Metabolism of Rhizobium bacteroids

Nitrogen fixation within legume nodules results from a complex metabolic exchange between bacteria of the family Rhizobiaciae and the plant host. Carbon is supplied to the differentiated bacterial cells, termed bacteroids, in the form of dicarboxylic acids to fuel nitrogen fixation. In exchange, fixed nitrogen is transferred to the plant. Both the bacteroid and the plant-derived peribacteroid membrane tightly regulate the exchange of metabolites. In the bacteroid oxidation of dicarboxylic acids via the TCA cycle occurs in an oxygen-limited environment. This restricts the TCA cycle at key points, such as the 2-oxoglutarate dehydrogenase complex, and requires that inputs of carbon and reductant are balanced with outputs from the TCA cycle. This may be achieved by metabolism through accessory pathways that can remove intermediates, reductant, or ATP from the cycle. These include synthesis of the carbon polymers PHB and glycogen and bypass pathways such as the recently identified 2-oxoglutarate decarboxylase reaction in soybean bacteroids. Recent labeling data have shown that bacteroids synthesize and secrete amino acids, which has led to controversy over the role of amino acids in nodule metabolism. Here we review bacteroid carbon metabolism in detail, evaluate the labeling studies that relate to amino acid metabolism by bacteroids, and place the work in context with the genome sequences of Mesorhizobium loti and Sinorhizobium meliloti. We also consider a wider range of metabolic pathways that are probably of great importance to rhizobia in the rhizosphere, during nodule initiation, infection thread development, and bacteroid development.

Comparative genomics suggests that an ancestral polyploidy event leads to enhanced root nodule symbiosis in the Papilionoideae

Root nodule symbiosis (RNS) is one of the most efficient biological systems for nitrogen fixation and it occurs in 90% of genera in the Papilionoideae, the largest subfamily of legumes. Most papilionoid species show evidence of a polyploidy event occurred approximately 58 million years ago. Although polyploidy is considered to be an important evolutionary force in plants, the role of this papilionoid polyploidy event, especially its association with RNS, is not understood. In this study, we explored this role using an integrated comparative genomic approach and conducted gene expression comparisons and gene ontology enrichment analyses. The results show the following: (1) approximately a quarter of the papilionoid-polyploidy-derived duplicate genes are retained; (2) there is a striking divergence in the level of expression of gene duplicate pairs derived from the polyploidy event; and (3) the retained duplicates are frequently involved in the processes crucial for RNS establishment, such as symbiotic signalling, nodule organogenesis, rhizobial infection and nutrient exchange and transport. Thus, we conclude that the papilionoid polyploidy event might have further refined RNS and induced a more robust and enhanced symbiotic system. This conclusion partly explains the widespread occurrence of the Papilionoideae.

A framework for designing multi-functional cover crops

Cover crops are sown to provide a number of ecosystem services including nutrient management, mitigation of diffuse pollution, improving soil structure and organic matter content, weed suppression, nitrogen fixation and provision of resources for biodiversity. Although the decision to sow a cover crop may be driven by a desire to achieve just one of these objectives, the diversity of cover crops species and mixtures available means that there is potential to combine a number of ecosystem services within the same crop and growing season. Designing multi-functional cover crops would potentially help to reconcile the often conflicting agronomic and environmental agendas and contribute to the optimal use of land. We present a framework for integrating multiple ecosystem services delivered by cover crops that aims to design a mixture of species with complementary growth habit and functionality. The optimal number and identity of species will depend on the services included in the analysis, the functional space represented by the available species pool and the community dynamics of the crop in terms of dominance and co-existence. Experience from a project that applied the framework to fertility building leys in organic systems demonstrated its potential and emphasised the importance of the initial choice of species to include in the analysis

SHIMMER (1.0): a novel mathematical model for microbial and biogeochemical dynamics in glacier forefield ecosystems

