Brucella melitensis MucR, an orthologue of Sinorhizobium meliloti MucR, is involved in resistance to oxidative, detergent, and saline stresses and cell envelope modifications.

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Mirabella, A; Terwagne, M; Zygmunt, M S; Cloeckaert, A; De Bolle, X; Letesson, J J

2013-02-01

Brucella spp. and Sinorhizobium meliloti are alphaproteobacteria that share not only an intracellular lifestyle in their respective hosts, but also a crucial requirement for cell envelope components and their timely regulation for a successful infectious cycle. Here, we report the characterization of Brucella melitensis mucR, which encodes a zinc finger transcriptional regulator that has previously been shown to be involved in cellular and mouse infections at early time points. MucR modulates the surface properties of the bacteria and their resistance to environmental stresses (i.e., oxidative stress, cationic peptide, and detergents). We show that B. melitensis mucR is a functional orthologue of S. meliloti mucR, because it was able to restore the production of succinoglycan in an S. meliloti mucR mutant, as detected by calcofluor staining. Similar to S. meliloti MucR, B. melitensis MucR also represses its own transcription and flagellar gene expression via the flagellar master regulator ftcR. More surprisingly, we demonstrate that MucR regulates a lipid A core modification in B. melitensis. These changes could account for the attenuated virulence of a mucR mutant. These data reinforce the idea that there is a common conserved circuitry between plant symbionts and animal pathogens that regulates the relationship they have with their hosts.

Crystallization and preliminary crystallographic studies of the recombinant dihydropyrimidinase from Sinorhizobium meliloti CECT4114

International Nuclear Information System (INIS)

Martínez-Rodríguez, Sergio; González-Ramírez, Luis Antonio; Clemente-Jiménez, Josefa María; Rodríguez-Vico, Felipe; Las Heras-Vázquez, Francisco Javier; Gavira, Jose A.; García-Ruíz, Juan Manuel

2006-01-01

The dihydropyrimidinase from S. meliloti CECT4114, with activity towards both hydantoin and dihydrouracil substrates, was crystallized, and diffraction data were collected to 1.85 Å resolution. Dihydropyrimidinases are involved in the reductive pathway of pyrimidine degradation, catalysing the hydrolysis of 5,6-dihydrouracil and 5,6-dihydrothymine to the corresponding N-carbamoyl β-amino acids. This enzyme has often been referred to as hydantoinase owing to its industrial application in the production of optically pure amino acids starting from racemic mixtures of 5-monosubstituted hydantoins. Recombinant dihydropyrimidinase from Sinorhizobium meliloti CECT4114 (SmelDhp) has been expressed, purified and crystallized. Crystallization was performed using the counter-diffusion method with capillaries of 0.3 mm inner diameter. Crystals of SmelDhp suitable for data collection and structure determination were grown in the presence of agarose at 0.1%(w/v) in order to ensure mass transport controlled by diffusion. X-ray data were collected to a resolution of 1.85 Å. The crystal belongs to the orthorhombic space group C222 1 , with unit-cell parameters a = 124.89, b = 126.28, c = 196.10 Å and two molecules in the asymmetric unit. A molecular-replacement solution has been determined and refinement is in progress

Comparative toxicity assessment of CeO{sub 2} and ZnO nanoparticles towards Sinorhizobium meliloti, a symbiotic alfalfa associated bacterium: Use of advanced microscopic and spectroscopic techniques

Energy Technology Data Exchange (ETDEWEB)

Bandyopadhyay, Susmita [Environmental Science and Engineering PhD Program, The University of Texas at El Paso, 500 West University Avenue, El Paso, TX 79968 (United States); University of California Center for Environmental Implications of Nanotechnology (UC CEIN), The University of Texas at El Paso (United States); Peralta-Videa, Jose R. [Department of Chemistry, The University of Texas at El Paso, 500 West University Avenue, El Paso, TX 79968 (United States); Plascencia-Villa, German; Jose-Yacaman, Miguel [Department of Physics and Astronomy, The University of Texas at San Antonio, One UTSA Circle, San Antonio, TX 78249 (United States); Gardea-Torresdey, Jorge L., E-mail: jgardea@utep.edu [Environmental Science and Engineering PhD Program, The University of Texas at El Paso, 500 West University Avenue, El Paso, TX 79968 (United States); Department of Chemistry, The University of Texas at El Paso, 500 West University Avenue, El Paso, TX 79968 (United States); University of California Center for Environmental Implications of Nanotechnology (UC CEIN), The University of Texas at El Paso (United States)

2012-11-30

Highlights: Black-Right-Pointing-Pointer First cytotoxicity study of CeO{sub 2} and ZnO nanoparticles to Sinorhizobium meliloti. Black-Right-Pointing-Pointer First report upon the mechanisms of CeO{sub 2} and ZnO NPs toxicity to S. meliloti. Black-Right-Pointing-Pointer ZnO NPs were found to be bactericidal in lower concentration. Black-Right-Pointing-Pointer CeO{sub 2} NPs had bacteriostatic effect on S. meliloti. - Abstract: Cerium oxide (CeO{sub 2}) and zinc oxide (ZnO) nanoparticles (NPs) are extensively used in a variety of instruments and consumer goods. These NPs are of great concern because of potential toxicity towards human health and the environment. The present work aimed to assess the toxic effects of 10 nm CeO{sub 2} and ZnO NPs towards the nitrogen fixing bacterium Sinorhizobium meliloti. Toxicological parameters evaluated included UV/Vis measurement of minimum inhibitory concentration, disk diffusion tests, and dynamic growth. Ultra high-resolution scanning transmission electron microscopy (STEM) and infrared spectroscopy (FTIR) were utilized to determine the spatial distribution of NPs and macromolecule changes in bacterial cells, respectively. Results indicate that ZnO NPs were more toxic than CeO{sub 2} NPs in terms of inhibition of dynamic growth and viable cells counts. STEM images revealed that CeO{sub 2} and ZnO NPs were found on bacterial cell surfaces and ZnO NPs were internalized into the periplasmic space of the cells. FTIR spectra showed changes in protein and polysaccharide structures of extra cellular polymeric substances present in bacterial cell walls treated with both NPs. The growth data showed that CeO{sub 2} NPs have a bacteriostatic effect, whereas ZnO NPs is bactericidal to S. meliloti. Overall, ZnO NPs were found to be more toxic than CeO{sub 2} NPs.

Symbiotic Nitrogen Fixation in Alfalfa (Medicago Sativa L.) by Sinorhizobium Meliloti at Al-Qassim Regions, Saudi Arabia

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Al-Barakah, F. N.; Mridha, M. A. U.

2016-01-01

The nodulation status in alfalfa (Medicago sativa L.) plants by Sinorhizobium meliloti under Saudi field condition was assessed in some selected farms in four seasons for two years. In the present study, we also monitored the introduced S. meliloti strains activity under Saudi soil conditions. The samples were collected at regular seasonal intervals from the selected farms. The total number of nodules, morphology of the nodules and the effectiveness of N/sub 2/-fixation was assessed. In general, it was revealed that soils in the selected areas in Saudi Arabia have sufficient bacteria of the proper types to nodulate the alfalfa plants. These nodules are high in number, small in size and white in color. The nodules obtained from most of the selected farms are ineffective for nitrogen fixation. Inoculation of alfalfa seeds with imported S. meliloti strains failed to fix the atmospheric nitrogen sufficiently and also the growth improvement of alfalfa plants. There was a wide variation in the occurrence of number of nodules among the four seasons in two years. It was also observed that summer season severely affected the nodulation making it nearly zero. This low number of nodules exerts a very slow recovery of nodule formation in the next year. The introduced strains were always over competing with the native strains but they did not survive because of hot and dry summer. Nitrogenase activity of the nodules collected from both the inoculated and non-inoculated farms were always very low in all the collected samples, which indicates that the ability of fixing nitrogen by S. meliloti strains in alfalfa under Saudi soils conditions is very low. (author)

Genome-engineered Sinorhizobium meliloti for the production of poly(lactic-co-3-hydroxybutyric) acid copolymer.

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Tran, Tam T; Charles, Trevor C

2016-02-01

Economically competitive commercial production of biodegradable bioplastics with desirable properties is an important goal. In this study, we demonstrate the use of chromosome engineering of an alternative bacterial host, Sinorhizobium meliloti, for production of the copolymer, poly(lactate-co-3-hydroxybutyrate). Codon-optimized genes for 2 previously engineered enzymes, Clostridium propionicum propionate CoA transferase (Pct532Cp) and Pseudomonas sp. strain MBEL 6-19 polyhydroxyalkanoate (PHA) synthase 1 (PhaC1400Ps6-19), were introduced into S. meliloti Rm1021 by chromosome integration, replacing the native phbC gene. On the basis of phenotypic analysis and detection of polymer product by gas chromatography analysis, synthesis and accumulation of the copolymer was confirmed. The chromosome integrant strain, with the introduced genes under the control of the native phbC promoter, is able to produce over 15% cell dry mass of poly(lactate-co-3-hydroxybutyrate), containing 30 mol% lactate, from growth on mannitol. We were also able to purify the polymer from the culture and confirm the structure by NMR and GC-MS. To our knowledge, this is the first demonstration of production of this copolymer in the Alphaproteobacteria. Further optimization of this system may eventually yield strains that are able to produce economically viable commercial product.

The DivJ, CbrA and PleC system controls DivK phosphorylation and symbiosis in Sinorhizobium meliloti

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Pini, Francesco; Frage, Benjamin; Ferri, Lorenzo; De Nisco, Nicole J.; Mohapatra, Saswat S.; Taddei, Lucilla; Fioravanti, Antonella; Dewitte, Frederique; Galardini, Marco; Brilli, Matteo; Villeret, Vincent; Bazzicalupo, Marco; Mengoni, Alessio; Walker, Graham C.; Becker, Anke; Biondi, Emanuele G.

2013-01-01

SUMMARY Sinorhizobium meliloti is a soil bacterium that invades the root nodules it induces on Medicago sativa, whereupon it undergoes an alteration of its cell cycle and differentiates into nitrogen-fixing, elongated and polyploid bacteroid with higher membrane permeability. In Caulobacter crescentus, a related alphaproteobacterium, the principal cell cycle regulator, CtrA, is inhibited by the phosphorylated response regulator DivK. The phosphorylation of DivK depends on the histidine kinase DivJ, while PleC is the principal phosphatase for DivK. Despite the importance of the DivJ in C. crescentus, the mechanistic role of this kinase has never been elucidated in other Alphaproteobacteria. We show here that the histidine kinases DivJ together with CbrA and PleC participate in a complex phosphorylation system of the essential response regulator DivK in S. meliloti. In particular, DivJ and CbrA are involved in DivK phosphorylation and in turn CtrA inactivation, thereby controlling correct cell cycle progression and the integrity of the cell envelope. In contrast, the essential PleC presumably acts as a phosphatase of DivK. Interestingly, we found that a DivJ mutant is able to elicit nodules and enter plant cells, but fails to establish an effective symbiosis suggesting that proper envelope and/or low CtrA levels are required for symbiosis. PMID:23909720

Polyamines contribute to salinity tolerance in the symbiosis Medicago truncatula-Sinorhizobium meliloti by preventing oxidative damage.

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López-Gómez, Miguel; Hidalgo-Castellanos, Javier; Muñoz-Sánchez, J Rubén; Marín-Peña, Agustín J; Lluch, Carmen; Herrera-Cervera, José A

2017-07-01

Polyamines (PAs) such as spermidine (Spd) and spermine (Spm) are small ubiquitous polycationic compounds that contribute to plant adaptation to salt stress. The positive effect of PAs has been associated to a cross-talk with other anti-stress hormones such as brassinosteroids (BRs). In this work we have studied the effects of exogenous Spd and Spm pre-treatments in the response to salt stress of the symbiotic interaction between Medicago truncatula and Sinorhizobium meliloti by analyzing parameters related to nitrogen fixation, oxidative damage and cross-talk with BRs in the response to salinity. Exogenous PAs treatments incremented the foliar and nodular Spd and Spm content which correlated with an increment of the nodule biomass and nitrogenase activity. Exogenous Spm treatment partially prevented proline accumulation which suggests that this polyamine could replace the role of this amino acid in the salt stress response. Additionally, Spd and Spm pre-treatments reduced the levels of H 2 O 2 and lipid peroxidation under salt stress. PAs induced the expression of genes involved in BRs biosynthesis which support a cross-talk between PAs and BRs in the salt stress response of M. truncatula-S. meliloti symbiosis. In conclusion, exogenous PAs improved the response to salinity of the M. truncatula-S. meliloti symbiosis by reducing the oxidative damage induced under salt stress conditions. In addition, in this work we provide evidences of the cross-talk between PAs and BRs in the adaptive responses to salinity. Copyright © 2017 Elsevier Masson SAS. All rights reserved.

Crystallization and preliminary crystallographic studies of an active-site mutant hydantoin racemase from Sinorhizobium meliloti CECT4114

International Nuclear Information System (INIS)

Martínez-Rodríguez, Sergio; González-Ramírez, Luis Antonio; Clemente-Jiménez, Josefa María; Rodríguez-Vico, Felipe; Las Heras-Vázquez, Francisco Javier; Gavira, Jose Antonio; García-Ruiz, Juan Ma.

2007-01-01

Crystals of an active-site mutated hydantoin racemase from S. meliloti have been obtained in the presence and absence of d,l-5-isopropyl-hydantoin and characterized by X-ray diffraction. A recombinant active-site mutant of hydantoin racemase (C76A) from Sinorhizobium meliloti CECT 4114 (SmeHyuA) has been crystallized in the presence and absence of the substrate d,l-5-isopropyl hydantoin. Crystals of the SmeHyuA mutant suitable for data collection and structure determination were grown using the counter-diffusion method. X-ray data were collected to resolutions of 2.17 and 1.85 Å for the free and bound enzymes, respectively. Both crystals belong to space group R3 and contain two molecules of SmeHyuA per asymmetric unit. The crystals of the free and complexed SmeHyuA have unit-cell parameters a = b = 85.43, c = 152.37 Å and a = b = 85.69, c = 154.38 Å, crystal volumes per protein weight (V M ) of 1.94 and 1.98 Å 3 Da −1 and solvent contents of 36.7 and 37.9%, respectively

Robustness encoded across essential and accessory replicons of the ecologically versatile bacterium Sinorhizobium meliloti

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Walker, Graham C.; Finan, Turlough M.; Mengoni, Alessio; Griffitts, Joel S.

2018-01-01

Bacterial genome evolution is characterized by gains, losses, and rearrangements of functional genetic segments. The extent to which large-scale genomic alterations influence genotype-phenotype relationships has not been investigated in a high-throughput manner. In the symbiotic soil bacterium Sinorhizobium meliloti, the genome is composed of a chromosome and two large extrachromosomal replicons (pSymA and pSymB, which together constitute 45% of the genome). Massively parallel transposon insertion sequencing (Tn-seq) was employed to evaluate the contributions of chromosomal genes to growth fitness in both the presence and absence of these extrachromosomal replicons. Ten percent of chromosomal genes from diverse functional categories are shown to genetically interact with pSymA and pSymB. These results demonstrate the pervasive robustness provided by the extrachromosomal replicons, which is further supported by constraint-based metabolic modeling. A comprehensive picture of core S. meliloti metabolism was generated through a Tn-seq-guided in silico metabolic network reconstruction, producing a core network encompassing 726 genes. This integrated approach facilitated functional assignments for previously uncharacterized genes, while also revealing that Tn-seq alone missed over a quarter of wild-type metabolism. This work highlights the many functional dependencies and epistatic relationships that may arise between bacterial replicons and across a genome, while also demonstrating how Tn-seq and metabolic modeling can be used together to yield insights not obtainable by either method alone. PMID:29672509

Succinoglycan Production Contributes to Acidic pH Tolerance in Sinorhizobium meliloti Rm1021.

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Hawkins, Justin P; Geddes, Barney A; Oresnik, Ivan J

2017-12-01

In this work, the hypothesis that exopolysaccharide plays a role in the survival of Sinorhizobium meliloti at low pH levels is addressed. When S. meliloti was grown at pH 5.75, synthesis of succinoglycan increased, whereas synthesis of galactoglucan decreased. Succinoglycan that was isolated from cultures grown at low pH had a lower degree of polymerization relative to that which was isolated from cultures grown at neutral pH, suggesting that low-molecular weight (LMW) succinoglycan might play a role in adaptation to low pH. Mutants unable to produce succinoglycan or only able to produce high-molecular weight polysaccharide were found to be sensitive to low pH. However, strains unable to produce LMW polysaccharide were 10-fold more sensitive. In response to low pH, transcription of genes encoding proteins for succinoglycan, glycogen, and cyclic β(1-2) glucans biosynthesis increased, while those encoding proteins necessary for the biosynthesis of galactoglucan decreased. While changes in pH did not affect the production of glycogen or cyclic β(1-2) glucan, it was found that the inability to produce cyclic β(1-2) glucan did contribute to pH tolerance in the absence of succinoglycan. Finally, in addition to being sensitive to low pH, a strain carrying mutations in exoK and exsH, which encode the glycanases responsible for the cleavage of succinoglycan to LMW succinoglycan, exhibited a delay in nodulation and was uncompetitive for nodule occupancy. Taken together, the data suggest that the role for LMW succinoglycan in nodule development may be to enhance survival in the colonized curled root hair.

Nodulation by Sinorhizobium meliloti originated from a mining soil alleviates Cd toxicity and increases Cd-phytoextraction in Medicago sativa L.

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Ghnaya, Tahar; Mnassri, Majda; Ghabriche, Rim; Wali, Mariem; Poschenrieder, Charlotte; Lutts, Stanley; Abdelly, Chedly

2015-01-01

Besides their role in nitrogen supply to the host plants as a result of symbiotic N fixation, the association between legumes and Rhizobium could be useful for the rehabilitation of metal-contaminated soils by phytoextraction. A major limitation presents the metal-sensitivity of the bacterial strains. The aim of this work was to explore the usefulness of Sinorhizobium meliloti originated from a mining site for Cd phytoextraction by Medicago sativa. Inoculated and non-inoculated plants were cultivated for 60 d on soils containing 50 and/or 100 mg Cd kg(-1) soil. The inoculation hindered the occurrence of Cd- induced toxicity symptoms that appeared in the shoots of non-inoculated plants. This positive effect of S. meliloti colonization was accompanied by an increase in biomass production and improved nutrient acquisition comparatively to non-inoculated plants. Nodulation enhanced Cd absorption by the roots and Cd translocation to the shoots. The increase of plant biomass concomitantly with the increase of Cd shoot concentration in inoculated plants led to higher potential of Cd-phytoextraction in these plants. In the presence of 50 mg Cd kg(-1) in the soil, the amounts of Cd extracted in the shoots were 58 and 178 μg plant(-1) in non-inoculated and inoculated plants, respectively. This study demonstrates that this association M. sativa-S. meliloti may be an efficient biological system to extract Cd from contaminated soils.

Hydrogen peroxide-regulated genes in the Medicago truncatula-Sinorhizobium meliloti symbiosis.

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Andrio, Emilie; Marino, Daniel; Marmeys, Anthony; de Segonzac, Marion Dunoyer; Damiani, Isabelle; Genre, Andrea; Huguet, Stéphanie; Frendo, Pierre; Puppo, Alain; Pauly, Nicolas

2013-04-01

Reactive oxygen species (ROS), particularly hydrogen peroxide (H(2)O(2)), play an important role in signalling in various cellular processes. The involvement of H(2)O(2) in the Medicago truncatula-Sinorhizobium meliloti symbiotic interaction raises questions about its effect on gene expression. A transcriptome analysis was performed on inoculated roots of M. truncatula in which ROS production was inhibited with diphenylene iodonium (DPI). In total, 301 genes potentially regulated by ROS content were identified 2 d after inoculation. These genes included MtSpk1, which encodes a putative protein kinase and is induced by exogenous H(2)O(2) treatment. MtSpk1 gene expression was also induced by nodulation factor treatment. MtSpk1 transcription was observed in infected root hair cells, nodule primordia and the infection zone of mature nodules. Analysis with a fluorescent protein probe specific for H(2)O(2) showed that MtSpk1 expression and H(2)O(2) were similarly distributed in the nodule infection zone. Finally, the establishment of symbiosis was impaired by MtSpk1 downregulation with an artificial micro-RNA. Several genes regulated by H(2)O(2) during the establishment of rhizobial symbiosis were identified. The involvement of MtSpk1 in the establishment of the symbiosis is proposed. © 2013 The Authors. New Phytologist © 2013 New Phytologist Trust.

Evaluation of SinoRhizobium meliloti Efficiency and Qualitative Traits of Alfalfa under Application of Molybdenum

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F Ahmadi Dana

2017-12-01

Full Text Available Introduction Agriculture depends heavily on nitrogen which is biologically fixed through the symbiotic association between rhizobia and legume plants in nodules located on plant roots. Alfalfa is a legume that should fix most of its own N requirement if it is sufficiently nodulated by viable Rhizobium meliloti inoculums. The process of nitrogen fixation is done by the help of an enzyme called nitrogenase and molybdenum which is an important element in the formation of this compound. Molybdenum is required by plants for protein synthesis and is especially important for legumes as it is needed for nitrogen fixation by rhizobia. Therefore the following research was done aimed on studying the effect of different amount of molybdenum and S. rhizobium bacteria on alfalfa’s yield. Material and Methods Alfalfa (Medicago sativa were grown in a field. The experiment was conducted at Karaj in 2013 in split plot arrangement based on completely randomized block design (RCBD, including 2 caring S. rhizobium inoculated seed and non-inoculated as the main plot factorand 3 levels of Molybdenum (0,5,10 kg ha-1 from ammonium molybdate (as the sub plot factor in three replications. Sinorhizobium meliloti bacteria were cultured on plates. Then half of the seeds were inoculated by Sinorhizobium meliloti. Nitrogen fertilizer was added only in one stage before planting up to 50 kg per hectare. Plants were grown until flowering. The data were analyzed by the SAS (9.1 software and mean comparisons were done by Duncan's MRT at the 1% and 5% probability level. Results and Discussion The results showed the effect of different levels of molybdenum and S. Rhizobium bacteria on dry matter yield, molybdenum concentrations in shoots and roots and the number of root nodules was significant. This treatment was significant in comparison to the control treatment with the14.27 ton per hectare. Increasing of molybdenum application, led to increasing of root nodules and showed a

A global analysis of protein expression profiles in Sinorhizobium meliloti: discovery of new genes for nodule occupancy and stress adaptation.

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Djordjevic, Michael A; Chen, Han Cai; Natera, Siria; Van Noorden, Giel; Menzel, Christian; Taylor, Scott; Renard, Clotilde; Geiger, Otto; Weiller, Georg F

2003-06-01

A proteomic examination of Sinorhizobium meliloti strain 1021 was undertaken using a combination of 2-D gel electrophoresis, peptide mass fingerprinting, and bioinformatics. Our goal was to identify (i) putative symbiosis- or nutrient-stress-specific proteins, (ii) the biochemical pathways active under different conditions, (iii) potential new genes, and (iv) the extent of posttranslational modifications of S. meliloti proteins. In total, we identified the protein products of 810 genes (13.1% of the genome's coding capacity). The 810 genes generated 1,180 gene products, with chromosomal genes accounting for 78% of the gene products identified (18.8% of the chromosome's coding capacity). The activity of 53 metabolic pathways was inferred from bioinformatic analysis of proteins with assigned Enzyme Commission numbers. Of the remaining proteins that did not encode enzymes, ABC-type transporters composed 12.7% and regulatory proteins 3.4% of the total. Proteins with up to seven transmembrane domains were identified in membrane preparations. A total of 27 putative nodule-specific proteins and 35 nutrient-stress-specific proteins were identified and used as a basis to define genes and describe processes occurring in S. meliloti cells in nodules and under stress. Several nodule proteins from the plant host were present in the nodule bacteria preparations. We also identified seven potentially novel proteins not predicted from the DNA sequence. Post-translational modifications such as N-terminal processing could be inferred from the data. The posttranslational addition of UMP to the key regulator of nitrogen metabolism, PII, was demonstrated. This work demonstrates the utility of combining mass spectrometry with protein arraying or separation techniques to identify candidate genes involved in important biological processes and niche occupations that may be intransigent to other methods of gene expression profiling.

Sinorhizobium meliloti sigma factors RpoE1 and RpoE4 are activated in stationary phase in response to sulfite.

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Bénédicte Bastiat

Full Text Available Rhizobia are soil bacteria able to establish a nitrogen-fixing symbiosis with legume plants. Both in soil and in planta, rhizobia spend non-growing periods resembling the stationary phase of in vitro-cultured bacteria. The primary objective of this work was to better characterize gene regulation in this biologically relevant growth stage in Sinorhizobium meliloti. By a tap-tag/mass spectrometry approach, we identified five sigma factors co-purifying with the RNA polymerase in stationary phase: the general stress response regulator RpoE2, the heat shock sigma factor RpoH2, and three extra-cytoplasmic function sigma factors (RpoE1, RpoE3 and RpoE4 belonging to the poorly characterized ECF26 subgroup. We then showed that RpoE1 and RpoE4 i are activated upon metabolism of sulfite-generating compounds (thiosulfate and taurine, ii display overlapping regulatory activities, iii govern a dedicated sulfite response by controlling expression of the sulfite dehydrogenase SorT, iv are activated in stationary phase, likely as a result of endogenous sulfite generation during bacterial growth. We showed that SorT is required for optimal growth of S. meliloti in the presence of sulfite, suggesting that the response governed by RpoE1 and RpoE4 may be advantageous for bacteria in stationary phase either by providing a sulfite detoxification function or by contributing to energy production through sulfite respiration. This paper therefore reports the first characterization of ECF26 sigma factors, the first description of sigma factors involved in control of sulphur metabolism, and the first indication that endogenous sulfite may act as a signal for regulation of gene expression upon entry of bacteria in stationary phase.

Inter-replicon Gene Flow Contributes to Transcriptional Integration in the Sinorhizobium meliloti Multipartite Genome.

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diCenzo, George C; Wellappili, Deelaka; Golding, G Brian; Finan, Turlough M

2018-05-04

Integration of newly acquired genes into existing regulatory networks is necessary for successful horizontal gene transfer (HGT). Ten percent of bacterial species contain at least two DNA replicons over 300 kilobases in size, with the secondary replicons derived predominately through HGT. The Sinorhizobium meliloti genome is split between a 3.7 Mb chromosome, a 1.7 Mb chromid consisting largely of genes acquired through ancient HGT, and a 1.4 Mb megaplasmid consisting primarily of recently acquired genes. Here, RNA-sequencing is used to examine the transcriptional consequences of massive, synthetic genome reduction produced through the removal of the megaplasmid and/or the chromid. Removal of the pSymA megaplasmid influenced the transcription of only six genes. In contrast, removal of the chromid influenced expression of ∼8% of chromosomal genes and ∼4% of megaplasmid genes. This was mediated in part by the loss of the ETR DNA region whose presence on pSymB is due to a translocation from the chromosome. No obvious functional bias among the up-regulated genes was detected, although genes with putative homologs on the chromid were enriched. Down-regulated genes were enriched in motility and sensory transduction pathways. Four transcripts were examined further, and in each case the transcriptional change could be traced to loss of specific pSymB regions. In particularly, a chromosomal transporter was induced due to deletion of bdhA likely mediated through 3-hydroxybutyrate accumulation. These data provide new insights into the evolution of the multipartite bacterial genome, and more generally into the integration of horizontally acquired genes into the transcriptome. Copyright © 2018 diCenzo, et al.

Inter-replicon Gene Flow Contributes to Transcriptional Integration in the Sinorhizobium meliloti Multipartite Genome

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George C. diCenzo

2018-05-01

Full Text Available Integration of newly acquired genes into existing regulatory networks is necessary for successful horizontal gene transfer (HGT. Ten percent of bacterial species contain at least two DNA replicons over 300 kilobases in size, with the secondary replicons derived predominately through HGT. The Sinorhizobium meliloti genome is split between a 3.7 Mb chromosome, a 1.7 Mb chromid consisting largely of genes acquired through ancient HGT, and a 1.4 Mb megaplasmid consisting primarily of recently acquired genes. Here, RNA-sequencing is used to examine the transcriptional consequences of massive, synthetic genome reduction produced through the removal of the megaplasmid and/or the chromid. Removal of the pSymA megaplasmid influenced the transcription of only six genes. In contrast, removal of the chromid influenced expression of ∼8% of chromosomal genes and ∼4% of megaplasmid genes. This was mediated in part by the loss of the ETR DNA region whose presence on pSymB is due to a translocation from the chromosome. No obvious functional bias among the up-regulated genes was detected, although genes with putative homologs on the chromid were enriched. Down-regulated genes were enriched in motility and sensory transduction pathways. Four transcripts were examined further, and in each case the transcriptional change could be traced to loss of specific pSymB regions. In particularly, a chromosomal transporter was induced due to deletion of bdhA likely mediated through 3-hydroxybutyrate accumulation. These data provide new insights into the evolution of the multipartite bacterial genome, and more generally into the integration of horizontally acquired genes into the transcriptome.

Mining the Sinorhizobium meliloti transportome to develop FRET biosensors for sugars, dicarboxylates and cyclic polyols.

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Alexandre Bourdès

Full Text Available Förster resonance energy transfer (FRET biosensors are powerful tools to detect biologically important ligands in real time. Currently FRET bisosensors are available for twenty-two compounds distributed in eight classes of chemicals (two pentoses, two hexoses, two disaccharides, four amino acids, one nucleobase, two nucleotides, six ions and three phytoestrogens. To expand the number of available FRET biosensors we used the induction profile of the Sinorhizobium meliloti transportome to systematically screen for new FRET biosensors.Two new vectors were developed for cloning genes for solute-binding proteins (SBPs between those encoding FRET partner fluorescent proteins. In addition to a vector with the widely used cyan and yellow fluorescent protein FRET partners, we developed a vector using orange (mOrange2 and red fluorescent protein (mKate2 FRET partners. From the sixty-nine SBPs tested, seven gave a detectable FRET signal change on binding substrate, resulting in biosensors for D-quinic acid, myo-inositol, L-rhamnose, L-fucose, β-diglucosides (cellobiose and gentiobiose, D-galactose and C4-dicarboxylates (malate, succinate, oxaloacetate and fumarate. To our knowledge, we describe the first two FRET biosensor constructs based on SBPs from Tripartite ATP-independent periplasmic (TRAP transport systems.FRET based on orange (mOrange2 and red fluorescent protein (mKate2 partners allows the use of longer wavelength light, enabling deeper penetration of samples at lower energy and increased resolution with reduced back-ground auto-fluorescence. The FRET biosensors described in this paper for four new classes of compounds; (i cyclic polyols, (ii L-deoxy sugars, (iii β-linked disaccharides and (iv C4-dicarboxylates could be developed to study metabolism in vivo.

Influence of different Sinorhizobium meliloti inocula on abundance of genes involved in nitrogen transformations in the rhizosphere of alfalfa (Medicago sativa L.).

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Babić, Katarina Huić; Schauss, Kristina; Hai, Brigitte; Sikora, Sanja; Redzepović, Sulejman; Radl, Viviane; Schloter, Michael

2008-11-01

Inoculation of leguminous seeds with selected rhizobial strains is practised in agriculture to ameliorate the plant yield by enhanced root nodulation and nitrogen uptake of the plant. However, effective symbiosis between legumes and rhizobia does not only depend on the capacity of nitrogen fixation but also on the entire nitrogen turnover in the rhizosphere. We investigated the influence of seed inoculation with two indigenous Sinorhizobium meliloti strains exhibiting different efficiency concerning plant growth promotion on nitrogen turnover processes in the rhizosphere during the growth of alfalfa. Quantification of six target genes (bacterial amoA, nirK, nirS, nosZ, nifH and archaeal amoA) within the nitrogen cycle was performed in rhizosphere samples before nodule formation, at bud development and at the late flowering stage. The results clearly demonstrated that effectiveness of rhizobial inocula is related to abundance of nifH genes in the late flowering phase of alfalfa. Moreover, other genes involved in nitrogen turnover had been affected by the inocula, e.g. higher numbers of amoA copies were observed during flowering when the more effective strain had been inoculated. However, the respective gene abundances differed overall to a greater extent between the three plant development stages than between the inoculation variants.

NCBI nr-aa BLAST: CBRC-DRER-26-0474 [SEVENS

Lifescience Database Archive (English)

Full Text Available CBRC-DRER-26-0474 ref|NP_384172.1| SENSOR HISTIDINE KINASE TRANSMEMBRANE PROTEIN [S...inorhizobium meliloti 1021] emb|CAC41453.1| SENSOR HISTIDINE KINASE TRANSMEMBRANE PROTEIN [Sinorhizobium meliloti] NP_384172.1 1e-159 68% ...

ORF Alignment: NC_003047 [GENIUS II[Archive

Lifescience Database Archive (English)

Full Text Available NC_003047 gi|15964777 >1v4aA 24 435 49 467 e-101 ... emb|CAC45596.1| PUTATIVE GLUTAMATE-AMMONIA...-LIGASE ADENYLYLTRANSFERASE PROTEIN ... [Sinorhizobium meliloti] ref|NP_385130.1| PUTATIVE ... GLUTAMATE-AMMO...NIA-LIGASE ADENYLYLTRANSFERASE PROTEIN ... [Sinorhizobium meliloti 1021] sp|

Transcriptome Response to Heavy Metals in Sinorhizobium meliloti CCNWSX0020 Reveals New Metal Resistance Determinants That Also Promote Bioremediation by Medicago lupulina in Metal-Contaminated Soil.

Science.gov (United States)

Lu, Mingmei; Jiao, Shuo; Gao, Enting; Song, Xiuyong; Li, Zhefei; Hao, Xiuli; Rensing, Christopher; Wei, Gehong

2017-10-15

The symbiosis of the highly metal-resistant Sinorhizobium meliloti CCNWSX0020 and Medicago lupulina has been considered an efficient tool for bioremediation of heavy metal-polluted soils. However, the metal resistance mechanisms of S. meliloti CCNWSX00200 have not been elucidated in detail. Here we employed a comparative transcriptome approach to analyze the defense mechanisms of S. meliloti CCNWSX00200 against Cu or Zn exposure. Six highly upregulated transcripts involved in Cu and Zn resistance were identified through deletion mutagenesis, including genes encoding a multicopper oxidase (CueO), an outer membrane protein (Omp), sulfite oxidoreductases (YedYZ), and three hypothetical proteins (a CusA-like protein, a FixH-like protein, and an unknown protein), and the corresponding mutant strains showed various degrees of sensitivity to multiple metals. The Cu-sensitive mutant (Δ cueO ) and three mutants that were both Cu and Zn sensitive (Δ yedYZ , Δ cusA -like, and Δ fixH -like) were selected for further study of the effects of these metal resistance determinants on bioremediation. The results showed that inoculation with the Δ cueO mutant severely inhibited infection establishment and nodulation of M. lupulina under Cu stress, while inoculation with the Δ yedYZ and Δ fixH -like mutants decreased just the early infection frequency and nodulation under Cu and Zn stresses. In contrast, inoculation with the Δ cusA -like mutant almost led to loss of the symbiotic capacity of M. lupulina to even grow in uncontaminated soil. Moreover, the antioxidant enzyme activity and metal accumulation in roots of M. lupulina inoculated with all mutants were lower than those with the wild-type strain. These results suggest that heavy metal resistance determinants may promote bioremediation by directly or indirectly influencing formation of the rhizobium-legume symbiosis. IMPORTANCE Rhizobium-legume symbiosis has been promoted as an appropriate tool for bioremediation of heavy

Three way interactions between Thymus vulgaris, Medicago truncatula and Sinorhizobium meliloti

DEFF Research Database (Denmark)

Grøndahl, Eva; Ehlers, Bodil Kirstine

2012-01-01

Thymus vulgaris is a dominating component of the Mediterranean garrigue vegetation. It produces aromatic oil, containing monoterpenes, which affects the performance (growth, survival) of other plants, and microorganisms. Annual plant species of the genus Medicago are commonly found in Mediterranean...... shows patterns of adaptation to its thyme neighbor, and 2) if any adaptive response was dependent on the rhizobium, and whether the rhizobium was either "experienced" or "naive" with respect to thyme monoterpenes. Using a G*G*E design, the fitness of 13 genotypes of Medicago truncatula was tested....... Of these genotypes, 7 were ”experienced”, and 6 were ”naive” to thyme. All genotypes were grown on soil either amended with thyme monoterpene or not. In addition, each plant received a rhizobium treatment, which was either: no rhizobium, a mix of thyme experienced Sinorhizobium genotypes, or a mix of thyme naive...

Evaluation of the Effects of Biological and Chemical Fertilizers on Some Physiological and Growth Characteristics of Fenugreek (Trigonella foenum-graecum

Directory of Open Access Journals (Sweden)

Samira Menbari

2017-12-01

Full Text Available Nowadays, the use of soil-born microorganisms as biological fertilizers is considered to be a natural and most desirable solution to maintain sustainability of agricultural soil system. Potassium releasing bacteria, nitrogen fixing and phosphorus dissolving bacteria make mentioned elements available to plants. In order to evaluate the effects of bio-fertilizers Potabarvar 2, Sinorhizobium meliloti, as well as urea fertilizer on physiological properties and yield of Fenugreek, an experiment as complete randomized block design was conducted with five treatments and three replications. Treatments included biofertilizer Potabarvar 2, S. meliloti, inoculation with a mixture of Sinorhizobium+Potabarvar 2, positive control (based on soil analysis and negative control (no fertilization and inoculation.The results showed that all morphological traits were significant at 1%. Most physiological traits except for carotenoid were significantly affected by S. meliloti, and a mixture of Sinorhizobium+Potabarvar 2. Seed inoculation with biofertilizer Sinorhizobium meliloti and Potabarvar 2 lead to increase in growth and eventually shoot yield. Separate application of these biofertilizers led to better results than the integrated application. Symbiotic relationship of Sinorhizobium with Fenugreek increased physiological indices data, especially the absorption of nitrogen and phosphorus, as well as the amount of phenolic antioxidant have been significantly affected. In general, application of S. meliloti resulted in better and more effective increase in yield, quality and plant growth than fertilizer Potabarvar 2 and a mixture of Sinorhizobium+Potabarvar 2.

ORF Alignment: NC_003037 [GENIUS II[Archive

Lifescience Database Archive (English)

Full Text Available NC_003037 gi|16263035 >1nyoA 11 162 8 158 4e-30 ... ref|NP_435828.1| Nex18 Symbiotica...lly induced conserved protein [Sinorhizobium ... meliloti 1021] gb|AAK65240.1| Nex18 Symbiotically ... ... ... induced conserved protein [Sinorhizobium meliloti 1021] ... pir||F95334 Nex18 Symbiotically

Transcriptome Analysis of Polyhydroxybutyrate Cycle Mutants Reveals Discrete Loci Connecting Nitrogen Utilization and Carbon Storage in Sinorhizobium meliloti.

Science.gov (United States)

D'Alessio, Maya; Nordeste, Ricardo; Doxey, Andrew C; Charles, Trevor C

2017-01-01

Polyhydroxybutyrate (PHB) and glycogen polymers are produced by bacteria as carbon storage compounds under unbalanced growth conditions. To gain insights into the transcriptional mechanisms controlling carbon storage in Sinorhizobium meliloti , we investigated the global transcriptomic response to the genetic disruption of key genes in PHB synthesis and degradation and in glycogen synthesis. Under both nitrogen-limited and balanced growth conditions, transcriptomic analysis was performed with genetic mutants deficient in PHB synthesis ( phbA , phbB , phbAB , and phbC ), PHB degradation ( bdhA , phaZ , and acsA2 ), and glycogen synthesis ( glgA1 ). Three distinct genomic regions of the pSymA megaplasmid exhibited altered expression in the wild type and the PHB cycle mutants that was not seen in the glycogen synthesis mutant. An Fnr family transcriptional motif was identified in the upstream regions of a cluster of genes showing similar transcriptional patterns across the mutants. This motif was found at the highest density in the genomic regions with the strongest transcriptional effect, and the presence of this motif upstream of genes in these regions was significantly correlated with decreased transcript abundance. Analysis of the genes in the pSymA regions revealed that they contain a genomic overrepresentation of Fnr family transcription factor-encoding genes. We hypothesize that these loci, containing mostly nitrogen utilization, denitrification, and nitrogen fixation genes, are regulated in response to the intracellular carbon/nitrogen balance. These results indicate a transcriptional regulatory association between intracellular carbon levels (mediated through the functionality of the PHB cycle) and the expression of nitrogen metabolism genes. IMPORTANCE The ability of bacteria to store carbon and energy as intracellular polymers uncouples cell growth and replication from nutrient uptake and provides flexibility in the use of resources as they are available to

Genome-Wide Transcriptional Changes and Lipid Profile Modifications Induced by Medicago truncatula N5 Overexpression at an Early Stage of the Symbiotic Interaction with Sinorhizobium meliloti

Directory of Open Access Journals (Sweden)

Chiara Santi

2017-12-01

Full Text Available Plant lipid-transfer proteins (LTPs are small basic secreted proteins, which are characterized by lipid-binding capacity and are putatively involved in lipid trafficking. LTPs play a role in several biological processes, including the root nodule symbiosis. In this regard, the Medicago truncatula nodulin 5 (MtN5 LTP has been proved to positively regulate the nodulation capacity, controlling rhizobial infection and nodule primordia invasion. To better define the lipid transfer protein MtN5 function during the symbiosis, we produced MtN5-downregulated and -overexpressing plants, and we analysed the transcriptomic changes occurring in the roots at an early stage of Sinorhizobium meliloti infection. We also carried out the lipid profile analysis of wild type (WT and MtN5-overexpressing roots after rhizobia infection. The downregulation of MtN5 increased the root hair curling, an early event of rhizobia infection, and concomitantly induced changes in the expression of defence-related genes. On the other hand, MtN5 overexpression favoured the invasion of the nodules by rhizobia and determined in the roots the modulation of genes that are involved in lipid transport and metabolism as well as an increased content of lipids, especially galactolipids that characterize the symbiosome membranes. Our findings suggest the potential participation of LTPs in the synthesis and rearrangement of membranes occurring during the formation of the infection threads and the symbiosome membrane.

Denitrification by Rhizobium meliloti

Energy Technology Data Exchange (ETDEWEB)

Rosen, A

1996-10-01

Rhizobium meliloti strains were investigated for their denitrification activity as free-living cells and in nodules on lucerne (Medicago sativa) roots. They were also investigated for presence of nitrous oxide reductase (nos) activity and for genes using a nosZ probe derived from the Pseudomonas stutzeri. To decide whether R. meliloti strains used as inoculants contribute to the total denitrification activity in a lucerne ley, strains with different denitrifying capacities were used in field and laboratory experiments. The nitrate reduction activity of R. meliloti during anaerobic respiration was compared with that of a strain of Pseudomonas aeruginosa. A great diversity in the denitrification activity was found within strains of R. meliloti, and four of thirteen investigated strains showed an obvious denitrification activity. Two denitrifying bacteria were used as references, one strain each of Bradyrhizobium japonicum and P. aeruginosa. All but one of the R. meliloti strains hybridized to the PstI-fragment of the nosZ-gene from P. stutzeri. Two sizes of the hybridizing fragment, 5 and 7 kb, were noticed. Nos activity was only shown in three R. meliloti strains, and these were all characterized by a high denitrification activity. The potential denitrification activity was about 20, 40, and 80 times higher than the actual denitrification activity for lucerne, fallow, and grass, respectively. The potential denitrification activity was almost the same in lucerne and grass planted soils. Compared with the unplanted soil, the presence of lucerne roots in the soil increased the actual denitrification activity, while roots of both plant species, grass and lucerne, increased the potential denitrification activity in the soil. 32 refs, 7 figs, 1 tab

Sinorhizobium arboris sp. nov. and Sinorhizobium kostiense sp. nov., isolated from leguminous trees in Sudan and Kenya.

