Londrina, Brazil
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Cardoso J.D.,State University Londrina | Cardoso J.D.,Instituto Agronomico Do Parana | Hungria M.,Embrapa Soja | Andrade D.S.,Instituto Agronomico Do Parana
Applied Microbiology and Biotechnology | Year: 2012

Common bean (Phaseolus vulgaris L.) is a legume that has been reported as highly promiscuous in nodulating with a variety of rhizobial strains, often with low effectiveness in fixing nitrogen. The aim of this work was to assess the symbiotic efficiency of rhizobial strains isolated from common bean seeds, nodules of Arachis hypogaea, Mucuna pruriens, and soils from various Brazilian agroecosystems, followed by the characterization of elite strains identified in the first screening. Forty-five elite strains were analyzed for symbiotic properties (nodulation, plant-growth, and nitrogen-fixation parameters) under greenhouse conditions in pots containing non-sterile soil, and variation in symbiotic performance was observed. Elite strains were also characterized in relation to morpho-physiological properties, genetic profiles of rep-polymerase chain reaction (PCR; BOX), and restriction fragment length polymorphism (RFLP)-PCR of the 16S rRNA. Sequence analyses of the 16S rRNA were obtained for 17 strains representative of the main groups resulting from all previous analyses. One of the most effective strains, IPR-Pv 2604, was clustered with Rhizobium tropici, whereas strain IPR-Pv 583, showing lower effectiveness in fixing N 2, was clustered with Herbaspirillum lusitanum. Surprisingly, effective strains were clustered with unusual symbiotic genera/species, including Leifsonia xyli, Stenotrophomonas maltophilia, Burkholderia, and Enterobacter. Some strains recognized in this study were outstanding in their nitrogen-fixing capacity and therefore, show high biotechnological potential for use in commercial inoculants. © 2011 Springer-Verlag.

Da Silva Batista J.S.,State University Londrina | Hungria M.,Embrapa Soja
Journal of Proteomics | Year: 2012

The rhizobia-legume symbiosis requires a coordinated molecular interaction between the symbionts, initiated by seed and root exudation of several compounds, mainly flavonoids, that trigger the expression of nodulation genes in the bacteria. Since the role of flavonoids seems to be broader than the induction of nodulation genes, we aimed at characterizing genistein-induced proteins of Bradyrhizobium japonicum CPAC 15 (=SEMIA 5079), used in commercial soybean inoculants in Brazil, and of two genetically related strains grown in vitro. Whole-cell proteins were extracted both from induced (1μM genistein) and from non-induced cultures of the three strains, and separated by two-dimensional electrophoresis. Spot profiles were compared between the two conditions and selected spots were excised and identified by mass spectrometry. Forty-seven proteins were significantly induced by genistein, including several hypothetical proteins, the cytoplasmic flagellar component FliG, periplasmic ABC transporters, a protein related to biosynthesis of exopolysaccharides (ExoN), and proteins involved in redox-state maintenance. Noteworthy was the induction of the PhyR-σ EcfG regulon, recently demonstrated to be involved in the symbiotic efficiency of, and general stress response in B. japonicum. Our results confirm that the role of flavonoids, such as genistein, can go far beyond the expression of nodulation-related proteins in B. japonicum. © 2011 Elsevier B.V.

Hungria M.,Embrapa Soja | Kaschuk G.,UNIPAR
Environmental and Experimental Botany | Year: 2014