SHIMMER (Soil biogeocHemIcal Model for Microbial Ecosystem Response) is a new numerical modelling framework designed to simulate microbial dynamics and biogeochemical cycling during initial ecosystem development in glacier forefield soils. However, it is also transferable to other extreme ecosystem types (such as desert soils or the surface of glaciers). The rationale for model development arises from decades of empirical observations in glacier forefields, and enables a quantitative and process focussed approach. Here, we provide a detailed description of SHIMMER, test its performance in two case study forefields: the Damma Glacier (Switzerland) and the Athabasca Glacier (Canada) and analyse sensitivity to identify the most sensitive and unconstrained model parameters. Results show that the accumulation of microbial biomass is highly dependent on variation in microbial growth and death rate constants, Q10 values, the active fraction of microbial biomass and the reactivity of organic matter. The model correctly predicts the rapid accumulation of microbial biomass observed during the initial stages of succession in the forefields of both the case study systems. Primary production is responsible for the initial build-up of labile substrate that subsequently supports heterotrophic growth. However, allochthonous contributions of organic matter, and nitrogen fixation, are important in sustaining this productivity. The development and application of SHIMMER also highlights aspects of these systems that require further empirical research: quantifying nutrient budgets and biogeochemical rates, exploring seasonality and microbial growth and cell death. This will lead to increased understanding of how glacier forefields contribute to global biogeochemical cycling and climate under future ice retreat.

Influence of organic fertilization on the number of culturable diazotrophic endophytic bacteria isolated from sugarcane

The numbers of culturable diazotrophic endophytic bacteria (CDEB) from roots stems and leaves of sugarcane submitted to organic inorganic or no fertilization were compared In order to determine the size of the N(2) fixing populations the Most Probable Number technique (MPN) was used The quantification of diazotrophic bacteria by using the acetylene reduction assay (ARA) was more accurate than observing the bacterial growth in the vials to confirm N(2) fixing capability the detection of gene nifH was performed on a sample of 105 Isolated bacteria The production of extracellular enzymes involved in the penetration of the plants by the bacteria was also studied The results showed that organic fertilization enhances the number of CDEB when compared with conventional fertilization used throughout the growing season The maximum number of bacteria was detected in the roots Roots and stems presented the greatest number of CDEB in the middle of the cropping season and in leaves numbers varied according to the treatment Using two pairs of primers and two different methods the nifH gene was found in 104 of the 105 tested isolates Larger amounts of pectinase were released by isolates from sugarcane treated with conventional fertilizers (66%) whereas larger amounts of cellulase were released by strains isolated from sugarcane treated with organic fertilizers (80%) (C) 2010 Elsevier Masson SAS All rights reserved

Fixação biológica do nitrogênio e crescimento de plantas de leucena (Leucaena leucocephala (Lam.) de wit.) sob salinidade

This study aimed to evaluate the effects of salinity in the growth and nitrogen fixation in leucaena. Plants were cultivated in Leonard pots containing clean and sterilized sand. The treatments were distributed in a completely randomized design, in a 3x3 factorial, with five replicates. The first factor corresponded to no inoculation and inoculation with Bradyrhizobium sp SEMIA 6070 and SEMIA 6153 strains. The second factor corresponded to NaCl concentrations: 0, 25 and 50 mol m(-3). Were analyzed the height and dry matter accumulation, the number of nodules and nitrogen in plants. The inoculation of plants with the strain SEMIA 6070 provided higher growth and greater nitrogen accumulation in plants thar were not subject to salinity. Nodulation was not affected by salinity. Inoculation with SEMIA 6153 strain provided greater tolerance to salinity.

Mecanismos de abertura de sulcos, inoculação e adubação nitrogenada em feijoeiro em sistema plantio direto

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

Nitrogênio em cobertura e molibdênio foliar no feijoeiro de inverno

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

Estirpes de Rhizobium tropici na inoculação do feijoeiro

Avaliando o comportamento do feijoeiro inoculado com cinco estirpes de Rhizobium tropici e a adubação mineral com nitrogênio, sobre alguns fatores relacionados à sua produtividade, utilizou-se um delineamento experimental de blocos ao acaso, com oito tratamentos constituídos pela inoculação do feijoeiro cultivar IAC Carioca com cinco estirpes de Rhizobium tropici (CIAT 899 - estirpe referência; F35; F54; F81 e CM255), dois controles sem inoculação sendo um adubado com N na semeadura e em cobertura e outro sem adubação e um cultivar não nodulante (NORH 54) adubado; com seis repetições. Avaliaram-se: número de nódulos por planta; massa de material seco da parte aérea; teor de N nas folhas; número de vagens por planta; número de grãos por planta; número de grãos por vagem; peso de 100 grãos e produtividade de grãos. A inoculação de estirpes eficientes de Rhizobium em cultivar nodulante de feijoeiro, ou o cultivo deste em solos com população nativa eficiente, pode possibilitar a não utilização de nitrogênio em cobertura na cultura do feijoeiro, sem afetar a produtividade.