Science.gov (United States)

Nick, G; de Lajudie, P; Eardly, B D; Suomalainen, S; Paulin, L; Zhang, X; Gillis, M; Lindström, K

1999-10-01

SDS-PAGE of total bacterial proteins was applied to the classification of 25 Sudanese and five Kenyan strains isolated from the root nodules of Acacia senegal and Prosopis chilensis. Twenty strains were also studied by multilocus enzyme electrophoresis (MLEE) and the whole 16S rRNA gene was sequenced from two strains representing the two major clusters. These results, together with the previously reported numerical taxonomy analysis, pulsed-field gel electrophoresis studies, DNA-DNA dot-blot hybridization, genomic fingerprinting using repetitive sequence-based PCR, DNA base composition analysis, DNA-DNA reassociation analysis, partial sequencing of the 16S rRNA gene and RFLP analysis of the amplified 16S rRNA gene, showed that all 30 strains belong to the genus Sinorhizobium. Two of the strains grouped with Sinorhizobium saheli and seven with Sinorhizobium terangae, while the rest did not cluster with any of the established species. The majority of the strains formed two phenotypically and genotypically distinct groups and we therefore propose that these strains should be classified as two new species, Sinorhizobium arboris sp. nov. and Sinorhizobium kostiense sp. nov.

Medicago sativa--Sinorhizobium meliloti Symbiosis Promotes the Bioaccumulation of Zinc in Nodulated Roots.

Science.gov (United States)

Zribi, Kais; Nouairi, Issam; Slama, Ines; Talbi-Zribi, Ons; Mhadhbi, Haythem

2015-01-01

In this study we investigated effects of Zn supply on germination, growth, inorganic solutes (Zn, Ca, Fe, and Mg) partitioning and nodulation of Medicago sativa This plant was cultivated with and without Zn (2 mM). Treatments were plants without (control) and with Zn tolerant strain (S532), Zn intolerant strain (S112) and 2 mM urea nitrogen fertilisation. Results showed that M. sativa germinates at rates of 50% at 2 mM Zn. For plants given nitrogen fertilisation, Zn increased plant biomass production. When grown with symbionts, Zn supply had no effect on nodulation. Moreover, plants with S112 showed a decrease of shoot and roots biomasses. However, in symbiosis with S532, an increase of roots biomass was observed. Plants in symbiosis with S. meliloti accumulated more Zn in their roots than nitrogen fertilised plants. Zn supply results in an increase of Ca concentration in roots of fertilised nitrogen plants. However, under Zn supply, Fe concentration decreased in roots and increased in nodules of plants with S112. Zn supply showed contrasting effects on Mg concentrations for plants with nitrogen fertilisation (increase) and plants with S112 (decrease). The capacity of M. sativa to accumulate Zn in their nodulated roots encouraged its use in phytostabilisation processes.

ORF Alignment: NC_003047 [GENIUS II[Archive

Lifescience Database Archive (English)

Full Text Available ... [Sinorhizobium meliloti 1021] ... Length = 101 ... Query: 1 ... MFAVIKTGGKQYRVAANDVITIEKLEGVAGDKIEF...TEILMVGVGADATIGAPFVEGAVVS 60 ... MFAVIKTGGKQYRVAANDVITIEKLEGVAGDKIEFTEILMV...GVGADATIGAPFVEGAVVS Sbjct: 1 ... MFAVIKTGGKQYRVAANDVITIEKLEGVAGDKIEFTEILMVGVGADATIGAPFVEGAVVS 60 ...

ORF Alignment: NC_003047 [GENIUS II[Archive

Lifescience Database Archive (English)

Full Text Available ... [Sinorhizobium meliloti 1021] ... Length = 118 ... Query: 249 ELTCADIMSRDVVTVPGDTTPDHARYLLLKHDIRT...LPVLDENEKLQGTVGLRELAGKEPG 308 ... ELTCADIMSRDVVTVPGDTTPDHARYLLLKHDIRTLP...VLDENEKLQGTVGLRELAGKEPG Sbjct: 1 ... ELTCADIMSRDVVTVPGDTTPDHARYLLLKHDIRTLPVLDENEKLQGTVGLRELAGKEPG 60 ...

The symbiotic biofilm of Sinorhizobium fredii SMH12, necessary for successful colonization and symbiosis of Glycine max cv Osumi, is regulated by Quorum Sensing systems and inducing flavonoids via NodD1.

Directory of Open Access Journals (Sweden)

Francisco Pérez-Montaño

Full Text Available Bacterial surface components, especially exopolysaccharides, in combination with bacterial Quorum Sensing signals are crucial for the formation of biofilms in most species studied so far. Biofilm formation allows soil bacteria to colonize their surrounding habitat and survive common environmental stresses such as desiccation and nutrient limitation. This mode of life is often essential for survival in bacteria of the genera Mesorhizobium, Sinorhizobium, Bradyrhizobium, and Rhizobium. The role of biofilm formation in symbiosis has been investigated in detail for Sinorhizobium meliloti and Bradyrhizobium japonicum. However, for S. fredii this process has not been studied. In this work we have demonstrated that biofilm formation is crucial for an optimal root colonization and symbiosis between S. fredii SMH12 and Glycine max cv Osumi. In this bacterium, nod-gene inducing flavonoids and the NodD1 protein are required for the transition of the biofilm structure from monolayer to microcolony. Quorum Sensing systems are also required for the full development of both types of biofilms. In fact, both the nodD1 mutant and the lactonase strain (the lactonase enzyme prevents AHL accumulation are defective in soybean root colonization. The impairment of the lactonase strain in its colonization ability leads to a decrease in the symbiotic parameters. Interestingly, NodD1 together with flavonoids activates certain quorum sensing systems implicit in the development of the symbiotic biofilm. Thus, S. fredii SMH12 by means of a unique key molecule, the flavonoid, efficiently forms biofilm, colonizes the legume roots and activates the synthesis of Nod factors, required for successfully symbiosis.

ORF Alignment: NC_003047 [GENIUS II[Archive

Lifescience Database Archive (English)

Full Text Available SE ... PROTEIN [Sinorhizobium meliloti 1021] ... Length = 130 ... Query: 751 AREDEGAAEASRLRMEIDELRSILETATDGVVVLGRDGDIRT...MNRSASALFDYDEADMRG 810 ... AREDEGAAEASRLRMEIDELRSILETATDGVVVLGRDGDIRT...MNRSASALFDYDEADMRG Sbjct: 1 ... AREDEGAAEASRLRMEIDELRSILETATDGVVVLGRDGDIRTMNRSASALFDYDEADMRG

ORF Alignment: NC_003047 [GENIUS II[Archive

Lifescience Database Archive (English)

Full Text Available ... [Sinorhizobium meliloti 1021] ... Length = 138 ... Query: 9 ... SKAPRSVLFMCGMNAIRSPMAEALARVALPKGTYVASAGVRQGERD...PFVDVVLEEVGLTI 68 ... SKAPRSVLFMCGMNAIRSPMAEALARVALPKGTYVASAGVRQGERD...PFVDVVLEEVGLTI Sbjct: 1 ... SKAPRSVLFMCGMNAIRSPMAEALARVALPKGTYVASAGVRQGERDPFVDVVLEEVGLTI 60 ... Query: 12

ORF Alignment: NC_003047 [GENIUS II[Archive

Lifescience Database Archive (English)

Full Text Available R ... PROTEIN [Sinorhizobium meliloti 1021] ... Length = 106 ... Query: 12 ... ELTVGEVAERSGLAVSTLHFYEAKGLIRSN...RSRGNQRRYPRSVLRRVAVIKVAQRTGIPL 71 ... ELTVGEVAERSGLAVSTLHFYEAKGLIRSN...RSRGNQRRYPRSVLRRVAVIKVAQRTGIPL Sbjct: 1 ... ELTVGEVAERSGLAVSTLHFYEAKGLIRSNRSRGNQRRYPRSVLRRVAVIKVAQRTGIPL 60 ...

ORF Alignment: NC_003078 [GENIUS II[Archive

Lifescience Database Archive (English)

Full Text Available lator, ArsR family protein [Sinorhizobium meliloti ... 1021] ... Length = 83 ... Query: 8 ... LSALADPTRRAIVARLAAGEATVNELAAPFEM...SLPAVSKHLKVLERAGLISRGRNAQWRP 67 ... LSALADPTRRAIVARLAAGEATVNELAAPFEMSLPAVSKHL...KVLERAGLISRGRNAQWRP Sbjct: 1 ... LSALADPTRRAIVARLAAGEATVNELAAPFEMSLPAVSKHLKVLERAGLISRGRNAQWRP 60 ...

ORF Sequence: NC_003078 [GENIUS II[Archive

Lifescience Database Archive (English)

Full Text Available etitiveness [Sinorhizobium meliloti 1021] MQLSACARRREAVRYRRRMARILILLFSLLSAFAFPVTPVP... NC_003078 gi|16264863 >gi|16264863|ref|NP_437655.1| probable membrane protein necessary for nodulation comp

ORF Sequence: NC_003047 [GENIUS II[Archive

Lifescience Database Archive (English)

Full Text Available NC_003047 gi|15964332 >gi|15964332|ref|NP_384685.1| PROBABLE PYRAZINAMIDASE/NICOTINAMIDAS...E (INCLUDES: PYRAZINAMIDASE, NICOTINAMIDASE) PROTEIN [Sinorhizobium meliloti 1021] MADAARPDLREAMADEAL

ORF Alignment: NC_003047 [GENIUS II[Archive

Lifescience Database Archive (English)

Full Text Available MOYL-PHOSPHATE SYNTHASE LARGE CHAIN (AMMONIA CHAIN ... ARGININE BIOSYNTHESIS) PROTEIN [Sinorhizobium... meliloti] ... ref|NP_385682.1| PROBABLE CARBAMOYL-PHOSPHATE SYNTHASE ... LARGE CHAIN (AMMONIA

Roles of Extracellular Polysaccharides and Biofilm Formation in Heavy Metal Resistance of Rhizobia

OpenAIRE

Natalia Nocelli; Pablo C. Bogino; Erika Banchio; Walter Giordano

2016-01-01

Bacterial surface components and extracellular compounds, particularly flagella, lipopolysaccharides (LPSs), and exopolysaccharides (EPSs), in combination with environmental signals and quorum-sensing signals, play crucial roles in bacterial autoaggregation, biofilm development, survival, and host colonization. The nitrogen-fixing species Sinorhizobium meliloti (S. meliloti) produces two symbiosis-promoting EPSs: succinoglycan (or EPS I) and galactoglucan (or EPS II). Studies of the S. melilo...

ORF Sequence: NC_003047 [GENIUS II[Archive

Lifescience Database Archive (English)

Full Text Available NC_003047 gi|15965329 >gi|15965329|ref|NP_385682.1| PROBABLE CARBAMOYL-PHOSPHATE SYNTHASE LARGE CHAIN (AMMO...NIA CHAIN ARGININE BIOSYNTHESIS) PROTEIN [Sinorhizobium meliloti 1021] MPKRQDIKSILI

ORF Alignment: NC_003047 [GENIUS II[Archive

Lifescience Database Archive (English)

Full Text Available NC_003047 gi|15966456 >1gkmA 2 501 3 499 e-107 ... emb|CAC47282.1| PUTATIVE HISTIDINE AMMONIA...-LYASE PROTEIN [Sinorhizobium meliloti] ... ref|NP_386809.1| PUTATIVE HISTIDINE AMMONIA-LYASE

Final report for DOE grant FG02-06ER15805

Energy Technology Data Exchange (ETDEWEB)

Gage, Daniel

2012-05-31

DOE funding was used to investigate the role of the phosphotransferase system (PTS) in the symbiotic, nodulating bacterium Sinorhizobium meliloti. This system is well studied in several bacterial species. However, it's organization and function in S. meliloti is substantially different than in the those other, well-studied bacteria. The S. meliloti PTS, through our DOE-funded work, has become a model for how this important signal transduction system works in the a-proteobacteria. We have found that the PTS is relatively simple, used for only signal transduction and not transport, and is involved in regulation of carbon metabolism in response to carbon availability and nitrogen availability.

Rhizobia from Lanzarote, the Canary Islands, that nodulate Phaseolus vulgaris have characteristics in common with LMW RNA group II Sinorhizobium meliloti of Medicago, Melilotus and Trigonella from soils of mainland Spain

Science.gov (United States)

Several isolates from nodules of Phaseolus vulgaris grown in soil of Lanzarote, an island of the Canaries, had electrophoretic LMW RNA patterns identical with a less common pattern within S. meliloti (assigned as group II) obtained from nodules of alfalfa and alfalfa-related legumes grown in northe...

"Ménage à trois"

DEFF Research Database (Denmark)

Ehlers, Bodil K; Grøndahl, Eva; Ronfort, Joëlle

2012-01-01

of both partners. So far these studies have been typically carried out in a single environment. Here, we ask if the genetic correlation between fitness of the host plant Medicago truncatula (Fabaceae) and its bacterial symbiont Sinorhizobium meliloti is affected by the presence/absence of a monoterpene...

Replicon-dependent differentiation of symbiosis-related genes in Sinorhizobium strains nodulating Glycine max.

Science.gov (United States)

Guo, Hui Juan; Wang, En Tao; Zhang, Xing Xing; Li, Qin Qin; Zhang, Yan Ming; Tian, Chang Fu; Chen, Wen Xin

2014-02-01

In order to investigate the genetic differentiation of Sinorhizobium strains nodulating Glycine max and related microevolutionary mechanisms, three housekeeping genes (SMc00019, truA, and thrA) and 16 symbiosis-related genes on the chromosome (7 genes), pSymA (6 genes), and pSymB (3 genes) were analyzed. Five distinct species were identified among the test strains by calculating the average nucleotide identity (ANI) of SMc00019-truA-thrA: Sinorhizobium fredii, Sinorhizobium sojae, Sinorhizobium sp. I, Sinorhizobium sp. II, and Sinorhizobium sp. III. These species assignments were also supported by population genetics and phylogenetic analyses of housekeeping genes and symbiosis-related genes on the chromosome and pSymB. Different levels of genetic differentiation were observed among these species or different replicons. S. sojae was the most divergent from the other test species and was characterized by its low intraspecies diversity and limited geographic distribution. Intergenic recombination dominated the evolution of 19 genes from different replicons. Intraspecies recombination happened frequently in housekeeping genes and symbiosis-related genes on the chromosome and pSymB, whereas pSymA genes showed a clear pattern of lateral-transfer events between different species. Moreover, pSymA genes were characterized by a lower level of polymorphism and recombination than those on the chromosome and pSymB. Taken together, genes from different replicons of rhizobia might be involved in the establishment of symbiosis with legumes, but these symbiosis-related genes might have evolved differently according to their corresponding replicons.

Effect of exogenous application of rhizopine on lucerne root nodulation

African Journals Online (AJOL)

Rhizopine, 3-0 -methyl scyllo-inosamine was applied to the roots of luceme seedling inoculated with either rhizopine synthesizing Sinorhizobium meliloti strain L530 or the non-rhizopine synthesizing strain Rm 1021 . There was an initial delay in nodule formation. A significant increase in the number of nodules formed in ...

Analysis of nodule meristem persistence and ENOD40 functioning in Medicago truncatula nodule formation

NARCIS (Netherlands)

Wan Xi,

2007-01-01

Medicago root nodules are formed as a result of the interaction of the plant with the soil-borne bacterium Sinorhizobium meliloti. Several plant genes are induced during nodule formation and MtENOD40 is one of the earliest genes activated. The precise function as well as the molecule

Biochemical and Molecular Phylogenetic Study of Agriculturally Useful Association of a Nitrogen-Fixing Cyanobacterium and Nodule Sinorhizobium with Medicago sativa L.

Directory of Open Access Journals (Sweden)

E. V. Karaushu

2015-01-01

Full Text Available Seed inoculation with bacterial consortium was found to increase legume yield, providing a higher growth than the standard nitrogen treatment methods. Alfalfa plants were inoculated by mono- and binary compositions of nitrogen-fixing microorganisms. Their physiological and biochemical properties were estimated. Inoculation by microbial consortium of Sinorhizobium meliloti T17 together with a new cyanobacterial isolate Nostoc PTV was more efficient than the single-rhizobium strain inoculation. This treatment provides an intensification of the processes of biological nitrogen fixation by rhizobia bacteria in the root nodules and an intensification of plant photosynthesis. Inoculation by bacterial consortium stimulates growth of plant mass and rhizogenesis and leads to increased productivity of alfalfa and to improving the amino acid composition of plant leaves. The full nucleotide sequence of the rRNA gene cluster and partial sequence of the dinitrogenase reductase (nifH gene of Nostoc PTV were deposited to GenBank (JQ259185.1, JQ259186.1. Comparison of these gene sequences of Nostoc PTV with all sequences present at the GenBank shows that this cyanobacterial strain does not have 100% identity with any organisms investigated previously. Phylogenetic analysis showed that this cyanobacterium clustered with high credibility values with Nostoc muscorum.

Detection and isolation of novel rhizopine-catabolizing bacteria from the environment

Science.gov (United States)

Gardener; de Bruijn FJ

1998-12-01

Microbial rhizopine-catabolizing (Moc) activity was detected in serial dilutions of soil and rhizosphere washes. The activity observed generally ranged between 10(6) and 10(7) catabolic units per g, and the numbers of nonspecific culture-forming units were found to be approximately 10 times higher. A diverse set of 37 isolates was obtained by enrichment on scyllo-inosamine-containing media. However, none of the bacteria that were isolated were found to contain DNA sequences homologous to the known mocA, mocB, and mocC genes of Sinorhizobium meliloti L5-30. Twenty-one of the isolates could utilize an SI preparation as the sole carbon and nitrogen source for growth. Partial sequencing of 16S ribosomal DNAs (rDNAs) amplified from these strains indicated that five distinct bacterial genera (Arthrobacter, Sinorhizobium, Pseudomonas, Aeromonas, and Alcaligenes) were represented in this set. Only 6 of these 21 isolates could catabolize 3-O-methyl-scyllo-inosamine under standard assay conditions. Two of these, strains D1 and R3, were found to have 16S rDNA sequences very similar to those of Sinorhizobium meliloti. However, these strains are not symbiotically effective on Medicago sativa, and DNA sequences homologous to the nodB and nodC genes were not detected in strains D1 and R3 by Southern hybridization analysis.

Genomic resources for identification of the minimal N2 -fixing symbiotic genome.

Science.gov (United States)

diCenzo, George C; Zamani, Maryam; Milunovic, Branislava; Finan, Turlough M

2016-09-01

The lack of an appropriate genomic platform has precluded the use of gain-of-function approaches to study the rhizobium-legume symbiosis, preventing the establishment of the genes necessary and sufficient for symbiotic nitrogen fixation (SNF) and potentially hindering synthetic biology approaches aimed at engineering this process. Here, we describe the development of an appropriate system by reverse engineering Sinorhizobium meliloti. Using a novel in vivo cloning procedure, the engA-tRNA-rmlC (ETR) region, essential for cell viability and symbiosis, was transferred from Sinorhizobium fredii to the ancestral location on the S. meliloti chromosome, rendering the ETR region on pSymB redundant. A derivative of this strain lacking both the large symbiotic replicons (pSymA and pSymB) was constructed. Transfer of pSymA and pSymB back into this strain restored symbiotic capabilities with alfalfa. To delineate the location of the single-copy genes essential for SNF on these replicons, we screened a S. meliloti deletion library, representing > 95% of the 2900 genes of the symbiotic replicons, for their phenotypes with alfalfa. Only four loci, accounting for < 12% of pSymA and pSymB, were essential for SNF. These regions will serve as our preliminary target of the minimal set of horizontally acquired genes necessary and sufficient for SNF. © 2016 Society for Applied Microbiology and John Wiley & Sons Ltd.

A comparative genomics screen identifies a Sinorhizobium meliloti 1021 sodM-like gene strongly expressed within host plant nodules

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Queiroux Clothilde

2012-05-01

Full Text Available Abstract Background We have used the genomic data in the Integrated Microbial Genomes system of the Department of Energy’s Joint Genome Institute to make predictions about rhizobial open reading frames that play a role in nodulation of host plants. The genomic data was screened by searching for ORFs conserved in α-proteobacterial rhizobia, but not conserved in closely-related non-nitrogen-fixing α-proteobacteria. Results Using this approach, we identified many genes known to be involved in nodulation or nitrogen fixation, as well as several new candidate genes. We knocked out selected new genes and assayed for the presence of nodulation phenotypes and/or nodule-specific expression. One of these genes, SMc00911, is strongly expressed by bacterial cells within host plant nodules, but is expressed minimally by free-living bacterial cells. A strain carrying an insertion mutation in SMc00911 is not defective in the symbiosis with host plants, but in contrast to expectations, this mutant strain is able to out-compete the S. meliloti 1021 wild type strain for nodule occupancy in co-inoculation experiments. The SMc00911 ORF is predicted to encode a “SodM-like” (superoxide dismutase-like protein containing a rhodanese sulfurtransferase domain at the N-terminus and a chromate-resistance superfamily domain at the C-terminus. Several other ORFs (SMb20360, SMc01562, SMc01266, SMc03964, and the SMc01424-22 operon identified in the screen are expressed at a moderate level by bacteria within nodules, but not by free-living bacteria. Conclusions Based on the analysis of ORFs identified in this study, we conclude that this comparative genomics approach can identify rhizobial genes involved in the nitrogen-fixing symbiosis with host plants, although none of the newly identified genes were found to be essential for this process.

Medicago truncatula DNF2 is a PI-PLC-XD-containing protein required for bacteroid persistence and prevention of nodule early senescence and defense-like reactions.

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Bourcy, Marie; Brocard, Lysiane; Pislariu, Catalina I; Cosson, Viviane; Mergaert, Peter; Tadege, Millon; Mysore, Kirankumar S; Udvardi, Michael K; Gourion, Benjamin; Ratet, Pascal

2013-03-01

Medicago truncatula and Sinorhizobium meliloti form a symbiotic association resulting in the formation of nitrogen-fixing nodules. Nodule cells contain large numbers of bacteroids which are differentiated, nitrogen-fixing forms of the symbiotic bacteria. In the nodules, symbiotic plant cells home and maintain hundreds of viable bacteria. In order to better understand the molecular mechanism sustaining the phenomenon, we searched for new plant genes required for effective symbiosis. We used a combination of forward and reverse genetics approaches to identify a gene required for nitrogen fixation, and we used cell and molecular biology to characterize the mutant phenotype and to gain an insight into gene function. The symbiotic gene DNF2 encodes a putative phosphatidylinositol phospholipase C-like protein. Nodules formed by the mutant contain a zone of infected cells reduced to a few cell layers. In this zone, bacteria do not differentiate properly into bacteroids. Furthermore, mutant nodules senesce rapidly and exhibit defense-like reactions. This atypical phenotype amongst Fix(-) mutants unravels dnf2 as a new actor of bacteroid persistence inside symbiotic plant cells. © 2012 CNRS. New Phytologist © 2012 New Phytologist Trust.

The trehalose utilization gene thuA ortholog in Mesorhizobium loti does not influence competitiveness for nodulation on Lotus spp.

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Ampomah, Osei Yaw; Jensen, John Beck

2014-03-01

Competitiveness for nodulation is a desirable trait in rhizobia strains used as inoculant. In Sinorhizobium meliloti 1021 mutation in either of the trehalose utilization genes thuA or thuB influences its competitiveness for root colonization and nodule occupancy depending on the interacting host. We have therefore investigated whether mutation in the thuA ortholog in Mesorhizobium loti MAFF303099 also leads to a similar competitive phenotype on its hosts. The results show that M. loti thuA mutant Ml7023 was symbiotically effective and was as competitive as the wild type in colonization and nodule occupancy on Lotus corniculatus and Lotus japonicus. The thuA gene in M. loti was not induced during root colonization or in the infection threads unlike in S. meliloti, despite its induction by trehalose and high osmolarity in in vitro assays.

The transcriptional activator LdtR from 'Candidatus Liberibacter asiaticus' mediates osmotic stress tolerance.

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Fernando A Pagliai

2014-04-01

Full Text Available The causal agent of Huanglongbing disease, 'Candidatus Liberibacter asiaticus', is a non-culturable, gram negative, phloem-limited α-proteobacterium. Current methods to control the spread of this disease are still limited to the removal and destruction of infected trees. In this study, we identified and characterized a regulon from 'Ca. L. asiaticus' involved in cell wall remodeling, that contains a member of the MarR family of transcriptional regulators (ldtR, and a predicted L,D-transpeptidase (ldtP. In Sinorhizobium meliloti, mutation of ldtR resulted in morphological changes (shortened rod-type phenotype and reduced tolerance to osmotic stress. A biochemical approach was taken to identify small molecules that modulate LdtR activity. The LdtR ligands identified by thermal shift assays were validated using DNA binding methods. The biological impact of LdtR inactivation by the small molecules was then examined in Sinorhizobium meliloti and Liberibacter crescens, where a shortened-rod phenotype was induced by growth in presence of the ligands. A new method was also developed to examine the effects of small molecules on the viability of 'Ca. Liberibacter asiaticus', using shoots from HLB-infected orange trees. Decreased expression of ldtRLas and ldtPLas was observed in samples taken from HLB-infected shoots after 6 h of incubation with the LdtR ligands. These results provide strong proof of concept for the use of small molecules that target LdtR, as a potential treatment option for Huanglongbing disease.

Regulation of Long-Chain N-Acyl-Homoserine Lactones in Agrobacterium vitis

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Hao, Guixia; Burr, Thomas J.

2006-01-01

Homologs of quorum-sensing luxR and luxI regulatory genes, avsR and avsI, were identified in Agrobacterium vitis strain F2/5. Compared to other LuxI proteins from related species, the deduced AvsI shows the greatest identity to SinI (71%) from Sinorhizobium meliloti Rm1021. AvsR possesses characteristic autoinducer binding and helix-turn-helix DNA binding domains and shares a high level of identity with SinR (38%) from Rm1021. Site-directed mutagenesis of avsR and avsI was performed, and both...

Identification of the rctA Gene, Which Is Required for Repression of Conjugative Transfer of Rhizobial Symbiotic Megaplasmids†

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Pérez-Mendoza, Daniel; Sepúlveda, Edgardo; Pando, Victoria; Muñoz, Socorro; Nogales, Joaquina; Olivares, José; Soto, Maria J.; Herrera-Cervera, José A.; Romero, David; Brom, Susana; Sanjuán, Juan

2005-01-01

An analysis of the conjugative transfer of pRetCFN42d, the symbiotic plasmid (pSym) of Rhizobium etli, has revealed a novel gene, rctA, as an essential element of a regulatory system for silencing the conjugative transfer of R. etli pSym by repressing the transcription of conjugal transfer genes in standard laboratory media. The rctA gene product lacks sequence conservation with other proteins of known function but may belong to the winged-helix DNA-binding subfamily of transcriptional regulators. Similar to that of many transcriptional repressors, rctA transcription seems to be positively autoregulated. rctA expression is greatly reduced upon overexpression of another gene, rctB, previously identified as a putative activator of R. etli pSym conjugal transfer. Thus, rctB seems to counteract the repressive action of rctA. rctA homologs are present in at least three other bacterial genomes within the order Rhizobiales, where they are invariably located adjacent to and divergently transcribed from putative virB-like operons. We show that similar to that of R. etli pSym, conjugative transfer of the 1.35-Mb symbiotic megaplasmid A of Sinorhizobium meliloti is also subjected to the inhibitory action of rctA. Our data provide strong evidence that the R. etli and S. meliloti pSym plasmids are indeed self-conjugative plasmids and that this property would only be expressed under optimal, as yet unknown conditions that entail inactivation of the rctA function. The rctA gene seems to represent novel but probably widespread regulatory systems controlling the transfer of conjugative elements within the order Rhizobiales. PMID:16237017

Genome sequence of Ensifer meliloti strain WSM1022; a highly effective microsymbiont of the model legume Medicago truncatula A17.

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Terpolilli, Jason; Hill, Yvette; Tian, Rui; Howieson, John; Bräu, Lambert; Goodwin, Lynne; Han, James; Liolios, Konstantinos; Huntemann, Marcel; Pati, Amrita; Woyke, Tanja; Mavromatis, Konstantinos; Markowitz, Victor; Ivanova, Natalia; Kyrpides, Nikos; Reeve, Wayne

2013-12-20

Ensifer meliloti WSM1022 is an aerobic, motile, Gram-negative, non-spore-forming rod that can exist as a soil saprophyte or as a legume microsymbiont of Medicago. WSM1022 was isolated in 1987 from a nodule recovered from the roots of the annual Medicago orbicularis growing on the Cyclades Island of Naxos in Greece. WSM1022 is highly effective at fixing nitrogen with M. truncatula and other annual species such as M. tornata and M. littoralis and is also highly effective with the perennial M. sativa (alfalfa or lucerne). In common with other characterized E. meliloti strains, WSM1022 will nodulate but fixes poorly with M. polymorpha and M. sphaerocarpos and does not nodulate M. murex. Here we describe the features of E. meliloti WSM1022, together with genome sequence information and its annotation. The 6,649,661 bp high-quality-draft genome is arranged into 121 scaffolds of 125 contigs containing 6,323 protein-coding genes and 75 RNA-only encoding genes, and is one of 100 rhizobial genomes sequenced as part of the DOE Joint Genome Institute 2010 Genomic Encyclopedia for Bacteria and Archaea-Root Nodule Bacteria (GEBA-RNB) project.

Competitive Advantage Provided by Bacterial Motility in the Formation of Nodules by Rhizobium meliloti

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Ames, Peter; Bergman, Kostia

1981-01-01

The effect of motility on the competitive success of Rhizobium meliloti in nodule production was investigated. A motile strain formed more nodules than expected when mixed at various unfavorable ratios with either flagellated or nonflagellated nonmotile derivatives. We conclude that motility confers a selective advantage on rhizobia when competing with nonmotile strains. PMID:7298580

Extended region of nodulation genes in Rhizobium meliloti 1021. II. Nucleotide sequence, transcription start sites and protein products

International Nuclear Information System (INIS)

Fisher, R.F.; Swanson, J.A.; Mulligan, J.T.; Long, S.R.

1987-01-01

The authors have established the DNA sequence and analyzed the transcription and translation products of a series of putative nodulation (nod) genes in Rhizobium meliloti strain 1021. Four loci have been designated nodF, nodE, nodG and nodH. The correlation of transposon insertion positions with phenotypes and open reading frames was confirmed by sequencing the insertion junctions of the transposons. The protein products of these nod genes were visualized by in vitro expression of cloned DNA segments in a R. meliloti transcription-translation system. In addition, the sequence for nodG was substantiated by creating translational fusions in all three reading frames at several points in the sequence; the resulting fusions were expressed in vitro in both E. coli and R. meliloti transcription-translation systems. A DNA segment bearing several open reading frames downstream of nodG corresponds to the putative nod gene mutated in strain nod-216. The transcription start sites of nodF and nodH were mapped by primer extension of RNA from cells induced with the plant flavone, luteolin. Initiation of transcription occurs approximately 25 bp downstream from the conserved sequence designated the nod box, suggesting that this conserved sequence acts as an upstream regulator of inducible nod gene expression. Its distance from the transcription start site is more suggestive of an activator binding site rather than an RNA polymerase binding site

Ensifer meliloti overexpressing Escherichia coli phytase gene ( appA) improves phosphorus (P) acquisition in maize plants

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Sharma, Vikas; Kumar, Ajit; Archana, G.; Kumar, G. Naresh

2016-10-01

The Escherichia coli phytase gene appA encoding enzyme AppA was cloned in a broad host range plasmid pBBR1MCS2 ( lac promoter), termed pVA1, and transformed into the Ensifer meliloti 1020. Transformation of pVA1 in Ensifer meliloti { E. m (pVA1)} increased its phosphatase and phytase activity by ˜9- and ˜50-fold, respectively, compared to the transformants containing empty plasmid as control { E. m (pBBR1MCS2)}. The western blot experiments using rabbit anti-AppA antibody showed that AppA is translocated into the periplasm of the host after its expression. Ensifer meliloti harboring AppA protein { E. m (pVA1)} and { E. m (pBBR1MCS2)} could acidify the unbuffered phytate minimal media (pH 8.0) containing Ca-phytate or Na-phytate as sole organic P (Po) source to below pH 5.0 and released P. However, both { E. m (pVA1)} and { E. m (pBBR1MCS2)} neither dropped pH of the medium nor released P when the medium was buffered at pH 8.0 using Tris-Cl, indicating that acidification of medium was important for the enzymatic hydrolysis of phytate. Further experiments proved that maize plants inoculated with { E. m. (pVA1)} showed increase in growth under sterile semi solid agar (SSA) medium containing Na-phytate as sole P source. The present study could be helpful in generating better transgenic bioinoculants harboring phosphate mineralization properties that ultimately promote plant growth.

Stable symbiotic nitrogen fixation under water-deficit field conditions by a stress-tolerant alfalfa microsymbiont and its complete genome sequence.

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Jozefkowicz, Cintia; Brambilla, Silvina; Frare, Romina; Stritzler, Margarita; Piccinetti, Carlos; Puente, Mariana; Berini, Carolina Andrea; Pérez, Pedro Reyes; Soto, Gabriela; Ayub, Nicolás

2017-12-10

We here characterized the stress-tolerant alfalfa microsymbiont Sinorhizobium meliloti B401. B401-treated plants showed high nitrogen fixation rates under humid and semiarid environments. The production of glycine betaine in isolated bacteroids positively correlated with low precipitation levels, suggesting that this compound acts as a critical osmoprotectant under field conditions. Genome analysis revealed that strain B401 contains alternative pathways for the biosynthesis and uptake of glycine betaine and its precursors. Such genomic information will offer substantial insight into the environmental physiology of this biotechnologically valuable nitrogen-fixing bacterium. Copyright © 2017 Elsevier B.V. All rights reserved.

Roles of Extracellular Polysaccharides and Biofilm Formation in Heavy Metal Resistance of Rhizobia

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Natalia Nocelli

2016-05-01

Full Text Available Bacterial surface components and extracellular compounds, particularly flagella, lipopolysaccharides (LPSs, and exopolysaccharides (EPSs, in combination with environmental signals and quorum-sensing signals, play crucial roles in bacterial autoaggregation, biofilm development, survival, and host colonization. The nitrogen-fixing species Sinorhizobium meliloti (S. meliloti produces two symbiosis-promoting EPSs: succinoglycan (or EPS I and galactoglucan (or EPS II. Studies of the S. meliloti/alfalfa symbiosis model system have revealed numerous biological functions of EPSs, including host specificity, participation in early stages of host plant infection, signaling molecule during plant development, and (most importantly protection from environmental stresses. We evaluated functions of EPSs in bacterial resistance to heavy metals and metalloids, which are known to affect various biological processes. Heavy metal resistance, biofilm production, and co-culture were tested in the context of previous studies by our group. A range of mercury (Hg II and arsenic (As III concentrations were applied to S. meliloti wild type strain and to mutant strains defective in EPS I and EPS II. The EPS production mutants were generally most sensitive to the metals. Our findings suggest that EPSs are necessary for the protection of bacteria from either Hg (II or As (III stress. Previous studies have described a pump in S. meliloti that causes efflux of arsenic from cells to surrounding culture medium, thereby protecting them from this type of chemical stress. The presence of heavy metals or metalloids in culture medium had no apparent effect on formation of biofilm, in contrast to previous reports that biofilm formation helps protect various microorganism species from adverse environmental conditions. In co-culture experiments, EPS-producing heavy metal resistant strains exerted a protective effect on AEPS-non-producing, heavy metal-sensitive strains; a phenomenon

Strigolactones in the Rhizobium-legume symbiosis: Stimulatory effect on bacterial surface motility and down-regulation of their levels in nodulated plants.

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Peláez-Vico, María A; Bernabéu-Roda, Lydia; Kohlen, Wouter; Soto, María J; López-Ráez, Juan A

2016-04-01

Strigolactones (SLs) are multifunctional molecules acting as modulators of plant responses under nutrient deficient conditions. One of the roles of SLs is to promote beneficial association with arbuscular mycorrhizal (AM) fungi belowground under such stress conditions, mainly phosphorus shortage. Recently, a role of SLs in the Rhizobium-legume symbiosis has been also described. While SLs' function in AM symbiosis is well established, their role in the Rhizobium-legume interaction is still emerging. Recently, SLs have been suggested to stimulate surface motility of rhizobia, opening the possibility that they could also act as molecular cues. The possible effect of SLs in the motility in the alfalfa symbiont Sinorhizobium meliloti was investigated, showing that the synthetic SL analogue GR24 stimulates swarming motility in S. meliloti in a dose-dependent manner. On the other hand, it is known that SL production is regulated by nutrient deficient conditions and by AM symbiosis. Using the model alfalfa-S. meliloti, the impact of phosphorus and nitrogen deficiency, as well as of nodulation on SL production was also assessed. The results showed that phosphorus starvation promoted SL biosynthesis, which was abolished by nitrogen deficiency. In addition, a negative effect of nodulation on SL levels was detected, suggesting a conserved mechanism of SL regulation upon symbiosis establishment. Copyright © 2016 Elsevier Ireland Ltd. All rights reserved.

Methods for the Isolation of Genes Encoding Novel PHA Metabolism Enzymes from Complex Microbial Communities.

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Cheng, Jiujun; Nordeste, Ricardo; Trainer, Maria A; Charles, Trevor C

2017-01-01

Development of different PHAs as alternatives to petrochemically derived plastics can be facilitated by mining metagenomic libraries for diverse PHA cycle genes that might be useful for synthesis of bio-plastics. The specific phenotypes associated with mutations of the PHA synthesis pathway genes in Sinorhizobium meliloti and Pseudomonas putida, allows the use of powerful selection and screening tools to identify complementing novel PHA synthesis genes. Identification of novel genes through their function rather than sequence facilitates the functional proteins that may otherwise have been excluded through sequence-only screening methodology. We present here methods that we have developed for the isolation of clones expressing novel PHA metabolism genes from metagenomic libraries.

Methods for the isolation of genes encoding novel PHB cycle enzymes from complex microbial communities.

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Nordeste, Ricardo F; Trainer, Maria A; Charles, Trevor C

2010-01-01

Development of different PHAs as alternatives to petrochemically derived plastics can be facilitated by mining metagenomic libraries for diverse PHA cycle genes that might be useful for synthesis of bioplastics. The specific phenotypes associated with mutations of the PHA synthesis pathway genes in Sinorhizobium meliloti allows for the use of powerful selection and screening tools to identify complementing novel PHA synthesis genes. Identification of novel genes through their function rather than sequence facilitates finding functional proteins that may otherwise have been excluded through sequence-only screening methodology. We present here methods that we have developed for the isolation of clones expressing novel PHA metabolism genes from metagenomic libraries.

Bacterial Molecular Signals in the Sinorhizobium fredii-Soybean Symbiosis

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Francisco J. López-Baena

2016-05-01

Full Text Available Sinorhizobium (Ensifer fredii (S. fredii is a rhizobial species exhibiting a remarkably broad nodulation host-range. Thus, S. fredii is able to effectively nodulate dozens of different legumes, including plants forming determinate nodules, such as the important crops soybean and cowpea, and plants forming indeterminate nodules, such as Glycyrrhiza uralensis and pigeon-pea. This capacity of adaptation to different symbioses makes the study of the molecular signals produced by S. fredii strains of increasing interest since it allows the analysis of their symbiotic role in different types of nodule. In this review, we analyze in depth different S. fredii molecules that act as signals in symbiosis, including nodulation factors, different surface polysaccharides (exopolysaccharides, lipopolysaccharides, cyclic glucans, and K-antigen capsular polysaccharides, and effectors delivered to the interior of the host cells through a symbiotic type 3 secretion system.

The Effects of Clinorotation on the Host Plant, Medicago truncatula, and Its Microbial Symbionts

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Ariel J.C. Dauzart

2016-02-01

Full Text Available Understanding the outcome of the plant-microbe symbiosis in altered gravity is vital to developing life support systems for long-distance space travel and colonization of other planets. Thus, the aim of this research was to understand mutualistic relationships between plants and endophytic microbes under the influence of altered gravity. This project utilized the model tripartite relationship among Medicago truncatula ¬– Sinorhizobium meliloti – Rhizophagus irregularis. Plants were inoculated with rhizobial bacteria (S. meliloti, arbuscular mycorrhizal fungi (R. irregularis, or both microbes, and placed on a rotating clinostat. Vertical and horizontal static controls were also performed. Clinorotation significantly reduced M. truncatula dry mass and fresh mass compared to the static controls. The addition of rhizobia treatments under clinorotation also altered total root length and root-to-shoot fresh mass ratio. Nodule size decreased under rhizobia + clinorotation treatment, and nodule density was significantly decreased compared to the vertical treatment. However, inoculation with arbuscular mycorrhizal fungi was shown to increase biomass accumulation and nodule size. Thus, clinorotation significantly affected M. truncatula and its symbiotic relationships with S. meliloti and R. irregularis. In the long term, the results observed in this clinostat study on the changes of plant-microbe mutualism need to be investigated in spaceflight experiments. Thus, careful consideration of the symbiotic microbes of plants should be included in the design of bioregenerative life support systems needed for space travel.

Exploring the plant-associated bacterial communities in Medicago sativa L

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Pini Francesco

2012-05-01

Full Text Available Abstract Background Plant-associated bacterial communities caught the attention of several investigators which study the relationships between plants and soil and the potential application of selected bacterial species in crop improvement and protection. Medicago sativa L. is a legume crop of high economic importance as forage in temperate areas and one of the most popular model plants for investigations on the symbiosis with nitrogen fixing rhizobia (mainly belonging to the alphaproteobacterial species Sinorhizobium meliloti. However, despite its importance, no studies have been carried out looking at the total bacterial community associated with the plant. In this work we explored for the first time the total bacterial community associated with M. sativa plants grown in mesocosms conditions, looking at a wide taxonomic spectrum, from the class to the single species (S. meliloti level. Results Results, obtained by using Terminal-Restriction Fragment Length Polymorphism (T-RFLP analysis, quantitative PCR and sequencing of 16 S rRNA gene libraries, showed a high taxonomic diversity as well as a dominance by members of the class Alphaproteobacteria in plant tissues. Within Alphaproteobacteria the families Sphingomonadaceae and Methylobacteriaceae were abundant inside plant tissues, while soil Alphaproteobacteria were represented by the families of Hyphomicrobiaceae, Methylocystaceae, Bradyirhizobiaceae and Caulobacteraceae. At the single species level, we were able to detect the presence of S. meliloti populations in aerial tissues, nodules and soil. An analysis of population diversity on nodules and soil showed a relatively low sharing of haplotypes (30-40% between the two environments and between replicate mesocosms, suggesting drift as main force shaping S. meliloti population at least in this system. Conclusions In this work we shed some light on the bacterial communities associated with M. sativa plants, showing that Alphaproteobacteria may

Diversity and numbers of root-nodule bacteria (rhizobia in Polish soils

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Stefan Martyniuk

2011-01-01

Full Text Available Using a sand pouch-plant infection method, populations of several species of root-nodule bacteria (rhizobia were enumerated in eighty soils collected throughout Poland. Rhizobium leguminosarum bv. viciae (symbionts of pea, faba bean, vetch and R. leguminosarum bv. trifolii (symbionts of clover were detected in 77 and 76 soils, respectively. Most of these soils contained moderate and high numbers of these species of the rhizobia. Symbionts of beans, R. leguminosarum bv. phaseoli, were assessed in 76 soils; of this number 15 soils had no detectable populations of bean rhizobia and in 40 soils high or moderate numbers of these bacteria were found. Bradyrhizobium sp. (Lupinus, root-nodule bacteria of lupine and serradella, were absent in 19 soils, out of 80 tested, and 34 soils were colonised by high or moderate populations of bradyrhizobia. Sinorhizobium meliloti, rhizobia nodulating alfalfa, were sparse in the examined soils; with 56 soil containing no detectable numbers of S. meliloti and only 6 soils harbouring high or moderate populations of this species. The estimated numbers of the rhizobia in the studied soils were also related to some physical and chemical properties of these soils.