Legumes need large amounts of N to grow satisfactorily. Under low NO3 - availability in the soil, many legumes meet their N requirements by N2 fixation in association with rhizobia. Both NO3 - uptake and N2 fixation decrease as temperature exceeds optimal growth conditions, but the mechanisms of regulation of N2 fixation and NO3 -/NH4 + assimilation under high temperature stress are not completely understood. We describe an experiment in which physiological mechanisms regulating N metabolism of common bean (Phaseolus vulgaris L.) are investigated in plants submitted to daily maximum temperatures of 28, 34 and 39°C. Common bean was grown in symbiosis with each of six rhizobial strains-belonging to four different species and varying in N2 fixation effectiveness-or fertilized with NO3 - until flowering. Harvest measurements included the activities of shoot, stem and root NO3 - reductase (NR), nodule glutamine synthetase (GS), NADH-dependent glutamate synthase (GOGAT), nitrogenase, phosphoenol pyruvate carboxylase (PEPcase), N-export rates by nodules and concentration of N compounds in the xylem sap. Higher temperatures inhibited N2 fixation resulting in lower proportion of ureide-N in nodules and xylem sap of nodulated plants in relation to amide-N and α-amino-N. Higher temperatures consistently reduced the activity of NR in leaves of N-fertilized plants. Higher temperatures also decreased N exported from nodules and activities of nitrogenase, GS, GOGAT and PEPcase. The rate of decreases varied in plants with different strains. Furthermore, the activities of GS and GOGAT were more strongly affected by high temperatures than the activity of nitrogenase. There was a remarkable increase in the concentration of NH4 +-N and ureide-N in the nodules when GS and GOGAT activities decreased. Therefore, the results provide evidence that N2 fixation in common bean submitted to heat stress is limited by NH4 + assimilation via GS-GOGAT rather than by decreased activity of nitrogenase. Rhizobial effectiveness determined the degree of down-regulation of GS-GOGAT activity in nodule tissues. © 2013 Elsevier B.V.

Menna P.,Embrapa Soja | Menna P.,Brazilian National Council for Scientific and Technological Development | Hungria M.,Embrapa Soja | Hungria M.,Brazilian National Council for Scientific and Technological Development
International Journal of Systematic and Evolutionary Microbiology | Year: 2011

Bacteria belonging to the genus Bradyrhizobium are capable of establishing symbiotic relationships with a broad range of plants belonging to the three subfamilies of the family Leguminosae (=Fabaceae), with the formation of specialized structures on the roots called nodules, where fixation of atmospheric nitrogen takes place. Symbiosis is under the control of finely tuned expression of common and host-specific nodulation genes and also of genes related to the assembly and activity of the nitrogenase, which, in Bradyrhizobium strains investigated so far, are clustered in a symbiotic island. Information about the diversity of these genes is essential to improve our current poor understanding of their origin, spread and maintenance and, in this study, we provide information on 40 Bradyrhizobium strains, mostly of tropical origin. For the nodulation trait, common (nodA), Bradyrhizobium-specific (nodY/K) and host-specific (nodZ) nodulation genes were studied, whereas for fixation ability, the diversity of nifH was investigated. In general, clustering of strains in all nod and nifH trees was similar and the Bradyrhizobium group could be clearly separated from other rhizobial genera. However, the congruence of nod and nif genes with ribosomal and housekeeping genes was low. nodA and nodY/K were not detected in three strains by amplification or hybridization with probes using Bradyrhizobium japonicum and Bradyrhizobium elkanii type strains, indicating the high diversity of these genes or that strains other than photosynthetic Bradyrhizobium must have alternative mechanisms to initiate the process of nodulation. For a large group of strains, the high diversity of nod genes (with an emphasis on nodZ), the low relationship between nod genes and the host legume, and some evidence of horizontal gene transfer might indicate strategies to increase host range. On the other hand, in a group of five symbionts of Acacia mearnsii, the high congruence between nod and ribosomal/housekeeping genes, in addition to shorter nodY/K sequences and the absence of nodZ, highlights a co-evolution process. Additionally, in a group of B. japonicum strains that were symbionts of soybean, vertical transfer seemed to represent the main genetic event. In conclusion, clustering of nodA and nifH gives additional support to the theory of monophyletic origin of the symbiotic genes in Bradyrhizobium and, in addition to the analysis of nodY/K and nodZ, indicates spread and maintenance of nod and nif genes through both vertical and horizontal transmission, apparently with the dominance of one or other of these events in some groups of strains. © 2011 IUMS.

