reactions. Electron microscopy analysis revealed that, despite membranes dismantling, most of the bacteroids seem to be structurally intact. Taken together, our results show that the carbohydrate starvation induced in soybean by dark stress triggers a profound metabolic and structural rearrangement in the infected cells of soybean nodule which is representative of symbiotic cessation.

Immunolocalization of glyoxysomal malate synthase from soybean cotyledons (Glycine max. L.)

Malate synthase (MS; EC 4.1.3.2), an enzyme specific to the glyoxylate cycle, was studied in cotyledons of dark-grown soybean (Glycine max L) seedlings with light and electron microscopy techniques. Immunogold localization confirmed biochemical evidence that MS from soybean is a glyoxysomal matrix enzyme.

Glyoxysomal malate-dehydrogenase and malate synthase from Soybean cotyledons (Glycine max. L.): Enzyme association, antibody-production and cDNA cloning

In order to investigate a possible association between soybean malate synthase (MS; L-malate glyoxylate-lyase, CoA-acetylating, EC 4.1.3.2) and glyoxysomal malate dehydrogenase (gMDH; (S)-malate: NAD(+) oxidoreductase, EC 1.1.1.37), two consecutive enzymes in the glyoxylate cycle, their elution profiles were analyzed on Superdex 200 HR fast protein liquid chromatography columns equilibrated in low- and high-ionic-strength buffers. Starting with soluble proteins extracted from the cotyledons of 5-d-old soybean seedlings and a 45% ammonium sulfate precipitation, MS and gMDH coeluted on Superdex 200 HR (low-ionic-strength buffer) as a complex with an approximate relative molecular mass (M(r)) of 670000. Dissociation was achieved in the presence of 50 mM KCl and 5 mM MgCl2, with the elution of MS as an octamer of M, 510 000 and of gMDH as a dimer of M, 73 000. Polyclonal antibodies raised to the native copurified enzymes recognized both denatured MS and gMDH on immunoblots, and their native forms after gel filtration. When these antibodies were used to screen a lambda ZAP II expression library containing cDNA from 3-d-old soybean cotyledons, they identified seven clones encoding gMDH, whereas ten clones encoding MS were identified using an antibody to SDS-PAGE-purified MS. Of these cDNA clones a 1.8 kb clone for MS and a 1.3-kb clone for gMDH were fully sequenced. While 88% identity was found between mature soybean gMDH and watermelon gMDH, the N-terminal transit peptides showed only 37% identity. Despite this low identity, the soybean gMDH transit peptide conserves the consensus R(X(6))HL motif also found in plant and mammalian thiolases.

Germination, senescence and pathogenic attack in soybean (Glycine max. L.): Identification of the cytosolic aconitase participating in the glyoxylate cycle

During our study of the glyoxylate cycle in soybean (Glycine max. L. var. Maple arrow), two mitochondrial and three cytosolic aconitase molecular species (EC 4.2.1.3) were detected, designated as M1, M2, C1, C2 and C3 isoforms, respectively, according to their intracellular locations and electrophoretic mobilities. Using the glyoxylate cycle marker enzymes isocitrate lyase (ICL, EC 4.1.3.1) and malate synthase (MS, EC 4.1.3.2), the activity of this pathway providing the essential link between P-oxidation and gluconeogenesis was confirmed during germination (cotyledons) and senescence (leaves). It was then established that, in both cases, the activity of the CI aconitase isoform developed concomitantly with the transcription and translation levels of the icl and ms genes. This strongly suggests that C1 aconitase is constitutive of the glyoxylate cycle. In addition, the same isoform was found to be active during pathogenic attack as well (hypocotyls). It might be assumed that in such a case the glyoxylate cycle is reinitiated as a part of a carbon reallocation system feeding on the diseased tissue cellular components.

Pathogenic attack and carbon reallocation in soybean leaves (Glycine max. L.): Reinitiation of the glyoxylate cycle as a defence reaction

Pathogenic attack by the fungus Botrytis cinerea (primary pathogen) on soybean leaves (Glycine max. L.; cv. Maple arrow) results in a hypersensitive response (necrotising infected leaves), in the establishment of local acquired resistance, as well as in the systemic induction of genes coding for pathogenesis-related proteins. It now appears that, concomitantly with these already well documented defence reactions, the pathogenic attack also induces the carbon reallocation mechanism based on the reinitiation of the glyoxylate cycle (pseudo-senescence of the infected leaves).