Symbiotic Properties of Sinorhizobium Fredii, J-TGS50 an Indonesian Soybean Nodule Forming Bacteria

International Nuclear Information System (INIS)

Setiyo Hadi Waluyo

2004-01-01

Green House experiments were conducted to study symbiotic properties of Sinorhizobium Fredii, J-TGS50. Sinorhizobium Fredii USDA 192, USDA 201, USDA 205, USDA 206, USDA 217 and Bradyrhizobium japonicum USDA 110 were used as references. Yeast extract mannitol broth culture of the bacteria were made and used as inoculation for several local and imported soybean varieties used in this study. Plants were harvested at 20 days after inoculation. Number of nodules were counted, fresh weight of nodules and shoot were determined. S. Fredii J-TGS50 and S. Fredii USDA 192, USDA 201, USDA 205, USDA 206, USDA 217 were found different in their symbiotic properties. S. Fredii J-TGS50 formed nodules on same imported soybean. While there were no nodules obtained from the plant inoculated with S. Fredii USDA 192, USDA 201, USDA 205, USDA 206, USDA 217. S. Fredii J-TGS50 and recommended B. Japonicum USDA 110 formed nodule on several local soybean varieties. There was no differences between those two bacteria either in nodulation efficiency or in the effectiveness of the formed nodules. Results of this study can be concluded that S. Fredii, J-TGS50 is a native to Indonesian soil and it is a promising soybean nodule forming bacteria in Indonesia. Using indigenous bacteria is valuable. Since they are mostly more tolerant and adaptable than the introduced ones. An important aspect for the success of Biological Nitrogen Fixation (BNF) is insight in the structure of indigenous soybean rhizobia populations. Study on the biodiversity of soybean rhizobia was important conducted. (author)

Production of nodulation factors by Rhizobium meliloti: fermentation, purification and characterization of glycolipids.

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Kohring, B; Baier, R; Niehaus, K; Pühler, A; Flaschel, E

1997-12-01

Lipooligosaccharides, synthesized by soil bacteria of the genera Rhizobium, are known to have multifunctional effects on a wide variety of plants as signal substances in symbiosis initiation, cell response elicitation and growth regulation. These so called nodulation (Nod-) factors represent interesting biotechnological products with respect to fundamental studies of symbiotic interactions as well as for potential applications. Therefore, a batch fermentation process on a scale of 30 l has been developed by means of the Rhizobium meliloti strain R.m. 1021 (pEK327) strongly overexpressing the genes for the synthesis of Nod factors. Induction by the flavone luteolin led to growth associated production of the lipooligosaccharides. Ultrafiltration was used for separating the biomass from the filtrate containing the extracellular Nod factors. Simultaneously, ultrafiltration reduced the amount of lipophilic substances, which would otherwise interfere with processes downstream. The second separation step consisted in adsorption on XAD-2, a nonspecific hydrophobic adsorptive resin. Adsorption of Nod factors was carried out by batch operation of a stirred tank. Desorption was performed by elution with methanol in a fixed bed column. A semi-preparative reversed phase HPLC (Polygoprep 100-30 C18) was chosen as the final purification step. The Nod factors were obtained after evaporation and lyophilization. Thus, about 600 mg of Nod factors were produced from 20 l of fermentation broth. The Nod factors produced by Rhizobium meliloti R.m. 1021 (pEK327) were identified by liquid secondary ion mass spectrometry and by reversed-phase HPLC as fluorescent derivatives of 2-aminobenzamide. The biological activity of the products was demonstrated by means of the root hair deformation (HAD-) assay.

Rhizobial symbiosis effect on the growth, metal uptake, and antioxidant responses of Medicago lupulina under copper stress.

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Kong, Zhaoyu; Mohamad, Osama Abdalla; Deng, Zhenshan; Liu, Xiaodong; Glick, Bernard R; Wei, Gehong

2015-08-01

The effects of rhizobial symbiosis on the growth, metal uptake, and antioxidant responses of Medicago lupulina in the presence of 200 mg kg(-1) Cu(2+) throughout different stages of symbiosis development were studied. The symbiosis with Sinorhizobium meliloti CCNWSX0020 induced an increase in plant growth and nitrogen content irrespective of the presence of Cu(2+). The total amount of Cu uptake of inoculated plants significantly increased by 34.0 and 120.4% in shoots and roots, respectively, compared with non-inoculated plants. However, although the rhizobial symbiosis promoted Cu accumulation both in shoots and roots, the increase in roots was much higher than in shoots, thus decreasing the translocation factor and helping Cu phytostabilization. The rate of lipid peroxidation was significantly decreased in both shoots and roots of inoculated vs. non-inoculated plants when measured either 8, 13, or 18 days post-inoculation. In comparison with non-inoculated plants, the activities of superoxide dismutase and ascorbate peroxidase of shoots of inoculated plants exposed to excess Cu were significantly elevated at different stages of symbiosis development; similar increases occurred in the activities of superoxide dismutase, catalase, and glutathione reductase of inoculated roots. The symbiosis with S. meliloti CCNWSX0020 also upregulated the corresponding genes involved in antioxidant responses in the plants treated with excess Cu. The results indicated that the rhizobial symbiosis with S. meliloti CCNWSX0020 not only enhanced plant growth and metal uptake but also improved the responses of plant antioxidant defense to excess Cu stress.

Molecular and genetic characterization of the rhizopine catabolism (mocABRC) genes of Rhizobium meliloti L5-30.

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Rossbach, S; Kulpa, D A; Rossbach, U; de Bruijn, F J

1994-10-17

Rhizopine (L-3-O-methyl-scyllo-inosamine, 3-O-MSI) is a symbiosis-specific compound, which is synthesized in nitrogen-fixing nodules of Medicago sativa induced by Rhizobium meliloti strain L5-30. 3-O-MSI is thought to function as an unusual growth substrate for R. meliloti L5-30, which carries a locus (mos) responsible for its synthesis closely linked to a locus (moc) responsible for its degradation. Here, the essential moc genes were delimited by Tn5 mutagenesis and shown to be organized into two regions, separated by 3 kb of DNA. The DNA sequence of a 9-kb fragment spanning the two moc regions was determined, and four genes were identified that play an essential role in rhizopine catabolism (mocABC and mocR). The analysis of the DNA sequence and the amino acid sequence of the deduced protein products revealed that MocA resembles NADH-dependent dehydrogenases. MocB exhibits characteristic features of periplasmic-binding proteins that are components of high-affinity transport systems. MocC does not share significant homology with any protein in the database. MocR shows homology with the GntR class of bacterial regulator proteins. These results suggest that the mocABC genes are involved in the uptake and subsequent degradation of rhizopine, whereas mocR is likely to play a regulatory role.

Host-secreted antimicrobial peptide enforces symbiotic selectivity in Medicago truncatula.

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Wang, Qi; Yang, Shengming; Liu, Jinge; Terecskei, Kata; Ábrahám, Edit; Gombár, Anikó; Domonkos, Ágota; Szűcs, Attila; Körmöczi, Péter; Wang, Ting; Fodor, Lili; Mao, Linyong; Fei, Zhangjun; Kondorosi, Éva; Kaló, Péter; Kereszt, Attila; Zhu, Hongyan

2017-06-27

Legumes engage in root nodule symbioses with nitrogen-fixing soil bacteria known as rhizobia. In nodule cells, bacteria are enclosed in membrane-bound vesicles called symbiosomes and differentiate into bacteroids that are capable of converting atmospheric nitrogen into ammonia. Bacteroid differentiation and prolonged intracellular survival are essential for development of functional nodules. However, in the Medicago truncatula - Sinorhizobium meliloti symbiosis, incompatibility between symbiotic partners frequently occurs, leading to the formation of infected nodules defective in nitrogen fixation (Fix - ). Here, we report the identification and cloning of the M. truncatula NFS2 gene that regulates this type of specificity pertaining to S. meliloti strain Rm41. We demonstrate that NFS2 encodes a nodule-specific cysteine-rich (NCR) peptide that acts to promote bacterial lysis after differentiation. The negative role of NFS2 in symbiosis is contingent on host genetic background and can be counteracted by other genes encoded by the host. This work extends the paradigm of NCR function to include the negative regulation of symbiotic persistence in host-strain interactions. Our data suggest that NCR peptides are host determinants of symbiotic specificity in M. truncatula and possibly in closely related legumes that form indeterminate nodules in which bacterial symbionts undergo terminal differentiation.

Toxic effects of arsenic on Sinorhizobium-Medicago sativa symbiotic interaction

Energy Technology Data Exchange (ETDEWEB)

Pajuelo, Eloisa [Department of Microbiology and Parasitology, Faculty of Pharmacy, University of Seville, 41012 Seville (Spain); Rodriguez-Llorente, Ignacio D. [Department of Microbiology and Parasitology, Faculty of Pharmacy, University of Seville, 41012 Seville (Spain)], E-mail: irodri@us.es; Dary, Mohammed; Palomares, Antonio J. [Department of Microbiology and Parasitology, Faculty of Pharmacy, University of Seville, 41012 Seville (Spain)

2008-07-15

Recently, the Rhizobium-legume symbiotic interaction has been proposed as an interesting tool in bioremediation. However, little is known about the effect of most common contaminants on this process. The phytotoxic effects of arsenic on nodulation of Medicago sativa have been examined in vitro using the highly arsenic resistant and symbiotically effective Sinorhizobium sp. strain MA11. The bacteria were able to grow on plates containing As concentrations as high as 10 mM. Nevertheless, as little as 25-35 {mu}M arsenite produced a 75% decrease in the total number of nodules, due to a 90% reduction in the number of rhizobial infections, as could be determined using the strain MA11 carrying a lacZ reporter gene. This effect was associated to root hair damage and a shorter infective root zone. However, once nodulation was established nodule development seemed to continue normally, although earlier senescence could be observed in nodules of arsenic-grown plants. - First steps of nodulation of alfalfa, in particular infection thread formation, are more sensitive to As than nitrogen fixation due to plant effects.

Three phylogenetic groups of nodA and nifH genes in Sinorhizobium and Mesorhizobium isolates from leguminous trees growing in Africa and Latin America.

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Haukka, K; Lindström, K; Young, J P

1998-02-01

The diversity and phylogeny of nodA and nifH genes were studied by using 52 rhizobial isolates from Acacia senegal, Prosopis chilensis, and related leguminous trees growing in Africa and Latin America. All of the strains had similar host ranges and belonged to the genera Sinorhizobium and Mesorhizobium, as previously determined by 16S rRNA gene sequence analysis. The restriction patterns and a sequence analysis of the nodA and nifH genes divided the strains into the following three distinct groups: sinorhizobia from Africa, sinorhizobia from Latin America, and mesorhizobia from both regions. In a phylogenetic tree also containing previously published sequences, the nodA genes of our rhizobia formed a branch of their own, but within the branch no correlation between symbiotic genes and host trees was apparent. Within the large group of African sinorhizobia, similar symbiotic gene types were found in different chromosomal backgrounds, suggesting that transfer of symbiotic genes has occurred across species boundaries. Most strains had plasmids, and the presence of plasmid-borne nifH was demonstrated by hybridization for some examples. The nodA and nifH genes of Sinorhizobium teranga ORS1009T grouped with the nodA and nifH genes of the other African sinorhizobia, but Sinorhizobium saheli ORS609T had a totally different nodA sequence, although it was closely related based on the 16S rRNA gene and nifH data. This might be because this S. saheli strain was originally isolated from Sesbania sp., which belongs to a different cross-nodulation group than Acacia and Prosopis spp. The factors that appear to have influenced the evolution of rhizobial symbiotic genes vary in importance at different taxonomic levels.

An antimicrobial peptide essential for bacterial survival in the nitrogen-fixing symbiosis.

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Kim, Minsoo; Chen, Yuhui; Xi, Jiejun; Waters, Christopher; Chen, Rujin; Wang, Dong

2015-12-08

In the nitrogen-fixing symbiosis between legume hosts and rhizobia, the bacteria are engulfed by a plant cell membrane to become intracellular organelles. In the model legume Medicago truncatula, internalization and differentiation of Sinorhizobium (also known as Ensifer) meliloti is a prerequisite for nitrogen fixation. The host mechanisms that ensure the long-term survival of differentiating intracellular bacteria (bacteroids) in this unusual association are unclear. The M. truncatula defective nitrogen fixation4 (dnf4) mutant is unable to form a productive symbiosis, even though late symbiotic marker genes are expressed in mutant nodules. We discovered that in the dnf4 mutant, bacteroids can apparently differentiate, but they fail to persist within host cells in the process. We found that the DNF4 gene encodes NCR211, a member of the family of nodule-specific cysteine-rich (NCR) peptides. The phenotype of dnf4 suggests that NCR211 acts to promote the intracellular survival of differentiating bacteroids. The greatest expression of DNF4 was observed in the nodule interzone II-III, where bacteroids undergo differentiation. A translational fusion of DNF4 with GFP localizes to the peribacteroid space, and synthetic NCR211 prevents free-living S. meliloti from forming colonies, in contrast to mock controls, suggesting that DNF4 may interact with bacteroids directly or indirectly for its function. Our findings indicate that a successful symbiosis requires host effectors that not only induce bacterial differentiation, but also that maintain intracellular bacteroids during the host-symbiont interaction. The discovery of NCR211 peptides that maintain bacterial survival inside host cells has important implications for improving legume crops.

The Effects of Clinorotation on the Host Plant, Medicago truncatula, and Its Microbial Symbionts

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Dauzart, Ariel J. C.; Vandenbrink, Joshua P.; Kiss, John Z., E-mail: jzkiss@olemiss.edu [Department of Biology, Graduate School, University of Mississippi, University, MS (United States)

2016-02-26

Understanding the outcome of the plant-microbe symbiosis in reduced or altered is vital to developing life support systems for long-distance space travel and colonization of other planets. Thus, the aim of this research was to understand mutualistic relationships between plants and endophytic microbes under the influence of altered gravity. This project utilized the model tripartite relationship among Medicago truncatula—Sinorhizobium meliloti—Rhizophagus irregularis. Plants were inoculated with rhizobial bacteria (S. meliloti), arbuscular mycorrhizal fungi (R. irregularis), or both microbes, and placed on a rotating clinostat. Vertical and horizontal static controls were also performed. Clinorotation significantly reduced M. truncatula dry mass and fresh mass compared to the static controls. The addition of rhizobia treatments under clinorotation also altered total root length and root-to-shoot fresh mass ratio. Nodule size decreased under rhizobia + clinorotation treatment, and nodule density was significantly decreased compared to the vertical treatment. However, inoculation with arbuscular mycorrhizal fungi was shown to increase biomass accumulation and nodule size. Thus, clinorotation significantly affected M. truncatula and its symbiotic relationships with S. meliloti and R. irregularis. In the long term, the results observed in this clinostat study on the changes of plant-microbe mutualism need to be investigated in spaceflight experiments. Thus, careful consideration of the symbiotic microbes of plants should be included in the design of bioregenerative life support systems needed for space travel.

Co-inoculation of arbusculr mycorrhizae and nitrogen fixing bacteria enhance alfalfa yield under saline conditions

International Nuclear Information System (INIS)

Zhu, R.; Tang, F.; Liu, F.; Chen, J.

2016-01-01

The study was to investigate the effects of combined inoculation of Glomus mosseae (arbusculr mycorrhizae fungi, AMF) and Sinorhizobium meliloti (nitrogen-fixing bacteria, i.e., an Rhizobium meliloti, RM) on yield, nutrient contents, nodulation and mycorrhizal colonization of different alfalfa cultivars under saline conditions. An experiment was conducted to test the efficacy of AMF and RM inoculation in development of salt tolerance in alfalfa cultivars (Zhaodong, Nongjing and Longmu) under different salinity levels (0, 60, 120 and 180 mM NaCl). We found that under non stress condition, double inoculation of alfalfa with rhizobium and AM increased the alfalfa yield, nodule weight and number, as well as shoot proline contents, the most when plants were double inoculated followed by AM and rhizobium inoculation, respectively. Whereas under salinity condition, double inoculation of alfalfa with rhizobium and AM increased alfalfa yield, mycorrhizal infection, nodule weight and number as well as increased in shoot proline content, the most followed by AM and rhizobium inoculation, respectively. The Results suggest that growth of alfalfa may be improved by combined inoculation of alfalfa with AM and rhizobium under salt and non-stress conditions. Alleviation of alfalfa growth under saline condition was perhaps due to an increase in mycorrhizal infection and nodule weight and number as well as an increased in shoot proline content by dual inoculation. (author)

The Effects of Clinorotation on the Host Plant, Medicago truncatula, and Its Microbial Symbionts

International Nuclear Information System (INIS)

Dauzart, Ariel J. C.; Vandenbrink, Joshua P.; Kiss, John Z.

2016-01-01

Understanding the outcome of the plant-microbe symbiosis in reduced or altered is vital to developing life support systems for long-distance space travel and colonization of other planets. Thus, the aim of this research was to understand mutualistic relationships between plants and endophytic microbes under the influence of altered gravity. This project utilized the model tripartite relationship among Medicago truncatula—Sinorhizobium meliloti—Rhizophagus irregularis. Plants were inoculated with rhizobial bacteria (S. meliloti), arbuscular mycorrhizal fungi (R. irregularis), or both microbes, and placed on a rotating clinostat. Vertical and horizontal static controls were also performed. Clinorotation significantly reduced M. truncatula dry mass and fresh mass compared to the static controls. The addition of rhizobia treatments under clinorotation also altered total root length and root-to-shoot fresh mass ratio. Nodule size decreased under rhizobia + clinorotation treatment, and nodule density was significantly decreased compared to the vertical treatment. However, inoculation with arbuscular mycorrhizal fungi was shown to increase biomass accumulation and nodule size. Thus, clinorotation significantly affected M. truncatula and its symbiotic relationships with S. meliloti and R. irregularis. In the long term, the results observed in this clinostat study on the changes of plant-microbe mutualism need to be investigated in spaceflight experiments. Thus, careful consideration of the symbiotic microbes of plants should be included in the design of bioregenerative life support systems needed for space travel.

The alternative Medicago truncatula defense proteome of ROS – defective transgenic roots during early microbial infection

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Leonard Muriithi Kiirika

2014-07-01

Full Text Available ROP-type GTPases of plants function as molecular switches within elementary signal transduction pathways such as the regulation of ROS synthesis via activation of NADPH oxidases (RBOH-respiratory burst oxidase homologue in plants. Previously, we reported that silencing of the Medicago truncatula GTPase MtROP9 led to reduced ROS production and suppressed induction of ROS-related enzymes in transgenic roots (MtROP9i infected with pathogenic (Aphanomyces euteiches and symbiotic microorganisms (Glomus intraradices, Sinorhizobium meliloti. While fungal infections were enhanced, S. meliloti infection was drastically impaired. In this study, we investigate the temporal proteome response of M. truncatula MtROP9i transgenic roots during the same microbial interactions under conditions of deprived potential to synthesize ROS. In comparison with control roots (Mtvector, we present a comprehensive proteomic analysis using sensitive MS protein identification. For four early infection time-points (1, 3, 5, 24 hpi, 733 spots were found to be different in abundance: 213 spots comprising 984 proteins (607 unique were identified after S. meliloti infection, 230 spots comprising 796 proteins (580 unique after G. intraradices infection, and 290 spots comprising 1240 proteins (828 unique after A. euteiches infection. Data evaluation by GelMap in combination with a heatmap tool allowed recognition of key proteome changes during microbial interactions under conditions of hampered ROS synthesis. Overall, the number of induced proteins in MtROP9i was low as compared with controls, indicating a dual function of ROS in defense signaling as well as alternative response patterns activated during microbial infection. Qualitative analysis of induced proteins showed that enzymes linked to ROS production and scavenging were highly induced in control roots, while in MtROP9i the majority of proteins were involved in alternative defense pathways such as cell wall and protein

Effectivenes of inoculation in alfalfa breeding in ecological conditions of the Bjelovar and Bilogora county

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Darko Uher

2012-09-01

Full Text Available Development and basic existence of animal production as well as production of high quality milk depends upon possibility of sufficient production of quality and protein sufficient forage. Forage crop that satisfies these demands is alfalfa which is one of the most important perennial forage crop legumes. The aim of this study was to enhance alfalfa production on acid soil by liming and alfalfa seed inoculation with efficient Sinorhizobium meliloti strains in order to reduce the use of mineral nitrogen fertilization and enable qualitative and cost effective production of forage on the dairy farms. Field trial was established at family farm in the area of Bjelovar and Bilogora county. During two years experimental period statistically significant influence of inoculation and liming on forage and dry matteryield was determined. Significantly the lowest yields were determined on untreated plots without liming material. In all untreated plots, significantly lower yields were determined, but significant differences in yields were also obtained by inoculation with different S. meliloti strains, emphasizing the importance of strains selection used for alfalfa inoculation. In both experimental years total forage yield were ranging from 34 t/ha (untreated plots without liming up to 60 t/ha on plots inoculated with strain 2011 and without liming. Values of total dry matter yield for both experimental years ranged from 6.5 t/ha (untreated plots without liming up to 15,7 t/ha on plots inoculated with strain 2011 without liming. Results of this study showed that application of liming materials for acidity removal had positive effect on alfalfa yields in both experimental years and significantly improved alfalfa production on acid soils. The results of this study clearly showed that inoculation with selected S. meliloti strains may improve alfalfa production on acid soils and may contribute to more efficient forage production for dairy farms under particular

Colorimetric Detection of Some Highly Hydrophobic Flavonoids Using Polydiacetylene Liposomes Containing Pentacosa-10,12-diynoyl Succinoglycan Monomers

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Yun, Deokgyu; Jeong, Daham; Cho, Eunae; Jung, Seunho

2015-01-01

Flavonoids are a group of plant secondary metabolites including polyphenolic molecules, and they are well known for antioxidant, anti-allergic, anti-inflammatory and anti-viral propertied. In general, flavonoids are detected with various non-colorimetric detection methods such as column liquid chromatography, thin-layer chromatography, and electrochemical analysis. For the first time, we developed a straightforward colorimetric detection system allowing recognition of some highly hydrophobic flavonoids such as alpha-naphthoflavone and beta-naphthoflavone, visually using 10,12-pentacosadiynoic acid (PCDA) derivatized with succinoglycan monomers isolated from Sinorhizobium meliloti. Besides changes in visible spectrum, we also demonstrate fluorescence changes using our detection system in the presence of those flavonoids. The succinoglycan monomers attached to PCDA molecules may function as an unstructured molecular capturer for some highly hydrophobic flavonoids by hydrophobic interactions, and transmit their molecular interactions as a color change throughout the PCDA liposome. PMID:26600071

NPR1 protein regulates pathogenic and symbiotic interactions between Rhizobium and legumes and non-legumes.

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Smadar Peleg-Grossman

Full Text Available BACKGROUND: Legumes are unique in their ability to establish symbiotic interaction with rhizobacteria from Rhizobium genus, which provide them with available nitrogen. Nodulation factors (NFs produced by Rhizobium initiate legume root hair deformation and curling that entrap the bacteria, and allow it to grow inside the plant. In contrast, legumes and non-legumes activate defense responses when inoculated with pathogenic bacteria. One major defense pathway is mediated by salicylic acid (SA. SA is sensed and transduced to downstream defense components by a redox-regulated protein called NPR1. METHODOLOGY/PRINCIPAL FINDINGS: We used Arabidopsis mutants in SA defense pathway to test the role of NPR1 in symbiotic interactions. Inoculation of Sinorhizobium meliloti or purified NF on Medicago truncatula or nim1/npr1 A. thaliana mutants induced root hair deformation and transcription of early and late nodulins. Application of S. meliloti or NF on M. truncatula or A. thaliana roots also induced a strong oxidative burst that lasted much longer than in plants inoculated with pathogenic or mutualistic bacteria. Transient overexpression of NPR1 in M. truncatula suppressed root hair curling, while inhibition of NPR1 expression by RNAi accelerated curling. CONCLUSIONS/SIGNIFICANCE: We show that, while NPR1 has a positive effect on pathogen resistance, it has a negative effect on symbiotic interactions, by inhibiting root hair deformation and nodulin expression. Our results also show that basic plant responses to Rhizobium inoculation are conserved in legumes and non-legumes.

The nitrate-reduction gene cluster components exert lineage-dependent contributions to optimization of Sinorhizobium symbiosis with soybeans.

Science.gov (United States)

Liu, Li Xue; Li, Qin Qin; Zhang, Yun Zeng; Hu, Yue; Jiao, Jian; Guo, Hui Juan; Zhang, Xing Xing; Zhang, Biliang; Chen, Wen Xin; Tian, Chang Fu

2017-12-01

Receiving nodulation and nitrogen fixation genes does not guarantee rhizobia an effective symbiosis with legumes. Here, variations in gene content were determined for three Sinorhizobium species showing contrasting symbiotic efficiency on soybeans. A nitrate-reduction gene cluster absent in S. sojae was found to be essential for symbiotic adaptations of S. fredii and S. sp. III. In S. fredii, the deletion mutation of the nap (nitrate reductase), instead of nir (nitrite reductase) and nor (nitric oxide reductase), led to defects in nitrogen-fixation (Fix - ). By contrast, none of these core nitrate-reduction genes were required for the symbiosis of S. sp. III. However, within the same gene cluster, the deletion of hemN1 (encoding oxygen-independent coproporphyrinogen III oxidase) in both S. fredii and S. sp. III led to the formation of nitrogen-fixing (Fix + ) but ineffective (Eff - ) nodules. These Fix + /Eff - nodules were characterized by significantly lower enzyme activity of glutamine synthetase indicating rhizobial modulation of nitrogen-assimilation by plants. A distant homologue of HemN1 from S. sojae can complement this defect in S. fredii and S. sp. III, but exhibited a more pleotropic role in symbiosis establishment. These findings highlighted the lineage-dependent optimization of symbiotic functions in different rhizobial species associated with the same host. © 2017 Society for Applied Microbiology and John Wiley & Sons Ltd.

High resolution imaging of surface patterns of single bacterial cells

International Nuclear Information System (INIS)

Greif, Dominik; Wesner, Daniel; Regtmeier, Jan; Anselmetti, Dario

2010-01-01

We systematically studied the origin of surface patterns observed on single Sinorhizobium meliloti bacterial cells by comparing the complementary techniques atomic force microscopy (AFM) and scanning electron microscopy (SEM). Conditions ranged from living bacteria in liquid to fixed bacteria in high vacuum. Stepwise, we applied different sample modifications (fixation, drying, metal coating, etc.) and characterized the observed surface patterns. A detailed analysis revealed that the surface structure with wrinkled protrusions in SEM images were not generated de novo but most likely evolved from similar and naturally present structures on the surface of living bacteria. The influence of osmotic stress to the surface structure of living cells was evaluated and also the contribution of exopolysaccharide and lipopolysaccharide (LPS) by imaging two mutant strains of the bacterium under native conditions. AFM images of living bacteria in culture medium exhibited surface structures of the size of single proteins emphasizing the usefulness of AFM for high resolution cell imaging.

High-quality forage production under salinity by using a salt-tolerant AtNXH1-expressing transgenic alfalfa combined with a natural stress-resistant nitrogen-fixing bacterium.

Science.gov (United States)

Stritzler, Margarita; Elba, Pagano; Berini, Carolina; Gomez, Cristina; Ayub, Nicolás; Soto, Gabriela

2018-06-20

Alfalfa, usually known as the "Queen of Forages", is the main source of vegetable protein to meat and milk production systems worldwide. This legume is extremely rich in proteins due to its highly efficient symbiotic association with nitrogen-fixing strains. In the last years, alfalfa culture has been displaced to saline environments by other important crops, including major cereals, a fact that has reduced its biomass production and symbiotic nitrogen fixation. In this short communication, we report the high forage production and nutrient quality of alfalfa under saline conditions by alfalfa transformation with the AtNHX1 Na + /H + antiporter and inoculation with the stress-resistant nitrogen-fixing strain Sinorhizobium meliloti B401. Therefore, the incorporation of transgenic traits into salt-sensitive legumes in association with the inoculation with natural stress-resistant isolates could be a robust approach to improve the productivity and quality of these important nitrogen-fixing crops. Copyright © 2018. Published by Elsevier B.V.

A bioassay for the detection of benzimidazoles reveals their presence in a range of environmental samples

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Terence S Crofts

2014-11-01

Full Text Available Cobamides are a family of enzyme cofactors that include vitamin B12 (cobalamin and are produced solely by prokaryotes. Structural variability in the lower axial ligand has been observed in cobamides produced by diverse organisms. Of the three classes of lower ligands, the benzimidazoles are uniquely found in cobamides, whereas the purine and phenolic bases have additional biological functions. Many organisms acquire cobamides by salvaging and remodeling cobamides or their precursors from the environment. These processes require free benzimidazoles for incorporation as lower ligands, though the presence of benzimidazoles in the environment has not been previously investigated. Here, we report a new purification method and bioassay to measure the total free benzimidazole content of samples from microbial communities and laboratory media components. The bioassay relies on the calcofluor-bright phenotype of a bluB mutant of the model cobalamin-producing bacterium Sinorhizobium meliloti. The concentrations of individual benzimidazoles in these samples were measured by liquid chromatography-tandem mass spectrometry. Several benzimidazoles were detected in subpicomolar to subnanomolar concentrations in host-associated and environmental samples. In addition, benzimidazoles were found to be common contaminants of laboratory media components. These results suggest that benzimidazoles present in the environment and in laboratory media have the potential to influence microbial metabolic activities.

Bacterial PerO Permeases Transport Sulfate and Related Oxyanions.

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Hoffmann, Marie-Christine; Pfänder, Yvonne; Tintel, Marc; Masepohl, Bernd

2017-07-15

Rhodobacter capsulatus synthesizes the high-affinity ABC transporters CysTWA and ModABC to specifically import the chemically related oxyanions sulfate and molybdate, respectively. In addition, R. capsulatus has the low-affinity permease PerO acting as a general oxyanion transporter, whose elimination increases tolerance to molybdate and tungstate. Although PerO-like permeases are widespread in bacteria, their function has not been examined in any other species to date. Here, we present evidence that PerO permeases from the alphaproteobacteria Agrobacterium tumefaciens , Dinoroseobacter shibae , Rhodobacter sphaeroides , and Sinorhizobium meliloti and the gammaproteobacterium Pseudomonas stutzeri functionally substitute for R. capsulatus PerO in sulfate uptake and sulfate-dependent growth, as shown by assimilation of radioactively labeled sulfate and heterologous complementation. Disruption of perO genes in A. tumefaciens , R. sphaeroides , and S. meliloti increased tolerance to tungstate and, in the case of R. sphaeroides , to molybdate, suggesting that heterometal oxyanions are common substrates of PerO permeases. This study supports the view that bacterial PerO permeases typically transport sulfate and related oxyanions and, hence, form a functionally conserved permease family. IMPORTANCE Despite the widespread distribution of PerO-like permeases in bacteria, our knowledge about PerO function until now was limited to one species, Rhodobacter capsulatus In this study, we showed that PerO proteins from diverse bacteria are functionally similar to the R. capsulatus prototype, suggesting that PerO permeases form a conserved family whose members transport sulfate and related oxyanions. Copyright © 2017 American Society for Microbiology.

Efecto de la inoculación con rizobios procedentes de Alberta, Canadá, en sorgo (Sorghum bicolor L. Moench, en condiciones de campo Effect of the inoculation with rhizobia from Alberta, Canada, in sorghum (Sorghum bicolor L. Moench, under field conditions

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C. J Bécquer

2011-09-01

Full Text Available Se realizó un experimento de campo con el objetivo de medir el efecto de cepas de rizobio en las variables agronómicas del sorgo, en las condiciones ambientales de Sancti Spíritus, Cuba. Se utilizaron 10 cepas de Sinorhizobium meliloti, procedentes de ecosistemas ganaderos de Alberta, Canadá; así como cuatro cepas de referencia pertenecientes a diferentes géneros y especies de rizobio, que procedían de la colección de Agriculture and AgriFood Canada. La confección de los inóculos y la inoculación de las semillas se realizaron por métodos estándar. El diseño experimental fue de bloques al azar, con 16 tratamientos y cuatro réplicas. Se evaluó el peso seco aéreo, la longitud del tallo y la longitud de la panoja; además, se calculó el incremento del peso seco aéreo en los tratamientos inoculados con relación al control absoluto. Los resultados demostraron la capacidad de las cepas estudiadas de influir en las variables agronómicas, ya que los tratamientos seleccionados igualaron sus valores a los del control fertilizado y presentaron un incremento de más del 100% del peso seco aéreo, comparado con el control absoluto.A field trial was conducted with the objective of measuring the effect of rhizobium strains on the agronomic variables of sorghum under the environmental conditions of Sancti Spiritus, Cuba. Ten Sinorhizobium meliloti strains, from livestock production ecosystems of Alberta, Canada, were used; as well as four reference strains belonging to different rhizobium genera and strains, which were from the collection of Agriculture and AgriFood Canada. The inoculi confection and seed inoculation were made by standard methods. The experimental design was randomized blocks, with 16 treatments and four replications. The dry aerial weight, stem length and ear length were evaluated; in addition, the increase of aerial dry weight was calculated in the inoculated treatments as compared to the absolute control. The results

Detección de actividad pectolítica en el cultivo de la cepa GR4 de rhyzobíum meliloti

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P. Martínez M.

2010-07-01

Full Text Available Se ensayaron varios métodos para la obtención y purificación parcial de pectinasas a partir de sobrenadante del cultivo de la cepa GR4 de Rhizobium meliloti. Se describe el método con el cual se obtuvo el sobrenadante en el que se logró detectar la presencia de actividad pectolítica. Empleando una muestra comercial de enzimas pécticas (Pectinex, Novo se estudió la estabilidad de la actividad enzimática durante el proceso de purificación parcial establecido; se observó una pérdida gradual de la actividad en función del tiempo de duración del proceso.

Molecular characterization of the pSinB plasmid of the arsenite oxidizing, metallotolerant Sinorhizobium sp. M14 - insight into the heavy metal resistome of sinorhizobial extrachromosomal replicons.

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Romaniuk, Krzysztof; Dziewit, Lukasz; Decewicz, Przemyslaw; Mielnicki, Sebastian; Radlinska, Monika; Drewniak, Lukasz

2017-01-01

Sinorhizobium sp. M14 is an As(III)-oxidizing, psychrotolerant strain, capable of growth in the presence of extremely high concentrations of arsenic and many other heavy metals. Metallotolerant abilities of the M14 strain depend upon the presence of two extrachromosomal replicons: pSinA (∼ 109 kb) and pSinB (∼ 300 kb). The latter was subjected to complex analysis. The performed analysis demonstrated that the plasmid pSinB is a narrow-host-range repABC-type replicon, which is fully stabilized by the phd-vapC-like toxin-antitoxin stabilizing system. In silico analysis showed that among the phenotypic gene clusters of the plasmid pSinB, eight modules are potentially involved in heavy metals resistance (HMR). These modules carry genes encoding efflux pumps, permeases, transporters and copper oxidases, which provide resistance to arsenic, cadmium, cobalt, copper, iron, mercury, nickel, silver and zinc. The functional analysis revealed that the HMR modules are active and have an effect on the minimal inhibitory concentration (MIC) values observed for the heterological host cells. The phenotype was manifested by an increase or decrease of the MICs of heavy metals and it was strain specific. The analysis of distribution of the heavy metal resistance genes, i.e. resistome, in Sinorhizobium spp. plasmids, revealed that the HMR modules are common in these replicons. © FEMS 2016. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com.

Host Factors Influencing the Retrohoming Pathway of Group II Intron RmInt1, Which Has an Intron-Encoded Protein Naturally Devoid of Endonuclease Activity.

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Rafael Nisa-Martínez

Full Text Available Bacterial group II introns are self-splicing catalytic RNAs and mobile retroelements that have an open reading frame encoding an intron-encoded protein (IEP with reverse transcriptase (RT and RNA splicing or maturase activity. Some IEPs carry a DNA endonuclease (En domain, which is required to cleave the bottom strand downstream from the intron-insertion site for target DNA-primed reverse transcription (TPRT of the inserted intron RNA. Host factors complete the insertion of the intron. By contrast, the major retrohoming pathway of introns with IEPs naturally lacking endonuclease activity, like the Sinorhizobium meliloti intron RmInt1, is thought to involve insertion of the intron RNA into the template for lagging strand DNA synthesis ahead of the replication fork, with possible use of the nascent strand to prime reverse transcription of the intron RNA. The host factors influencing the retrohoming pathway of such introns have not yet been described. Here, we identify key candidates likely to be involved in early and late steps of RmInt1 retrohoming. Some of these host factors are common to En+ group II intron retrohoming, but some have different functions. Our results also suggest that the retrohoming process of RmInt1 may be less dependent on the intracellular free Mg2+ concentration than those of other group II introns.

A model for the catabolism of rhizopine in Rhizobium leguminosarum involves a ferredoxin oxygenase complex and the inositol degradative pathway.

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Bahar, M; de Majnik, J; Wexler, M; Fry, J; Poole, P S; Murphy, P J

1998-11-01

Rhizopines are nodule-specific compounds that confer an intraspecies competitive nodulation advantage to strains that can catabolize them. The rhizopine (3-O-methyl-scyllo-inosamine, 3-O-MSI) catabolic moc gene cluster mocCABRDE(F) in Rhizobium leguminosarum bv. viciae strain 1a is located on the Sym plasmid. MocCABR are homologous to the mocCABR gene products from Sinorhizobium meliloti. MocD and MocE contain motifs corresponding to a TOL-like oxygenase and a [2Fe-2S] Rieske-like ferredoxin, respectively. The mocF gene encodes a ferredoxin reductase that would complete the oxygenase system, but is not essential for rhizopine catabolism. We propose a rhizopine catabolic model whereby MocB transports rhizopine into the cell and MocDE and MocF (or a similar protein elsewhere in the genome), under the regulation of MocR, act in concert to form a ferredoxin oxygenase system that demethylates 3-O-MSI to form scyllo-inosamine (SI). MocA, an NAD(H)-dependent dehydrogenase, and MocC continue the catabolic process. Compounds formed then enter the inositol catabolic pathway.

Characterization of Crude Oil Degrading Bacteria Isolated from Contaminated Soils Surrounding Gas Stations.

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Abou-Shanab, Reda A I; Eraky, Mohamed; Haddad, Ahmed M; Abdel-Gaffar, Abdel-Rahman B; Salem, Ahmed M

2016-11-01

A total of twenty bacterial cultures were isolated from hydrocarbon contaminated soil. Of the 20 isolates, RAM03, RAM06, RAM13, and RAM17 were specifically chosen based on their relatively higher growth on salt medium amended with 4 % crude oil, emulsion index, surface tension, and degradation percentage. These bacterial cultures had 16S rRNA gene sequences that were most similar to Ochrobactrum cytisi (RAM03), Ochrobactrum anthropi (RAM06 and RAM17), and Sinorhizobium meliloti (RAM13) with 96 %, 100 % and 99 %, and 99 % similarity. The tested strains revealed a promising potential for bioremediation of petroleum oil contamination as they could degrade >93 % and 54 % of total petroleum hydrocarbons (TPHs) in a liquid medium and soil amended with 4 % crude oil, respectively, after 30 day incubation. These bacteria could effectively remove both aliphatic and aromatic petroleum hydrocarbons. In conclusion, these strains could be considered as good prospects for their application in bioremediation of hydrocarbon contaminated environment.

Genomic analyses of metal resistance genes in three plant growth promoting bacteria of legume plants in Northwest mine tailings, China.

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Xie, Pin; Hao, Xiuli; Herzberg, Martin; Luo, Yantao; Nies, Dietrich H; Wei, Gehong

2015-01-01

To better understand the diversity of metal resistance genetic determinant from microbes that survived at metal tailings in northwest of China, a highly elevated level of heavy metal containing region, genomic analyses was conducted using genome sequence of three native metal-resistant plant growth promoting bacteria (PGPB). It shows that: Mesorhizobium amorphae CCNWGS0123 contains metal transporters from P-type ATPase, CDF (Cation Diffusion Facilitator), HupE/UreJ and CHR (chromate ion transporter) family involved in copper, zinc, nickel as well as chromate resistance and homeostasis. Meanwhile, the putative CopA/CueO system is expected to mediate copper resistance in Sinorhizobium meliloti CCNWSX0020 while ZntA transporter, assisted with putative CzcD, determines zinc tolerance in Agrobacterium tumefaciens CCNWGS0286. The greenhouse experiment provides the consistent evidence of the plant growth promoting effects of these microbes on their hosts by nitrogen fixation and/or indoleacetic acid (IAA) secretion, indicating a potential in-site phytoremediation usage in the mining tailing regions of China. Copyright © 2014. Published by Elsevier B.V.

A FIELD STUDY WITH GENETICALLY ENGINEERED ALFALFA INOCULATED WITH RECOMBINANT SINORHIZOBIUM MELILOTI: EFFECTS ON THE SOIL ECOSYSTEM

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The agricultural use of genetically engineered plants and microorganisms has become increasingly common. Because genetically engineered plants and microorganisms can produce compounds foreign to their environment, there is concern that they may become established outside of thei...

Polyphosphate-dependent synthesis of ATP and ADP by the family-2 polyphosphate kinases in bacteria.

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Nocek, Boguslaw; Kochinyan, Samvel; Proudfoot, Michael; Brown, Greg; Evdokimova, Elena; Osipiuk, Jerzy; Edwards, Aled M; Savchenko, Alexei; Joachimiak, Andrzej; Yakunin, Alexander F

2008-11-18

Inorganic polyphosphate (polyP) is a linear polymer of tens or hundreds of phosphate residues linked by high-energy bonds. It is found in all organisms and has been proposed to serve as an energy source in a pre-ATP world. This ubiquitous and abundant biopolymer plays numerous and vital roles in metabolism and regulation in prokaryotes and eukaryotes, but the underlying molecular mechanisms for most activities of polyP remain unknown. In prokaryotes, the synthesis and utilization of polyP are catalyzed by 2 families of polyP kinases, PPK1 and PPK2, and polyphosphatases. Here, we present structural and functional characterization of the PPK2 family. Proteins with a single PPK2 domain catalyze polyP-dependent phosphorylation of ADP to ATP, whereas proteins containing 2 fused PPK2 domains phosphorylate AMP to ADP. Crystal structures of 2 representative proteins, SMc02148 from Sinorhizobium meliloti and PA3455 from Pseudomonas aeruginosa, revealed a 3-layer alpha/beta/alpha sandwich fold with an alpha-helical lid similar to the structures of microbial thymidylate kinases, suggesting that these proteins share a common evolutionary origin and catalytic mechanism. Alanine replacement mutagenesis identified 9 conserved residues, which are required for activity and include the residues from both Walker A and B motifs and the lid. Thus, the PPK2s represent a molecular mechanism, which potentially allow bacteria to use polyP as an intracellular energy reserve for the generation of ATP and survival.

Rhizobial peptidase HrrP cleaves host-encoded signaling peptides and mediates symbiotic compatibility.

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Price, Paul A; Tanner, Houston R; Dillon, Brett A; Shabab, Mohammed; Walker, Graham C; Griffitts, Joel S

2015-12-08

Legume-rhizobium pairs are often observed that produce symbiotic root nodules but fail to fix nitrogen. Using the Sinorhizobium meliloti and Medicago truncatula symbiotic system, we previously described several naturally occurring accessory plasmids capable of disrupting the late stages of nodule development while enhancing bacterial proliferation within the nodule. We report here that host range restriction peptidase (hrrP), a gene found on one of these plasmids, is capable of conferring both these properties. hrrP encodes an M16A family metallopeptidase whose catalytic activity is required for these symbiotic effects. The ability of hrrP to suppress nitrogen fixation is conditioned upon the genotypes of both the host plant and the hrrP-expressing rhizobial strain, suggesting its involvement in symbiotic communication. Purified HrrP protein is capable of degrading a range of nodule-specific cysteine-rich (NCR) peptides encoded by M. truncatula. NCR peptides are crucial signals used by M. truncatula for inducing and maintaining rhizobial differentiation within nodules, as demonstrated in the accompanying article [Horváth B, et al. (2015) Proc Natl Acad Sci USA, 10.1073/pnas.1500777112]. The expression pattern of hrrP and its effects on rhizobial morphology are consistent with the NCR peptide cleavage model. This work points to a symbiotic dialogue involving a complex ensemble of host-derived signaling peptides and bacterial modifier enzymes capable of adjusting signal strength, sometimes with exploitative outcomes.