Kaschuk G.,Embrapa Soja | Alberton O.,Embrapa Soja | Hungria M.,Embrapa Soja
Soil Biology and Biochemistry | Year: 2010

Soil microbial biomass plays important roles in nutrient cycling, plant-pathogen suppression, decomposition of residues and degradation of pollutants; therefore, it is often regarded as a good indicator of soil quality. We reviewed more than a hundred studies in which microbial biomass-C (MB-C), microbial quotient (MB-C/TSOC, total soil organic carbon) and metabolic quotient (qCO2) were evaluated with the objective of understanding MB-C responses to various soil-management practices in Brazilian ecosystems. These practices included tillage systems, crop rotations, pastures, organic farming, inputs of industrial residues and urban sewage sludge, applications of agrochemicals and burning. With a meta-analysis of 233 data points, we confirmed the benefits of no-tillage in preserving MB-C and reducing qCO2 in comparison to conventional tillage. A large number of studies described increases in MB-C and MB-C/TSOC due to permanent organic farming, also benefits from crop rotations particularly with several species involved, whereas application of agrochemicals and burning severely disturbed soil microbial communities. The MB-C decreased in overgrazed pastures, but increased in pastures rotated with well-managed crops. Responses of MB-C, MB-C/TSOC and qCO2 to amendment with organic industrial residues varied with residue type, dose applied and soil texture. In conclusion, MB-C and related parameters were, indeed, useful indicators of soil quality in various Brazilian ecosystems. However, direct relationships between MB-C and nutrient-cycling dynamics, microbial diversity and functionality are still unclear. Further studies are needed to develop strategies to maximize beneficial effects of microbial communities on soil fertility and crop productivity. © 2009 Elsevier Ltd. All rights reserved.

Crop livestock integration systems (CLIS) involving pastures of Brachiaria brizantha in autumn/winter and soybeans in summer have been considered an option to increase and diversify crop production and improve no-tillage system. This work aimed to evaluate the effect of grass desiccation timings or cattle grazing pressures on soil physical attributes and soybean yields in a crop-livestock integration system. The experiment was carried out on a dark red Latosol (Rhodic Eutrudox, US Soil Taxonomy) in southern Brazil. In the major plots, grazing pressures of 6.9, 9.4, and 16.5 animal unities per hectare were applied during 19 days to a pasture of Brachiaria brizantha cv. 'Xaraes', which phytomass was desiccated 30 days before soybean sowing (DBS). Ungrazed plots were desiccated 30, 44 and 52 DBS. Remaining phytomass of the pasture; soil bulk density, porosity and soil resistance to penetration; and yield of soybean cultivars 'BRS 294RR' and 'BRS 255RR' (sowed in subplots), were determined. Changes in soil physical attributes by animal trampling were concentrated on topsoil layer (0.0- 0.05m), and soil compaction level was increased by the highest grazing pressure down the 0.2-0.3 layer. The yield of soybean cv. 'BRS 294RR' was increased with rising grazing pressures, whereas the yield of soybean cv. 'BRS 255RR' was not affected, showing that soybean genotypes had different responses to soil compaction level and, mainly, to remaining phytomass of pasture. In CLIS with high production of B. brizantha dry mass, soybean yields are closely associated to time interval between grass desiccation and soybean planting.

de Oliveira Jr. A.,Embrapa Soja | Prochnow L.I.,International Plant Nutrition Institute IPNI | Klepker D.,Embrapa Soja Setor Experimental de Balsas
Scientia Agricola | Year: 2011