Intensidade da ferrugem asiática (Phakopsora pachyrhizi H. Sydow & P. Sydow) da soja [Glycine max (L.) Merr.] nas cultivares Conquista, Savana e Suprema sob diferentes temperaturas e períodos de molhamento foliar

A Ferrugem Asiática (Phakopsora pachyrhizi H. Sydow & P. Sydow), relatada em diversas regiões do globo terrestre de climas tropicais e subtropicais, causa redução significativa na produtividade da soja [Glycine max (L.) Merr.]. Fatores bióticos como interação patógeno-hospedeiro e abióticos influenciam o progresso da doença. Objetivou-se neste trabalho estudar os efeitos da temperatura e de períodos de molhamento foliar no progresso da Ferrugem Asiática nas cultivares Conquista, Savana e Suprema. O experimento foi conduzido no Departamento de Fitopatologia da Universidade Federal de Lavras, em junho de 2004, em câmaras de crescimento vegetal nas temperaturas de 15, 20, 25 e 30 °C e períodos de molhamento foliar de 0, 6, 12, 18 e 24 horas. A inoculação foi realizada pulverizando-se as plantas com suspensão de 10(4) uredósporos de P. pachyrhizi.mL-1 de água. Dados da incidência e da severidade foram utilizados para avaliar o progresso da doença e integrados por meio da área abaixo da curva de progresso da incidência (AACPI) e da severidade (AACPS). Modelos de regressão não-linear foram ajustados para a AACPI e AACPS. Foi calculado o volume abaixo da superfície de resposta para incidência (VASRI) e severidade (VASRS) em relação à temperatura e molhamentos foliares com o objetivo de detectar diferenças entre cultivares. Molhamentos foliares acima de 15 horas e temperaturas próximas a 20 ºC, nas 3 cultivares avaliadas, determinaram maior intensidade da Ferrugem Asiática. Temperaturas próximas a 30 e 15 ºC ocasionaram menor intensidade da doença. Períodos de molhamento foliar abaixo de 6 horas reduziram a intensidade da doença. Todas as cultivares testadas foram suscetíveis à doença, entretanto, a cultivar Conquista apresentou maior VASRI e VASRS da Ferrugem Asiática comparada às cultivares Savana e Suprema, as quais não diferiram estatisticamente. Houve diferença entre as cultivares para AACPI em cada temperatura e molhamento foliar.

Aiotteko tutkijaksi vai poliitikoksi? : Max Weberin analyysi tieteestä ja politiikasta ammatteina

kirjallisuusarvostelu

Efeitos de sistemas de preparo do solo na cultura da soja (Glycine max (L.) Merrill)

Com o objetivo de avaliar o efeito de cinco equipamentos de preparo do solo sobre o desenvolvimento e a produtividade da cultura da soja (Glycine max L. (Merril)), foi conduzida uma pesquisa em Latossolo Vermelho distrófico, no município de Uberaba-MG, em delineamento experimental em blocos casualizados, com cinco tratamentos (escarificador, enxada-rotativa, arado de aivecas, arado de discos e grade aradora) e quatro repetições. Os sistemas de preparo do solo não mostraram diferenças para o número médio de dias para a emergência das plântulas, estande final, altura da primeira vagem e produtividade da cultura da soja. Os tratamentos com arado de disco, arado de aivecas e grade aradora proporcionaram as maiores populações iniciais da cultura. A altura das plantas foi afetada pelos sistemas de preparo utilizados, destacando-se o preparo com o arado de aivecas e com a enxada-rotativa, com os maiores e menores valores absolutos, respectivamente.

Mapeamento de semeaduras de soja (Glycine max (L.)Merr.) mediante dados MODIS/Terra E TM/Landsat 5: um comparativo

O objetivo deste trabalho foi comparar mapeamentos de semeadura da cultura da soja na região oeste do Paraná, realizados com imagens MODIS/Terra e TM/Landsat 5. Primeiramente, construiu-se máscara de referência, considerando seis imagens TM ao longo do ciclo da cultura, utilizando-se dos algoritmos Paralelepípedo e MaxVer com posterior análise visual. As imagens MODIS foram classificadas com o algorítimo Paralelepípedo, em quatro passagens referentes ao pico vegetativo. O desempenho das classificações foi avaliado por meio de Matrizes de Erros, calculadas pela análise de 100 pontos amostrais (soja ou não-soja), aleatoriamente distribuídos em cada um dos oito municípios da área de estudo. Os principais resultados mostraram que a Exatidão Global (EG) e o Índice Kappa (IK), que variaram entre 0,55 e 0,80, em ambos os sensores, são considerados bons a muito bons. Quando EG e IK dos sensores TM e MODIS foram comparados, não se encontrou diferença significativa. O mapeamento da soja utilizando o sensor MODIS produziu 70% de confiabilidade sob o ponto de vista do usuário. A principal conclusão é a viabilidade de mapear a soja pelo sensor MODIS com as vantagens de que as imagens MODIS têm melhor resolução temporal e são disponibilizadas gratuitamente na Internet.