A multicopper oxidase is essential for manganese oxidation and laccase-like activity in Pedomicrobium sp. ACM 3067.

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Ridge, Justin P; Lin, Marianne; Larsen, Eloise I; Fegan, Mark; McEwan, Alastair G; Sly, Lindsay I

2007-04-01

Pedomicrobium sp. ACM 3067 is a budding-hyphal bacterium belonging to the alpha-Proteobacteria which is able to oxidize soluble Mn2+ to insoluble manganese oxide. A cosmid, from a whole-genome library, containing the putative genes responsible for manganese oxidation was identified and a primer-walking approach yielded 4350 bp of novel sequence. Analysis of this sequence showed the presence of a predicted three-gene operon, moxCBA. The moxA gene product showed homology to multicopper oxidases (MCOs) and contained the characteristic four copper-binding motifs (A, B, C and D) common to MCOs. An insertion mutation of moxA showed that this gene was essential for both manganese oxidation and laccase-like activity. The moxB gene product showed homology to a family of outer membrane proteins which are essential for Type I secretion in Gram-negative bacteria. moxBA has not been observed in other manganese-oxidizing bacteria but homologues were identified in the genomes of several bacteria including Sinorhizobium meliloti 1021 and Agrobacterium tumefaciens C58. These results suggest that moxBA and its homologues constitute a family of genes encoding an MCO and a predicted component of the Type I secretion system.

Tolerância ao sal e às altas temperaturas de estirpes de Sinorhizobium provenientes de zonas secas do Alentejo Salt and temperature tolerance of Sinorhizobium strains isolated from dry environments in Alentejo

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P. Fareleira

2007-07-01

Full Text Available O trabalho teve como principal objectivo a obtenção de estirpes de rizóbio adequadas à nodulação de luzernas anuais e adaptadas a sobreviver nas condições ambientais susceptíveis de ocorrer em solos degradados. Dado que as populações de rizóbio apresentam variabilidade considerável no que respeita à tolerância a factores ambientais, os estudos focaram-se na pesquisa e na selecção de estirpes resistentes a condições adversas. Efectuaram-se colheitas de solos em diversas zonas no sul do País, seleccionando-se locais afectados por secura, temperaturas elevadas e, pontualmente, salinidade. A partir destes solos, isolaram-se estirpes de rizóbio, usando como planta hospedeira a luzerna anual Medicago polymorpha. Estudaram-se os efeitos de condições de stresse ambiental, como a salinidade e as altas temperaturas, no crescimento das estirpes isoladas. Dos 41 isolamentos analisados, 11 apresentaram crescimento em meio con-tendo 1,4 M de cloreto de sódio e suplementado com 10% de extracto de terra, e 22 cresceram quando incubadas a 45 ºC em meio sem aditivos. Três estirpes mostraram ter capacidade para crescer sob os efeitos conjuntos da salinidade e da alta temperatura. A análise de extractos etanólicos de estirpes tolerantes à salinidade revelou, na maior parte dos casos, a acumulação, induzida pelo sal, dos solutos compatíveis de Sinorhizobium: o dipéptido N - acetilglutaminilglutamina amida, vários tipos de betaínas, trealose, glutamato e prolina. A observação, por NMR de 31P in vivo, de uma estirpe tolerante ao sal, proveniente de um solo xistoso de baixo teor em fósforo, mostrou a presença de níveis elevados de reservas intracelulares de fosfato inorgânico (polifosfato, sugerindo um bom potencial para utilização em solos onde os riscos de salinização se conjuguem com deficiências em fósforo assimilável.The main objective of this work was to obtain rhizobial strains able to nodulate annual medics and

Plant-fed versus chemicals-fed rhizobacteria of Lucerne: Plant-only teabags culture media not only increase culturability of rhizobacteria but also recover a previously uncultured Lysobacter sp., Novosphingobium sp. and Pedobacter sp.

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Hegazi, Nabil A; Sarhan, Mohamed S; Fayez, Mohamed; Patz, Sascha; Murphy, Brian R; Ruppel, Silke

2017-01-01

In an effort to axenically culture the previously uncultivable populations of the rhizobacteria of Lucerne (Medicago sativa L.), we propose plant-only teabags culture media to mimic the nutritional matrix available in the rhizosphere. Here, we show that culture media prepared from Lucerne powder teabags substantially increased the cultivability of Lucerne rhizobacteria compared with a standard nutrient agar, where we found that the cultivable populations significantly increased by up to 60% of the total bacterial numbers as estimated by Quantitative Real-time Polymerase Chain Reaction (qRT-PCR). Cluster analysis of 16S rDNA Polymerase Chain Reaction-Denaturing Gradient Gel Electrophoresis (PCR-DGGE) of cultivable Colony-Forming Units (CFUs) revealed a more distinct composition and separation of bacterial populations recovered on the plant-only teabags culture media than those developed on a standard nutrient agar. Further, the new plant medium gave preference to the micro-symbiont Sinorhizobium meliloti, and succeeded in isolating a number of not-yet-cultured bacteria, most closely matched to Novosphingobium sp., Lysobacter sp. and Pedobacter sp. The present study may encourage other researchers to consider moving from the well-established standard culture media to the challenging new plant-only culture media. Such a move may reveal previously hidden members of rhizobacteria, and help to further explore their potential environmental impacts.

Rapid and simple colorimetric method for the quantification of AI-2 produced from Salmonella Typhimurium.

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Wattanavanitchakorn, Siriluck; Prakitchaiwattana, Cheunjit; Thamyongkit, Patchanita

2014-04-01

The aim of this study was to evaluate the feasibility of Fe(III) ion reduction for the simple and rapid quantification of autoinducer-2 (AI-2) produced from bacteria using Salmonella Typhimurium as a model. Since the molecular structure of AI-2 is somewhat similar to ascorbic acid it was expected that AI-2 would also act as a reducing agent and reduce Fe(III) ions in the presence of 1,10-phenanthroline to form the colored [(o-phen)3 Fe(II)]SO4 ferroin complex that could be quantified colorimetrically. In support of this, colony rinses and cell free supernatants from cultures of all tested AI-2 producing strains, but not the AI-2 negative Sinorhizobium meliloti, formed a colored complex with a λmax of 510nm. The OD510 values of these culture supernatants or colony rinses were in broad agreement with the % activity observed in the same samples using the standard Vibrio harveyi bioluminescence assay for AI-2 detection, and with previously reported results. This methodology could potentially be developed as an alternative method for the simple and rapid quantification of AI-2 levels produced in bacterial cultures. Copyright © 2014 Elsevier B.V. All rights reserved.

A Lux-like Quorum Sensing System in the Extreme Acidophile Acidithiobacillus ferrooxidans

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MARIELLA RIVAS

2005-01-01

Full Text Available The genome of the acidophilic, proteobacterium Acidithiobacillus ferrooxidans, contains linked but divergently oriented genes, termed afeI and afeR, whose predicted protein products are significantly similar to the LuxI and LuxR families of proteins. A possible promoter and Lux box are predicted upstream of afeI. A cloned copy of afeI, expressed in E. coli, encodes an enzyme that catalyzes the production of a diffusible compound identified by gas chromatography and mass spectrometry as an unsubstituted N-acyl homoserine lactone (AHL of chain length C14. This AHL can be detected by a reporter strain of Sinorhizobium meliloti Rm41 suggesting that it is biologically active. The reporter strain also responds to extracts of the supernatant of A. ferrooxidans grown to early stationary phase in sulfur medium indicating that a diffusible AHL is produced by this microorganism. Semi-quantitative RT-PCR experiments indicate that afeI and afeR are expressed maximally in early stationary phase and are more expressed when A. ferrooxidans is grown in sulfur- rather than iron-containing medium. Given the predicted amino acid sequence and functional properties of AfeI and AfeR it is proposed that A. ferrooxidans has a quorum sensing system similar to the LuxI-LuxR paradigm.

Interplay of Pathogen-Induced Defense Responses and Symbiotic Establishment in Medicago truncatula

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Tao Chen

2017-05-01

Full Text Available Suppression of host innate immunity appears to be required for the establishment of symbiosis between rhizobia and host plants. In this study, we established a system that included a host plant, a bacterial pathogen and a symbiotic rhizobium to study the role of innate immunity during symbiotic interactions. A pathogenic bacterium, Pseudomonas syringae pv. tomato strain DC3000 (Pst DC3000, was shown to cause chlorosis in Medicago truncatula A17. Sinorhizobium meliloti strain Sm2011 (Sm2011 and Pst DC3000 strain alone induced similar defense responses in M. truncatula. However, when co-inoculated, Sm2011 specifically suppressed the defense responses induced by Pst DC3000, such as MAPK activation and ROS production. Inoculation with Sm2011 suppressed the transcription of defense-related genes triggered by Pst DC3000 infection, including the receptor of bacterial flagellin (FLS2, pathogenesis-related protein 10 (PR10, and the transcription factor WRKY33. Interestingly, inoculation with Pst DC3000 specifically inhibited the expression of the symbiosis marker genes nodule inception and nodulation pectate lyase and reduced the numbers of infection threads and nodules on M. truncatula A17 roots, indicating that Pst DC3000 inhibits the establishment of symbiosis in M. truncatula. In addition, defense-related genes, such as MAPK3/6, RbohC, and WRKY33, exhibited a transient increase in their expression in the early stage of symbiosis with Sm2011, but the expression dropped down to normal levels at later symbiotic stages. Our results suggest that plant innate immunity plays an antagonistic role in symbiosis by directly reducing the numbers of infection threads and nodules.

Polychlorinated biphenyl rhizoremediation by Pseudomonas fluorescens F113 derivatives, using a Sinorhizobium meliloti nod system to drive bph gene expression

Czech Academy of Sciences Publication Activity Database

Villacieros, M.; Whelan, C.; Macková, Martina; Molgaard, J.; Sánchez-Contreras, M.; Lloret, J.; Cárcer de, D.; Oruezábal, R. I.; Bolanos, L.; Macek, Tomáš; Karlson, U.; Dowling, D. N.; Martín, M.; Rivilla, R.

2005-01-01

Roč. 71, č. 5 (2005), 2687-2694 ISSN 0099-2240 Grant - others:5thFW(XE) QLK3-CT-2001-00101; European Commission(XE) BIO4-CT97-2227 Institutional research plan: CEZ:AV0Z4055905 Keywords : polychlorinated biphenyls * rhizoremediation Subject RIV: CC - Organic Chemistry Impact factor: 3.818, year: 2005

Concomitant degradation of bisphenol A during ultrasonication and Fenton oxidation and production of biofertilizer from wastewater sludge.

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Mohapatra, D P; Brar, S K; Tyagi, R D; Surampalli, R Y

2011-09-01

Degradation of bisphenol A (BPA), an endocrine disruptor, from wastewater sludge (WWS) has attracted great interest recently. In the present study, the effects of different pre-treatment methods, including ultrasonication (US), Fenton's oxidation (FO) and ferro-sonication (FS) was assessed in terms of increase in solubilization of WWS and simultaneous degradation of BPA. Among US, FO and FS pre-treatment, higher suspended solids (SS), volatile suspended solids (VSS), chemical oxygen demand (COD) and soluble organic carbon (SOC) solubilization (39.7%, 51.2%, 64.5% and 17.6%, respectively) was observed during a ferro-sonication pre-treatment process carried out for 180 min, resulting in higher degradation of BPA (82.7%). In addition, the effect of rheological parameters (viscosity and particle size) and zeta potential on the degradation of BPA in raw and different pre-treated sludges were also investigated. The results showed that a decrease in viscosity and particle size and an increase in zeta potential resulted in higher degradation of BPA. BPA degradation by laccases produced by Sinorhizobium meliloti in raw and pre-treated sludge was also determined. Higher activity of laccases (207.9 U L(-1)) was observed in ferro-sonicated pre-treated sludge (180 min ultrasonic time), resulting in higher removal of BPA (0.083 μg g(-1)), suggesting concomitant biological degradation of BPA. Copyright © 2011 Elsevier B.V. All rights reserved.

The stress-related, rhizobial small RNA RcsR1 destabilizes the autoinducer synthase encoding mRNA sinI in Sinorhizobium meliloti

Czech Academy of Sciences Publication Activity Database

Baumgardt, K.; Šmídová, Klára; Rahn, H.; Lochnit, G.; Robledo, M.; Evguenieva-Hackenberg, E.

2016-01-01

Roč. 13, č. 5 (2016), s. 486-499 ISSN 1547-6286 Institutional support: RVO:61388971 Keywords : Agrobacterium * autoinducer synthase * degradosome Subject RIV: EE - Microbiology, Virology Impact factor: 3.900, year: 2016

Acacia senegal and Prosopis chilensis-nodulating rhizobia Sinorhizobium arboris HAMBI 2361 and S. kostiense HAMBI 2362 produce tetra- and pentameric LCOs that are N-methylated, O-6-carbamoylated and partially sulfated.

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Nowak, Petri; Soupas, Laura; Thomas-Oates, Jane; Lindström, Kristina

2004-04-28

Sinorhizobium arboris and S. kostiense are rhizobia that nodulate the tropical leguminous trees Acacia senegal and Prosopis chilensis. The lipochito-oligosaccharidic signalling molecules (LCOs) of S. arboris HAMBI 2361 and S. kostiense HAMBI 2362 were analyzed by mass spectrometry. The major LCOs produced by the strains were shown to be pentameric, acylated with common fatty acids, N-methylated, O-6-carbamoylated and partially sulfated, as are the LCOs characterized to date for other Acacia-nodulating rhizobia. Besides the major LCOs the two strains produced (i) tetrameric LCOs, (ii) LCOs acylated with fatty acids other than those commonly found, (iii) LCOs with only an acyl substituent and (iv) noncarbamoylated LCOs. Production of LCOs (i) to (iii) are novel among Acacia-nodulating rhizobia. The roles of the different structural characteristics of LCOs in the rhizobium-A. senegal symbiosis are discussed. Specific structural features of the LCOs are proposed to be important in the selection of effective nitrogen-fixing rhizobia by A. senegal.

Enhanced resistance to nanoparticle toxicity is conferred by overproduction of extracellular polymeric substances

International Nuclear Information System (INIS)

Joshi, Nimisha; Ngwenya, Bryne T.; French, Christopher E.

2012-01-01

Highlights: ► Demonstration that bacteria engineered for EPS overproduction have better survival against Ag nanotoxicity. ► EPS destabilises Ag nanoparticles and promotes their aggregation. ► TEM demonstration that EPS traps the Ag nanoparticles outside the cell. ► EPS from overexpressing strains offers protection to non-EPS strains of bacteria. ► EPS polymer analogues such as xanthan also produce a similar response. - Abstract: The increasing production and use of engineered nanoparticles, coupled with their demonstrated toxicity to different organisms, demands the development of a systematic understanding of how nanoparticle toxicity depends on important environmental parameters as well as surface properties of both cells and nanomaterials. We demonstrate that production of the extracellular polymeric substance (EPS), colanic acid by engineered Escherichia coli protects the bacteria against silver nanoparticle toxicity. Moreover, exogenous addition of EPS to a control strain results in an increase in cell viability, as does the addition of commercial EPS polymer analogue xanthan. Furthermore, we have found that an EPS producing strain of Sinorhizobium meliloti shows higher survival upon exposure to silver nanoparticles than the parent strain. Transmission electron microscopy (TEM) observations showed that EPS traps the nanoparticles outside the cells and reduces the exposed surface area of cells to incoming nanoparticles by inducing cell aggregation. Nanoparticle size characterization in the presence of EPS and xanthan indicated a marked tendency towards aggregation. Both are likely effective mechanisms for reducing nanoparticle toxicity in the natural environment.

Genome sequence of Ensifer adhaerens OV14 provides insights into its ability as a novel vector for the genetic transformation of plant genomes.

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Rudder, Steven; Doohan, Fiona; Creevey, Christopher J; Wendt, Toni; Mullins, Ewen

2014-04-07

Recently it has been shown that Ensifer adhaerens can be used as a plant transformation technology, transferring genes into several plant genomes when equipped with a Ti plasmid. For this study, we have sequenced the genome of Ensifer adhaerens OV14 (OV14) and compared it with those of Agrobacterium tumefaciens C58 (C58) and Sinorhizobium meliloti 1021 (1021); the latter of which has also demonstrated a capacity to genetically transform crop genomes, albeit at significantly reduced frequencies. The 7.7 Mb OV14 genome comprises two chromosomes and two plasmids. All protein coding regions in the OV14 genome were functionally grouped based on an eggNOG database. No genes homologous to the A. tumefaciens Ti plasmid vir genes appeared to be present in the OV14 genome. Unexpectedly, OV14 and 1021 were found to possess homologs to chromosomal based genes cited as essential to A. tumefaciens T-DNA transfer. Of significance, genes that are non-essential but exert a positive influence on virulence and the ability to genetically transform host genomes were identified in OV14 but were absent from the 1021 genome. This study reveals the presence of homologs to chromosomally based Agrobacterium genes that support T-DNA transfer within the genome of OV14 and other alphaproteobacteria. The sequencing and analysis of the OV14 genome increases our understanding of T-DNA transfer by non-Agrobacterium species and creates a platform for the continued improvement of Ensifer-mediated transformation (EMT).

Volatile compounds from beneficial or pathogenic bacteria differentially regulate root exudation, transcription of iron transporters, and defense signaling pathways in Sorghum bicolor.

Science.gov (United States)

Hernández-Calderón, Erasto; Aviles-Garcia, Maria Elizabeth; Castulo-Rubio, Diana Yazmín; Macías-Rodríguez, Lourdes; Ramírez, Vicente Montejano; Santoyo, Gustavo; López-Bucio, José; Valencia-Cantero, Eduardo

2018-02-01

Our results show that Sorghum bicolor is able to recognize bacteria through its volatile compounds and differentially respond to beneficial or pathogens via eliciting nutritional or defense adaptive traits. Plants establish beneficial, harmful, or neutral relationships with bacteria. Plant growth promoting rhizobacteria (PGPR) emit volatile compounds (VCs), which may act as molecular cues influencing plant development, nutrition, and/or defense. In this study, we compared the effects of VCs produced by bacteria with different lifestyles, including Arthrobacter agilis UMCV2, Bacillus methylotrophicus M4-96, Sinorhizobium meliloti 1021, the plant pathogen Pseudomonas aeruginosa PAO1, and the commensal rhizobacterium Bacillus sp. L2-64, on S. bicolor. We show that VCs from all tested bacteria, except Bacillus sp. L2-64, increased biomass and chlorophyll content, and improved root architecture, but notheworthy A. agilis induced the release of attractant molecules, whereas P. aeruginosa activated the exudation of growth inhibitory compounds by roots. An analysis of the expression of iron-transporters SbIRT1, SbIRT2, SbYS1, and SbYS2 and genes related to plant defense pathways COI1 and PR-1 indicated that beneficial, pathogenic, and commensal bacteria could up-regulate iron transporters, whereas only beneficial and pathogenic species could induce a defense response. These results show how S. bicolor could recognize bacteria through their volatiles profiles and highlight that PGPR or pathogens can elicit nutritional or defensive traits in plants.

Enhanced resistance to nanoparticle toxicity is conferred by overproduction of extracellular polymeric substances

Energy Technology Data Exchange (ETDEWEB)

Joshi, Nimisha, E-mail: joshi.nimisha@gmail.com [School of GeoSciences, Microbial Geochemistry Laboratory, University of Edinburgh, West Mains Road, Edinburgh EH9 3JW (United Kingdom); Ngwenya, Bryne T. [School of GeoSciences, Microbial Geochemistry Laboratory, University of Edinburgh, West Mains Road, Edinburgh EH9 3JW (United Kingdom); French, Christopher E. [School of Biological Sciences, Institute of Cell Biology, Darwin Building, University of Edinburgh, Mayfield Road, Edinburgh EH9 3JR (United Kingdom)

2012-11-30

Highlights: Black-Right-Pointing-Pointer Demonstration that bacteria engineered for EPS overproduction have better survival against Ag nanotoxicity. Black-Right-Pointing-Pointer EPS destabilises Ag nanoparticles and promotes their aggregation. Black-Right-Pointing-Pointer TEM demonstration that EPS traps the Ag nanoparticles outside the cell. Black-Right-Pointing-Pointer EPS from overexpressing strains offers protection to non-EPS strains of bacteria. Black-Right-Pointing-Pointer EPS polymer analogues such as xanthan also produce a similar response. - Abstract: The increasing production and use of engineered nanoparticles, coupled with their demonstrated toxicity to different organisms, demands the development of a systematic understanding of how nanoparticle toxicity depends on important environmental parameters as well as surface properties of both cells and nanomaterials. We demonstrate that production of the extracellular polymeric substance (EPS), colanic acid by engineered Escherichia coli protects the bacteria against silver nanoparticle toxicity. Moreover, exogenous addition of EPS to a control strain results in an increase in cell viability, as does the addition of commercial EPS polymer analogue xanthan. Furthermore, we have found that an EPS producing strain of Sinorhizobium meliloti shows higher survival upon exposure to silver nanoparticles than the parent strain. Transmission electron microscopy (TEM) observations showed that EPS traps the nanoparticles outside the cells and reduces the exposed surface area of cells to incoming nanoparticles by inducing cell aggregation. Nanoparticle size characterization in the presence of EPS and xanthan indicated a marked tendency towards aggregation. Both are likely effective mechanisms for reducing nanoparticle toxicity in the natural environment.

Modeling Bacteria-Water Interactions in Soil: EPS Dynamics Under Evaporative Conditions

Science.gov (United States)

Furrer, J.; Hinestroza, H. F.; Guo, Y. S.; Gage, D. J.; Cho, Y. K.; Shor, L. M.

2017-12-01

The soil habitat represents a major linkage between the water and carbon cycles: the ability of soils to sequester or release carbon is determined primarily by soil moisture. Water retention and distribution in soils controls the abundance and activity of soil microbes. Microbes in turn impact water retention by creating biofilms, composed of extracellular polymeric substances (EPS). We model the effects of bacterial EPS on water retention at the pore scale. We use the lattice Boltzmann method (LBM), a well-established fluid dynamics modeling platform, and modify it to include the effects of water uptake and release by the swelling/shrinking EPS phase. The LB model is implemented in 2-D, with a non-ideal gas equation of state that allows condensation and evaporation of fluid in pore spaces. Soil particles are modeled according to experimentally determined particle size distributions and include realistic pore geometries, in contrast to many soil models which use spherical soil particles for simplicity. Model results are compared with evaporation experiments in soil micromodels and other simpler experimental systems, and model parameters are tuned to match experimental results. Drying behavior and solid-gel contact angle of EPS produced by the soil bacteria Sinorhizobium meliloti has been characterized and compared to the behavior of deionized water under the same conditions. The difference in behavior between the fluids is used to parameterize the model. The model shows excellent qualitative agreement for soil micromodels with both aggregated and non-aggregated particle arrangements under no-EPS conditions, and reproduces realistic drying behavior for EPS. This work represents a multi-disciplinary approach to understanding microbe-soil interactions at the pore scale.

Low vapour pressure deficit reduces the beneficial effect of elevated CO{sub 2} on growth of N{sub 2}-fixing alfalfa plants

Energy Technology Data Exchange (ETDEWEB)

Luis, I. De; Irigoyen, J.J.; Sanchez-Diaz, M. [Univ. de Navarra, Dept. de Fisioligia Vegetal, Pamplona (Spain)

2002-11-01

Plant responses to elevated CO{sub 2} can be modified by many environmental factors, but very little attention has been paid to the interaction between CO{sub 2} and changes in vapour pressure deficit (VPD). Thirty-day-old alfalfa plants (Medicago sativa L. cv. Aragon), which were inoculated with Sinorhizobium meliloti 102F78 strain, were grown for 1 month in controlled environment chambers at 25/15 deg C, 14 h photoperiod, and 600 mol m{sup -2} s{sup -1} photosynthetic photon flux (PPF), using a factorial combination of CO{sub 2} concentration (400 mol mol{sup -1} or 700 mol mol{sup -1}) and vapour pressure deficit (0.48 kPa or 1.74 kPa, which corresponded to relative humidities of 85% and 45% at 25 deg C, respectively). Elevated CO{sub 2} strongly stimulated plant growth under high VPD conditions, but this beneficial effect was not observed under low VPD. Under low VPD, elevated CO{sub 2} also did not enhance plant photosynthesis, and plant water stress was greatest for plants grown at elevated CO{sub 2} and low VPD. Moreover, plants grown under elevated CO{sub 2} and low VPD had a lower leaf soluble protein and photosynthetic activity (photosynthetic rate and carboxylation efficiency) than plants grown under elevated CO{sub 2} and high VPD. Elevated CO{sub 2} significantly increased leaf adaxial and abaxial temperatures. Because the effects of elevated CO{sub 2} were dependent on vapour pressure deficit, VPD needs to be controlled in experiments studying the effect of elevated CO{sub 2} as well as considered in the extrapolations of results to a warmer, high-CO{sub 2} world. (au)

utilization of bio fertilizers and organic sources in arable soils under saline conditions using tracer technique

International Nuclear Information System (INIS)

Salama, O.A.E.

2011-01-01

Recently, more attention has been paid to conserve and save surrounding environment via minimizing the excessive use of chemical fertilizers and, in general, the agrochemicals applied in heavy quantities in agricultural agroecosystems. Therefore, the attention of most of agronomists was turned towards the use of so called clean agriculture or organic farming. Many of organic systems was pointed out such as the recycling of farm wastes i.e. crop residues, animal manure, organic conditioners for reclamation of soil and in the same time enhancement of plant growth and improving yield quality. The application of organic wastes combined with or without microbial inoculants to plant media are considered as a good management practice in any agricultural production system because it improves, plant quality and soil fertility. Therefore, we have the opportunity to conduct some experiments for achieving the clean agriculture approach, combating the adverse effects of salinity and avoiding the environmental pollution. Series of laboratory and greenhouse experiments were carried out to evaluate the impact of (1) potent isolated fungi (Aspergillus oryzae and Aspergillus terreus) on degrading plant residues (Leucaena and Acacia green parts), and (2) biofertilizers (Sinorhizobium meliloti, Azospirillum brasilense, and Pseudomonas aeruginosa) in assessing barley and spinach plants to combat salinity of soil and irrigation water. 15 N-tracer technique that considered unique and more reliable technique may benefits in clarifying the responsible mechanisms related to plant growth and gave us the opportunity to quantify the exact amounts of N derived from the different sources of nitrogen available to spinach and barley plants grown on sandy saline soil and irrigated with saline water.

Bacterial exopolysaccharides as a modern biotechnological tool for modification of fungal laccase properties and metal ion binding.

Science.gov (United States)

Osińska-Jaroszuk, Monika; Jaszek, Magdalena; Starosielec, Magdalena; Sulej, Justyna; Matuszewska, Anna; Janczarek, Monika; Bancerz, Renata; Wydrych, Jerzy; Wiater, Adrian; Jarosz-Wilkołazka, Anna

2018-03-26

Four bacterial EPSs extracted from Rhizobium leguminosarum bv. trifolii Rt24.2, Sinorhizobium meliloti Rm1021, Bradyrhizobium japonicum USDA110, and Bradyrhizobium elkanii USDA76 were determined towards their metal ion adsorption properties and possible modification of Cerrena unicolor laccase properties. The highest magnesium and iron ion-sorption capacity (~ 42 and ~ 14.5%, respectively) was observed for EPS isolated from B. japonicum USDA110. An evident influence of EPSs on the stability of laccase compared to the control values (without EPSs) was shown after 30-day incubation at 25 °C. The residual activity of laccases was obtained in the presence of Rh76EPS and Rh1021EPS, i.e., 49.5 and 41.5% of the initial catalytic activity, respectively. This result was confirmed by native PAGE electrophoresis. The EPS effect on laccase stability at different pH (from 3.8 to 7.0) was also estimated. The most significant changes at the optimum pH value (pH 5.8) was observed in samples of laccase stabilized by Rh76EPS and Rh1021EPS. Cyclic voltamperometry was used for analysis of electrochemical parameters of laccase stabilized by bacterial EPS and immobilized on single-walled carbon nanotubes (SWCNTs) with aryl residues. Laccases with Rh76EPS and Rh1021EPS had an evident shift of the value of the redox potential compared to the control without EPS addition. In conclusion, the results obtained in this work present a new potential use of bacterial EPSs as a metal-binding component and a modulator of laccase properties especially stability of enzyme activity, which can be a very effective tool in biotechnology and industrial applications.

A cheZ-Like Gene in Azorhizobium caulinodans Is a Key Gene in the Control of Chemotaxis and Colonization of the Host Plant.

Science.gov (United States)

Liu, Xiaolin; Liu, Wei; Sun, Yu; Xia, Chunlei; Elmerich, Claudine; Xie, Zhihong

2018-02-01

Chemotaxis can provide bacteria with competitive advantages for survival in complex environments. The CheZ chemotaxis protein is a phosphatase, affecting the flagellar motor in Escherichia coli by dephosphorylating the response regulator phosphorylated CheY protein (CheY∼P) responsible for clockwise rotation. A cheZ gene has been found in Azorhizobium caulinodans ORS571, in contrast to other rhizobial species studied so far. The CheZ protein in strain ORS571 has a conserved motif similar to that corresponding to the phosphatase active site in E. coli The construction of a cheZ deletion mutant strain and of cheZ mutant strains carrying a mutation in residues of the putative phosphatase active site showed that strain ORS571 participates in chemotaxis and motility, causing a hyperreversal behavior. In addition, the properties of the cheZ deletion mutant revealed that ORS571 CheZ is involved in other physiological processes, since it displayed increased flocculation, biofilm formation, exopolysaccharide (EPS) production, and host root colonization. In particular, it was observed that the expression of several exp genes, involved in EPS synthesis, was upregulated in the cheZ mutant compared to that in the wild type, suggesting that CheZ negatively controls exp gene expression through an unknown mechanism. It is proposed that CheZ influences the Azorhizobium -plant association by negatively regulating early colonization via the regulation of EPS production. This report established that CheZ in A. caulinodans plays roles in chemotaxis and the symbiotic association with the host plant. IMPORTANCE Chemotaxis allows bacteria to swim toward plant roots and is beneficial to the establishment of various plant-microbe associations. The level of CheY phosphorylation (CheY∼P) is central to the chemotaxis signal transduction. The mechanism of the signal termination of CheY∼P remains poorly characterized among Alphaproteobacteria , except for Sinorhizobium meliloti , which

Comparative study of the fungicide Benomyl toxicity on some plant growth promoting bacteria and some fungi in pure cultures

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Elslahi Randa H.

2014-03-01

Full Text Available Six laboratory experiments were carried out to investigate the effect of the fungicide Benomyl on pure cultures of some plant growth promoting bacteria (PGPB and some fungi. The highest LD50 was recorded for Bacillus circulans and proved to be the most resistant to the fungicide, followed by Azospirillum braziliense, while Penicillium sp. was the most affected microorganism. LD50 values for the affected microorganisms were in 21-240 orders of magnitude lower in comparison with the LD50 value for Azospirillum braziliense. The results indicate a strong selectivity for Benomyl against Rhizobium meliloti and Penicillium sp. when compared to other microorganisms tested. The highest safety coefficient was recorded for Bacillus circulans followed by Azospirillum braziliense, while Rhizobium meliloti, showed the lowest safety coefficient value compared to other bacteria. The lowest toxicity index was recorded for Bacillus circulans and Azospirillum braziliense. The slope of the curves for Bacillus sp. and Rhizobium meliloti was steeper than that of the other curves, suggesting that even a slight increase of the dose of the fungicide can cause a very strong negative effect. In conclusion, Benomyl could be applied without restriction when using inocula based on growth promoting bacteria such as symbiotic nitrogen fixers (Rhizobium meliloti, non-symbiotic nitrogen fixers (Azospirillum braziliense or potassium solibilizers (Bacillus circulans, given that the fungicide is applied within the range of the recommended field dose.

Comparative study of the fungicide Benomyl toxicity on some plant growth promoting bacteria and some fungi in pure cultures.

Science.gov (United States)

Elslahi, Randa H; Osman, Awad G; Sherif, Ashraf M; Elhussein, Adil A

2014-03-01

Six laboratory experiments were carried out to investigate the effect of the fungicide Benomyl on pure cultures of some plant growth promoting bacteria (PGPB) and some fungi. The highest LD50 was recorded for Bacillus circulans and proved to be the most resistant to the fungicide, followed by Azospirillum braziliense, while Penicillium sp. was the most affected microorganism. LD50 values for the affected microorganisms were in 21-240 orders of magnitude lower in comparison with the LD50 value for Azospirillum braziliense. The results indicate a strong selectivity for Benomyl against Rhizobium meliloti and Penicillium sp. when compared to other microorganisms tested. The highest safety coefficient was recorded for Bacillus circulans followed by Azospirillum braziliense, while Rhizobium meliloti, showed the lowest safety coefficient value compared to other bacteria. The lowest toxicity index was recorded for Bacillus circulans and Azospirillum braziliense. The slope of the curves for Bacillus sp. and Rhizobium meliloti was steeper than that of the other curves, suggesting that even a slight increase of the dose of the fungicide can cause a very strong negative effect. In conclusion, Benomyl could be applied without restriction when using inocula based on growth promoting bacteria such as symbiotic nitrogen fixers (Rhizobium meliloti), non-symbiotic nitrogen fixers (Azospirillum braziliense) or potassium solibilizers (Bacillus circulans), given that the fungicide is applied within the range of the recommended field dose.

Prevalence, Host Range, and Comparative Genomic Analysis of Temperate Ochrobactrum Phages

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Claudia Jäckel

2017-06-01

Full Text Available Ochrobactrum and Brucella are closely related bacteria that populate different habitats and differ in their pathogenic properties. Only little is known about mobile genetic elements in these genera which might be important for survival and virulence. Previous studies on Brucella lysogeny indicated that active phages are rare in this genus. To gain insight into the presence and nature of prophages in Ochrobactrum, temperate phages were isolated from various species and characterized in detail. In silico analyses disclosed numerous prophages in published Ochrobactrum genomes. Induction experiments showed that Ochrobactrum prophages can be induced by various stress factors and that some strains released phage particles even under non-induced conditions. Sixty percent of lysates prepared from 125 strains revealed lytic activity. The host range and DNA similarities of 19 phages belonging to the families Myoviridae, Siphoviridae, or Podoviridae were determined suggesting that they are highly diverse. Some phages showed relationship to the temperate Brucella inopinata phage BiPB01. The genomic sequences of the myovirus POA1180 (41,655 bp and podovirus POI1126 (60,065 bp were analyzed. Phage POA1180 is very similar to a prophage recently identified in a Brucella strain isolated from an exotic frog. The POA1180 genome contains genes which may confer resistance to chromate and the ability to take up sulfate. Phage POI1126 is related to podoviruses of Sinorhizobium meliloti (PCB5, Erwinia pyrifoliae (Pep14, and Burkholderia cenocepacia (BcepIL02 and almost identical to an unnamed plasmid of the Ochrobactrum intermedium strain LMG 3301. Further experiments revealed that the POI1126 prophage indeed replicates as an extrachromosomal element. The data demonstrate for the first time that active prophages are common in Ochrobactrum and suggest that atypical brucellae also may be a reservoir for temperate phages.

Transcription reprogramming during root nodule development in Medicago truncatula.

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Sandra Moreau

Full Text Available Many genes which are associated with root nodule development and activity in the model legume Medicago truncatula have been described. However information on precise stages of activation of these genes and their corresponding transcriptional regulators is often lacking. Whether these regulators are shared with other plant developmental programs also remains an open question. Here detailed microarray analyses have been used to study the transcriptome of root nodules induced by either wild type or mutant strains of Sinorhizobium meliloti. In this way we have defined eight major activation patterns in nodules and identified associated potential regulatory genes. We have shown that transcription reprogramming during consecutive stages of nodule differentiation occurs in four major phases, respectively associated with (i early signalling events and/or bacterial infection; plant cell differentiation that is either (ii independent or (iii dependent on bacteroid differentiation; (iv nitrogen fixation. Differential expression of several genes involved in cytokinin biosynthesis was observed in early symbiotic nodule zones, suggesting that cytokinin levels are actively controlled in this region. Taking advantage of databases recently developed for M. truncatula, we identified a small subset of gene expression regulators that were exclusively or predominantly expressed in nodules, whereas most other regulators were also activated under other conditions, and notably in response to abiotic or biotic stresses. We found evidence suggesting the activation of the jasmonate pathway in both wild type and mutant nodules, thus raising questions about the role of jasmonate during nodule development. Finally, quantitative RT-PCR was used to analyse the expression of a series of nodule regulator and marker genes at early symbiotic stages in roots and allowed us to distinguish several early stages of gene expression activation or repression.

Identification of Antimony- and Arsenic-Oxidizing Bacteria Associated with Antimony Mine Tailing

Science.gov (United States)

Hamamura, Natsuko; Fukushima, Koh; Itai, Takaaki

2013-01-01

Antimony (Sb) is a naturally occurring toxic element commonly associated with arsenic (As) in the environment and both elements have similar chemistry and toxicity. Increasing numbers of studies have focused on microbial As transformations, while microbial Sb interactions are still not well understood. To gain insight into microbial roles in the geochemical cycling of Sb and As, soils from Sb mine tailing were examined for the presence of Sb- and As-oxidizing bacteria. After aerobic enrichment culturing with AsIII (10 mM) or SbIII (100 μM), pure cultures of Pseudomonas- and Stenotrophomonas-related isolates with SbIII oxidation activities and a Sinorhizobium-related isolate capable of AsIII oxidation were obtained. The AsIII-oxidizing Sinorhizobium isolate possessed the aerobic arsenite oxidase gene (aioA), the expression of which was induced in the presence of AsIII or SbIII. However, no SbIII oxidation activity was detected from the Sinorhizobium-related isolate, suggesting the involvement of different mechanisms for Sb and As oxidation. These results demonstrate that indigenous microorganisms associated with Sb mine soils are capable of Sb and As oxidation, and potentially contribute to the speciation and mobility of Sb and As in situ. PMID:23666539

Host range, symbiotic effectiveness and nodulation competitiveness ...

African Journals Online (AJOL)

SERVER

2008-04-17

Apr 17, 2008 ... ERIC-PCR DNA fingerprinting patterns were used to identify the ... Apart from cowpea where all the isolates were effective, there were significant ..... aminocyclopropane-1-carboxylate deaminase gene in Sinorhizobium.

[Symbiotic matching between soybean cultivar Luhuang No. 1 and different rhizobia].

Science.gov (United States)

Ji, Zhao-jun; Wang, Fei-meng; Wang, Su-ge; Yang, Sheng-hui; Guo, Rui; Tang, Ru-you; Chen, Wen-xin; Chen, Wen-feng

2014-12-01

Soybean plants could establish symbiosis and fix nitrogen with different rhizobial species in the genera of Sinorhizobium and Bradyrhizobium. Studies on the symbiotic matching between soybean cultivars and different rhizobial species are theoretically and practically important for selecting effective strains used to inoculate the plants and improve the soybean production and quality. A total of 27 strains were isolated and purified from a soil sample of Huanghuaihai area by using the soybean cultivar Luhang No. 1, a protein-rich cultivar grown in that area, as the trapping plants. These strains were identified as members of Sinorhizobium (18 strains) and Bradyrhizobium (9 strains) based on the sequence analysis of housekeeping gene recA. Two representative strains (Sinorhizobium fredii S6 and Bradyrhizobium sp. S10) were used to inoculate the seeds of Luhang No. 1 alone or mixed, in pots filled with vermiculite or soil, and in the field trial to investigate their effects on soybean growth, nodulation, nitrogen fixation activity, yield, contents of protein and oil in seeds. The results demonstrated that strain S6 showed better effects on growth-promotion, yield of seeds and seed quality than strain S10. Thus strain S6 was finally regarded as the effective rhizobium matching to soybean Luhuang No. 1, which could be the candidate as a good inoculant for planting the soybean Luhuang No. 1 at a large scale in the Huanghuaihai area.

Interaction of arbuscular mycorrhizal fungi and rhizobia: Effects on flax yield in spoil-bank clay

Czech Academy of Sciences Publication Activity Database

Rydlová, Jana; Püschel, David; Sudová, Radka; Gryndler, M.; Mikanová, O.; Vosátka, Miroslav

2011-01-01

Roč. 174, č. 1 (2011), s. 128-134 ISSN 1436-8730 R&D Projects: GA MŠk 1M0571 Institutional research plan: CEZ:AV0Z60050516 Keywords : Linum usitatissimum * Glomus * Sinorhizobium Subject RIV: EF - Botanics Impact factor: 1.596, year: 2011

Genomic characterization of Ensifer aridi, a proposed new species of nitrogen-fixing rhizobium recovered from Asian, African and American deserts.

Science.gov (United States)

Le Quéré, Antoine; Tak, Nisha; Gehlot, Hukam Singh; Lavire, Celine; Meyer, Thibault; Chapulliot, David; Rathi, Sonam; Sakrouhi, Ilham; Rocha, Guadalupe; Rohmer, Marine; Severac, Dany; Filali-Maltouf, Abdelkarim; Munive, Jose-Antonio

2017-01-14

Nitrogen fixing bacteria isolated from hot arid areas in Asia, Africa and America but from diverse leguminous plants have been recently identified as belonging to a possible new species of Ensifer (Sinorhizobium). In this study, 6 strains belonging to this new clade were compared with Ensifer species at the genome-wide level. Their capacities to utilize various carbon sources and to establish a symbiotic interaction with several leguminous plants were examined. Draft genomes of selected strains isolated from Morocco (Merzouga desert), Mexico (Baja California) as well as from India (Thar desert) were produced. Genome based species delineation tools demonstrated that they belong to a new species of Ensifer. Comparison of its core genome with those of E. meliloti, E. medicae and E. fredii enabled the identification of a species conserved gene set. Predicted functions of associated proteins and pathway reconstruction revealed notably the presence of transport systems for octopine/nopaline and inositol phosphates. Phenotypic characterization of this new desert rhizobium species showed that it was capable to utilize malonate, to grow at 48 °C or under high pH while NaCl tolerance levels were comparable to other Ensifer species. Analysis of accessory genomes and plasmid profiling demonstrated the presence of large plasmids that varied in size from strain to strain. As symbiotic functions were found in the accessory genomes, the differences in symbiotic interactions between strains may be well related to the difference in plasmid content that could explain the different legumes with which they can develop the symbiosis. The genomic analysis performed here confirms that the selected rhizobial strains isolated from desert regions in three continents belong to a new species. As until now only recovered from such harsh environment, we propose to name it Ensifer aridi. The presented genomic data offers a good basis to explore adaptations and functionalities that enable them

Expression of nodule-specific genes in alfalfa root nodules blocked at an early stage of development.

NARCIS (Netherlands)

Dickstein, R.; Bisseling, T.; Reinhold, V.N.; Ausubel, F.M.

1988-01-01

To help dissect the molecular basis of the Rhizobium-legume symbiosis, we used in vitro translation and Northern blot analysis of nodule RNA to examine alfalfa-specific genes (nodulins) expressed in two types of developmentally defective root nodules elicited by Rhizobium meliloti. Fix- nodules were

Nitrous oxide reductase genes (nosZ) of denitrifying populations in soil and the earthworm gut are phylogenetically similar

DEFF Research Database (Denmark)

Horn, Marcus A.; Drake, Harold L.; Schramm, Andreas

2006-01-01

-derived sequences, or were related to N2O reductases of the genera Bradyrhizobium, Brucella, Dechloromonas, Flavobacterium, Pseudomonas, Ralstonia, and Sinorhizobium. Although the numbers of estimators for genotype richness of sequence data from the gut were higher than those of soil, only one gut-derived nos...