Nodulação e atividade da nitrato redutase em função da aplicação de molibdênio em soja

Among the nutrients that are essential for the biological nitrogen fixation by soybean plants, molybdenum stands out for being a cofactor of the nitrate reductase, affecting enzymatic activity and, consequently, the nodulation process. The research had as objective to evaluate the effects of molybdenum application on soybean nodulation and nitrate reductase activity. The experiment was conduced in greenhouse, sowing soybean in 12 L pots, with two plants per plot. The treatments consisted of two application via (with the seeds and leaf dressing) and two molybdenum doses (12 and 24 g ha(-1) with the seeds; 30 and 60 g ha(-1) leaf dressing) in ammonium molybdate form, plus the control. The number and dry mass of nodules and nitrogen content in soybean leaves were evaluated. Samples of leaves for the evaluation of nitrate reductase activity were taken at 10 a.m. and 10 p.m. It was concluded that soybean nodulation is affected by Mo dose and application via, resulting in higher number and weight of nodules when it is applied with the seeds. The enzymatic activity of the nitrate reductase is influenced by Mo fertilization and it is higher for leaf dressing with the double of the recommended dose.

COEVOLUTION BETWEEN ATTINE ANTS and ACTINOMYCETE BACTERIA: A REEVALUATION

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Nodule ultrastructure and initial growth of Anadenanthera peregrina (L.) Speg. var. falcata (Benth.) Altschul plants infected with rhizobia

The anatomy and ultrastructure of root nodules of Anadenanthera peregrina var. falcata (Leguminosae-Mimosoideae) were analysed, as was plant growth. To ensure that nodules developed, seedlings were inoculated with a mixture of six strains of rhizobia. Nodules were produced that differed in appearance-and probably also effectiveness-but their structure was similar and they showed characteristics typical of indeterminate nodules, such as persistent meristematic tissue and a gradient of cells at different stages of development. Many starch grains were present in inner cortex cells and interstitial cells of infected tissue. Infected cells were densely packed with bacteroids, which contained many poly-beta-hydroxybutyrate granules. The high incidence of these granules, together with high levels of starch accumulation in interstitial cells, suggested low N-2-fixation efficiency of the rhizobia isolates used for inoculation. In the symbiosomes of early-senescent infected cells, reticulum-like structures, small vesicles and a fibrillar material were observed; these may be related to bacteroid degradation. In the cytoplasm of late-senescent infected cells, many vesicles and membrane-like structures were observed, probably associated with membrane degradation of bacteroids and peribacteroids. The total biomass of plants inoculated with rhizobia was low and their xylopodia and shoots had low levels of N compared with non-inoculated plants fertilized with ammonium nitrate. However, inoculated plants did not show N-deficiency symptoms and grew better than non-inoculated plants without N fertilization. These growth results, together with ultrastructural observations of nodules, suggest that nitrogen fixation of rhizobia isolates associated with Anadenanthera peregrina var. falcata roots is poor. (C) 2002 Annals of Botany Company.

Doses de inoculante e nitrogênio na semeadura da soja em área de primeiro cultivo

The inoculation of soybean crop with Bradyrhizobium bacteria is a widespread practice, due to excellent results achieved in productivity, however, some doubt still arise in relation the need of application of mineral nitrogen at sowing. In that mean, this work aimed to evaluate the response of the doses at inoculant and mineral nitrogen in the sowing of crop soybean in area of first cultivation. The experiments were carried in the experimental field of the University of the Estate of Mato Grosso, Campus of Alta Floresta - MT, Brazil, in the growing seasons of 2006/2007 and 2007/2008. The experimental design was randomized blocks, in a factorial escheme 2 x 4, with four replications. The factors consisted in the inoculation of seeds with Bradyrhizobium japonicum *(3 e 6 mL kg(-1) of seeds), and nitrogen fertilizer at sowing *(0, 10, 20 e 40 kg ha(-1)). The following variables were evaluated: plant height, insertion height of first pod, number of pod per plant, number of grain per pod, mass of 100 grain and grain productivity. In the two growing seasons there was no significant difference for doses of inoculants applied in first year of cultivation with soybean. To nitrogen application at sowing of soybean crop there was quadratic effect to plant height and of insertion height of first pod and growing linear to number of pod per plant in the second year of cultivation, however, without significant increase the productivity.

Genetical rearrangements in Azospirillum brasilense induced by the plasmid PJB4JI

The present work was conducted with the initial purpose of looking for the characterization of Azospirillum brasilense strain ATCC 29.145 mutants induced by insertions of the transposon Tn-5. The strategy used to transfer such mobile element was based on the vector named pJB4JI