Soybean (Glycine max L. Merrill) crop started to be planted in the Brazilian Cerrado in the 1970's, and this region currently contributes with 57% of total soybean production in Brazil. Under natural conditions in this region, the soils present chemical limitations such as low pH, low Cation Exchange Capacity, low nutrient availability, and moreover, clayey soils have a high P fixation capacity mainly due to high contents of Fe/Al oxides. Since P is the most limiting nutrient is this region, a study was performed in the state of Maranhão, Brazil, in a Typic Hapludox, with clayey texture and low available P (extracted by resin). Treatments were defined to evaluate soybean response to broadcast Arad phosphate rock (PR) plus banded triple superphosphate (TSP) and to evaluate the soybean response to three proportions of PR and TSP. The experiment was established in October 2004 and was carried out for three consecutive crop years (2004/05 to 2006/07). The associated use of PR and TSP, in several situations, resulted in yields at least similar to that obtained with the use of the water soluble P source and, in some cases, even using lower P rates. Regarding the "mixtures", a linear response was observed when they were banded; however, when they were broadcasted, no increase in yield was observed above 50% of relative solubility. In conclusion, the association of sources differing in solubility may be a feasible agronomic option for P fertilizer management of soybeans.

The weed, pest and disease infestation, often occur simultaneously in a farm. In such a situation, the most economical alternative to control and, therefore, the more commonly adopted by the producers is the pesticide spray mixture in the tank. In this study, field practices adopted by farmers related to mixtures of pesticides in tank in Brazilian farms are described and quantified. To obtain such information, a questionnaire was developed and implemented with the professionals in different regions of the country. It was found that 97% of respondents use tank mixtures and most of the time 95% using 2-5 products. Most often using the highest recommended doses. The applications of glyphosate in soybean are carried out simultaneously with insecticides, fungicides and other herbicides, in 86% of the times. Most respondents (72%) reported not knowing about the tank mix prectices and 99% indicated interest in receiving such data. Whereas the tank in pesticide mixture is common practice among farmers and that this practice has its advantages and disadvantages, it is concluded that the issue should be discussed urgently by government agencies and by other segments of the soybean production chain. © 2015 Sociedade Brasileira da Ciencia das Plantas Daninha. All Rights Received.

The objective of this work was to evaluate the soybean neotropical brown stink bug (Euschistus heros) resistance to methamidophos through laboratory bioassays. Bioassays were carried out using populations of adults and nymphs of Euschistus heros in the last instar collected in soybean crops in Paraná State. Highest frequencies of genotypes resistant to methamidophos were observed in Alvorada do Sul, Toledo and Nova Santa Rosa, PR. Populations from Bela Vista do Paraíso, Araruna, Campo Mourão, Mariluz, Cascavel, São João do Ivaí, and Mamboré were more susceptible to methamidophos. However, an increase on resistant genotypes in this locations cannot be discarded.

Kaschuk G.,Paranaense University | Alberton O.,Paranaense University | Hungria M.,Embrapa Soja
Plant and Soil | Year: 2011

Maintenance of soil quality is a key component of agriculture sustainability and a main goal of most farmers, environmentalists and government policymakers. However, as there are no parameters or methods to evaluate soil quality directly, some attributes of relevant soil functions are taken as indicators; lately, an increase in the use of soil microbial parameters has occurred, and their viability as indicators of proper land use has been highlighted. In this study we performed a meta-analysis of the response ratios of several microbial and chemical parameters to soil disturbance by different land uses in the Brazilian biomes. The studies included native forests, pastures and perennial and annual cropping systems. The introduction of agricultural practices in all biomes covered previously with natural vegetation profoundly affected microbial biomass-C (MB-C)-with an overall decrease of 31%. Annual crops most severely reduced microbial biomass and soil organic C, with an average decrease of 53% in the MB-C. In addition, the MB-C/TSOC (total soil organic carbon) ratio was significantly decreased with the transformation of forests to perennial plantation (25%), pastures (26%), and annual cropping (20%). However, each biome reacted differently to soil disturbance, i. e., decreases in MB-C followed the order of Cerrado>Amazon>Caatinga>Atlantic Forest. In addition, the Cerrado appeared to have the most fragile soil ecosystem because of lower MB-C/TSOC and higher qCO2. Unfortunately, the Cerrado and the Amazon, demonstrated by our study as the most fragile biomes, have been subjected to the highest agronomic pressure. The results reported here may help to infer the best land-use strategies to improve soil quality and achieve agriculture sustainability. The approach can also be very useful to monitor soil quality in other tropical and subtropical biomes. © 2010 Springer Science+Business Media B.V.

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