CONDITIONING MICROBIAL PRODUCTS CONTAINING NITROGEN FIXING BACTERIA WITH DIFFERENT SOLID EXCIPIENTS

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VINTILĂ T.

2007-01-01

Full Text Available The stability in real time of two strains of Rhizobium (Rhizobium meliloti andRhizobium japonicum mixed with different excipients was evaluated during a6-months period. The excipients studied were: peat, peat and calciumcarbonate, zeolite, and ceramic. Liquid cultures and excipients mixtures weredried (12-14% humidity, sealed in plastic bags and preserved at +4oC. Thecells were activated periodically by suspending aliquots from dry products in0.9% saline solution. The viability of Rhizobium cells was evaluated bycultivation of diluted suspensions in YMA plates. The number of viable cells isdecreasing during drying in all cases, increase in the first month of storage,and remains constant or decrease very slowly during storage for all obtaineddry products containing rhizobia mixed with solid dry excipients. The highestnumber of viable cells at the end of the experiment was obtained in ceramicwith Rhizobium japonicum (8x105 cells/gram, and the lowest number ofviable cells was obtained in zeolite with Rhizobium meliloti (1,1x103cells/gram.

Genetic Diversity among Rhizobium leguminosarum bv. Trifolii Strains Revealed by Allozyme and Restriction Fragment Length Polymorphism Analyses

Science.gov (United States)

Demezas, David H.; Reardon, Terry B.; Watson, John M.; Gibson, Alan H.

1991-01-01

Allozyme electrophoresis and restriction fragment length polymorphism (RFLP) analyses were used to examine the genetic diversity of a collection of 18 Rhizobium leguminosarum bv. trifolii, 1 R. leguminosarum bv. viciae, and 2 R. meliloti strains. Allozyme analysis at 28 loci revealed 16 electrophoretic types. The mean genetic distance between electrophoretic types of R. leguminosarum and R. meliloti was 0.83. Within R. leguminosarum, the single strain of bv. viciae differed at an average of 0.65 from strains of bv. trifolii, while electrophoretic types of bv. trifolii differed at a range of 0.23 to 0.62. Analysis of RFLPs around two chromosomal DNA probes also delineated 16 unique RFLP patterns and yielded genetic diversity similar to that revealed by the allozyme data. Analysis of RFLPs around three Sym (symbiotic) plasmid-derived probes demonstrated that the Sym plasmids reflect genetic divergence similar to that of their bacterial hosts. The large genetic distances between many strains precluded reliable estimates of their genetic relationships. PMID:16348600

CONDITIONING MICROBIAL PRODUCTS CONTAINING NITROGEN FIXING BACTERIA WITH DIFFERENT SOLID EXCIPIENTS

Directory of Open Access Journals (Sweden)

DANIELA VINTILĂ

2007-05-01

Full Text Available The stability in real time of two strains of Rhizobium (Rhizobium meliloti and Rhizobium japonicum mixed with different excipients was evaluated during a 6- months period. The excipients studied were: peat, peat and calcium carbonate, zeolite, and ceramic. Liquid cultures and excipients mixtures were dried (12-14% humidity, sealed in plastic bags and preserved at +4oC. The cells were activated periodically by suspending aliquots from dry products in 0.9% saline solution. The viability of Rhizobium cells was evaluated by cultivation of diluted suspensions in YMA plates. The number of viable cells is decreasing during drying in all cases, increase in the first month of storage, and remains constant or decrease very slowly during storage for all obtained dry products containing rhizobia mixed with solid dry excipients. The highest number of viable cells at the end of the experiment was obtained in ceramic with Rhizobium japonicum (8x105 cells/gram, and the lowest number of viable cells was obtained in zeolite with Rhizobium meliloti (1,1x103 cells/gram.

Biosynthesis of compatible solutes in rhizobial strains isolated from Phaseolus vulgaris nodules in Tunisian fields

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Nieto Joaquín J

2010-07-01

Full Text Available Abstract Background Associated with appropriate crop and soil management, inoculation of legumes with microbial biofertilizers can improve food legume yield and soil fertility and reduce pollution by inorganic fertilizers. Rhizospheric bacteria are subjected to osmotic stress imposed by drought and/or NaCl, two abiotic constraints frequently found in semi-arid lands. Osmostress response in bacteria involves the accumulation of small organic compounds called compatible solutes. Whereas most studies on rhizobial osmoadaptation have focussed on the model species Sinorhizobium meliloti, little is known on the osmoadaptive mechanisms used by native rhizobia, which are good sources of inoculants. In this work, we investigated the synthesis and accumulations of compatible solutes by four rhizobial strains isolated from root nodules of Phaseolus vulgaris in Tunisia, as well as by the reference strain Rhizobium tropici CIAT 899T. Results The most NaCl-tolerant strain was A. tumefaciens 10c2, followed (in decreasing order by R. tropici CIAT 899, R. leguminosarum bv. phaseoli 31c3, R. etli 12a3 and R. gallicum bv. phaseoli 8a3. 13C- and 1H-NMR analyses showed that all Rhizobium strains synthesized trehalose whereas A. tumefaciens 10c2 synthesized mannosucrose. Glutamate synthesis was also observed in R. tropici CIAT 899, R. leguminosarum bv. phaseoli 31c3 and A. tumefaciens 10c2. When added as a carbon source, mannitol was also accumulated by all strains. Accumulation of trehalose in R. tropici CIAT 899 and of mannosucrose in A. tumefaciens 10c2 was osmoregulated, suggesting their involvement in osmotolerance. The phylogenetic analysis of the otsA gene, encoding the trehalose-6-phosphate synthase, suggested the existence of lateral transfer events. In vivo 13C labeling experiments together with genomic analysis led us to propose the uptake and conversion pathways of different carbon sources into trehalose. Collaterally, the β-1,2-cyclic glucan from R

Efficacy of wild plant in combination with microbial antagonists for the control of root rot fungi on mungbean and cowpea

International Nuclear Information System (INIS)

Ikram, N.; Dawar, S.

2015-01-01

Present work was carried out to investigate the efficacy of Aerva javanica in combination with different microbial antagonists namely Rhizobium meliloti, Pseudomonas aeruginosa, Trichoderma harzianum and Aspergillus niger. Soil amended with A. javanica stem, leaves, flower powder at the rate1% w/w and seeds of cowpea (Vigna unguiculata L.) and mungbean (Vigna radiata L.) were coated with microbial antagonists for the control of root infecting fungi like Macrophomina phaseolina (Tassi) Goid, Fusarium spp. and Rhizoctonia solani Kiihn. Infection of M. phaseolina and R. solani were completely suppressed when seeds were coated with P. aeruginosa, T. harzianum, A. niger, R. meliloti and A. javanica leaves powder mixed in soil at the rate 1% w/w. All antagonists showed reduction in combination with A. javanica leaves powder at the rate1% but T. harzianum and P. aeruginosa in combination with A. javanica leaves showed promising results in complete reduction of R. solani and M. phaseolina on both crops. All growth parameters were maximum when soil was amended with A. javanica leaves powder at the rate 1% w/w and seeds were coated with T. harzianum and P. aeruginosa. (author)

Genetic identification and symbiotic efficiency of Sinorhizobium ...

African Journals Online (AJOL)

STORAGESEVER

2008-08-18

Aug 18, 2008 ... numerical analysis (diversity data base for DNA v2.1.1) of gel data (Figure .... strains nodulating small senegalese legumes by 16S-23S rRNA intergenic gene ... 169-174. Gao J, Terefework Z, Chen W-X, Lindstrom K (2001).

Tolerância de rizóbios de diferentes procedências ao zinco, cobre e cádmio Tolerance of rhizobia genera from different origins to zinc, copper and cadmium

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Alexandre Matsuda

2002-03-01

Full Text Available Sessenta estirpes/isolados dos gêneros Bradyrhizobium, Rhizobium, Sinorhizobium, Mesorhizobium e Azorhizobium, procedentes de diferentes locais (Mata Atlântica, Amazônia, culturas agrícolas e experimentos com metais pesados e de espécies hospedeiras pertencentes às subfamílias Papilionoideae, Mimosoideae e Caesalpinoideae, foram avaliadas quanto à tolerância a Zn, Cu e Cd em meio YMA modificado pela adição de tampões biológicos (HEPES e MES e suplementados com Cu (0 a 60 mg L-1, Cd (0 a 60 mg L-1 e Zn (0 a 1.000 mg L-1. Mediante padrões de crescimento atribuídos às culturas nas diferentes concentrações dos metais, avaliaram-se as concentrações máximas toleradas e as doses tóxicas destes metais para redução de crescimento em 25% (DT25 e 50% (DT50. Não houve influência da procedência na concentração máxima de metal tolerada. A ordem de sensibilidade aos metais, considerando-se as concentrações máximas toleradas, foi Azorhizobium > Rhizobium = Mesorhizobium = Sinorhizobium > Bradyrhizobium. A DT25 e a DT50 foram úteis para diferenciarem estirpes/isolados de um mesmo gênero, que atingiram a mesma concentração máxima tolerada a Zn, Cu e Cd. A ordem de toxicidade dos metais estudados foi Cu > Cd > Zn.Sixty strains/isolates of the genera Bradyrhizobium, Rhizobium, Sinorhizobium, Mesorhizobium and Azorhizobium, isolated from different hosts (legume subfamilies: Papilionoideae, Mimosoideae and Caesalpinoideae and location (Atlantic Forest, Amazon region, crop plantings and heavy metal experiments, were evaluated for Zn, Cu and Cd tolerance in YMA medium modified by the addition of biological buffers (HEPES and MES and supplemented with Cu (0 to 60 mg L-1, Cd (0 to 60 mg L-1, and Zn (0 to 1,000 mg L-1sulphates. Growth standards were applied to evaluate rhizobia cultures growth at different metal concentrations, allowing evaluation of highest tolerated concentrations of Zn, Cu, and Cd and the toxic doses

Degradation of the Herbicide Glyphosate by Members of the Family Rhizobiaceae

OpenAIRE

Liu, C.-M.; McLean, P. A.; Sookdeo, C. C.; Cannon, F. C.

1991-01-01

Several strains of the family Rhizobiaceae were tested for their ability to degrade the phosphonate herbicide glyphosate (isopropylamine salt of N-phosphonomethylglycine). All organisms tested (seven Rhizobium meliloti strains, Rhizobium leguminosarum, Rhizobium galega, Rhizobium trifolii, Agrobacterium rhizogenes, and Agrobacterium tumefaciens) were able to grow on glyphosate as the sole source of phosphorus in the presence of the aromatic amino acids, although growth on glyphosate was not a...

Effects of rhizobial bacteria on K, Ca and Na concentration of wheat (Triticum aestivum L. in saline soils

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S homayoon

2016-05-01

Full Text Available Introduction Soil salinity is one of the major agricultural problems and it is limiting crop productivity in many parts of the cultivated areas all over the world. Saline soils are differentiated by the presence of great ratios of Na/Ca, Na/K, Ca2+, Mg2+, and Cl/NO3 (Gratan & Catherine, 1993 and high levels of neutral salts in the surface layers, which are resulting from the capillary action (Al-Falih, 2002. Osmotic stress occurs when soluble salts increase in the soils and then results in specific ion toxicity (Agarwal & Ahmad, 2010. Therefore, one of the most important side effects of salinity is nutritional disorders. High concentration of NaCl in the root medium usually reduces nutrients uptake and affects the transportation of potassium and calcium ions in plant. (Gratan & Catherine, 1993 reported that the salinity of soils changes ionic strength of the substrate and it can influence mineral nutrient uptake and translocation. Salinity also changes the mineral nutrient availability and disrupts the mineral relations of plants. Hence, the main purpose of this research is to evaluate the effects of rhizobial bacteria inoculation on K, Ca and Na concentration of wheat (Triticum aestivum L. in saline soils. Material and methods Soil sample was collected from Astan Ghodse Razavi farm, Mashhad Iran, and then was dried and passed through a 12-mesh (approximately 2 mm screen. Soil sample was divided into three parts and then was placed into three containers. Each container was watered by a different proportion of saline water (EC= 10 dS.m-1. Salinity of soils was regularly monitored until three salinities (2, 6 and 10 dS.m-1 came out. Then, a completely randomized design with a factorial arrangement was carried out in a greenhouse condition. The experimental factors included four levels of inoculation (Sinorhizobium meliloti, Bradyrhizobium japonicum and Rhizobium leguminosarum and control and three levels of soil salinity (2, 6 and 10 dS.m-1 with

Geographically structured genetic variation in the Medicago lupulina-Ensifer mutualism.

Science.gov (United States)

Harrison, Tia L; Wood, Corlett W; Heath, Katy D; Stinchcombe, John R

2017-07-01

Gene flow between genetically differentiated populations can maintain variation in species interactions, especially when population structure is congruent between interacting species. However, large-scale empirical comparisons of the population structure of interacting species are rare, particularly in positive interspecific interactions (mutualisms). One agriculturally and ecologically important mutualism is the partnership between legume plants and rhizobia. Through characterizing and comparing the population genomic structure of the legume Medicago lupulina and two rhizobial species (Ensifer medicae and E. meliloti), we explored the spatial scale of population differentiation between interacting partners in their introduced range in North America. We found high proportions of E. meliloti in southeastern populations and high proportions of E. medicae in northwestern populations. Medicago lupulina and the Ensifer genus showed similar patterns of spatial genetic structure (isolation by distance). However, we detected no evidence of isolation by distance or population structure within either species of bacteria. Genome-wide nucleotide diversity within each of the two Ensifer species was low, suggesting limited introduction of strains, founder events, or severe bottlenecks. Our results suggest that there is potential for geographically structured coevolution between M. lupulina and the Ensifer genus, but not between M. lupulina and either Ensifer species. © 2017 The Author(s). Evolution © 2017 The Society for the Study of Evolution.

Aislamiento y caracterización de cepas de rizobios aisladas de diferentes leguminosas en la región de Cascajal, Villa Clara Isolation and characterization of rhizobia strains isolated from different legumes in the Cascajal region, Villa Clara

Directory of Open Access Journals (Sweden)

Pérez Guianeya

2008-06-01

Full Text Available Se aislaron 19 cepas de rizobio a partir de los nódulos de las leguminosas Canavalia ensiformis, Stylosanthes guianensis, Centrosema molle, Pueraria phaseoloides y Macroptilium atropurpureum, con el objetivo de obtener inoculantes efectivos para dichos cultivos en suelos ácidos. Para la caracterización de los aislados bacterianos se estudiaron sus características micromorfológicas, tintoriales y culturales, así como algunas respuestas fisiológico-bioquímicas, tales como la excreción de ácido o base al medio de cultivo y el ensayo de la cetolactasa. Se hizo un ensayo de nodulación in vitro con cada una de las leguminosas para determinar la efectividad de los aislados. De acuerdo con las características culturales, la tasa de crecimiento en medio LMA y la producción de ácido o base, los aislados obtenidos pudieran pertenecer a los géneros Bradyrhizobium y Rhizobium y/o Sinorhizobium. La prueba de la cetolactasa permitió conocer que ninguno de los aislados en estudio pertenece al género Agrobacterium. El ensayo con plantas inoculadas mostró que todas las cepas aisladas fueron efectivas en la nodulación, ya que nodularon en las leguminosas probadas. Se concluye que de las cepas aisladas 11 pudieran pertenecer al género Bradyrhizobium, mientras que las ocho restantes comparten características similares con los géneros Rhizobium y/o Sinorhizobium; todos los aislados fueron efectivos en la nodulación y pueden considerarse cepas promisorias para la obtención de inoculantes para las leguminosas forrajeras en estudio.Nineteen rhizobium strains were isolated from the nodules of the legumes Canavalia ensiformis, Stylosanthes guianensis, Centrosema molle, Pueraria phaseoloides and Macroptilium atropurpureum, with the objective of obtaining effective inoculants for such crops in acid soils. For the characterization of the bacterial isolates, their micromorphological, staining and cultural characteristics were studied, as well as some

Practical use of CMC-amended rhizobial inoculant for Mucuna pruriens cultivation to enhance the growth and protection against Macrophomina phaseolina.

Science.gov (United States)

Aeron, Abhinav; Khare, Ekta; Kumar Arora, Naveen; Kumar Maheshwari, Dinesh

2012-01-01

In many parts of the world Mucuna pruriens is used as an important medicinal, forage and green manure crop. In the present investigation the effect of the addition of CMC in carrier during development of bioformulation on shelflife, plant growth promotive and biocontrol activity against Macrophomina phaseolina was screened taking M. pruriens as a test crop. Ensifer meliloti RMP6(Ery+Kan+) and Bradyrhizobium sp. BMP7(Tet+Kan+) (kanamycin resistance engineered by Tn5 transposon mutagenesis) used in the study showed production of siderophore, IAA, solubilizing phosphate and biocontrol of M. phaseolina. RMP6(Ery+Kan+) also showed ACC deaminase activity. The survival of both the strains in sawdust-based bioformulation was enhanced with an increase in the concentration of CMC from 0 to 1%. At 0% CMC Bradyrhizobium sp. BMP7(Tet+Kan+) showed more increase in nodule number/plant (500.00%) than E. meliloti RMP6(Ery+Kan+) (52.38%), over the control in M. phaseolina-infested soil. There was 185.94% and 59.52% enhancement in nodule number/plant by RMP6(Ery+Kan+) and BMP7(Tet+Kan+) with an increase in the concentration of CMC from 0% to 1% in the bioformulations. However further increase in concentration of CMC did not result in enhancement in survival of either the strains or nodule number/plant.

STABILITY IN REAL TIME OF SOME CRYOPRESERVED MICROBIAL STRAINS WITH REFERENCE TO GENETICALLY MODIFIED MICROORGANISMS

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DANIELA VINTILĂ

2007-05-01

Full Text Available The aim of this work is to analyze the viability of microorganisms from Collection of Industrial Microorganisms from Faculty of Animal Science and Biotechnology – Timisoara, during freezing and thawing as part of cryopreservation technique. The stability in real time of 19 strains cryopreserved in 16% glycerol was evaluated during a 6-months period. The strains studied were: Escherichia coli, Lactobacillus acidophilus, Rhizobium meliloti, Saccharomyces cerevisiae, Aspergillus oryzae, Aspergillus niger, Trichoderma viride, Bacillus globigii, Bacillus licheniformis, and 9 strains of Bacillus subtilis. The strains cryopreserved at -20oC and -70oC were activated using the fast thawing protocol. A better cell recovery was achieved with the -70oC protocol reaching an average viability for E. coli of 86,3%, comparing with 78,6% in -20oC protocol. The cell recovery percentages for the other strains were: 92,4% for L. acidophilus, 93,9% for A.niger, 89% for A. oryzae, 86,7% for T. viride, 94,2% for R. meliloti, 82,1% for S. cerevisiae, 89,9% for B. licheniformis. Regarding the viability of genetically modified microorganisms, the values shows a good recovering after freezing and thawing, even after 180 days of cryopreservation. With the -20oC protocol lower viability was observed due probably to the formation of eutectic mixtures and recrystalization processes.

Concurrent synthesis and release of nod-gene-inducing flavonoids from alfalfa roots

International Nuclear Information System (INIS)

Maxwell, C.A.; Phillips, D.A.

1990-01-01

Flavonoid signals from alfalfa (Medicago sativa L.) induce transcription of nodulation (nod) genes in Rhizobium meliloti. Alfalfa roots release three major nod-gene inducers: 4',7-dihydroxyflavanone, 4',7-dihydroxyflavone, and 4,4'-dihydroxy-2'-methoxychalcone. The objective of the present study was to define temporal relationships between synthesis and exudation for those flavonoids. Requirements for concurrent flavonoid biosynthesis were assessed by treating roots of intact alfalfa seedlings with [U- 14 C]-L-phenylalanine in the presence or absence of the phenylalanine ammonia-lyase inhibitor L-2-aminoxy-3-phenylpropionic acid (AOPP). In the absence of AOPP, each of the three flavonoids in exudates contained 14 C. In the presence of AOPP, 14 C labeling and release of all the exuded nod-gene inducers were reduced significantly. AOPP inhibited labeling and release of the strongest nod-gene inducer, methoxychalcone, by more than 90%. The release process responsible for exudation of nod-gene inducers appears to be specific rather than a general phenomenon such as a sloughing off of cells during root growth

Effect of vanadium and tungsten on nitrogen fixation and the growth of Medicago sativa

Energy Technology Data Exchange (ETDEWEB)

Jha, K K

1969-01-01

In sand culture, it was found that vanadium had no stimulatory effect on nitrogen content or the growth of Medicago sativa inoculated with an effective strain of Rhizobium meliloti or supplied with ammonium nitrate. At the level of 500 ppm it reduced the plant growth, the inhibitory effect being particularly severe on the root. On the other hand tungsten increased nitrogen fixation and the dry matter yield of the inoculated plants. The results are suggestive of a direct role of tungsten in symbiotic nitrogen fixation. 4 references, 2 tables.

The abundance and diversity of legume-nodulating rhizobia in 28-year-old plantations of tropical, subtropical, and exotic tree species: a case study from the Forest Reserve of Bandia, Senegal.

Science.gov (United States)

Sene, Godar; Thiao, Mansour; Samba-Mbaye, Ramatoulaye; Khasa, Damase; Kane, Aboubacry; Mbaye, Mame Samba; Beaulieu, Marie-Ève; Manga, Anicet; Sylla, Samba Ndao

2013-01-01

Several fast-growing and multipurpose tree species have been widely used in West Africa to both reverse the tendency of land degradation and restore soil productivity. Although beneficial effects have been reported on soil stabilization, there still remains a lack of information about their impact on soil microorganisms. Our investigation has been carried out in exotic and native tree plantations of 28 years and aimed to survey and compare the abundance and genetic diversity of natural legume-nodulating rhizobia (LNR). The study of LNR is supported by the phylogenetic analysis which clustered the isolates into three genera: Bradyrhizobium, Mesorhizobium, and Sinorhizobium. The results showed close positive correlations between the sizes of LNR populations estimated both in the dry and rainy seasons and the presence of legume tree hosts. There were significant increases in Rhizobium spp. population densities in response to planting with Acacia spp., and high genetic diversities and richness of genotypes were fittest in these tree plantations. This suggests that enrichment of soil Rhizobium spp. populations is host specific. The results indicated also that species of genera Mesorhizobium and Sinorhizobium were lacking in plantations of non-host species. By contrast, there was a widespread distribution of Bradyrhizobium spp. strains across the tree plantations, with no evident specialization in regard to plantation type. Finally, the study provides information about the LNR communities associated with a range of old tree plantations and some aspects of their relationships to soil factors, which may facilitate the management of man-made forest systems that target ecosystem rehabilitation and preservation of soil biota.

Informe científico de investigador: Balatti, Pedro Alberto (2012)

OpenAIRE

Balatti, Pedro Alberto

2012-01-01

Se analizó la diversidad genética existente en los rizobios de crecimiento rápido que nodulan la soja. Por medio de la reacción en cadena de la polimerasa y con diversos primers se generaron marcadores ERIC, REP y RAPD. Además, se amplificó el RNA de 16S el que restringión con endonucleasas. Por medio de estos marcadores se estableció, entre los representantes de Sinorhizobium fredii aislados de la China y Vietnam y Rhizobium sp. NGR234, la diversidad genética existente entre ellos. Además se...

Assessing Requirements Quality through Requirements Coverage

Science.gov (United States)

Rajan, Ajitha; Heimdahl, Mats; Woodham, Kurt

2008-01-01

In model-based development, the development effort is centered around a formal description of the proposed software system the model. This model is derived from some high-level requirements describing the expected behavior of the software. For validation and verification purposes, this model can then be subjected to various types of analysis, for example, completeness and consistency analysis [6], model checking [3], theorem proving [1], and test-case generation [4, 7]. This development paradigm is making rapid inroads in certain industries, e.g., automotive, avionics, space applications, and medical technology. This shift towards model-based development naturally leads to changes in the verification and validation (V&V) process. The model validation problem determining that the model accurately captures the customer's high-level requirements has received little attention and the sufficiency of the validation activities has been largely determined through ad-hoc methods. Since the model serves as the central artifact, its correctness with respect to the users needs is absolutely crucial. In our investigation, we attempt to answer the following two questions with respect to validation (1) Are the requirements sufficiently defined for the system? and (2) How well does the model implement the behaviors specified by the requirements? The second question can be addressed using formal verification. Nevertheless, the size and complexity of many industrial systems make formal verification infeasible even if we have a formal model and formalized requirements. Thus, presently, there is no objective way of answering these two questions. To this end, we propose an approach based on testing that, when given a set of formal requirements, explores the relationship between requirements-based structural test-adequacy coverage and model-based structural test-adequacy coverage. The proposed technique uses requirements coverage metrics defined in [9] on formal high-level software

Tripartite symbiosis of Sophora tomentosa, rhizobia and arbuscular mycorhizal fungi.

Science.gov (United States)

Toma, Maíra Akemi; Soares de Carvalho, Teotonio; Azarias Guimarães, Amanda; Martins da Costa, Elaine; Savana da Silva, Jacqueline; de Souza Moreira, Fatima Maria

Sophora tomentosa is a pantropical legume species with potential for recovery of areas degraded by salinization, and for stabilization of sand dunes. However, few studies on this species have been carried out, and none regarding its symbiotic relationship with beneficial soil microorganisms. Therefore, this study aimed to evaluate the diversity of nitrogen-fixing bacteria isolated from nodules of Sophora tomentosa, and to analyze the occurrence of colonization of arbuscular mycorrhizal fungi on the roots of this legume in seafront soil. Thus, seeds, root nodules, and soil from the rhizosphere of Sophora tomentosa were collected. From the soil samples, trap cultures with this species were established to extract spores and to evaluate arbuscular mycorhizal fungi colonization in legume roots, as well as to capture rhizobia. Rhizobia strains were isolated from nodules collected in the field or from the trap cultures. Representative isolates of the groups obtained in the similarity dendrogram, based on phenotypic characteristics, had their 16S rRNA genes sequenced. The legume species showed nodules with indeterminate growth, and reddish color, distributed throughout the root. Fifty-one strains of these nodules were isolated, of which 21 were classified in the genus Bacillus, Brevibacillus, Paenibacillus, Rhizobium and especially Sinorhizobium. Strains closely related to Sinorhizobium adhaerens were the predominant bacteria in nodules. The other genera found, with the exception of Rhizobium, are probably endophytic bacteria in the nodules. Arbuscular mycorrhizal fungi was observed colonizing the roots, but arbuscular mycorhizal fungi spores were not found in the trap cultures. Therefore Sophora tomentosa is associated with both arbuscular mycorhizal fungi and nodulating nitrogen-fixing bacteria. Copyright © 2017 Sociedade Brasileira de Microbiologia. Published by Elsevier Editora Ltda. All rights reserved.

Software requirements

CERN Document Server

Wiegers, Karl E

2003-01-01

Without formal, verifiable software requirements-and an effective system for managing them-the programs that developers think they've agreed to build often will not be the same products their customers are expecting. In SOFTWARE REQUIREMENTS, Second Edition, requirements engineering authority Karl Wiegers amplifies the best practices presented in his original award-winning text?now a mainstay for anyone participating in the software development process. In this book, you'll discover effective techniques for managing the requirements engineering process all the way through the development cy

Seleção de bactérias endofíticas de tomateiro como potenciais agentes de biocontrole e de promoção de crescimento Screening of endophytic bacteria isolated from tomato plants as potencial biocontrol agents and growth promotion

Directory of Open Access Journals (Sweden)

Patrícia Baston Barretti

2009-01-01

Full Text Available Quarenta isolados bacterianos endofíticos de plantas sadias de tomateiro foram avaliados quanto à sua potencialidade como agentes de biocontrole de doenças do tomateiro. Foi realizada, em casa de vegetação, uma seleção massal utilizando-se Pseudomonas syringae pv. tomato e Alternaria solani, como patógenos desafiantes. Com base na média do número de lesões por planta, quatro isolados foram selecionados como potenciais agentes de biocontrole dessas enfermidades fúngica e bacteriana do tomateiro. Esses isolados foram identificados, por meio do sequenciamento do gene 16S do DNA ribossômico, como Acinetobacter johnsonii (UFV-E05, Serratia marcescens (UFV-E13, Sinorhizobium sp. (UFV-E25 e Bacillus megaterium (UFV-E26. Os mesmos isolados selecionados para o biocontrole também foram avaliados quanto à sua capacidade de promover o crescimento em plantas e somente S. marcescens (UFV-E13 proporcionou aumento na altura das plantas.Forty isolates of endophytic bacteria obtained from healthy tomato plants were tested for their potential as biocontrol agents of tomato diseases. A massal screening was performed at greenhouse using Pseudomonas syringae pv. tomato and Alternaria solani as challenging pathogens. Based on the average number of lesions per plant, four isolates were selected as potential agents of biocontrol of these tomato diseases caused by fungi and bacteria. These isolates were identified by 16S ribosomal DNA sequence analysis as Acinetobacter johnsonii (UFV-E05, Serratia marcescens (UFV-E13, Sinorhizobium sp. (UFV-E25 and Bacillus megaterium (UFV-E26. The four endophytes selected for biocontrol were also evaluated for their ability of promoting plant growth and only S. marcescens (UFV-E13 presented increase in the height of the plants.

Anticipating requirements changes-using futurology in requirements elicitation

OpenAIRE

Pimentel, João Henrique; Santos, Emanuel; Castro, Jaelson; Franch Gutiérrez, Javier

2012-01-01

It is well known that requirements changes in a later phase of software developments is a major source of software defects and costs. Thus, the need of techniques to control or reduce the amount of changes during software development projects. The authors advocate the use of foresight methods as a valuable input to requirements elicitation, with the potential to decrease the number of changes that would be required after deployment, by anticipating them. In this paper, the authors define a pr...

Evaluation of nitrogen fixation rates of soybean and species of rhizobia

International Nuclear Information System (INIS)

Ping Shuzhen; You Chongbiao

1993-01-01

Using 1 '5N dilution technique the nitrogen fixing rates were estimated from symbiosis between 20 varieties of soybean and 2 species of rhizobia: Bradyrhizobium japonicum and Sinorhizobium fredii. The nodulation status such as size and number of nodules differed among the varieties and species of bacteria. The Ndfa% of these varieties ranged from 40% ∼ 59% for S. fredii, 38% ∼ 62% for B. japonicum and 32% ∼ 56% for inoculant of the mixture of the species, respectively. Among 3 inoculants the B. japonicum is the best one. The variety of soybean, however, plays a significant role in the symbiosis. Therefore, improving the soybean and selecting a good partner for raising the yield and nitrogen fixation are important

Capturing Requirements for Autonomous Spacecraft with Autonomy Requirements Engineering

Science.gov (United States)

Vassev, Emil; Hinchey, Mike

2014-08-01

The Autonomy Requirements Engineering (ARE) approach has been developed by Lero - the Irish Software Engineering Research Center within the mandate of a joint project with ESA, the European Space Agency. The approach is intended to help engineers develop missions for unmanned exploration, often with limited or no human control. Such robotics space missions rely on the most recent advances in automation and robotic technologies where autonomy and autonomic computing principles drive the design and implementation of unmanned spacecraft [1]. To tackle the integration and promotion of autonomy in software-intensive systems, ARE combines generic autonomy requirements (GAR) with goal-oriented requirements engineering (GORE). Using this approach, software engineers can determine what autonomic features to develop for a particular system (e.g., a space mission) as well as what artifacts that process might generate (e.g., goals models, requirements specification, etc.). The inputs required by this approach are the mission goals and the domain-specific GAR reflecting specifics of the mission class (e.g., interplanetary missions).

Azospirillum IV

International Nuclear Information System (INIS)

Klingmuller, W.

1988-01-01

This book's contents include: Advances in the genetics of Azospirillum brasilense Sp7: Use of Tn5 mutagenesis for gene mapping and identification; Characterization of DNA segments adjacent to the nifHDK genes of Azospirillum brasilense by Sp7 Tn5 site-directed mutagenesis; Selection at the chemostat of Azospirillum brasilense Cd N 2 -fixing at high O 2 pressure. Root hair deformation induced on maize and medicago by an Azospirillum transconjugant containing a Rhizobium meliloti nodulation region. Azospirilla are bacteria that live in association with the roots of many grain crops. Since these bacteria bind molecular nitrogen from the air and excrete plant growth substances, interest has focussed on their potential to increase crop yields

Tool-based requirement traceability between requirement and design artifacts

CERN Document Server

Turban, Bernhard

2013-01-01

Processes for developing safety-critical systems impose special demands on ensuring requirements traceability. Achieving valuable traceability information, however, is especially difficult concerning the transition from requirements to design. Bernhard Turban analyzes systems and software engineering theories cross-cutting the issue (embedded systems development, systems engineering, software engineering, requirements engineering and management, design theory and processes for safety-critical systems). As a solution, the author proposes a new tool approach to support designers in their thinkin

The center for plant and microbial complex carbohydrates at the University of Georgia Complex Carbohydrate Research Center

Energy Technology Data Exchange (ETDEWEB)

Albersheim, P.; Darvill, A.

1991-08-01

Research from the Complex Carbohydrates Research Center at the University of Georgia is presented. Topics include: Structural determination of soybean isoflavones which specifically induce Bradyrhizobium japonicum nodD1 but not the nodYABCSUIJ operon; structural analysis of the lipopolysaccharides (LPSs) from symbiotic mutants of Bradyrhizobium japonicum; structural characterization of lipooligosaccharides from Bradyrhizobium japonicum that are required for the specific nodulation of soybean; structural characterization of the LPSs from R. Leguminosarum biovar phaseoli, the symbiont of bean; characterization of bacteroid-specific LPS epitopes in R. leguminosarum biovar viciae; analysis of the surface polysaccharides of Rhizobium meliloti mutants whose lipopolysaccharides and extracellular polysaccharides can have the same function in symbiosis; characterization of a polysaccharide produced by certain Bradyrhizobium japonicum strains within soybean nodules; structural analysis of a streptococcal adhesin polysaccharide receptor; conformational studies of xyloglucan, the role of the fucosylated side chain in surface-specific cellulose-xyloglucan interactions; the structure of an acylated glucosamine oligosaccharide signal molecule (nod factor) involved in the symbiosis of Rhizobium leguminosarum biovar viciae with its host Vicia sativa; investigating membrane responses induced by oligogalacturonides in cultured cells; the polygalacturonase inhibitor protein; characterization of the self-incompatability glycoproteins from Petunia hybrida; investigation of the cell wall polysaccharide structures of Arabidopsis thaliana; and the glucan inhibition of virus infection of tabacco.

The center for plant and microbial complex carbohydrates at the University of Georgia Complex Carbohydrate Research Center. Annual report, September 15, 1990--December 31, 1991

Energy Technology Data Exchange (ETDEWEB)

Albersheim, P.; Darvill, A.

1991-08-01

Research from the Complex Carbohydrates Research Center at the University of Georgia is presented. Topics include: Structural determination of soybean isoflavones which specifically induce Bradyrhizobium japonicum nodD1 but not the nodYABCSUIJ operon; structural analysis of the lipopolysaccharides (LPSs) from symbiotic mutants of Bradyrhizobium japonicum; structural characterization of lipooligosaccharides from Bradyrhizobium japonicum that are required for the specific nodulation of soybean; structural characterization of the LPSs from R. Leguminosarum biovar phaseoli, the symbiont of bean; characterization of bacteroid-specific LPS epitopes in R. leguminosarum biovar viciae; analysis of the surface polysaccharides of Rhizobium meliloti mutants whose lipopolysaccharides and extracellular polysaccharides can have the same function in symbiosis; characterization of a polysaccharide produced by certain Bradyrhizobium japonicum strains within soybean nodules; structural analysis of a streptococcal adhesin polysaccharide receptor; conformational studies of xyloglucan, the role of the fucosylated side chain in surface-specific cellulose-xyloglucan interactions; the structure of an acylated glucosamine oligosaccharide signal molecule (nod factor) involved in the symbiosis of Rhizobium leguminosarum biovar viciae with its host Vicia sativa; investigating membrane responses induced by oligogalacturonides in cultured cells; the polygalacturonase inhibitor protein; characterization of the self-incompatability glycoproteins from Petunia hybrida; investigation of the cell wall polysaccharide structures of Arabidopsis thaliana; and the glucan inhibition of virus infection of tabacco.

Environmental Requirements Management

Energy Technology Data Exchange (ETDEWEB)

Cusack, Laura J.; Bramson, Jeffrey E.; Archuleta, Jose A.; Frey, Jeffrey A.

2015-01-08

CH2M HILL Plateau Remediation Company (CH2M HILL) is the U.S. Department of Energy (DOE) prime contractor responsible for the environmental cleanup of the Hanford Site Central Plateau. As part of this responsibility, the CH2M HILL is faced with the task of complying with thousands of environmental requirements which originate from over 200 federal, state, and local laws and regulations, DOE Orders, waste management and effluent discharge permits, Comprehensive Environmental Response, Compensation, and Liability Act (CERCLA) response and Resource Conservation and Recovery Act (RCRA) corrective action documents, and official regulatory agency correspondence. The challenge is to manage this vast number of requirements to ensure they are appropriately and effectively integrated into CH2M HILL operations. Ensuring compliance with a large number of environmental requirements relies on an organization’s ability to identify, evaluate, communicate, and verify those requirements. To ensure that compliance is maintained, all changes need to be tracked. The CH2M HILL identified that the existing system used to manage environmental requirements was difficult to maintain and that improvements should be made to increase functionality. CH2M HILL established an environmental requirements management procedure and tools to assure that all environmental requirements are effectively and efficiently managed. Having a complete and accurate set of environmental requirements applicable to CH2M HILL operations will promote a more efficient approach to: • Communicating requirements • Planning work • Maintaining work controls • Maintaining compliance

Feed tank transfer requirements

International Nuclear Information System (INIS)

Freeman-Pollard, J.R.

1998-01-01

This document presents a definition of tank turnover; DOE responsibilities; TWRS DST permitting requirements; TWRS Authorization Basis (AB) requirements; TWRS AP Tank Farm operational requirements; unreviewed safety question (USQ) requirements; records and reporting requirements, and documentation which will require revision in support of transferring a DST in AP Tank Farm to a privatization contractor for use during Phase 1B

CH2M Hill Hanford Group, Inc., Standards and Requirements Identification Document (SRID) Requirements Management System and Requirements Specification

International Nuclear Information System (INIS)

JOHNSON, A.L.

2000-01-01

The current Tank Farm Contractor (TFC) for the U. S. Department of Energy, Office of River Protection (ORP), River Protection Project (RPP), CH2M Hill Hanford Group, Inc. (CHG), will use a computer based requirements management system. The system will serve as a tool to assist in identifying, capturing, and maintaining the Standards/Requirements Identification Document (S/RID) requirements and links to implementing procedures and other documents. By managing requirements as one integrated set, CHG will be able to carry out its mission more efficiently and effectively. CHG has chosen the Dynamic Object Oriented Requirements System (DOORS(trademark)) as the preferred computer based requirements management system. Accordingly, the S/RID program will use DOORS(trademark). DOORS(trademark) will replace the Environmental Requirements Management Interface (ERMI) system as the tool for S/RID data management. The DOORS(trademark) S/RID test project currently resides on the DOORSTM test server. The S/RID project will be migrated to the DOORS(trademark) production server. After the migration the S/RID project will be considered a production project and will no longer reside on the test server

Feed tank transfer requirements

Energy Technology Data Exchange (ETDEWEB)

Freeman-Pollard, J.R.

1998-09-16

This document presents a definition of tank turnover. Also, DOE and PC responsibilities; TWRS DST permitting requirements; TWRS Authorization Basis (AB) requirements; TWRS AP Tank Farm operational requirements; unreviewed safety question (USQ) requirements are presented for two cases (i.e., tank modifications occurring before tank turnover and tank modification occurring after tank turnover). Finally, records and reporting requirements, and documentation which will require revision in support of transferring a DST in AP Tank Farm to a privatization contractor are presented.

Feed tank transfer requirements

International Nuclear Information System (INIS)

Freeman-Pollard, J.R.

1998-01-01

This document presents a definition of tank turnover. Also, DOE and PC responsibilities; TWRS DST permitting requirements; TWRS Authorization Basis (AB) requirements; TWRS AP Tank Farm operational requirements; unreviewed safety question (USQ) requirements are presented for two cases (i.e., tank modifications occurring before tank turnover and tank modification occurring after tank turnover). Finally, records and reporting requirements, and documentation which will require revision in support of transferring a DST in AP Tank Farm to a privatization contractor are presented

Azospirillum IV

Energy Technology Data Exchange (ETDEWEB)

Klingmuller, W.

1988-01-01

This book's contents include: Advances in the genetics of Azospirillum brasilense Sp7: Use of Tn5 mutagenesis for gene mapping and identification; Characterization of DNA segments adjacent to the nifHDK genes of Azospirillum brasilense by Sp7 Tn5 site-directed mutagenesis; Selection at the chemostat of Azospirillum brasilense Cd N/sub 2/-fixing at high O/sub 2/ pressure. Root hair deformation induced on maize and medicago by an Azospirillum transconjugant containing a Rhizobium meliloti nodulation region. Azospirilla are bacteria that live in association with the roots of many grain crops. Since these bacteria bind molecular nitrogen from the air and excrete plant growth substances, interest has focussed on their potential to increase crop yields.

Transportation System Requirements Document

International Nuclear Information System (INIS)

1993-09-01

This Transportation System Requirements Document (Trans-SRD) describes the functions to be performed by and the technical requirements for the Transportation System to transport spent nuclear fuel (SNF) and high-level radioactive waste (HLW) from Purchaser and Producer sites to a Civilian Radioactive Waste Management System (CRWMS) site, and between CRWMS sites. The purpose of this document is to define the system-level requirements for Transportation consistent with the CRWMS Requirement Document (CRD). These requirements include design and operations requirements to the extent they impact on the development of the physical segments of Transportation. The document also presents an overall description of Transportation, its functions, its segments, and the requirements allocated to the segments and the system-level interfaces with Transportation. The interface identification and description are published in the CRWMS Interface Specification

Replacing reserve requirements

OpenAIRE

Edward J. Stevens

1993-01-01

An examination of the fading significance of the Federal Reserve System's reserve requirements and the recent flowering of required clearing balances, a rapidly growing feature of Reserve Bank operations.

TRANSPORTATION SYSTEM REQUIREMENTS DOCUMENT

International Nuclear Information System (INIS)

2004-01-01

This document establishes the Transportation system requirements for the U.S. Department of Energy's (DOE's) Civilian Radioactive Waste Management System (CRWMS). These requirements are derived from the Civilian Radioactive Waste Management System Requirements Document (CRD). The Transportation System Requirements Document (TSRD) was developed in accordance with LP-3.1Q-OCRWM, Preparation, Review, and Approval of Office of National Transportation Level-2 Baseline Requirements. As illustrated in Figure 1, the TSRD forms a part of the DOE Office of Civilian Radioactive Waste Management (OCRWM) Technical Baseline

Closure requirements

International Nuclear Information System (INIS)

Hutchinson, I.P.G.; Ellison, R.D.

1992-01-01

Closure of a waste management unit can be either permanent or temporary. Permanent closure may be due to: economic factors which make it uneconomical to mine the remaining minerals; depletion of mineral resources; physical site constraints that preclude further mining and beneficiation; environmental, regulatory or other requirements that make it uneconomical to continue to develop the resources. Temporary closure can occur for a period of several months to several years, and may be caused by factors such as: periods of high rainfall or snowfall which prevent mining and waste disposal; economic circumstances which temporarily make it uneconomical to mine the target mineral; labor problems requiring a cessation of operations for a period of time; construction activities that are required to upgrade project components such as the process facilities and waste management units; and mine or process plant failures that require extensive repairs. Permanent closure of a mine waste management unit involves the provision of durable surface containment features to protect the waters of the State in the long-term. Temporary closure may involve activities that range from ongoing maintenance of the existing facilities to the installation of several permanent closure features in order to reduce ongoing maintenance. This paper deals with the permanent closure features

Hanford analytical services quality assurance requirements documents. Volume 1: Administrative Requirements

International Nuclear Information System (INIS)

Hyatt, J.E.

1997-01-01

Hanford Analytical Services Quality Assurance Requirements Document (HASQARD) is issued by the Analytical Services, Program of the Waste Management Division, US Department of Energy (US DOE), Richland Operations Office (DOE-RL). The HASQARD establishes quality requirements in response to DOE Order 5700.6C (DOE 1991b). The HASQARD is designed to meet the needs of DOE-RL for maintaining a consistent level of quality for sampling and field and laboratory analytical services provided by contractor and commercial field and laboratory analytical operations. The HASQARD serves as the quality basis for all sampling and field/laboratory analytical services provided to DOE-RL through the Analytical Services Program of the Waste Management Division in support of Hanford Site environmental cleanup efforts. This includes work performed by contractor and commercial laboratories and covers radiological and nonradiological analyses. The HASQARD applies to field sampling, field analysis, and research and development activities that support work conducted under the Hanford Federal Facility Agreement and Consent Order Tri-Party Agreement and regulatory permit applications and applicable permit requirements described in subsections of this volume. The HASQARD applies to work done to support process chemistry analysis (e.g., ongoing site waste treatment and characterization operations) and research and development projects related to Hanford Site environmental cleanup activities. This ensures a uniform quality umbrella to analytical site activities predicated on the concepts contained in the HASQARD. Using HASQARD will ensure data of known quality and technical defensibility of the methods used to obtain that data. The HASQARD is made up of four volumes: Volume 1, Administrative Requirements; Volume 2, Sampling Technical Requirements; Volume 3, Field Analytical Technical Requirements; and Volume 4, Laboratory Technical Requirements. Volume 1 describes the administrative requirements

Conversion of dependability deterministic requirements into probabilistic requirements

International Nuclear Information System (INIS)

Bourgade, E.; Le, P.

1993-02-01

This report concerns the on-going survey conducted jointly by the DAM/CCE and NRE/SR branches on the inclusion of dependability requirements in control and instrumentation projects. Its purpose is to enable a customer (the prime contractor) to convert into probabilistic terms dependability deterministic requirements expressed in the form ''a maximum permissible number of failures, of maximum duration d in a period t''. The customer shall select a confidence level for each previously defined undesirable event, by assigning a maximum probability of occurrence. Using the formulae we propose for two repair policies - constant rate or constant time - these probabilized requirements can then be transformed into equivalent failure rates. It is shown that the same formula can be used for both policies, providing certain realistic assumptions are confirmed, and that for a constant time repair policy, the correct result can always be obtained. The equivalent failure rates thus determined can be included in the specifications supplied to the contractors, who will then be able to proceed to their previsional justification. (author), 8 refs., 3 annexes

Requirements in engineering projects

CERN Document Server

Fernandes, João M

2016-01-01

This book focuses on various topics related to engineering and management of requirements, in particular elicitation, negotiation, prioritisation, and documentation (whether with natural languages or with graphical models). The book provides methods and techniques that help to characterise, in a systematic manner, the requirements of the intended engineering system.  It was written with the goal of being adopted as the main text for courses on requirements engineering, or as a strong reference to the topics of requirements in courses with a broader scope. It can also be used in vocational courses, for professionals interested in the software and information systems domain.   Readers who have finished this book will be able to: - establish and plan a requirements engineering process within the development of complex engineering systems; - define and identify the types of relevant requirements in engineering projects; - choose and apply the most appropriate techniques to elicit the requirements of a giv...

An Engineering Method of Civil Jet Requirements Validation Based on Requirements Project Principle

Science.gov (United States)

Wang, Yue; Gao, Dan; Mao, Xuming

2018-03-01

A method of requirements validation is developed and defined to meet the needs of civil jet requirements validation in product development. Based on requirements project principle, this method will not affect the conventional design elements, and can effectively connect the requirements with design. It realizes the modern civil jet development concept, which is “requirement is the origin, design is the basis”. So far, the method has been successfully applied in civil jet aircraft development in China. Taking takeoff field length as an example, the validation process and the validation method of the requirements are detailed introduced in the study, with the hope of providing the experiences to other civil jet product design.

From requirements to Java in a snap model-driven requirements engineering in practice

CERN Document Server

Smialek, Michal

2015-01-01

This book provides a coherent methodology for Model-Driven Requirements Engineering which stresses the systematic treatment of requirements within the realm of modelling and model transformations. The underlying basic assumption is that detailed requirements models are used as first-class artefacts playing a direct role in constructing software. To this end, the book presents the Requirements Specification Language (RSL) that allows precision and formality, which eventually permits automation of the process of turning requirements into a working system by applying model transformations and co

Getting Grip on Security Requirements Elicitation by Structuring and Reusing Security Requirements Sources

Directory of Open Access Journals (Sweden)

Christian Schmitt

2015-07-01

Full Text Available This paper presents a model for structuring and reusing security requirements sources. The model serves as blueprint for the development of an organization-specific repository, which provides relevant security requirements sources, such as security information and knowledge sources and relevant compliance obligations, in a structured and reusable form. The resulting repository is intended to be used by development teams during the elicitation and analysis of security requirements with the goal to understand the security problem space, incorporate all relevant requirements sources, and to avoid unnecessary effort for identifying, understanding, and correlating applicable security requirements sources on a project-wise basis. We start with an overview and categorization of important security requirements sources, followed by the description of the generic model. To demonstrate the applicability and benefits of the model, the instantiation approach and details of the resulting repository of security requirements sources are presented.

Subsurface Contamination Focus Area technical requirements. Volume 1: Requirements summary

International Nuclear Information System (INIS)

Nickelson, D.; Nonte, J.; Richardson, J.

1996-10-01

This document summarizes functions and requirements for remediation of source term and plume sites identified by the Subsurface Contamination Focus Area. Included are detailed requirements and supporting information for source term and plume containment, stabilization, retrieval, and selective retrieval remedial activities. This information will be useful both to the decision-makers within the Subsurface Contamination Focus Area (SCFA) and to the technology providers who are developing and demonstrating technologies and systems. Requirements are often expressed as graphs or charts, which reflect the site-specific nature of the functions that must be performed. Many of the tradeoff studies associated with cost savings are identified in the text

Waste management system requirements document

International Nuclear Information System (INIS)

1991-02-01

This volume defines the top level requirements for the Mined Geologic Disposal System (MGDS). It is designed to be used in conjunction with Volume 1 of the WMSR, General System Requirements. It provides a functional description expanding the requirements allocated to the MGDS in Volume 1 and elaborates on each requirement by providing associated performance criteria as appropriate. Volumes 1 and 4 of the WMSR provide a minimum set of requirements that must be satisfied by the final MGDS design. This document sets forth specific requirements that must be fulfilled. It is not the intent or purpose of this top level document to describe how each requirement is to be satisfied in the final MGDS design. Each subsequent level of the technical document hierarchy must provide further guidance and definition as to how each of these requirements is to be implemented in the design. It is expected that each subsequent level of requirements will be significantly more detailed. Section 2 of this volume provides a functional description of the MGDS. Each function is addressed in terms of requirements, and performance criteria. Section 3 provides a list of controlling documents. Each document cited in a requirement of Chapter 2 is included in this list and is incorporated into this document as a requirement on the final system. The WMSR addresses only federal requirements (i.e., laws, regulations and DOE orders). State and local requirements are not addressed. However, it will be specifically noted at the potentially affected WMSR requirements that there could be additional or more stringent regulations imposed by a state or local requirements or administering agency over the cited federal requirements

User Requirements for Wireless

DEFF Research Database (Denmark)

in the elicitation process. Cases and user requirement elements discussed in the book include: User requirements elicitation processes for children, construction workers, and farmers User requirements for personalized services of a broadcast company Variations in user involvement Practical elements of user...

Future Home Network Requirements

DEFF Research Database (Denmark)

Charbonnier, Benoit; Wessing, Henrik; Lannoo, Bart

This paper presents the requirements for future Home Area Networks (HAN). Firstly, we discuss the applications and services as well as their requirements. Then, usage scenarios are devised to establish a first specification for the HAN. The main requirements are an increased bandwidth (towards 1...

Requirements model generation to support requirements elicitation: The Secure Tropos experience

NARCIS (Netherlands)

Kiyavitskaya, N.; Zannone, N.

2008-01-01

In recent years several efforts have been devoted by researchers in the Requirements Engineering community to the development of methodologies for supporting designers during requirements elicitation, modeling, and analysis. However, these methodologies often lack tool support to facilitate their

Molecular and phenotypic characterization of endophytic bacteria isolated from sulla nodules.

Science.gov (United States)

Beghalem, Hamida; Aliliche, Khadidja; Chriki, Ali; Landoulsi, Ahmed

2017-10-01

In the current study, bacterial diversity was investigated in root nodules of Sulla pallida and Sulla capitata. The isolates were analyzed on the basis of their phenotypic and molecular characteristics. The phylogenetic analysis based on 16S rRNA and housekeeping genes (recA and atpD) showed that the isolated bacteria related to Sinorhizobium, Neorhizobium, Phyllobacterium, Arthrobacter, Variovorax and Pseudomonas genera. This is the first report of Neorhizobium genus associated with Hedysarum genus. Phenotypically, all strains tolerate the elevated temperature of 40 °C, and salt stress at a concentration of 2%. In addition, the isolates failed to induce nodulation on their original host; and the symbiotic genes could not be amplified, suggesting that these strains are endophytic bacteria. Copyright © 2017 Elsevier Ltd. All rights reserved.

Root and Nodulation Phenotypes of the Ethylene-Insensitive Sickle Mutant of Medicago truncatula

Directory of Open Access Journals (Sweden)

JOKO PRAYITNO

2010-09-01

Full Text Available The sickle (skl mutant of the model legume Medicago truncatula is an ethylene-sensitive mutant that have a ten-fold increase in nodule numbers. The nodulation and root phenotypes of the skl mutant were investigated and further characterised. The skl mutant had longer roots than the wild type, but when inoculated with Sinorhizobium, its root length was reduced to the level of wild type. Furthermore, lateral root numbers in uninoculated skl were similar to those in uninoculated wild type. However, when the root tips were decapitated, fewer lateral roots formed in skl than in wild type. Nodule numbers of the skl mutant were significantly reduced by low nitrate concentration (2.5 mM. These results suggest that skl mutant has alterations in both root and nodule development.

System requirements and design description for the environmental requirements management interface (ERMI)

International Nuclear Information System (INIS)

Biebesheimer, E.

1997-01-01

This document describes system requirements and the design description for the Environmental Requirements Management Interface (ERMI). The ERMI database assists Tank Farm personnel with scheduling, planning, and documenting procedure compliance, performance verification, and selected corrective action tracking activities for Tank Farm S/RID requirements. The ERMI database was developed by Science Applications International Corporation (SAIC). This document was prepared by SAIC and edited by LMHC

12 CFR 564.3 - Appraisals required; transactions requiring a State certified or licensed appraiser.

Science.gov (United States)

2010-01-01

... 12 Banks and Banking 5 2010-01-01 2010-01-01 false Appraisals required; transactions requiring a State certified or licensed appraiser. 564.3 Section 564.3 Banks and Banking OFFICE OF THRIFT SUPERVISION, DEPARTMENT OF THE TREASURY APPRAISALS § 564.3 Appraisals required; transactions requiring a State...

PIT Coating Requirements Analysis

International Nuclear Information System (INIS)

MINTEER, D.J.

2000-01-01

This study identifies the applicable requirements for procurement and installation of a coating intended for tank farm valve and pump pit interior surfaces. These requirements are intended to be incorporated into project specification documents and design media. This study also evaluates previously recommended coatings and identifies requirement-compliant coating products

PIT Coating Requirements Analysis

Energy Technology Data Exchange (ETDEWEB)

MINTEER, D.J.

2000-10-20

This study identifies the applicable requirements for procurement and installation of a coating intended for tank farm valve and pump pit interior surfaces. These requirements are intended to be incorporated into project specification documents and design media. This study also evaluates previously recommended coatings and identifies requirement-compliant coating products.

Waste Management System Requirement document

International Nuclear Information System (INIS)

1990-04-01

This volume defines the top level technical requirements for the Monitored Retrievable Storage (MRS) facility. It is designed to be used in conjunction with Volume 1, General System Requirements. Volume 3 provides a functional description expanding the requirements allocated to the MRS facility in Volume 1 and, when appropriate, elaborates on requirements by providing associated performance criteria. Volumes 1 and 3 together convey a minimum set of requirements that must be satisfied by the final MRS facility design without unduly constraining individual design efforts. The requirements are derived from the Nuclear Waste Policy Act of 1982 (NWPA), the Nuclear Waste Policy Amendments Act of 1987 (NWPAA), the Environmental Protection Agency's (EPA) Environmental Standards for the Management and Disposal of Spent Nuclear Fuel (40 CFR 191), NRC Licensing Requirements for the Independent Storage of Spent Nuclear and High-Level Radioactive Waste (10 CFR 72), and other federal statutory and regulatory requirements, and major program policy decisions. This document sets forth specific requirements that will be fulfilled. Each subsequent level of the technical document hierarchy will be significantly more detailed and provide further guidance and definition as to how each of these requirements will be implemented in the design. Requirements appearing in Volume 3 are traceable into the MRS Design Requirements Document. Section 2 of this volume provides a functional breakdown for the MRS facility. 1 tab

Electrochemical selective detection of dopamine on microbial carbohydrate-doped multiwall carbon nanotube-modified electrodes.

Science.gov (United States)

Jin, Joon-Hyung; Cho, Eunae; Jung, Seunho

2010-03-01

Microbial carbohydrate-doped multiwall carbon nanotube (MWNT)-modified electrodes were prepared for the purpose of determining if 4-(2-aminoethyl)benzene-1,2-diol (3,4-dihydroxyphenylalanine; dopamine) exists in the presence of 0.5 mM ascorbic acid, a representative interfering agent in neurotransmitter detection. The microbial carbohydrate dopants were alpha-cyclosophorohexadecaose (alpha-C16) from Xanthomonas oryzae and cyclic-(1 --> 2)-beta-d-glucan (Cys) from Rhizobium meliloti. The cyclic voltammetric responses showed that the highest sensitivity (5.8 x 10(-3) mA cm(-2) microM(-1)) is attained with the Cys-doped MWNT-modified ultra-trace carbon electrode, and that the alpha-C16-doped MWNT-modified glassy carbon electrode displays the best selectivity to dopamine (the approximate peak potential separation is 310 mV).

Functional requirements of road lighting.

NARCIS (Netherlands)

Schreuder, D.A.

1975-01-01

The functional, technical and visual requirements for public lighting are discussed. The improvement of the presentation of information to the road user is the main functional requirement. The visual requirements can be deduced from the functional requirement of enabling drivers to follow the

Utility requirements for fusion

International Nuclear Information System (INIS)

Vondrasek, R.J.

1982-02-01

This report describes work done and results obtained during performance of Task 1 of a study of Utility Requirements and Criteria for Fusion Options. The work consisted of developing a list of utility requirements for fusion optics containing definition of the requirements and showing their relative importance to the utility industry. The project team members developed a preliminary list which was refined by discussions and literature searches. The refined list was recast as a questionnaire which was sent to a substantial portion of the utility industry in this country. Forty-three questionnaire recipients responded including thirty-two utilities. A workshop was held to develop a revised requirements list using the survey responses as a major input. The list prepared by the workshop was further refined by a panel consisting of vice presidents of the three project team firms. The results of the study indicate that in addition to considering the cost of energy for a power plant, utilities consider twenty-three other requirements. Four of the requirements were judged to be vital to plant acceptability: Plant Capital Cost, Financial Liability, Plant Safety and Licensability

Engineering Requirements for crowds

Directory of Open Access Journals (Sweden)

Rogeiro Silva

2015-12-01

Full Text Available In the software project the interested parts are highly distributed and form numerous and heterogeneous groups, online or face, constituting what could be called crowds. The development of social applications and cloud computing and mobile has generated a marked increase in environments based requirements in crowds. Technical Requirements Engineering (RE traditional face these scalability issues, and require the co-presence of interested and engineers in joint meetings that can not be made in common physical environments. While different approaches have been introduced to partially automate RE in these contexts, still is required a multi-method approach to semi-automate all activities related to work with crowds. In this paper is propose an approach that integrates existing elicitation techniques and requirements analysis and is complemented by introducing new concepts. The information is collected through direct interaction and social collaboration, and through data mining techniques.

Neutrosophy for software requirement prioritization

Directory of Open Access Journals (Sweden)

Ronald Barriga Dias

2017-09-01

Full Text Available Software engineers are involved in complex decisions that require multiples viewpoints. A specific case is the requirement prioritization process. This process is used to decide which software requirement to develop in certain release from a group of candidate requirements. Criteria involved in this process can involve indeterminacy. In this paper a software requirement prioritization model is develop based SVN numbers. Finally, an illustrative example is presented in order to show the proposed model.

Postmarket Requirements and Commitments

Data.gov (United States)

U.S. Department of Health & Human Services — Provides information to the public on postmarket requirements and commitments. The phrase postmarket requirements and commitments refers to studies and clinical...

40 CFR 63.2343 - What are my requirements for emission sources not requiring control?

Science.gov (United States)

2010-07-01

... 40 Protection of Environment 12 2010-07-01 2010-07-01 true What are my requirements for emission sources not requiring control? 63.2343 Section 63.2343 Protection of Environment ENVIRONMENTAL PROTECTION... (Non-Gasoline) What This Subpart Covers § 63.2343 What are my requirements for emission sources not...

Requirements for existing buildings

DEFF Research Database (Denmark)

Thomsen, Kirsten Engelund; Wittchen, Kim Bjarne

This report collects energy performance requirements for existing buildings in European member states by June 2012.......This report collects energy performance requirements for existing buildings in European member states by June 2012....

Safety Requirements and Modern Technical Requirements in Human Information Systems in Amman Hotels

OpenAIRE

Farouq Ahmad Alazzam; Sattam Rakan Allahawiah; Mohammad Nayef Alsarayreh; Kafa Hmoud Abdallah al Nawaiseh

2015-01-01

This study aimed to demonstrate the availability of Safety requirements and modern technical requirements in human information systems in Amman hotels. an the most important results of this study is the availability of security and safety requirements in human information systems In Amman hotels and The adequacy of the information that it provided .and show that all departments are not connected by appropriate and effective communication networks in adequate form . Also sophisticated operatin...

Autonomous Real Time Requirements Tracing

Science.gov (United States)

Plattsmier, George; Stetson, Howard

2014-01-01

One of the more challenging aspects of software development is the ability to verify and validate the functional software requirements dictated by the Software Requirements Specification (SRS) and the Software Detail Design (SDD). Insuring the software has achieved the intended requirements is the responsibility of the Software Quality team and the Software Test team. The utilization of Timeliner-TLX(sup TM) Auto- Procedures for relocating ground operations positions to ISS automated on-board operations has begun the transition that would be required for manned deep space missions with minimal crew requirements. This transition also moves the auto-procedures from the procedure realm into the flight software arena and as such the operational requirements and testing will be more structured and rigorous. The autoprocedures would be required to meet NASA software standards as specified in the Software Safety Standard (NASASTD- 8719), the Software Engineering Requirements (NPR 7150), the Software Assurance Standard (NASA-STD-8739) and also the Human Rating Requirements (NPR-8705). The Autonomous Fluid Transfer System (AFTS) test-bed utilizes the Timeliner-TLX(sup TM) Language for development of autonomous command and control software. The Timeliner-TLX(sup TM) system has the unique feature of providing the current line of the statement in execution during real-time execution of the software. The feature of execution line number internal reporting unlocks the capability of monitoring the execution autonomously by use of a companion Timeliner-TLX(sup TM) sequence as the line number reporting is embedded inside the Timeliner-TLX(sup TM) execution engine. This negates I/O processing of this type data as the line number status of executing sequences is built-in as a function reference. This paper will outline the design and capabilities of the AFTS Autonomous Requirements Tracker, which traces and logs SRS requirements as they are being met during real-time execution of the

Utility requirements for fusion power

International Nuclear Information System (INIS)

DeBellis, R.J.

1977-03-01

A four-man month study was undertaken to identify utility requirements of fusion power and define a role for the utilities in the fusion development process during the 1980s. This report, preliminary in nature, serves mainly as a planning document for future requirements analyses. A requirements organization was defined to consist of three major chronological phases: research and development, plant installation, and plant operation. Thirty-seven requirements were identified, covering all categories. In addition, training, environment, safety, licensing, and utility model were identified as five matrix-type requirements. As the requirement definition process continued during the study period, comments received from utility representatives revealed a consistency of key issues in the fusion development process. These issues form the basis for the eventual establishment of definitive roles for the utilities during the 1980s. The issues are not meant to reflect a negative view of fusion, but are items that must be solved before fusion can be introduced commercially as an electrical power source. As a result of this requirements study, preliminary candidate roles for the utilities in the fusion development process during the 1980s were identified as public education, commercialization studies, industry investment analyses, training plan implementation, alternate reactor concept development, ERDA concept design review, and requirements refinement

Creativity in Requirement Identification

DEFF Research Database (Denmark)

Sørensen, Lene Tolstrup; Olesen, Henning

Traditional requirements engineering focuses mainly on analysis and elicitation. However, current trends in new system, device and software are towards involving all stakeholders in the early stages of the engineering process to define the user requirements. Creativity is here seen as a major...... keystone in this process in order to open up stakeholder's mind to new technologies, which do not yet exist. This paper dis-cusses the application of creativity in the requirements process and illustrate through cases from the MAGNET and MAGNET Beyond projects....

Capital Requirements and Banks' Leniency

DEFF Research Database (Denmark)

Dietrich, J. Kimball; Wihlborg, Clas

2003-01-01

We investigate the effect of changes in capital regulation on the strictness(leniency) of loan terms using a simple model of bank capital requirements andasset quality examinations. Banks offer different levels of `leniency' in the senseof willingness to offer automatic extensions of loans...... rates. As capital requirements increase thedifference between initial capital levels and between interest rates of strict andlenient banks decrease. Thus, higher capital requirements in recessions tend toreduce the interest rate premium paid for leniency. If a recession is interpreted asan increase...... in the required return, the interest rate premium paid for leniency isincreased in recession at a given level of required capital....

Capturing security requirements for software systems.

Science.gov (United States)

El-Hadary, Hassan; El-Kassas, Sherif

2014-07-01

Security is often an afterthought during software development. Realizing security early, especially in the requirement phase, is important so that security problems can be tackled early enough before going further in the process and avoid rework. A more effective approach for security requirement engineering is needed to provide a more systematic way for eliciting adequate security requirements. This paper proposes a methodology for security requirement elicitation based on problem frames. The methodology aims at early integration of security with software development. The main goal of the methodology is to assist developers elicit adequate security requirements in a more systematic way during the requirement engineering process. A security catalog, based on the problem frames, is constructed in order to help identifying security requirements with the aid of previous security knowledge. Abuse frames are used to model threats while security problem frames are used to model security requirements. We have made use of evaluation criteria to evaluate the resulting security requirements concentrating on conflicts identification among requirements. We have shown that more complete security requirements can be elicited by such methodology in addition to the assistance offered to developers to elicit security requirements in a more systematic way.

Capturing security requirements for software systems

Directory of Open Access Journals (Sweden)

Hassan El-Hadary

2014-07-01

Full Text Available Security is often an afterthought during software development. Realizing security early, especially in the requirement phase, is important so that security problems can be tackled early enough before going further in the process and avoid rework. A more effective approach for security requirement engineering is needed to provide a more systematic way for eliciting adequate security requirements. This paper proposes a methodology for security requirement elicitation based on problem frames. The methodology aims at early integration of security with software development. The main goal of the methodology is to assist developers elicit adequate security requirements in a more systematic way during the requirement engineering process. A security catalog, based on the problem frames, is constructed in order to help identifying security requirements with the aid of previous security knowledge. Abuse frames are used to model threats while security problem frames are used to model security requirements. We have made use of evaluation criteria to evaluate the resulting security requirements concentrating on conflicts identification among requirements. We have shown that more complete security requirements can be elicited by such methodology in addition to the assistance offered to developers to elicit security requirements in a more systematic way.

Capturing security requirements for software systems

Science.gov (United States)

El-Hadary, Hassan; El-Kassas, Sherif

2014-01-01

Security is often an afterthought during software development. Realizing security early, especially in the requirement phase, is important so that security problems can be tackled early enough before going further in the process and avoid rework. A more effective approach for security requirement engineering is needed to provide a more systematic way for eliciting adequate security requirements. This paper proposes a methodology for security requirement elicitation based on problem frames. The methodology aims at early integration of security with software development. The main goal of the methodology is to assist developers elicit adequate security requirements in a more systematic way during the requirement engineering process. A security catalog, based on the problem frames, is constructed in order to help identifying security requirements with the aid of previous security knowledge. Abuse frames are used to model threats while security problem frames are used to model security requirements. We have made use of evaluation criteria to evaluate the resulting security requirements concentrating on conflicts identification among requirements. We have shown that more complete security requirements can be elicited by such methodology in addition to the assistance offered to developers to elicit security requirements in a more systematic way. PMID:25685514

Rapid quality assurance with requirements smells

OpenAIRE

Femmer, Henning; Méndez Fernández, Daniel; Wagner, Stefan; Eder, Sebastian

2016-01-01

Context: Bad requirements quality can cause expensive consequences during the software development lifecycle, especially if iterations are long and feedback comes late. Objectives: We aim at a light-weight static requirements analysis approach that allows for rapid checks immediately when requirements are written down. Method: We transfer the concept of code smells to Requirements Engineering as Requirements Smells. To evaluate the benefits and limitations, we define Requirements Smells, real...

National Ignition Facility site requirements

International Nuclear Information System (INIS)

1996-07-01

The Site Requirements (SR) provide bases for identification of candidate host sites for the National Ignition Facility (NIF) and for the generation of data regarding potential actual locations for the facilities. The SR supplements the NIF Functional Requirements (FR) with information needed for preparation of responses to queries for input to HQ DOE site evaluation. The queries are to include both documents and explicit requirements for the potential host site responses. The Sr includes information extracted from the NIF FR (for convenience), data based on design approaches, and needs for physical and organization infrastructure for a fully operational NIF. The FR and SR describe requirements that may require new construction or may be met by use or modification of existing facilities. The SR do not establish requirements for NIF design or construction project planning. The SR document does not constitute an element of the NIF technical baseline

Writing testable software requirements

Energy Technology Data Exchange (ETDEWEB)

Knirk, D. [Sandia National Labs., Albuquerque, NM (United States)

1997-11-01

This tutorial identifies common problems in analyzing requirements in the problem and constructing a written specification of what the software is to do. It deals with two main problem areas: identifying and describing problem requirements, and analyzing and describing behavior specifications.

Utility requirements for fusion power

International Nuclear Information System (INIS)

DeBellis, R.J.

1977-03-01

A four-man-month study, jointly funded by EPRI and McDonnell Douglas Astronautics Company-EAST, was undertaken to identify the utility requirements of fusion power and define a role for the utilities in the fusion development process during the 1980's. This report, preliminary in nature, serves mainly as a planning document for future requirements analyses. A requirements organization was defined to consist of three major chronological phases: research and development, plant installation, and plant operation. Thirty-seven requirements were identified, covering all categories. In addition, training, environment, safety, licensing, and utility model were identified as five matrix-type requirements. As the requirement definition process continued during the study period, comments received from utility representatives revealed a consistency of key issues in the fusion development process. These issues form the basis for the eventual establishment of definitive roles for the utilities during the 1980's. The issues are not meant to reflect a negative view of fusion, but are items which must be solved before fusion can be introduced commercially as an electrical power source. As a result of this requirements study, preliminary candidate roles for the utilities in the fusion development process during the 1980's were identified as public education, commercialization studies, industry investment analyses, training plan implementation, alternate reactor concept development, ERDA concept design review, and requirements refinement

Physician Requirements-1990. For Cardiology.

Science.gov (United States)

Tracy, Octavious; Birchette-Pierce, Cheryl

Professional requirements for physicians specializing in cardiology were estimated to assist policymakers in developing guidelines for graduate medical education. The determination of physician requirements was based on an adjusted needs rather than a demand or utilization model. For each illness, manpower requirements were modified by the…

Requirements Elicitation Problems: A Literature Analysis

Directory of Open Access Journals (Sweden)

Bill Davey

2015-06-01

Full Text Available Requirements elicitation is the process through which analysts determine the software requirements of stakeholders. Requirements elicitation is seldom well done, and an inaccurate or incomplete understanding of user requirements has led to the downfall of many software projects. This paper proposes a classification of problem types that occur in requirements elicitation. The classification has been derived from a literature analysis. Papers reporting on techniques for improving requirements elicitation practice were examined for the problem the technique was designed to address. In each classification the most recent or prominent techniques for ameliorating the problems are presented. The classification allows the requirements engineer to be sensitive to problems as they arise and the educator to structure delivery of requirements elicitation training.

Estimating ISABELLE shielding requirements

International Nuclear Information System (INIS)

Stevens, A.J.; Thorndike, A.M.

1976-01-01

Estimates were made of the shielding thicknesses required at various points around the ISABELLE ring. Both hadron and muon requirements are considered. Radiation levels at the outside of the shield and at the BNL site boundary are kept at or below 1000 mrem per year and 5 mrem/year respectively. Muon requirements are based on the Wang formula for pion spectra, and the hadron requirements on the hadron cascade program CYLKAZ of Ranft. A muon shield thickness of 77 meters of sand is indicated outside the ring in one area, and hadron shields equivalent to from 2.7 to 5.6 meters in thickness of sand above the ring. The suggested safety allowance would increase these values to 86 meters and 4.0 to 7.2 meters respectively. There are many uncertainties in such estimates, but these last figures are considered to be rather conservative

Range Flight Safety Requirements

Science.gov (United States)

Loftin, Charles E.; Hudson, Sandra M.

2018-01-01

The purpose of this NASA Technical Standard is to provide the technical requirements for the NPR 8715.5, Range Flight Safety Program, in regards to protection of the public, the NASA workforce, and property as it pertains to risk analysis, Flight Safety Systems (FSS), and range flight operations. This standard is approved for use by NASA Headquarters and NASA Centers, including Component Facilities and Technical and Service Support Centers, and may be cited in contract, program, and other Agency documents as a technical requirement. This standard may also apply to the Jet Propulsion Laboratory or to other contractors, grant recipients, or parties to agreements to the extent specified or referenced in their contracts, grants, or agreements, when these organizations conduct or participate in missions that involve range flight operations as defined by NPR 8715.5.1.2.2 In this standard, all mandatory actions (i.e., requirements) are denoted by statements containing the term “shall.”1.3 TailoringTailoring of this standard for application to a specific program or project shall be formally documented as part of program or project requirements and approved by the responsible Technical Authority in accordance with NPR 8715.3, NASA General Safety Program Requirements.

Top-tier requirements for KNGR

International Nuclear Information System (INIS)

Sung-Jae, Ch.; Kwangho, L.; Dong Wook, J.

1996-01-01

In 1992, Korea Electric Power Corporation (KEPCO) has launched the next generation reactor project to develop the standard design of an advanced pressurized water reactor by 2000. This advanced reactor aims to have the sufficient capability to be a safe, environmentally sound and economical energy source for 2000's in Korea. In conjunction with the project development, the program phase I is studied and it is in the Korean Next Generation Reactor (KNGR) first phase project that the requirements of this specification called ''Top-tier'' have been established. These functional requirements are of the first importance for the design, construction and operation of a nuclear power plant. These requirements are divided into safety requirements, serious accidents control, design base requirements, definition of the system characteristics, performance, construction feasibility, economical objectives, site parameters and design processes. The ''Top-tier'' requirements are concentrated on the improvement of the safety and reliability. Safety is one of the first priorities. In particular, the requirements for the design of the next reactors generation must include the capacity to control serious accidents because when an accident occurs, the protection degree is crucial. The KNGR requirements include the existing nuclear power plants competitiveness as well as those of the coal thermal plants. Moreover, when safety is reinforced, the economic competitiveness can be assured. At the present time, a subsequent specification for the KNGR considering the bases of the domestic technology and experimenting the running. (O.M.)

World enrichment requirements to 2005

International Nuclear Information System (INIS)

Anon.

1991-01-01

The primary enrichment suppliers-Eurodif, Techsnabexport, Urenco, and the US DOE - are positioning themselves to take advantage of the post - 1995 market. Overall, unfilled requirements represent about 40 percent of world requirements in the year 2000. The USA will be the primary market, as US utilities' unfilled enrichment requirements account for over 60 percent of the world's total unfilled requirements. The enrichment market is moving toward more global competition, as each supplier tries to maintain its current regional market base and then to capture additional market share in other regions

The NLC Software Requirements Methodology

Energy Technology Data Exchange (ETDEWEB)

Shoaee, Hamid

2002-08-20

We describe the software requirements and development methodology developed for the NLC control system. Given the longevity of that project, and the likely geographical distribution of the collaborating engineers, the planned requirements management process is somewhat more formal than the norm in high energy physics projects. The short term goals of the requirements process are to accurately estimate costs, to decompose the problem, and to determine likely technologies. The long term goal is to enable a smooth transition from high level functional requirements to specific subsystem and component requirements for individual programmers, and to support distributed development. The methodology covers both ends of that life cycle. It covers both the analytical and documentary tools for software engineering, and project management support. This paper introduces the methodology, which is fully described in [1].

Requirements for Ion Sources

International Nuclear Information System (INIS)

Scrivens, R

2013-01-01

Ion sources produce beams for a large variety of different physical experiments, industrial processes and medical applications. In order to characterize the beam delivered by them, a list of requirements is necessary. In this chapter the list of principal requirements is specified and definitions for them are given. (author)

Entrepreneurial learning requires action

DEFF Research Database (Denmark)

Brink, Tove; Madsen, Svend Ole

2014-01-01

that is enhanced by essential large-scale industry players and other SME managers are required to create action and value in learning. An open-mindedness to new learning approaches by SME managers and an open-mindedness to multi- and cross-disciplinary collaboration with SME managers by facilitators is required....

Loss of the nodule-specific cysteine rich peptide, NCR169, abolishes symbiotic nitrogen fixation in the Medicago truncatula dnf7 mutant.

Science.gov (United States)

Horváth, Beatrix; Domonkos, Ágota; Kereszt, Attila; Szűcs, Attila; Ábrahám, Edit; Ayaydin, Ferhan; Bóka, Károly; Chen, Yuhui; Chen, Rujin; Murray, Jeremy D; Udvardi, Michael K; Kondorosi, Éva; Kaló, Péter

2015-12-08

Host compatible rhizobia induce the formation of legume root nodules, symbiotic organs within which intracellular bacteria are present in plant-derived membrane compartments termed symbiosomes. In Medicago truncatula nodules, the Sinorhizobium microsymbionts undergo an irreversible differentiation process leading to the development of elongated polyploid noncultivable nitrogen fixing bacteroids that convert atmospheric dinitrogen into ammonia. This terminal differentiation is directed by the host plant and involves hundreds of nodule specific cysteine-rich peptides (NCRs). Except for certain in vitro activities of cationic peptides, the functional roles of individual NCR peptides in planta are not known. In this study, we demonstrate that the inability of M. truncatula dnf7 mutants to fix nitrogen is due to inactivation of a single NCR peptide, NCR169. In the absence of NCR169, bacterial differentiation was impaired and was associated with early senescence of the symbiotic cells. Introduction of the NCR169 gene into the dnf7-2/NCR169 deletion mutant restored symbiotic nitrogen fixation. Replacement of any of the cysteine residues in the NCR169 peptide with serine rendered it incapable of complementation, demonstrating an absolute requirement for all cysteines in planta. NCR169 was induced in the cell layers in which bacteroid elongation was most pronounced, and high expression persisted throughout the nitrogen-fixing nodule zone. Our results provide evidence for an essential role of NCR169 in the differentiation and persistence of nitrogen fixing bacteroids in M. truncatula.

Specifying semantic information on functional requirements

OpenAIRE

YAO, WUPING

2012-01-01

Requirements engineering is a challenging process in software development projects. Requirements, in general, are documented in natural language. They often have issues related to ambiguity, completeness and consistency. How to improve the quality of requirements documentation remains a classic research topic. This research aims at improving the way of editing and documenting functional requirements. We propose a meta-model to specify the semantic information of functional requirements, and d...

Requirements management at Westinghouse Electric Company

International Nuclear Information System (INIS)

Gustavsson, Henrik

2014-01-01

Field studies and surveys made in various industry branches support the Westinghouse opinion that qualitative systems engineering and requirements management have a high value in the development of complex systems and products. Two key issues causing overspending and schedule delays in projects are underestimation of complexity and misunderstandings between the different sub-project teams. These issues often arise when a project jumps too early into detail design. Good requirements management practice before detail design helps the project teams avoid such issues. Westinghouse therefore puts great effort into requirements management. The requirements management methodology at Westinghouse rests primarily on four key cornerstones: 1 - Iterative team work when developing requirements specifications, 2 - Id number tags on requirements, 3 - Robust change routine, and 4 - Requirements Traceability Matrix. (authors)

Design requirement on HYPER blanket fuel assembly

International Nuclear Information System (INIS)

Hwang, Woan; Lee, B. O.; Nam, C.; Ryu, W. S.; Lee, B. S.; Park, W. S.

2000-07-01

This document describes design requirements which are needed for designing the blanket assembly of the HYPER as design guidance. The blanket assembly of the HYPER consists of blanket fuel rods, mounting rail, spacer, upper nozzle with handling socket, bottom nozzle with mounting rail and skeleton structure. The blanket fuel rod consists of top end plug, bottom end plug with key way, blanket fuel slug, and cladding. In the assembly, the rods are in a triangular pitch array. This report contains functional requirements, performance and operational requirements, interfacing systems requirements, core restraint and interface requirements, design limits and strength requirements, system configuration and essential feature requirements, seismic requirements, structural requirements, environmental requirements, reliability and safety requirements, standard and codes, QA programs, and other requirements for the blanket fuel assembly of the HYPER

40 CFR 141.70 - General requirements.

Science.gov (United States)

2010-07-01

...) NATIONAL PRIMARY DRINKING WATER REGULATIONS Filtration and Disinfection § 141.70 General requirements. (a... regulations establish criteria under which filtration is required as a treatment technique for public water... filtration requirements in § 141.73 and the disinfection requirements in § 141.72(b). (c) Each public water...

20 CFR 655.152 - Advertising requirements.

Science.gov (United States)

2010-04-01

... 20 Employees' Benefits 3 2010-04-01 2010-04-01 false Advertising requirements. 655.152 Section 655... Employment in the United States (H-2A Workers) Post-Acceptance Requirements § 655.152 Advertising requirements. All advertising conducted to satisfy the required recruitment activities under § 655.151 must...

ENVIRONMENTAL SPECIFICATION REQUIREMENTS

International Nuclear Information System (INIS)

TIFFT, S.R.

2003-01-01

Through regulations, permitting or binding negotiations, Regulators establish requirements, limits, permit conditions and Notice of Construction (NOC) conditions with which the Office of River Protection (ORP) and the Tank Farm Contractor (TFC) must comply. Operating Specifications are technical limits which are set on a process to prevent injury to personnel, or damage to the facility or environment. The main purpose of this document is to provide specification limits and recovery actions for the TFC Environmental Surveillance Program at the Hanford Site. Specification limits are given for monitoring frequencies and permissible variation of readings from an established baseline or previous reading. The requirements in this document are driven by environmental considerations and data analysis issues, rather than facility design or personnel safety issues. This document is applicable to all SST and DST waste tanks, and the associated catch tanks and receiver tanks, and transfer systems. This Tank Farm ESD implements environmental-regulatory limits on the configuration and operation of the Hanford Tank Farms facility that have been established by Regulators. This ESD contains specific field operational limits and recovery actions for compliance with airborne effluent regulations and agreements, liquid effluents regulations and agreements, and environmental tank system requirements. The scope of this ESD is limited to conditions that have direct impact on Operations/Projects or that Operations/Projects have direct impact upon. This document does not supercede or replace any DOE Orders, regulatory permits, notices of construction, or Regulatory agency agreements binding on the ORP or the TFC. Refer to the appropriate regulation, permit, or NOC for an inclusive listing of requirements

Data Crosscutting Requirements Review

Energy Technology Data Exchange (ETDEWEB)

Kleese van Dam, Kerstin [Pacific Northwest National Lab. (PNNL), Richland, WA (United States); Shoshani, Arie [Pacific Northwest National Lab. (PNNL), Richland, WA (United States); Plata, Charity [Pacific Northwest National Lab. (PNNL), Richland, WA (United States)

2013-04-01

In April 2013, a diverse group of researchers from the U.S. Department of Energy (DOE) scientific community assembled to assess data requirements associated with DOE-sponsored scientific facilities and large-scale experiments. Participants in the review included facilities staff, program managers, and scientific experts from the offices of Basic Energy Sciences, Biological and Environmental Research, High Energy Physics, and Advanced Scientific Computing Research. As part of the meeting, review participants discussed key issues associated with three distinct aspects of the data challenge: 1) processing, 2) management, and 3) analysis. These discussions identified commonalities and differences among the needs of varied scientific communities. They also helped to articulate gaps between current approaches and future needs, as well as the research advances that will be required to close these gaps. Moreover, the review provided a rare opportunity for experts from across the Office of Science to learn about their collective expertise, challenges, and opportunities. The "Data Crosscutting Requirements Review" generated specific findings and recommendations for addressing large-scale data crosscutting requirements.

46 CFR 11.707 - Examination requirements.

Science.gov (United States)

2010-10-01

... 46 Shipping 1 2010-10-01 2010-10-01 false Examination requirements. 11.707 Section 11.707 Shipping... OFFICER ENDORSEMENTS Professional Requirements for Pilots § 11.707 Examination requirements. (a) An... required to pass the examination described in subpart I of this part. (b) An applicant for an extension of...

21 CFR 1304.11 - Inventory requirements.

Science.gov (United States)

2010-04-01

... the inventory of the registered location to which they are subject to control or to which the person... 21 Food and Drugs 9 2010-04-01 2010-04-01 false Inventory requirements. 1304.11 Section 1304.11... REGISTRANTS Inventory Requirements § 1304.11 Inventory requirements. (a) General requirements. Each inventory...

Non-functional Avionics Requirements

Science.gov (United States)

Paulitsch, Michael; Ruess, Harald; Sorea, Maria

Embedded systems in aerospace become more and more integrated in order to reduce weight, volume/size, and power of hardware for more fuel-effi ciency. Such integration tendencies change architectural approaches of system ar chi tec tures, which subsequently change non-functional requirements for plat forms. This paper provides some insight into state-of-the-practice of non-func tional requirements for developing ultra-critical embedded systems in the aero space industry, including recent changes and trends. In particular, formal requi re ment capture and formal analysis of non-functional requirements of avionic systems - including hard-real time, fault-tolerance, reliability, and per for mance - are exemplified by means of recent developments in SAL and HiLiTE.

General review of quality assurance system requirements. The utility or customer requirement

International Nuclear Information System (INIS)

Fowler, J.L.

1976-01-01

What are the customer's Quality Assurance requirements and how does he convey these to his contractor, or apply them to himself. Many documents have been prepared mostly by countries with high technology availability and it is significant to note that many of the documents, particularly those of the United States of America, were prepared for nuclear safety related plant, but the logic of these documents equally applied to heavy engineering projects that are cost effective, and this is the current thinking and practice within the CEGB (Central Electricity Generating Board). Some documents have legislative backing, others rely on contractual disciplines, but they all appear to repeat the same basic requirements, so why does one continue to write more documents. The basic problem is that customers have to satisfy differing national legislative, economic and commercial requirements and, like all discerning customers, wish to reserve the right to satisfy their own needs, which are very often highly specialized. The CEGB are aware of this problem and are actively co-operating with most of the national and international authorities who are leading in this field, with a view to obtaining compatibility of requirements, but now there still remains the problem of satisfying national custom and practice. (author)

Advanced Scientific Computing Research Network Requirements: ASCR Network Requirements Review Final Report

Energy Technology Data Exchange (ETDEWEB)

Bacon, Charles [Argonne National Lab. (ANL), Argonne, IL (United States); Bell, Greg [ESnet, Berkeley, CA (United States); Canon, Shane [Lawrence Berkeley National Lab. (LBNL), Berkeley, CA (United States); Dart, Eli [ESnet, Berkeley, CA (United States); Dattoria, Vince [Dept. of Energy (DOE), Washington DC (United States). Office of Science. Advanced Scientific Computing Research (ASCR); Goodwin, Dave [Dept. of Energy (DOE), Washington DC (United States). Office of Science. Advanced Scientific Computing Research (ASCR); Lee, Jason [Lawrence Berkeley National Lab. (LBNL), Berkeley, CA (United States); Hicks, Susan [Oak Ridge National Lab. (ORNL), Oak Ridge, TN (United States); Holohan, Ed [Argonne National Lab. (ANL), Argonne, IL (United States); Klasky, Scott [Oak Ridge National Lab. (ORNL), Oak Ridge, TN (United States); Lauzon, Carolyn [Dept. of Energy (DOE), Washington DC (United States). Office of Science. Advanced Scientific Computing Research (ASCR); Rogers, Jim [Oak Ridge National Lab. (ORNL), Oak Ridge, TN (United States); Shipman, Galen [Oak Ridge National Lab. (ORNL), Oak Ridge, TN (United States); Skinner, David [Lawrence Berkeley National Lab. (LBNL), Berkeley, CA (United States); Tierney, Brian [ESnet, Berkeley, CA (United States)

2013-03-08

The Energy Sciences Network (ESnet) is the primary provider of network connectivity for the U.S. Department of Energy (DOE) Office of Science (SC), the single largest supporter of basic research in the physical sciences in the United States. In support of SC programs, ESnet regularly updates and refreshes its understanding of the networking requirements of the instruments, facilities, scientists, and science programs that it serves. This focus has helped ESnet to be a highly successful enabler of scientific discovery for over 25 years. In October 2012, ESnet and the Office of Advanced Scientific Computing Research (ASCR) of the DOE SC organized a review to characterize the networking requirements of the programs funded by the ASCR program office. The requirements identified at the review are summarized in the Findings section, and are described in more detail in the body of the report.

Design requirements for the new reactor

International Nuclear Information System (INIS)

Koski, S.

2005-01-01

This presentation deals with the safety related design requirements specified for the new nuclear power plant to be built in Finland (FINS). The legislation, codes and standards, on which the design requirements are based, can be arranged into a hierarchical pyramid as follows: The safety related design criteria are based on the three uppermost hierarchical levels: Finnish legislation (e.g. decisions of the State Council) Basic Regulations (75-INSAG-3, USNRC General Design Criteria) Process oriented nuclear documents (YVL- guides or corresponding US/German rules). The European Utility Requirements (EUR) document was used as the starting point for the writing of the design requirements document. The structure and headlines of EUR could be kept, but in many cases the contents had to be deleted and rewritten to correspond to the requirement level of the above codes and standards. This was the case, for example, with the requirements concerning safety classification or application of failure criteria. In the presentation, the most important safety related design criteria are reviewed, with an emphasis on those requirements which exceed the requirement level applied on the existing plant units. Some hints are also given on the main differences between Finnish and international safety requirements. (orig.)

Capital Requirements and Credit Rationing

OpenAIRE

Itai Agur

2010-01-01

This paper analyzes the trade-off between financial stability and credit rationing that arises when increasing capital requirements. It extends the Stiglitz-Weiss model of credit rationing to allow for bank default. Bank capital structure then matters for lending incentives. With default and rationing endogenous, optimal capital requirements can be analyzed. Introducing bank financiers, the paper also shows that uninsured funding raises the sensitivity of rationing to capital requirements. In...

National Land Imaging Requirements (NLIR) Pilot Project summary report: summary of moderate resolution imaging user requirements

Science.gov (United States)

Vadnais, Carolyn; Stensaas, Gregory

2014-01-01

Under the National Land Imaging Requirements (NLIR) Project, the U.S. Geological Survey (USGS) is developing a functional capability to obtain, characterize, manage, maintain and prioritize all Earth observing (EO) land remote sensing user requirements. The goal is a better understanding of community needs that can be supported with land remote sensing resources, and a means to match needs with appropriate solutions in an effective and efficient way. The NLIR Project is composed of two components. The first component is focused on the development of the Earth Observation Requirements Evaluation System (EORES) to capture, store and analyze user requirements, whereas, the second component is the mechanism and processes to elicit and document the user requirements that will populate the EORES. To develop the second component, the requirements elicitation methodology was exercised and refined through a pilot project conducted from June to September 2013. The pilot project focused specifically on applications and user requirements for moderate resolution imagery (5–120 meter resolution) as the test case for requirements development. The purpose of this summary report is to provide a high-level overview of the requirements elicitation process that was exercised through the pilot project and an early analysis of the moderate resolution imaging user requirements acquired to date to support ongoing USGS sustainable land imaging study needs. The pilot project engaged a limited set of Federal Government users from the operational and research communities and therefore the information captured represents only a subset of all land imaging user requirements. However, based on a comparison of results, trends, and analysis, the pilot captured a strong baseline of typical applications areas and user needs for moderate resolution imagery. Because these results are preliminary and represent only a sample of users and application areas, the information from this report should only

ENVIRONMENTAL SPECIFICATION REQUIREMENTS

International Nuclear Information System (INIS)

TIFFT, S.R.

2003-01-01

Through regulations, permitting or binding negotiations, Regulators establish requirements, limits, permit conditions and Notice of Construction (NOC) conditions with which the Office of River Protection (ORP) and the Tank Farm Contractor (TFC) must comply. Operating Specifications are technical limits which are set on a process to prevent injury to personnel, or damage to the facility or environment. The main purpose of this document is to provide specification limits and recovery actions for the TFC Environmental Surveillance Program at the Hanford Site. Specification limits are given for monitoring frequencies and permissible variation of readings from an established baseline or previous reading. The requirements in this document are driven by environmental considerations and data analysis issues, rather than facility design or personnel safety issues. This document is applicable to all single-shell tank (SST) and double-shell tank (DST) waste tanks, and the associated catch tanks and receiver tanks, and transfer systems. This Tank Farm Environmental Specifications Document (ESD) implements environmental-regulatory limits on the configuration and operation of the Hanford Tank Farms facility that have been established by Regulators. This ESD contains specific field operational limits and recovery actions for compliance with airborne effluent regulations and agreements, liquid effluents regulations and agreements, and environmental tank system requirements. The scope of this ESD is limited to conditions that have direct impact on Operations Projects or that Operations/Projects have direct impact upon. This document does not supercede or replace any DOE Orders, regulatory permits, notices of construction, or Regulatory agency agreements binding on the ORP or the TFC. Refer to the appropriate regulation, permit, or NOC for an inclusive listing of requirements

Requirements Analysis in the Value Methodology

Energy Technology Data Exchange (ETDEWEB)

Conner, Alison Marie

2001-05-01

The Value Methodology (VM) study brings together a multidisciplinary team of people who own the problem and have the expertise to identify and solve it. With the varied backgrounds and experiences the team brings to the study, come different perspectives on the problem and the requirements of the project. A requirements analysis step can be added to the Information and Function Analysis Phases of a VM study to validate whether the functions being performed are required, either regulatory or customer prescribed. This paper will provide insight to the level of rigor applied to a requirements analysis step and give some examples of tools and techniques utilized to ease the management of the requirements and functions those requirements support for highly complex problems.

UTM TCL2 Software Requirements

Science.gov (United States)

Smith, Irene S.; Rios, Joseph L.; McGuirk, Patrick O.; Mulfinger, Daniel G.; Venkatesan, Priya; Smith, David R.; Baskaran, Vijayakumar; Wang, Leo

2017-01-01

The Unmanned Aircraft Systems (UAS) Traffic Management (UTM) Technical Capability Level (TCL) 2 software implements the UTM TCL 2 software requirements described herein. These software requirements are linked to the higher level UTM TCL 2 System Requirements. Each successive TCL implements additional UTM functionality, enabling additional use cases. TCL 2 demonstrated how to enable expanded multiple operations by implementing automation for beyond visual line-of-sight, tracking operations, and operations flying over sparsely populated areas.

Information requirements for enterprise systems

OpenAIRE

Sommerville, Ian; Lock, Russell; Storer, Tim

2012-01-01

In this paper, we discuss an approach to system requirements engineering, which is based on using models of the responsibilities assigned to agents in a multi-agency system of systems. The responsibility models serve as a basis for identifying the stakeholders that should be considered in establishing the requirements and provide a basis for a structured approach, described here, for information requirements elicitation. We illustrate this approach using a case study drawn from civil emergenc...

Dosimeter characteristics and service performance requirements

International Nuclear Information System (INIS)

Ambrosi, P.; Bartlett, D.T.

1999-01-01

The requirements for personal dosimeters and dosimetry services given by ICRP 26, ICRP 35, ICRP 60 and ICRP 75 are summarised and compared with the requirements given in relevant international standards. Most standards could be made more relevant to actual workplace conditions. In some standards, the required tests of energy and angular dependence of the response are not sufficient, or requirements on overall uncertainty are lacking. (author)

PFP requirements development planning guide

International Nuclear Information System (INIS)

SINCLAIR, J.C.

1999-01-01

The PFP Requirements Development Planning Guide presents the strategy and process used for the identification, allocation, and maintenance of requirements within the Plutonium Finishing Plant (PFP) integrated project baseline. Future revisions to this document will be included as attachments (e.g., results of the PFP Requirements Analysis attributable to this approach). This document is intended be a Project-owned management tool. As such, this document will periodically require revisions resulting from improvements of the information, processes, and techniques as now described. Future updates may be made to this document by PFP management and final approval of the content will be accomplished in a Baseline Change Request as it impacts the Multi-Year Work Plan, or baseline information managed in the Hanford Site Systems Engineering Baseline

Requirements for Space Settlement Design

Science.gov (United States)

Gale, Anita E.; Edwards, Richard P.

2004-02-01

When large space settlements are finally built, inevitably the customers who pay for them will start the process by specifying requirements with a Request for Proposal (RFP). Although we are decades away from seeing the first of these documents, some of their contents can be anticipated now, and provide insight into the variety of elements that must be researched and developed before space settlements can happen. Space Settlement Design Competitions for High School students present design challenges in the form of RFPs, which predict basic requirements for space settlement attributes in the future, including structural features, infrastructure, living conveniences, computers, business areas, and safety. These requirements are generically summarized, and unique requirements are noted for specific space settlement locations and applications.

Waste Acceptance System Requirements document (WASRD)

International Nuclear Information System (INIS)

1993-01-01

This Waste Acceptance System Requirements document (WA-SRD) describes the functions to be performed and the technical requirements for a Waste Acceptance System for accepting spent nuclear fuel (SNF) and high-level radioactive waste (HLW) into the Civilian Radioactive Waste Management System (CRWMS). This revision of the WA-SRD addresses the requirements for the acceptance of HLW. This revision has been developed as a top priority document to permit DOE's Office of Environmental Restoration and Waste Management (EM) to commence waste qualification runs at the Savannah River Site's (SRS) Defense Waste Processing Facility (DWPF) in a timely manner. Additionally, this revision of the WA-SRD includes the requirements from the Physical System Requirements -- Accept Waste document for the acceptance of SNF. A subsequent revision will fully address requirements relative to the acceptance of SNF

Mined Geologic Disposal System Requirements Document

International Nuclear Information System (INIS)

1993-01-01

This Mined Geologic Disposal System Requirements document (MGDS-RD) describes the functions to be performed by, and the requirements for, a Mined Geologic Disposal System (MGDS) for the permanent disposal of spent nuclear fuel (SNF) and commercial and defense high level radioactive waste (HLW) in support of the Civilian Radioactive Waste Management System (CRWMS). The development and control of the MGDS-RD is quality-affecting work and is subject to the Department of Energy (DOE) Office of Civilian Radioactive Waste Management (OCRWM) Quality Assurance Requirements Document (QARD). As part of the technical requirements baseline, it is also subject to Baseline Management Plan controls. The MGDS-RD and the other program-level requirements documents have been prepared and managed in accordance with the Technical Document Preparation Plan (TDPP) for the Preparation of System Requirements Documents

Requirements Modeling with the Aspect-oriented User Requirements Notation (AoURN): A Case Study

Science.gov (United States)

Mussbacher, Gunter; Amyot, Daniel; Araújo, João; Moreira, Ana

The User Requirements Notation (URN) is a recent ITU-T standard that supports requirements engineering activities. The Aspect-oriented URN (AoURN) adds aspect-oriented concepts to URN, creating a unified framework that allows for scenario-based, goal-oriented, and aspect-oriented modeling. AoURN is applied to the car crash crisis management system (CCCMS), modeling its functional and non-functional requirements (NFRs). AoURN generally models all use cases, NFRs, and stakeholders as individual concerns and provides general guidelines for concern identification. AoURN handles interactions between concerns, capturing their dependencies and conflicts as well as the resolutions. We present a qualitative comparison of aspect-oriented techniques for scenario-based and goal-oriented requirements engineering. An evaluation carried out based on the metrics adapted from literature and a task-based evaluation suggest that AoURN models are more scalable than URN models and exhibit better modularity, reusability, and maintainability.

7 CFR 4284.18 - Audit requirements.

Science.gov (United States)

2010-01-01

... Grant Programs § 4284.18 Audit requirements. Grantees must comply with the audit requirements of 7 CFR part 3052. The audit requirements apply to the years in which grant funds are received and years in...

Puberty menorrhagia Requiring Inpatient Admission

Directory of Open Access Journals (Sweden)

AH Khosla

2010-06-01

Full Text Available INTRODUCTION: Puberty menorrhagia is a significant health problem in adolescent age group and severe cases may require admission and blood transfusion. Aim of this study was to evaluate the causes, associated complications and management of puberty menorrhagia. METHODS: Hospital records of all patients of puberty menorrhagia requiring admission were analyzed for etiology, duration since menarche, duration of bleeding, investigation profile and management. RESULTS: There were 18 patients of puberty menorrhagia requiring hospital admission. Etiology was anovulatory bleeding in 11 patients, bleeding disorders in five which included idiopathic thrombocytopenia purpura in three and one each with Von-Willebrand disease and leukemia. Two patients had hypothyroidism as the cause. Fourteen patients presented with severe anaemia and required blood transfusion. All except one responded to oral hormonal therapy. CONCLUSIONS: Puberty menorrhagia can be associated with severe complications and requiring blood transfusion. Although most common cause is anovulation but bleeding disorder, other medical condition and other organic causes must be ruled out in any patient of Puberty menorrhagia. KEYWORDS: anovulation, bleeding disorder, puberty, menorrhagia, anaemia.

The JPL functional requirements tool

Science.gov (United States)

Giffin, Geoff; Skinner, Judith; Stoller, Richard

1987-01-01

Planetary spacecraft are complex vehicles which are built according to many thousands of requirements. Problems encountered in documenting and maintaining these requirements led to the current attempt to reduce or eliminate these problems by a computer automated data base Functional Requirements Tool. The tool developed at JPL and in use on several JPL Projects is described. The organization and functionality of the Tool, together with an explanation of the data base inputs, their relationships, and use are presented. Methods of interfacing with external documents, representation of tables and figures, and methods of approval and change processing are discussed. The options available for disseminating information from the Tool are identified. The implementation of the Requirements Tool is outlined, and the operation is summarized. The conclusions drawn from this work is that the Requirements Tool represents a useful addition to the System Engineer's Tool kit, it is not currently available elsewhere, and a clear development path exists to expand the capabilities of the Tool to serve larger and more complex projects.

NP Science Network Requirements

Energy Technology Data Exchange (ETDEWEB)

Dart, Eli [Lawrence Berkeley National Laboratory (LBNL), Berkeley, CA (United States); Rotman, Lauren [Lawrence Berkeley National Laboratory (LBNL), Berkeley, CA (United States); Tierney, Brian [Lawrence Berkeley National Laboratory (LBNL), Berkeley, CA (United States)

2011-08-26

The Energy Sciences Network (ESnet) is the primary provider of network connectivity for the U.S. Department of Energy (DOE) Office of Science (SC), the single largest supporter of basic research in the physical sciences in the United States. To support SC programs, ESnet regularly updates and refreshes its understanding of the networking requirements of the instruments, facilities, scientists, and science programs it serves. This focus has helped ESnet to be a highly successful enabler of scientific discovery for over 20 years. In August 2011, ESnet and the Office of Nuclear Physics (NP), of the DOE SC, organized a workshop to characterize the networking requirements of the programs funded by NP. The requirements identified at the workshop are summarized in the Findings section, and are described in more detail in the body of the report.

Next Generation Microbiology Requirements

Science.gov (United States)

Ott, C. M.; Oubre, C. M.; Elliott, T. F.; Castro, V. A.; Pierson, D. L.

2012-01-01

As humans continue to explore deep into space, microorganisms will travel with them. The primary means to mitigate the risk of infectious disease are a combination of prudent spacecraft design and rigorous operational controls. The effectiveness of these methods are evaluated by microbiological monitoring of spacecraft, food, water, and the crew that is performed preflight, in-flight, and post-flight. Current NASA requirements associated with microbiological monitoring are based on culture-based methodology where microorganisms are grown on a semi-solid growth medium and enumerated. Subsequent identification of the organisms requires specialized labor and large equipment, which historically has been performed on Earth. Requirements that rely strictly on culture-based units limit the use of non-culture based monitoring technology. Specifically, the culture-based "measurement criteria" are Colony Forming Units (CFU, representing the growth of one microorganism at a single location on the agar medium) per a given volume, area, or sample size. As the CFU unit by definition is culture-based, these requirements limit alternative technologies for spaceflight applications. As spaceflight missions such as those to Mars extend further into space, culture-based technology will become difficult to implement due to the (a) limited shelf life of the culture media, (b) mass/volume necessary to carry these consumables, and (c) problems associated with the production of biohazardous material in the habitable volume of the spacecraft. In addition, an extensive amount of new knowledge has been obtained during the Space Shuttle, NASA-Mir, and International Space Station Programs, which gave direction for new or modified microbial control requirements for vehicle design and mission operations. The goal of this task is to develop and recommend a new set of requirements for vehicle design and mission operations, including microbiological monitoring, based upon "lessons learned" and new

Crewed Space Vehicle Battery Safety Requirements

Science.gov (United States)

Jeevarajan, Judith A.; Darcy, Eric C.

2014-01-01

This requirements document is applicable to all batteries on crewed spacecraft, including vehicle, payload, and crew equipment batteries. It defines the specific provisions required to design a battery that is safe for ground personnel and crew members to handle and/or operate during all applicable phases of crewed missions, safe for use in the enclosed environment of a crewed space vehicle, and safe for use in launch vehicles, as well as in unpressurized spaces adjacent to the habitable portion of a space vehicle. The required provisions encompass hazard controls, design evaluation, and verification. The extent of the hazard controls and verification required depends on the applicability and credibility of the hazard to the specific battery design and applicable missions under review. Evaluation of the design and verification program results shall be completed prior to certification for flight and ground operations. This requirements document is geared toward the designers of battery systems to be used in crewed vehicles, crew equipment, crew suits, or batteries to be used in crewed vehicle systems and payloads (or experiments). This requirements document also applies to ground handling and testing of flight batteries. Specific design and verification requirements for a battery are dependent upon the battery chemistry, capacity, complexity, charging, environment, and application. The variety of battery chemistries available, combined with the variety of battery-powered applications, results in each battery application having specific, unique requirements pertinent to the specific battery application. However, there are basic requirements for all battery designs and applications, which are listed in section 4. Section 5 includes a description of hazards and controls and also includes requirements.

Mined Geologic Disposal System Requirements Document

International Nuclear Information System (INIS)

1994-03-01

This Mined Geologic Disposal System Requirements Document (MGDS-RD) describes the functions to be performed by, and the requirements for, a Mined Geologic Disposal System (MGDS) for the permanent disposal of spent nuclear fuel (SNF) (including SNF loaded in multi-purpose canisters (MPCs)) and commercial and defense high-level radioactive waste (HLW) in support of the Civilian Radioactive Waste Management System (CRWMS). The purpose of the MGDS-RD is to define the program-level requirements for the design of the Repository, the Exploratory Studies Facility (ESF), and Surface Based Testing Facilities (SBTF). These requirements include design, operation, and decommissioning requirements to the extent they impact on the physical development of the MGDS. The document also presents an overall description of the MGDS, its functions (derived using the functional analysis documented by the Physical System Requirements (PSR) documents as a starting point), its segments as described in Section 3.1.3, and the requirements allocated to the segments. In addition, the program-level interfaces of the MGDS are identified. As such, the MGDS-RD provides the technical baseline for the design of the MGDS

Management system requirements for small reactors

Energy Technology Data Exchange (ETDEWEB)

Jones, K.A., E-mail: kenneth.jones@cnsc-ccsn.gc.ca [Canadian Nuclear Safety Commission, Ottawa, Ontario (Canada)

2013-07-01

This abstract identifies the management system requirements for the life cycle of small reactors from initial conception through completion of decommissioning. For small reactors, the requirements for management systems remain the same as those for 'large' reactors regardless of the licensee' business model and objectives. The CSA N-Series of standards provides an interlinked set of requirements for the management of nuclear facilities. CSA N286 provides overall direction to management to develop and implement sound management practices and controls, while other CSA nuclear standards provide technical requirements and guidance that support the management system. CSA N286 is based on a set of principles. The principles are then supported by generic requirements that are applicable to the life cycle of nuclear facilities. CNSC regulatory documents provide further technical requirements and guidance. (author)

ATLAS Future Framework Requirements Group Report

CERN Document Server

The ATLAS collaboration

2016-01-01

The Future Frameworks Requirements Group was constituted in Summer 2013 to consider and summarise the framework requirements from trigger and offline for configuring, scheduling and monitoring the data processing software needed by the ATLAS experiment. The principal motivation for such a re-examination arises from the current and anticipated evolution of CPUs, where multiple cores, hyper-threading and wide vector registers require a shift to a concurrent programming model. Such a model requires extensive changes in the current Gaudi/Athena frameworks and offers the opportunity to consider how HLT and offline processing can be better accommodated within the ATLAS framework. This note contains the report of the Future Frameworks Requirements Group.

Gamified Requirements Engineering: Model and Experimentation

NARCIS (Netherlands)

Lombriser, Philipp; Dalpiaz, Fabiano; Lucassen, Garm; Brinkkemper, Sjaak

2016-01-01

[Context & Motivation] Engaging stakeholders in requirements engineering (RE) influences the quality of the requirements and ultimately of the system to-be. Unfortunately, stakeholder engagement is often insufficient, leading to too few, low-quality requirements. [Question/problem] We aim to

48 CFR 9903.202 - Disclosure requirements.

Science.gov (United States)

2010-10-01

... ACCOUNTING STANDARDS CONTRACT COVERAGE CAS Program Requirements 9903.202 Disclosure requirements. ... 48 Federal Acquisition Regulations System 7 2010-10-01 2010-10-01 false Disclosure requirements. 9903.202 Section 9903.202 Federal Acquisition Regulations System COST ACCOUNTING STANDARDS BOARD...

Specifying Functional Requirements Dependency in the REWiki

OpenAIRE

ZHANG, ZHANG

2013-01-01

Most of the individual requirements cannot be treated in isolation. Requirements may affect each other in various ways. The dependency between requirements impacts a number of software development aspects and activities. How to classify and specify requirements dependency remains a classic research topic. This research aims at providing an approach of specifying functional requirements dependency. In this thesis we generalize a classification of functional requirements dependency. We also pro...

Rapid quality assurance with Requirements Smells

OpenAIRE

Femmer, H.; Fernández, D. Méndez; Wagner, S.; Eder, S.

2016-01-01

Bad requirements quality can cause expensive consequences during the software development lifecycle, especially if iterations are long and feedback comes late. %-- the faster a problem is found, the cheaper it is to fix. This makes explicit the need of a lightweight detection mechanism of requirements quality violations. We aim at a light-weight static requirements analysis approach that allows for rapid checks immediately when requirements are written down. We transfer the concept of code sm...

Airspace Operations Demo Functional Requirements Matrix

Science.gov (United States)

2005-01-01

The Flight IPT assessed the reasonableness of demonstrating each of the Access 5 Step 1 functional requirements. The functional requirements listed in this matrix are from the September 2005 release of the Access 5 Functional Requirements Document. The demonstration mission considered was a notional Western US mission (WUS). The conclusion of the assessment is that 90% of the Access 5 Step 1 functional requirements can be demonstrated using the notional Western US mission.

Reserve requirement systems in OECD countries

OpenAIRE

Yueh-Yun C. O’Brien

2007-01-01

This paper compares the reserve requirements of OECD countries. Reserve requirements are the minimum percentages or amounts of liabilities that depository institutions are required to keep in cash or as deposits with their central banks. To facilitate monetary policy implementation, twenty-four of the thirty OECD countries impose reserve requirements to influence their banking systems’ demand for liquidity. These include twelve OECD countries that are also members of the European Economic and...

Deaf mobile application accessibility requirements

Science.gov (United States)

Nathan, Shelena Soosay; Hussain, Azham; Hashim, Nor Laily

2016-08-01

Requirement for deaf mobile applications need to be analysed to ensure the disabilities need are instilled into the mobile applications developed for them. Universal design is understandable to comply every user needs, however specific disability is argued by the authors to have different need and requirements. These differences are among the reasons for these applications being developed to target for a specific group of people, however they are less usable and later abandoned. This study focuses on deriving requirements that are needed by the deaf in their mobile applications that are meant specifically for them. Studies on previous literature was conducted it can be concluded that graphic, text, multimedia and sign language interpreter are among mostly required features to be included in their mobile application to ensure the applications are usable for this community.

LH2 airport requirements study

Science.gov (United States)

Brewer, G. D. (Editor)

1976-01-01

A preliminary assessment of the facilities and equipment which will be required at a representative airport is provided so liquid hydrogen LH2 can be used as fuel in long range transport aircraft in 1995-2000. A complete facility was conceptually designed, sized to meet the projected air traffic requirement. The facility includes the liquefaction plant, LH2, storage capability, and LH2 fuel handling system. The requirements for ground support and maintenance for the LH2 fueled aircraft were analyzed. An estimate was made of capital and operating costs which might be expected for the facility. Recommendations were made for design modifications to the reference aircraft, reflecting results of the analysis of airport fuel handling requirements, and for a program of additional technology development for air terminal related items.

Organizing Performance Requirements For Dynamical Systems

Science.gov (United States)

Malchow, Harvey L.; Croopnick, Steven R.

1990-01-01

Paper describes methodology for establishing performance requirements for complicated dynamical systems. Uses top-down approach. In series of steps, makes connections between high-level mission requirements and lower-level functional performance requirements. Provides systematic delineation of elements accommodating design compromises.

48 CFR 1430.202 - Disclosure requirements.

Science.gov (United States)

2010-10-01

... 48 Federal Acquisition Regulations System 5 2010-10-01 2010-10-01 false Disclosure requirements. 1430.202 Section 1430.202 Federal Acquisition Regulations System DEPARTMENT OF THE INTERIOR GENERAL CONTRACTING REQUIREMENTS COST ACCOUNTING STANDARDS ADMINISTRATION CAS Program Requirements 1430.202 Disclosure...

48 CFR 430.202 - Disclosure requirements.

Science.gov (United States)

2010-10-01

... 48 Federal Acquisition Regulations System 4 2010-10-01 2010-10-01 false Disclosure requirements. 430.202 Section 430.202 Federal Acquisition Regulations System DEPARTMENT OF AGRICULTURE GENERAL CONTRACTING REQUIREMENTS COST ACCOUNTING STANDARDS ADMINISTRATION CAS Program Requirements 430.202 Disclosure...

48 CFR 30.202 - Disclosure requirements.

Science.gov (United States)

2010-10-01

... 48 Federal Acquisition Regulations System 1 2010-10-01 2010-10-01 false Disclosure requirements. 30.202 Section 30.202 Federal Acquisition Regulations System FEDERAL ACQUISITION REGULATION GENERAL CONTRACTING REQUIREMENTS COST ACCOUNTING STANDARDS ADMINISTRATION CAS Program Requirements 30.202 Disclosure...

48 CFR 1330.202 - Disclosure requirements.

Science.gov (United States)

2010-10-01

... 48 Federal Acquisition Regulations System 5 2010-10-01 2010-10-01 false Disclosure requirements. 1330.202 Section 1330.202 Federal Acquisition Regulations System DEPARTMENT OF COMMERCE GENERAL CONTRACTING REQUIREMENTS COST ACCOUNTING STANDARDS ADMINISTRATION CAS Program Requirements 1330.202 Disclosure...

Front Loaded Accurate Requirements Engineering (FLARE): A Requirements Analysis Concept for the 21st Century

National Research Council Canada - National Science Library

Leonard, Anthony

1997-01-01

This thesis focuses on ways to apply requirements engineering techniques and methods during the development and evolution of DoD software systems in an effort to reduce changes to system requirements...

14 CFR 125.265 - Flight engineer requirements.

Science.gov (United States)

2010-01-01

... 14 Aeronautics and Space 3 2010-01-01 2010-01-01 false Flight engineer requirements. 125.265... Requirements § 125.265 Flight engineer requirements. (a) No person may operate an airplane for which a flight engineer is required by the type certification requirements without a flight crewmember holding a current...

IRET: requirements for service platforms

OpenAIRE

Baresi, Luciano; Ripa, Gianluca; Pasquale, Liliana

2013-01-01

peer-reviewed This paper describes IRENE (Indenica Requirements ElicitatioN mEthod), a methodology to elicit and model the requirements of service platforms, and IRET (IREne Tool), the Eclipse-based modeling framework we developed for IRENE

Understand the Design Requirement in Companies

DEFF Research Database (Denmark)

Li, Xuemeng; Ahmed-Kristensen, Saeema

2015-01-01

requirements can lead to inappropriate products (Hall, et al., 2002). Understanding the nature of design requirements and the sources, from where they can or should be generated, is critical to before developing methods and processes to support this process. Requirement Engineering research, originated from...

76 FR 50881 - Required Scale Tests

Science.gov (United States)

2011-08-17

... RIN 0580-AB10 Required Scale Tests AGENCY: Grain Inspection, Packers and Stockyards Administration... required scale tests. Those documents defined ``limited seasonal basis'' incorrectly. This document... 20, 2011 (76 FR 3485) and on April 4, 2011 (76 FR 18348), concerning required scale tests. Those...

20 CFR 302.6 - Publication requirements.

Science.gov (United States)

2010-04-01

... 20 Employees' Benefits 1 2010-04-01 2010-04-01 false Publication requirements. 302.6 Section 302.6 Employees' Benefits RAILROAD RETIREMENT BOARD REGULATIONS UNDER THE RAILROAD UNEMPLOYMENT INSURANCE ACT QUALIFIED EMPLOYEE § 302.6 Publication requirements. (a) Publication of base year compensation requirement...

49 CFR 383.111 - Required knowledge.

Science.gov (United States)

2010-10-01

... importance of proper visual search, and proper visual search methods. (6) Communication. The principles and procedures for proper communications and the hazards of failure to signal properly. (7) Speed management. The... STANDARDS; REQUIREMENTS AND PENALTIES Required Knowledge and Skills § 383.111 Required knowledge. All...

Quality Assurance Requirements and Description

International Nuclear Information System (INIS)

Ram Murthy

2002-01-01

The Quality Assurance Requirements and Description (QARD) is the principal Quality Assurance (QA) document for the Civilian Radioactive Waste Management Program (Program). It establishes the minimum requirements for the QA program [INTRODUCTION :1p2s (NOT A REQUIREMENT)]. The QARD contains regulatory requirements and program commitments necessary for the development of an effective QA program [INTRODUCTION :1p3s (NOT A REQUIREMENT)]. Implementing documents must be based on, and be consistent with the QARD. The QARD applies to the following: (1) Acceptance of spent nuclear fuel and high-level waste. (2) Transport of spent nuclear fuel and high-level waste. (3) Storage of spent nuclear fuel through receipt of storage cask certification or a facility operating license. (4) Monitored Geologic Repository, including the site characterization activities [Exploratory Studies Facility (ESF) and surface based testing], through receipt of an operating license. (5) High-level waste form development through qualification, production, and acceptance. (6) Characterization of DOE spent nuclear fuel, and conditioning through acceptance of DOE spent nuclear fuel. Section 2.0, Quality Assurance Program, defines in greater detail criteria for determining work subject to the QARD

Identification of Salt-Tolerant Sinorhizobium sp Strain BL3 Membrane Proteins Based on Proteomics

DEFF Research Database (Denmark)

Tanthanuch, Waraporn; Mohammed, Shabaz; Matthiesen, Rune

2010-01-01

functional categories, the two biggest of which were energy production and conversion, and proteins not in clusters of orthologous groups (COGs). In addition, a comparative analysis of membrane proteins between salt-stressed and non-stressed BL3 cells was conducted using a membrane enrichment method and off-line...... SCX fractionation coupled to nanoLC-MS/MS. These techniques would be useful for further comparative analysis of membrane proteins that function in the response to environmental stress....

TRACER - TRACING AND CONTROL OF ENGINEERING REQUIREMENTS

Science.gov (United States)

Turner, P. R.

1994-01-01

TRACER (Tracing and Control of Engineering Requirements) is a database/word processing system created to document and maintain the order of both requirements and descriptive material associated with an engineering project. A set of hierarchical documents are normally generated for a project whereby the requirements of the higher level documents levy requirements on the same level or lower level documents. Traditionally, the requirements are handled almost entirely by manual paper methods. The problem with a typical paper system, however, is that requirements written and changed continuously in different areas lead to misunderstandings and noncompliance. The purpose of TRACER is to automate the capture, tracing, reviewing, and managing of requirements for an engineering project. The engineering project still requires communications, negotiations, interactions, and iterations among people and organizations, but TRACER promotes succinct and precise identification and treatment of real requirements separate from the descriptive prose in a document. TRACER permits the documentation of an engineering project's requirements and progress in a logical, controllable, traceable manner. TRACER's attributes include the presentation of current requirements and status from any linked computer terminal and the ability to differentiate headers and descriptive material from the requirements. Related requirements can be linked and traced. The program also enables portions of documents to be printed, individual approval and release of requirements, and the tracing of requirements down into the equipment specification. Requirement "links" can be made "pending" and invisible to others until the pending link is made "binding". Individuals affected by linked requirements can be notified of significant changes with acknowledgement of the changes required. An unlimited number of documents can be created for a project and an ASCII import feature permits existing documents to be incorporated

Managing Requirements-Documents to Data

Science.gov (United States)

Orr, Kevin; Hudson, Abe

2017-01-01

Managing Requirements on long term projects like International Space Station (ISS) can go thru many phases, from initial product development to almost over 20 years of operations and sustainment. Over that time many authorized changes have been made to the requirement set, that apply to any new systems that would visit the ISS today, like commercial cargo/crew vehicles or payloads. Explore the benefits of managing requirements in a database while satisfying traditional documents needs for contracts and stakeholder/user consumption that are not tied into the database.

27 CFR 7.28 - General requirements.

Science.gov (United States)

2010-04-01

... statements of alcoholic content, all mandatory information required on labels by this part shall be in script.... Except for statements of alcoholic content, all mandatory information required on labels by this part... OF THE TREASURY LIQUORS LABELING AND ADVERTISING OF MALT BEVERAGES Labeling Requirements for Malt...

47 CFR 80.1081 - Functional requirements.

Science.gov (United States)

2010-10-01

... 47 Telecommunication 5 2010-10-01 2010-10-01 false Functional requirements. 80.1081 Section 80... STATIONS IN THE MARITIME SERVICES Global Maritime Distress and Safety System (GMDSS) Equipment Requirements for Ship Stations § 80.1081 Functional requirements. Ships, while at sea, must be capable: (a) Except...

7 CFR 1942.117 - General requirements.

Science.gov (United States)

2010-01-01

... § 1942.117 General requirements. (a) Reserve requirements. Loans under this subpart are subject to the... 7 Agriculture 13 2010-01-01 2009-01-01 true General requirements. 1942.117 Section 1942.117 Agriculture Regulations of the Department of Agriculture (Continued) RURAL HOUSING SERVICE, RURAL BUSINESS...

Requirements Engineering for Pervasive Services

NARCIS (Netherlands)

Kolos, L.; Poulisse, Gert-Jan; van Eck, Pascal; Videira lopes, C.; Schaefer, S.; Clarke, S.; Elrad, T.; Jahnke, J.

2005-01-01

Developing pervasive mobile services for a mass market of end customers entails large up-front investments and therefore a good understanding of customer requirements is of paramount importance. This paper presents an approach for developing requirements engineering method that takes distinguishing

10 CFR 600.331 - Requirements.

Science.gov (United States)

2010-01-01

..., State or local authority as may have proper jurisdiction. Reports and Records ... Requirements. The following requirements pertain to recipients' procurements funded in whole or in part with Federal funds or with recipients' cost-share or match: (a) Reasonable cost. Recipients' procurement...

40 CFR 205.55 - Requirements.

Science.gov (United States)

2010-07-01

... 40 Protection of Environment 24 2010-07-01 2010-07-01 false Requirements. 205.55 Section 205.55 Protection of Environment ENVIRONMENTAL PROTECTION AGENCY (CONTINUED) NOISE ABATEMENT PROGRAMS TRANSPORTATION EQUIPMENT NOISE EMISSION CONTROLS Medium and Heavy Trucks § 205.55 Requirements. ...

Long-Term Stewardship Science and Technology Requirements

International Nuclear Information System (INIS)

McDonald, J.K.; Nickelson, R.A.

2002-01-01

To ensure technology developed for long-term stewardship will meet existing requirements, a review of requirements was performed. In addition to identifying existing science and technology related requirements, gaps and conflicts of requirements were identified

Advanced light water reactor utility requirements document: Volume 1--ALWR policy and summary of top-tier requirements

International Nuclear Information System (INIS)

Anon.

1990-01-01

The U.S. utilities are leading an industry wide effort to establish the technical foundation for the design of the Advanced Light Water Reactor (ALWR). This effort, the ALWR Program, is being managed for the U.S. electric utility industry by the Electric Power Research Institute (EPRI) and includes participation and sponsorship of several international utility companies and close cooperation with the U.S. Department of Energy (DOE). The cornerstone of the ALWR Program is a set of utility design requirements which are contained in the ALWR Requirements Document. The purpose of the Requirement Document is to present a clear, complete statement of utility desires for their next generation of nuclear plants. The Requirements Document covers the entire plant up to the grid interface. It therefore is the basis for an integrated plant design, i.e., nuclear steam supply system and balance of plant, and it emphasizes those areas which are most important to the objective of achieving an ALWR which is excellent with respect to safety, performance, constructibility, and economics. The document applies to both Pressurized Water Reactors (PWRs) and Boiling Water Reactors (BWRs). The Requirements Document is organized in three volumes. Volume 1 summarizes AlWR Program policy statements and top-tier requirements. The top-tier design requirements are categorized by major functions, including safety and investment protection, performance, and design process and constructibility. There is also a set of general design requirements, such as simplification and proven technology, which apply broadly to the ALWR design, and a set of economic goals for the ALWR program. The top-tier design requirements are described further in Volume 1 and are formally invoked as requirements in Volumes 2 and 3

42 CFR 424.36 - Signature requirements.

Science.gov (United States)

2010-10-01

... 42 Public Health 3 2010-10-01 2010-10-01 false Signature requirements. 424.36 Section 424.36... (CONTINUED) MEDICARE PROGRAM CONDITIONS FOR MEDICARE PAYMENT Claims for Payment § 424.36 Signature requirements. (a) General rule. The beneficiary's own signature is required on the claim unless the beneficiary...

40 CFR 141.80 - General requirements.

Science.gov (United States)

2010-07-01

... service line replacement, and public education. These requirements are triggered, in some cases, by lead... requirements. (1) All water systems shall install and operate optimal corrosion control treatment as defined in... specified by the State under § 141.83. (f) Lead service line replacement requirements. Any system exceeding...

40 CFR 63.347 - Reporting requirements.

Science.gov (United States)

2010-07-01

... for Chromium Emissions From Hard and Decorative Chromium Electroplating and Chromium Anodizing Tanks... required each time that an affected source becomes subject to the requirements of this subpart. (2) If the... time a notification of compliance status is required under this part, the owner or operator of an...

A vulnerability-centric requirements engineering framework : Analyzing security attacks, countermeasures, and requirements based on vulnerabilities

NARCIS (Netherlands)

Elahi, G.; Yu, E.; Zannone, N.

2010-01-01

Many security breaches occur because of exploitation of vulnerabilities within the system. Vulnerabilities are weaknesses in the requirements, design, and implementation, which attackers exploit to compromise the system. This paper proposes a methodological framework for security requirements

Spent fuel storage requirements

International Nuclear Information System (INIS)

Fletcher, J.

1982-06-01

Spent fuel storage requirements, as projected through the year 2000 for U.S. LWRs, were calculated using information supplied by the utilities reflecting plant status as of December 31, 1981. Projections through the year 2000 combined fuel discharge projections of the utilities with the assumed discharges of typical reactors required to meet the nuclear capacity of 165 GWe projected by the Energy Information Administration (EIA) for the year 2000. Three cases were developed and are summarized. A reference case, or maximum at-reactor (AR) capacity case, assumes that all reactor storage pools are increased to their maximum capacities as estimated by the utilities for spent fuel storage utilizing currently licensed technologies. The reference case assumes no transshipments between pools except as currently licensed by the Nuclear Regulatory Commission (NRC). This case identifies an initial requirement for 13 MTU of additional storage in 1984, and a cumulative requirement for 14,490 MTU additional storage in the year 2000. The reference case is bounded by two alternative cases. One, a current capacity case, assumes that only those pool storage capacity increases currently planned by the operating utilities will occur. The second, or maximum capacity with transshipment case, assumes maximum development of pool storage capacity as described above and also assumes no constraints on transshipment of spent fuel among pools of reactors of like type (BWR, PWR) within a given utility. In all cases, a full core discharge capability (full core reserve or FCR) is assumed to be maintained for each reactor, except that only one FCR is maintained when two reactors share a common pool. For the current AR capacity case the indicated storage requirements in the year 2000 are indicated to be 18,190 MTU; for the maximum capacity with transshipment case they are 11,320 MTU

Nuclear energy and investment requirements

International Nuclear Information System (INIS)

Voeltzel, D.

1978-01-01

The author assesses the investment requirements of the French nuclear programme within the framework of the national economy. He then evokes the means of financing these requirements as well as drawing attention to certain constraints which must be taken into account [fr

7 CFR 718.9 - Signature requirements.

Science.gov (United States)

2010-01-01

... 7 Agriculture 7 2010-01-01 2010-01-01 false Signature requirements. 718.9 Section 718.9... MULTIPLE PROGRAMS General Provisions § 718.9 Signature requirements. (a) When a program authorized by this chapter or Chapter XIV of this title requires the signature of a producer; landowner; landlord; or tenant...

28 CFR 80.12 - Accounting requirements.

Science.gov (United States)

2010-07-01

... 28 Judicial Administration 2 2010-07-01 2010-07-01 false Accounting requirements. 80.12 Section 80... PROCEDURE § 80.12 Accounting requirements. Neither the submission of a request for an FCPA Opinion, its... comply with the accounting requirements of 15 U.S.C. 78m(b)(2) and (3). ...

29 CFR 1620.32 - Recordkeeping requirements.

Science.gov (United States)

2010-07-01

... Regulations Relating to Labor (Continued) EQUAL EMPLOYMENT OPPORTUNITY COMMISSION THE EQUAL PAY ACT § 1620.32 Recordkeeping requirements. (a) Employers having employees subject to the Act are required to keep records in... subject to the equal pay provisions of the Act shall maintain and preserve all records required by the...

7 CFR 1948.96 - Audit requirements.

Science.gov (United States)

2010-01-01

... Program § 1948.96 Audit requirements. (a) Audit requirements for Site Development and Acquisition Grants will be made in accordance with FmHA Instruction 1942-G. (b) Audits for planning grants made in... 7 Agriculture 13 2010-01-01 2009-01-01 true Audit requirements. 1948.96 Section 1948.96...

22 CFR 126.13 - Required information.

Science.gov (United States)

2010-04-01

... 22 Foreign Relations 1 2010-04-01 2010-04-01 false Required information. 126.13 Section 126.13... PROVISIONS § 126.13 Required information. (a) All applications for licenses (DSP-5, DSP-61, DSP-73, and DSP... are multiple consignors, consignees or freight forwarders, and all the required information cannot be...

49 CFR 7.4 - Publication required.

Science.gov (United States)

2010-10-01

... 49 Transportation 1 2010-10-01 2010-10-01 false Publication required. 7.4 Section 7.4 Transportation Office of the Secretary of Transportation PUBLIC AVAILABILITY OF INFORMATION Information Required To Be Made Public by DOT § 7.4 Publication required. (a) General. The material described in § 7.3...

Monitored Retrievable Storage System Requirements Document

International Nuclear Information System (INIS)

1994-03-01

This Monitored Retrievable Storage System Requirements Document (MRS-SRD) describes the functions to be performed and technical requirements for a Monitored Retrievable Storage (MRS) facility subelement and the On-Site Transfer and Storage (OSTS) subelement. The MRS facility subelement provides for temporary storage, at a Civilian Radioactive Waste Management System (CRWMS) operated site, of spent nuclear fuel (SNF) contained in an NRC-approved Multi-Purpose Canister (MPC) storage mode, or other NRC-approved storage modes. The OSTS subelement provides for transfer and storage, at Purchaser sites, of spent nuclear fuel (SNF) contained in MPCs. Both the MRS facility subelement and the OSTS subelement are in support of the CRWMS. The purpose of the MRS-SRD is to define the top-level requirements for the development of the MRS facility and the OSTS. These requirements include design, operation, and decommissioning requirements to the extent they impact on the physical development of the MRS facility and the OSTS. The document also presents an overall description of the MRS facility and the OSTS, their functions (derived by extending the functional analysis documented by the Physical System Requirements (PSR) Store Waste Document), their segments, and the requirements allocated to the segments. In addition, the top-level interface requirements of the MRS facility and the OSTS are included. As such, the MRS-SRD provides the technical baseline for the MRS Safety Analysis Report (SAR) design and the OSTS Safety Analysis Report design

Space station data system analysis/architecture study. Task 1: Functional requirements definition, DR-5. Appendix: Requirements data base

Science.gov (United States)

1985-01-01

Appendix A contains data that characterize the system functions in sufficient depth as to determine the requirements for the Space Station Data System (SSDS). This data is in the form of: (1) top down traceability report; (2) bottom up traceability report; (3) requirements data sheets; and (4) cross index of requirements paragraphs of the source documents and the requirements numbers. A data base users guide is included that interested parties can use to access the requirements data base and get up to date information about the functions.

The nutritional requirements of exercising dogs.

Science.gov (United States)

Hill, R C

1998-12-01

The nutrient requirements of canine athletes are unique. Dogs have a greater capacity for fat oxidation than humans both at rest and during exercise. In dogs undertaking endurance exercise, such as sled dogs, high fat (>50% of energy) diets increase stamina and maximize energy production, and high protein (>30% of energy) diets prevent training-induced anemia. Nutrient requirements differ, however, for sprint racing dogs, such as greyhounds. Greyhounds run faster when fed moderately increased dietary fat but run more slowly when dietary protein is increased. Sled dogs have similar energy requirements to other breeds at rest in a thermoneutral environment ( approximately 550W0.75 kJ/d where W is body weight in kg) but may require as much as 4200W0.75 kJ/d during a race. The energy requirement of greyhounds in training, however, is only approximately 600W0.75 kJ/d. There is little information, however, concerning the vitamin, mineral or other nutrient requirements of athletic dogs; most sled dogs and greyhounds are fed "homemade" recipes. These recipes usually include raw meat and represent a health risk. More studies are required to improve the health and performance of working and racing dogs.

42 CFR 417.934 - Reserve requirement.

Science.gov (United States)

2010-10-01

... 42 Public Health 3 2010-10-01 2010-10-01 false Reserve requirement. 417.934 Section 417.934 Public... PLANS Administration of Outstanding Loans and Loan Guarantees § 417.934 Reserve requirement. (a) Timing... section 1305 of the PHS Act was required to establish a restricted reserve account on the earlier of the...

Consensus standard requirements and guidance

International Nuclear Information System (INIS)

Putman, V.L.

1995-01-01

This report presents information from the ANS Criticality Alarm System Workshop relating to the consensus standard requirements and guidance. Topics presented include: definition; nomenclature; requirements and recommendations; purpose of criticality alarms; design criteria; signal characteristics; reliability, dependability and durability; tests; and emergency preparedness and planning

Security and trust requirements engineering

NARCIS (Netherlands)

Giorgini, P.; Massacci, F.; Zannone, N.; Aldini, A.; Gorrieri, R.; Martinelli, F.

2005-01-01

Integrating security concerns throughout the whole software development process is one of today’s challenges in software and requirements engineering research. A challenge that so far has proved difficult to meet. The major difficulty is that providing security does not only require to solve

29 CFR 96.12 - Audit requirements.

Science.gov (United States)

2010-07-01

.... (b) The audit requirements contained in 29 CFR part 99 shall be followed for audits of all fiscal... 29 Labor 1 2010-07-01 2010-07-01 true Audit requirements. 96.12 Section 96.12 Labor Office of the Secretary of Labor AUDIT REQUIREMENTS FOR GRANTS, CONTRACTS, AND OTHER AGREEMENTS Audits of States, Local...

77 FR 6704 - Exemptions From Entry Requirements and Report of Arrival Requirements for Certain Department of...

Science.gov (United States)

2012-02-09

... requirements and thus also subject to advanced electronic presentation of cargo information requirements. Under..., Aircraft, Airports, Alcohol and alcoholic beverages, Cigars and cigarettes, Cuba, Customs duties and...

Systematic handling of requirements and conditions (in compliance with waste acceptance requirements for a radioactive waste disposal facility)

International Nuclear Information System (INIS)

Keyser, Peter; Helander, Anita

2012-01-01

This Abstract and presentation will demonstrate the need for a structured requirement management and draw upon experiences and development from SKB requirements data base and methodology, in addition to international guidelines and software tools. The presentation will include a discussion on how requirement management can be applied for the decommissioning area. The key issue in the decommissioning of nuclear facilities is the progressive removal of hazards, by stepwise decontamination and dismantling activities that have to be carried out safely and within the boundaries of an approved safety case. For decommissioning there exists at least two safety cases, one for the pre-disposal activities and one for the disposal facility, and a need for a systematic handling of requirements and conditions to safely manage the radioactive waste in the long term. The decommissioning safety case is a collection of arguments and evidence to demonstrate the safety of a decommissioning project. It also includes analyzing and updating the decommissioning safety case in accordance with the waste acceptance criteria's and the expected output, i.e. waste packages. It is a continuous process to confirm that all requirements have been met. On the other hand there is the safety case for a radioactive waste disposal facility, which may include the following processes and requirements: i) Integrating relevant scientific (and other) information in a structured, traceable and transparent way and, thereby, developing and demonstrating an understanding of the potential behavior and performance of the disposal system; ii) Identifying uncertainties in the behavior and performance of the disposal system, describing the possible significance of the uncertainties, and identifying approaches for the management of significant uncertainties; iii) Demonstrating long-term safety and providing reasonable assurance that the disposal facility will perform in a manner that protects human health and the

16 CFR 303.4 - English language requirement.

Science.gov (United States)

2010-01-01

... 16 Commercial Practices 1 2010-01-01 2010-01-01 false English language requirement. 303.4 Section... AND REGULATIONS UNDER THE TEXTILE FIBER PRODUCTS IDENTIFICATION ACT § 303.4 English language requirement. All required information shall be set out in the English language. If the required information...

Meeting Quay 2k30's requirements

NARCIS (Netherlands)

Wijnants, G.H.; Toorn, A. van der; Schuylenburg, M.; Heijnen, H.P.J.; Gijt, J.G. de; Molenaar, W.F.; Ligteringen, H.; Krom, A.H.M.

2005-01-01

The requirements that a quay design should meet in order to yield a viable port infrastructure, vary widely from flexibility due to future customers requirements to durability due to owners requirements. In a Port of Rotterdam backed project, current and future requirements have been aggregated by

12 CFR 204.7 - Supplemental reserve requirement.

Science.gov (United States)

2010-01-01

... 12 Banks and Banking 2 2010-01-01 2010-01-01 false Supplemental reserve requirement. 204.7 Section... RESERVE REQUIREMENTS OF DEPOSITORY INSTITUTIONS (REGULATION D) § 204.7 Supplemental reserve requirement... reserve requirement on every depository institution of not more than 4 percent of its total transaction...

25 CFR 276.8 - Financial reporting requirements.

Science.gov (United States)

2010-04-01

... 25 Indians 1 2010-04-01 2010-04-01 false Financial reporting requirements. 276.8 Section 276.8... ASSISTANCE ACT PROGRAM UNIFORM ADMINISTRATIVE REQUIREMENTS FOR GRANTS § 276.8 Financial reporting requirements. Requirements for grantees to report financial information to the Bureau, and to request advances...

Security Requirements – Analysis of the Issue

Directory of Open Access Journals (Sweden)

Jhon Vincent

2013-12-01

Full Text Available Needs about security are matters little taken into account when managing requirements engineering , and when considered in the life cycle of the system , they tend to become a general list of functions, as password of protection , firewalls , virus detection tools , and other similar. But in fact, they cannot be considered as requirements of security, because they are implementation mechanisms to try to meet unspecified requirements, as an authenticated access. As a result, the security requirements for the system are ignored, which are required to protect essential services and assets, besides, when are specified, is not considered the prospect of future attacks. This paper describes the need for a systematic approach to managing security requirements engineering, in order to help avoid the problem of generic lists and take into account the future perspective. Several related approaches are described and also are provided references additional material that can help requirements engineers to ensure that their products be taken into account, effectively , the security requirements.

Requirements Engineering for Software Integrity and Safety

Science.gov (United States)

Leveson, Nancy G.

2002-01-01

Requirements flaws are the most common cause of errors and software-related accidents in operational software. Most aerospace firms list requirements as one of their most important outstanding software development problems and all of the recent, NASA spacecraft losses related to software (including the highly publicized Mars Program failures) can be traced to requirements flaws. In light of these facts, it is surprising that relatively little research is devoted to requirements in contrast with other software engineering topics. The research proposed built on our previous work. including both criteria for determining whether a requirements specification is acceptably complete and a new approach to structuring system specifications called Intent Specifications. This grant was to fund basic research on how these ideas could be extended to leverage innovative approaches to the problems of (1) reducing the impact of changing requirements, (2) finding requirements specification flaws early through formal and informal analysis, and (3) avoiding common flaws entirely through appropriate requirements specification language design.

Safety of Research Reactors. Safety Requirements

International Nuclear Information System (INIS)

2010-01-01

The main objective of this Safety Requirements publication is to provide a basis for safety and a basis for safety assessment for all stages in the lifetime of a research reactor. Another objective is to establish requirements on aspects relating to regulatory control, the management of safety, site evaluation, design, operation and decommissioning. Technical and administrative requirements for the safety of research reactors are established in accordance with these objectives. This Safety Requirements publication is intended for use by organizations engaged in the site evaluation, design, manufacturing, construction, operation and decommissioning of research reactors as well as by regulatory bodies

78 FR 38555 - Importer Permit Requirements for Tobacco Products and Processed Tobacco, and Other Requirements...

Science.gov (United States)

2013-06-27

..., and Other Requirements for Tobacco Products, Processed Tobacco, and Cigarette Papers and Tubes AGENCY... administration and enforcement of importer permits over the past decade, TTB believes that it can gain... minimum manufacturing and marking requirements for tobacco products and cigarette papers and tubes, and...

7 CFR 2900.4 - Natural gas requirements.

Science.gov (United States)

2010-01-01

... 7 Agriculture 15 2010-01-01 2010-01-01 false Natural gas requirements. 2900.4 Section 2900.4..., DEPARTMENT OF AGRICULTURE ESSENTIAL AGRICULTURAL USES AND VOLUMETRIC REQUIREMENTS-NATURAL GAS POLICY ACT § 2900.4 Natural gas requirements. For purposes of Section 401(c), NGPA, the natural gas requirements for...

12 CFR 204.9 - Emergency reserve requirement.

Science.gov (United States)

2010-01-01

... 12 Banks and Banking 2 2010-01-01 2010-01-01 false Emergency reserve requirement. 204.9 Section... RESERVE REQUIREMENTS OF DEPOSITORY INSTITUTIONS (REGULATION D) § 204.9 Emergency reserve requirement. (a..., additional reserve requirements on depository institutions at any ratio on any liability upon a finding by at...

Civilian Radioactive Waste Management System Requirements Document

International Nuclear Information System (INIS)

1992-12-01

This document specifies the top-level requirements for the Civilian Radioactive Waste Management System (CRWMS). The document is referred to herein as the CRD, for CRWMS Requirements document. The OCRWM System Engineering Management Plan (SEMP) establishes the technical document hierarchy (hierarchy of technical requirements and configuration baseline documents) for the CRWMS program. The CRD is the top-level document in this hierarchy. The immediate subordinate documents are the System Requirements Documents (SRDS) for the four elements of the CRWMS and the Interface Specification (IFS). The four elements of the CRWMS are the Waste Acceptance System, the Transportation System, the Monitored Retrievable Storage (MRS) System and the Mined Geologic Disposal System (MGDS). The Interface Specification describes the six inter-element interfaces between the four elements. This hierarchy establishes the requirements to be addressed by the design of the system elements. Many of the technical requirements for the CRWMS are documented in a variety of Federal regulations, DOE directives and other Government documentation. It is the purpose of the CRD to establish the technical requirements for the entire program. In doing so, the CRD summarizes source documentation for requirements that must be addressed by the program, specifies particular requirements, and documents derived requirements that are not covered in regulatory and other Government documentation, but are necessary to accomplish the mission of the CRWMS. The CRD defines the CRWMS by identifying the top-level functions the elements must perform (These top-level functions were derived using functional analysis initially documented in the Physical System Requirements (PSR) documents). The CRD also defines the top-level physical architecture of the system and allocates the functions and requirements to the architectural elements of the system

13 CFR 120.851 - CDC ethical requirements.

Science.gov (United States)

2010-01-01

... 13 Business Credit and Assistance 1 2010-01-01 2010-01-01 false CDC ethical requirements. 120.851... Company Loan Program (504) Other Cdc Requirements § 120.851 CDC ethical requirements. CDCs and their Associates must act ethically and exhibit good character. They must meet all of the ethical requirements of...

29 CFR 1602.19 - Additional reporting requirements.

Science.gov (United States)

2010-07-01

....19 Additional reporting requirements. The Commission reserves the right to require reports, other... necessary to accomplish the purpose of title VII, the ADA, or GINA. Any system for the requirement of such... 29 Labor 4 2010-07-01 2010-07-01 false Additional reporting requirements. 1602.19 Section 1602.19...

29 CFR 1602.11 - Additional reporting requirements.

Science.gov (United States)

2010-07-01

... Additional reporting requirements. The Commission reserves the right to require reports, other than that... accomplish the purposes of title VII, the ADA, or GINA. Any system for the requirement of such reports will... 29 Labor 4 2010-07-01 2010-07-01 false Additional reporting requirements. 1602.11 Section 1602.11...

29 CFR 1602.26 - Additional reporting requirements.

Science.gov (United States)

2010-07-01

... Report § 1602.26 Additional reporting requirements. The Commission reserves the right to require reports... to accomplish the purposes of title VII, the ADA, or GINA. Any system for requirement of such reports... 29 Labor 4 2010-07-01 2010-07-01 false Additional reporting requirements. 1602.26 Section 1602.26...

12 CFR 932.8 - Minimum liquidity requirements.

Science.gov (United States)

2010-01-01

... 12 Banks and Banking 7 2010-01-01 2010-01-01 false Minimum liquidity requirements. 932.8 Section... CAPITAL STANDARDS FEDERAL HOME LOAN BANK CAPITAL REQUIREMENTS § 932.8 Minimum liquidity requirements. In addition to meeting the deposit liquidity requirements contained in § 965.3 of this chapter, each Bank...

Development of transportation operations requirements

International Nuclear Information System (INIS)

Grady, S.T.; Best, R.E.; Danese, F.L.; Peterson, R.W.; Pope, R.B.

1990-01-01

Transport conditions at various utility sties vary dramatically in terms of characteristics at and near the site, requirements, administrative procedures, and other factors. Continuation of design efforts for the OCRWM transportation operations system requires that the operating requirements for the transportation system -- quantity of fuel per unit time per site -- be identified so that the effect the variations have on the system can be accommodated. The approach outlined in this paper provides for an identification of specific sites, evaluation of shipment capabilities at each site, and integration of the sites into multi-site shipping campaigns to scope the logistics management problem for the transportation operations system. 1 fig., 1 tab

Design requirement on KALIMER control rod assembly duct

International Nuclear Information System (INIS)

Hwang, W.; Kang, H. Y.; Nam, C.; Kim, J. O.; Kim, Y. J.

1998-03-01

This document establishes the design guidelines which are needs for designing the control rod assembly duct of the KALIMER as design requirements. it describes control rod assembly duct of the KALIMER and its requirements that includes functional requirements, performance requirements, interfacing systems, design limits and strength requirements, seismic requirements, structural requirements, environmental requirements, reliability and safety requirements, standard and codes, QA programs, and other requirements. The control rod system consists of three parts, which are drive mechanism, drive-line, and absorber bundle. This report deals with the absorber bundle and its outer duct only because the others are beyond the scope of fuel system design. The guidelines for design requirements intend to be used for an improved design of the control rod assembly duct of the KALIMER. (author). 19 refs

Design requirement on KALIMER control rod assembly duct

Energy Technology Data Exchange (ETDEWEB)

Hwang, W.; Kang, H. Y.; Nam, C.; Kim, J. O.; Kim, Y. J

1998-03-01

This document establishes the design guidelines which are needs for designing the control rod assembly duct of the KALIMER as design requirements. it describes control rod assembly duct of the KALIMER and its requirements that includes functional requirements, performance requirements, interfacing systems, design limits and strength requirements, seismic requirements, structural requirements, environmental requirements, reliability and safety requirements, standard and codes, QA programs, and other requirements. The control rod system consists of three parts, which are drive mechanism, drive-line, and absorber bundle. This report deals with the absorber bundle and its outer duct only because the others are beyond the scope of fuel system design. The guidelines for design requirements intend to be used for an improved design of the control rod assembly duct of the KALIMER. (author). 19 refs.

EPR compared to international requirements (Mainly EUR)

International Nuclear Information System (INIS)

Broecker, B.

1996-01-01

A number of European Utilities have entered an agreement to write common requirements dedicated to future light water nuclear power plants to be built in Europe. The activities are known under the sign EUR (European Utilities Requirements). EPR, the future European Pressurized water Reactor, is the first installation of this type which will be operational from the year 2000 onwards, must fulfill the European requirements. EPR will serve as a test whether these requirements are realistic and well balanced. At the basic design stage of EPR, this paper concentrates on four main topics: the requirements which are new compared with existing reactors and which put a major challenge to the designer; the requirements today still open and the way they can be met by the EPR or not; the points for which already today the EPR special requirements exceed the EUR; the examples where the design of the EPR has given feedback which has led to a change of the EUR. EPR and EUR are different approaches to the reactor of the future. EUR is a set of requirements which leaves a flexibility to the designer while EPR is a real project which defines the technical solutions. EPR will fulfill the EUR and will at the same time serve as a test whether these requirements are realistic. EPR will also fulfill international requirements with minor changes. (J.S.). 7 figs

40 CFR 161.340 - Toxicology data requirements.

Science.gov (United States)

2010-07-01

... 40 Protection of Environment 23 2010-07-01 2010-07-01 false Toxicology data requirements. 161.340... Toxicology data requirements. (a) Table. Sections 161.100 through 161.102 describe how to use this table to determine the toxicology data requirements and the substance to be tested. Kind of data required (b) Notes...

Safety requirements for the Pu carriers

International Nuclear Information System (INIS)

Mishima, H.

1993-01-01

Ministry of Transport of Japan has now set about studying requirements for Pu carriers to ensure safety. It was first studied what the basic concept of safe carriage of Pu should be, and the basic ideas have been worked out. Next the requirements for the Pu carriers were studied based on the above. There are at present no international requirements of construction and equipment for the nuclear-material carriers, but MOT of Japan has so far required special construction and equipment for the nuclear-material carriers which carry a large amount of radioactive material, such as spent fuel or low level radioactive waste, corresponding to the level of the respective potential hazard. The requirements of construction and equipment of the Pu carriers have been established considering the difference in heat generation between Pu and spent fuel, physical protection, and so forth, in addition to the above basic concept. (J.P.N.)

42 CFR 84.256 - Quality control requirements.

Science.gov (United States)

2010-10-01

... 42 Public Health 1 2010-10-01 2010-10-01 false Quality control requirements. 84.256 Section 84.256... § 84.256 Quality control requirements. (a) In addition to the construction and performance requirements specified in §§ 84.251, 84.252, 84.253, 84.254, and 84.255, the quality control requirements in paragraphs...

Formal Verification of Real-Time System Requirements

Directory of Open Access Journals (Sweden)

Marcin Szpyrka

2000-01-01

Full Text Available The methodology of system requirements verification presented in this paper is a proposition of a practical procedure for reducing some negatives of the specification of requirements. The main problem that is considered is to create a complete description of the system requirements without any negatives. Verification of the initially defined requirements is based on the coloured Petri nets. Those nets are useful for testing some properties of system requirements such as completeness, consistency and optimality. An example ofthe litt controller is presented.

Training Requirements and Information Management System

Energy Technology Data Exchange (ETDEWEB)

Cillan, T.F.; Hodgson, M.A.

1992-05-01

This is the software user's guide for the Training Requirements and Information Management System. This guide defines and describes the software operating procedures as they apply to the end user of the software program. This guide is intended as a reference tool for the user who already has an indepth knowledge of the Training Requirements and Information Management System functions and data reporting requirement.

Improvement of Requirement Elicitation Process through Cognitive Psychology

Directory of Open Access Journals (Sweden)

Sana Fatima

2017-06-01

Full Text Available Proper requirement elicitation is necessary for client satisfaction along with the overall project success, but requirement engineers face problems in understanding user requirements and the users of the required system fail to make requirement engineering team understand what they actually want. It is then responsibility of requirement engineers to extract proper requirements. This paper discusses how to use cognitive psychology and learning style models (LSM to understand the psychology of clients. Moreover, it also discusses usage of proper elicitation technique according to one’s learning style and gather the right requirements.

Humidity requirements in WSCF Laboratories

International Nuclear Information System (INIS)

Evans, R.A.

1994-01-01

The purpose of this paper is to develop and document a position on Relative Humidity (RH) requirements in the WSCF Laboratories. A current survey of equipment vendors for Organic, Inorganic and Radiochemical laboratories indicate that 25% - 80% relative humidity may meet the environmental requirements for safe operation and protection of all the laboratory equipment

FAA Financial Requirements

Science.gov (United States)

1997-06-04

In June 1995, the FAA developed a "total requirements" estimate for the period : FY 97-FY 02 to help explain the difficulty of supporting a dynamic, growing : aviation industry under a federal budget picture which projected flat or reduced : funding ...

Design requirement on KALIMER blanket fuel assembly duct

International Nuclear Information System (INIS)

Hwang, Woan; Kang, H. Y.; Nam, C.; Kim, J. O.

1998-03-01

This document describes design requirements which are needed for designing the blanket fuel assembly duct of the KALIMER as design guidance. The blanket fuel assembly duct of the KALIMER consists of fuel rods, mounting rail, nosepiece, duct with pad, handling socket with pad. Blanket fuel rod consists of top end plug, bottom end plug with solid ferritic-martensitic steel rod and key way blanket fuel slug, cladding, and wire wrap. In the assembly, the rods are in a triangular pitch array, and the rod bundle is attached to the nosepiece with mounting rails. The bottom end of the assembly duct is formed by a long nosepiece which provides the lower restraint function and the paths for coolant inlet. This report contains functional requirements, performance and operational requirements, interfacing systems requirements, core restraint and interface requirements, design limits and strength requirements, system configuration and essential feature requirements, seismic requirements, structural requirements, environmental requirements, reliability and safety requirements, standard and codes, QA programs, and other requirements. (author). 20 refs., 4 figs

Identified adjustability dimensions when generating a product specific requirements specification by requirements reuse

DEFF Research Database (Denmark)

Hauksdóttir, Dagný; Mortensen, Niels Henrik; Nielsen, Poul Erik

2014-01-01

. An extensive state of the art is included to introduce the presented methods related to each adjustability dimensions. The options for implementing each adjustability dimensions in a requirement reuse approach are illustrated along with a discussion regarding the benefits and issues resulting from each option....... This discussion should help practitioners to better understand the possible methods that can be implemented and to design a user friendly and sustainable approach. A case study, describing how the dimensions are incorporated in two requirements reuse approaches, for Danfoss Solar Inverters (SI) and Danfoss...

A Requirement Specification Language for AADL

Science.gov (United States)

2016-06-01

models. The objective of ReqSpec is to support the elicitation , definition, and modeling of requirements for real-time embedded systems in an iterative...A Requirement Specification Language for AADL Peter H. Feiler Julien Delange Lutz Wrage June 2016 TECHNICAL REPORT CMU/SEI-2016-TR-008...is required for any other external and/or commercial use. Requests for permission should be directed to the Software En- gineering Institute at

Fiscal year 1999 waste information requirements document

International Nuclear Information System (INIS)

Adams, M.R.

1998-01-01

The Waste Information Requirements Document (WIRD) has the following purposes: To describe the overall drivers that require characterization information and to document their source; To define how characterization is going to satisfy the drivers, close issues, and measure and report progress; and To describe deliverables and acceptance criteria for characterization. Characterization information is required to maintain regulatory compliance, perform operations and maintenance, resolve safety issues, and prepare for disposal of waste. Commitments addressing these requirements are derived from the Hanford Federal Facility Agreement and Consent Order, also known as the Tri-Party Agreement; the Recommendation 93-5 Implementation Plan (DOE-RL 1996a) to the Defense Nuclear Facilities Safety Board (DNFSB); and other requirement sources listed in Section 2.0. The Waste Information Requirements Document replaces the tank waste analysis plans and the tank characterization plan previously required by the Tri-Party Agreement, Milestone M-44-01 and M-44-02 series

Cold vacuum drying facility design requirements

Energy Technology Data Exchange (ETDEWEB)

IRWIN, J.J.

1999-07-01

This document provides the detailed design requirements for the Spent Nuclear Fuel Project Cold Vacuum Drying Facility. Process, safety, and quality assurance requirements and interfaces are specified.

Cold vacuum drying facility design requirements

International Nuclear Information System (INIS)

IRWIN, J.J.

1999-01-01

This document provides the detailed design requirements for the Spent Nuclear Fuel Project Cold Vacuum Drying Facility. Process, safety, and quality assurance requirements and interfaces are specified

PABRE-Proj: applying patterns in requirements elicitation

OpenAIRE

Palomares Bonache, Cristina; Quer Bosor, Maria Carme; Franch Gutiérrez, Javier

2013-01-01

Software requirement patterns have been proposed as a type of artifact for fostering requirements reuse. In this paper, we present PABRE-Proj, a tool aimed at supporting requirements elicitation and specification. Peer Reviewed

38 CFR 39.10 - Application requirements.

Science.gov (United States)

2010-07-01

...-10) Grant Requirements and Procedures § 39.10 Application requirements. (a) For a project to be..., etc., (10) Economic activities, (11) Cultural resources, (12) Aesthetics, (13) Residential population...

24 CFR 266.105 - Application requirements.

Science.gov (United States)

2010-04-01

... HOUSING FINANCE AGENCY RISK-SHARING PROGRAM FOR INSURED AFFORDABLE MULTIFAMILY PROJECT LOANS Housing Finance Agency Requirements § 266.105 Application requirements. (a) Applications for approval as a HUD...

Cloud Security Requirements - A checklist with security and privacy requirements for public cloud services

OpenAIRE

Bernsmed, Karin; Meland, Per Håkon; Jaatun, Martin Gilje

2015-01-01

- This document contains a checklist that can be used to develop or evaluate security and privacy requirements for Cloud computing services. The content has been gathered from established industry standards and best practices, supplemented with requirements from European data protection legislation, and taking into account security issues identified in recent research on Cloud security. The document is intended to be used by potential cloud customers that need to assess the security of a c...

From document to database: modernizing requirements management

International Nuclear Information System (INIS)

Giajnorio, J.; Hamilton, S.

2007-01-01

The creation, communication, and management of design requirements are central to the successful completion of any large engineering project, both technically and commercially. Design requirements in the Canadian nuclear industry are typically numbered lists in multiple documents created using word processing software. As an alternative, GE Nuclear Products implemented a central requirements management database for a major project at Bruce Power. The database configured the off-the-shelf software product, Telelogic Doors, to GE's requirements structure. This paper describes the advantages realized by this scheme. Examples include traceability from customer requirements through to test procedures, concurrent engineering, and automated change history. (author)

Are functional foods redefining nutritional requirements?

Science.gov (United States)

Jones, Peter J; Varady, Krista A

2008-02-01

Functional foods are increasing in popularity owing to their ability to confer health and physiological benefits. Nevertheless, the notion that functional foods improve health when providing nutrients at levels above and beyond existing recommended intakes is inconsistent with the definition of requirement. This disparity highlights the need for an alternative definition of nutrient requirement. The present objective is to examine distinctions between optimization of health, as defined by what we currently deem as required intakes, versus adding physiological benefit using bioactive agents found in functional foods. Presently, requirement is defined as the lowest amount of intake of a nutrient that will maintain a defined level of nourishment for a specific indicator of adequacy. In contrast, functional foods are described as ingredients that are not necessary for body function, yet provide added physiological benefit that confer better overall health. Plant sterols are one example of such an ingredient. Plant sterols lower plasma cholesterol concentrations, and may thus be considered essential nutrients in physiological situations where circulating cholesterol concentrations are high. Similarly, intakes of omega-3 fats beyond existing requirement may confer additional health benefits such as hypolipidemic and anti-diabetic effects. These examples underscore the inconsistencies between what is defined as a nutrient requirement versus what is identified as a health benefit of a functional food. Such discrepancies emphasize the need for a more all-encompassing definition of a nutrient requirement; that is, one that moves beyond the prevention of overt deficiency to encompass improved health and disease risk reduction.

Legal requirements governing proxy voting in Denmark

DEFF Research Database (Denmark)

Werlauff, Erik

2008-01-01

The requirements in Danish company law concerning proxy voting in companies whose shares have been accepted for listing on a regulated market have been successively tightened in recent years, and corporate governance principles have also led to the introduction of several requirements concerning...... proxy holders. A thorough knowledge of these requirements is important not only for the listed companies but also for their advisers and investors in Denmark and abroad. This article considers these requirements as well as the additional requirements which will derive from Directive 2007....../36 on the exercise of shareholders' rights in listed companies, which must be implemented by 3 August 2009. It is pointed out that companies may provide with advantage in their articles of association for both the existing and the forthcoming requirements at this early stage....

Disposal of Radioactive Waste. Specific Safety Requirements

International Nuclear Information System (INIS)

2011-01-01

This publication establishes requirements applicable to all types of radioactive waste disposal facility. It is linked to the fundamental safety principles for each disposal option and establishes a set of strategic requirements that must be in place before facilities are developed. Consideration is also given to the safety of existing facilities developed prior to the establishment of present day standards. The requirements will be complemented by Safety Guides that will provide guidance on good practice for meeting the requirements for different types of waste disposal facility. Contents: 1. Introduction; 2. Protection of people and the environment; 3. Safety requirements for planning for the disposal of radioactive waste; 4. Requirements for the development, operation and closure of a disposal facility; 5. Assurance of safety; 6. Existing disposal facilities; Appendices.

24 CFR 266.110 - Reserve requirements.

Science.gov (United States)

2010-04-01

... HOUSING FINANCE AGENCY RISK-SHARING PROGRAM FOR INSURED AFFORDABLE MULTIFAMILY PROJECT LOANS Housing Finance Agency Requirements § 266.110 Reserve requirements. (a) HFAs with top-tier designation or overall...

Towards a Scope Management of Non-Functional Requirements in Requirements Engineering

NARCIS (Netherlands)

Kassab, M.; Ormandjieva, O.; Daneva, Maia; Ebert, C.; Herrmann, A.; Rupp, C.

2007-01-01

Getting business stakeholders’ goals formulated clearly and project scope defined realistically increases the chance of success for any application development process. As a consequence, stakeholders at early project stages acquire as much as possible knowledge about the requirements, their risk

47 CFR 10.540 - Attestation requirement. [Reserved

Science.gov (United States)

2010-10-01

... 47 Telecommunication 1 2010-10-01 2010-10-01 false Attestation requirement. [Reserved] 10.540 Section 10.540 Telecommunication FEDERAL COMMUNICATIONS COMMISSION GENERAL COMMERCIAL MOBILE ALERT SYSTEM Equipment Requirements § 10.540 Attestation requirement. [Reserved] ...

Goals, requirements and prerequisites for teleradiology

International Nuclear Information System (INIS)

Walz, M.; Wein, B.; Lehmann, K.J.; Bolte, R.; Kilbinger, M.; Loose, R.; Guenther, R.W.; Georgi, M.

1997-01-01

Specific radiological requirements have to be considered for the realization of telemedicine. In this article the goals and requirements for an extensive introduction of teleradiology will be defined from the radiological user's point of view. Necessary medical, legal and professional prerequisites for teleradiology are presented. Essential requirements, such as data security maintenance of personal rights and standardization, must be realized. Application-specific requirements, e.g. quality and extent of teleradiological functions, as well as technological alternatives, are discussed. Each project must be carefully planned in relation to one's own needs, extent of functions and system selection. Topics, such as acknowledgement of electronic documentation, reimbursement of teleradiology and liability, must be clarified. Legal advice and the observance of quality guidelines are recommended. (orig.) [de

Advanced Extravehicular Activity Pressure Garment Requirements Development

Science.gov (United States)

Ross, Amy

2014-01-01

The NASA Johnson Space Center advanced pressure garment technology development team is addressing requirements development for exploration missions. Lessons learned from the Z-2 high fidelity prototype development have reiterated that clear low-level requirements and verification methods reduce risk to the government, improve efficiency in pressure garment design efforts, and enable the government to be a smart buyer. The expectation is to provide requirements at the specification level that are validated so that their impact on pressure garment design is understood. Additionally, the team will provide defined verification protocols for the requirements. However, in reviewing exploration space suit high level requirements there are several gaps in the team's ability to define and verify related lower level requirements. This paper addresses the efforts in requirement areas such as mobility/fit/comfort and environmental protection (dust, radiation, plasma, secondary impacts) to determine the by what method the requirements can be defined and use of those methods for verification. Gaps exist at various stages. In some cases component level work is underway, but no system level effort has begun, in other cases no effort has been initiated to close the gap. Status of ongoing efforts and potential approaches to open gaps are discussed.

A Design Theory for Requirements Mining Systems

OpenAIRE

Meth, Hendrik

2013-01-01

Software requirements are often communicated in unstructured text documents, which need to be analyzed in order to identify and classify individual needs. This process is referred to as requirements mining in the context of this thesis. It is known to be time-consuming and error-prone when performed manually by a requirements engineer. Thus, there is a demand to support requirements mining through information technology. However, little research has been conducted to conceptualize theoretical...

TANK FARM ENVIRONMENTAL REQUIREMENTS

International Nuclear Information System (INIS)

TIFFT, S.R.

2003-01-01

Through regulations, permitting or binding negotiations, Regulators establish requirements, limits, permit conditions and Notice of Construction (NOC) conditions with which the Office of River Protection (ORP) and the Tank Farm Contractor (TFC) must comply. Operating Specifications are technical limits which are set on a process to prevent injury to personnel, or damage to the facility or environment, The main purpose of this document is to provide specification limits and recovery actions for the TFC Environmental Surveillance Program at the Hanford Site. Specification limits are given for monitoring frequencies and permissible variation of readings from an established baseline or previous reading. The requirements in this document are driven by environmental considerations and data analysis issues, rather than facility design or personnel safety issues. This document is applicable to all single-shell tank (SST) and double-shell tank (DST) waste tanks, and the associated catch tanks and receiver tanks, and transfer systems. This Tank Farm Environmental Specifications Document (ESD) implements environmental-regulatory limits on the configuration and operation of the Hanford Tank Farms facility that have been established by Regulators. This ESD contains specific field operational limits and recovery actions for compliance with airborne effluent regulations and agreements, liquid effluents regulations and agreements, and environmental tank system requirements. The scope of this ESD is limited to conditions that have direct impact on Operations/Projects or that Operations Projects have direct impact upon. This document does not supercede or replace any Department of Energy (DOE) Orders, regulatory permits, notices of construction, or Regulatory agency agreements binding on the ORP or the TFC. Refer to the appropriate regulation, permit, or Notice of Construction for an inclusive listing of requirements

12 CFR 932.4 - Credit risk capital requirement.

Science.gov (United States)

2010-01-01

... 12 Banks and Banking 7 2010-01-01 2010-01-01 false Credit risk capital requirement. 932.4 Section... CAPITAL STANDARDS FEDERAL HOME LOAN BANK CAPITAL REQUIREMENTS § 932.4 Credit risk capital requirement. (a) General requirement. Each Bank's credit risk capital requirement shall be equal to the sum of the Bank's...

21 CFR 369.3 - Warnings required on drugs exempted from prescription-dispensing requirements of section 503(b)(1...

Science.gov (United States)

2010-04-01

... 21 Food and Drugs 5 2010-04-01 2010-04-01 false Warnings required on drugs exempted from prescription-dispensing requirements of section 503(b)(1)(C). 369.3 Section 369.3 Food and Drugs FOOD AND DRUG... subject to the labeling requirements prescribed in § 310.201(a) of this chapter. Although, for convenience...

TWRS configuration management requirement source document

International Nuclear Information System (INIS)

Vann, J.M.

1997-01-01

The TWRS Configuration Management (CM) Requirement Source document prescribes CM as a basic product life-cycle function by which work and activities are conducted or accomplished. This document serves as the requirements basis for the TWRS CM program. The objective of the TWRS CM program is to establish consistency among requirements, physical/functional configuration, information, and documentation for TWRS and TWRS products, and to maintain this consistency throughout the life-cycle of TWRS and the product, particularly as changes are being made

Safety of magnetic fusion facilities: Requirements

International Nuclear Information System (INIS)

1996-05-01

This Standard identifies safety requirements for magnetic fusion facilities. Safety functions are used to define outcomes that must be achieved to ensure that exposures to radiation, hazardous materials, or other hazards are maintained within acceptable limits. Requirements applicable to magnetic fusion facilities have been derived from Federal law, policy, and other documents. In addition to specific safety requirements, broad direction is given in the form of safety principles that are to be implemented and within which safety can be achieved

8 CFR 1216.2 - Notification requirements.

Science.gov (United States)

2010-01-01

... second time of the requirement that the alien and the petitioning spouse or alien entrepreneur must file... does not relieve the alien and the petitioning spouse, or alien entrepreneur of the requirement to file...

Authorization basis requirements comparison report

Energy Technology Data Exchange (ETDEWEB)

Brantley, W.M.

1997-08-18

The TWRS Authorization Basis (AB) consists of a set of documents identified by TWRS management with the concurrence of DOE-RL. Upon implementation of the TWRS Basis for Interim Operation (BIO) and Technical Safety Requirements (TSRs), the AB list will be revised to include the BIO and TSRs. Some documents that currently form part of the AB will be removed from the list. This SD identifies each - requirement from those documents, and recommends a disposition for each to ensure that necessary requirements are retained when the AB is revised to incorporate the BIO and TSRs. This SD also identifies documents that will remain part of the AB after the BIO and TSRs are implemented. This document does not change the AB, but provides guidance for the preparation of change documentation.

Authorization basis requirements comparison report

International Nuclear Information System (INIS)

Brantley, W.M.

1997-01-01

The TWRS Authorization Basis (AB) consists of a set of documents identified by TWRS management with the concurrence of DOE-RL. Upon implementation of the TWRS Basis for Interim Operation (BIO) and Technical Safety Requirements (TSRs), the AB list will be revised to include the BIO and TSRs. Some documents that currently form part of the AB will be removed from the list. This SD identifies each - requirement from those documents, and recommends a disposition for each to ensure that necessary requirements are retained when the AB is revised to incorporate the BIO and TSRs. This SD also identifies documents that will remain part of the AB after the BIO and TSRs are implemented. This document does not change the AB, but provides guidance for the preparation of change documentation

A Metamodeling Approach for Reasoning about Requirements

NARCIS (Netherlands)

Göknil, Arda; Ivanov, Ivan; van den Berg, Klaas; Schieferdecker, I.; Hartman, A.

In requirements engineering, there are several approaches for requirements modeling such as goal-oriented, aspect-driven, and system requirements modeling. In practice, companies often customize a given approach to their specific needs. Thus, we seek a solution that allows customization in a

Vehicle systems and payload requirements evaluation. [computer programs for identifying launch vehicle system requirements

Science.gov (United States)

Rea, F. G.; Pittenger, J. L.; Conlon, R. J.; Allen, J. D.

1975-01-01

Techniques developed for identifying launch vehicle system requirements for NASA automated space missions are discussed. Emphasis is placed on development of computer programs and investigation of astrionics for OSS missions and Scout. The Earth Orbit Mission Program - 1 which performs linear error analysis of launch vehicle dispersions for both vehicle and navigation system factors is described along with the Interactive Graphic Orbit Selection program which allows the user to select orbits which satisfy mission requirements and to evaluate the necessary injection accuracy.

46 CFR 403.300 - Financial reporting requirements.

Science.gov (United States)

2010-10-01

... 46 Shipping 8 2010-10-01 2010-10-01 false Financial reporting requirements. 403.300 Section 403... UNIFORM ACCOUNTING SYSTEM Reporting Requirements § 403.300 Financial reporting requirements. (a) General: (1) The financial statements shall list each active account, including subsidiary accounts. (2) The...

Critical Review of NOAA's Observation Requirements Process

Science.gov (United States)

LaJoie, M.; Yapur, M.; Vo, T.; Templeton, A.; Bludis, D.

2017-12-01

NOAA's Observing Systems Council (NOSC) maintains a comprehensive database of user observation requirements. The requirements collection process engages NOAA subject matter experts to document and effectively communicate the specific environmental observation measurements (parameters and attributes) needed to produce operational products and pursue research objectives. User observation requirements documented using a structured and standardized manner and framework enables NOAA to assess its needs across organizational lines in an impartial, objective, and transparent manner. This structure provides the foundation for: selecting, designing, developing, acquiring observing technologies, systems and architectures; budget and contract formulation and decision-making; and assessing in a repeatable fashion the productivity, efficiency and optimization of NOAA's observing system enterprise. User observation requirements are captured independently from observing technologies. Therefore, they can be addressed by a variety of current or expected observing capabilities and allow flexibility to be remapped to new and evolving technologies. NOAA's current inventory of user observation requirements were collected over a ten-year period, and there have been many changes in policies, mission priorities, and funding levels during this time. In light of these changes, the NOSC initiated a critical, in-depth review to examine all aspects of user observation requirements and associated processes during 2017. This presentation provides background on the NOAA requirements process, major milestones and outcomes of the critical review, and plans for evolving and connecting observing requirements processes in the next year.

46 CFR 111.95-3 - General requirements.

Science.gov (United States)

2010-10-01

... 46 Shipping 4 2010-10-01 2010-10-01 false General requirements. 111.95-3 Section 111.95-3 Shipping COAST GUARD, DEPARTMENT OF HOMELAND SECURITY (CONTINUED) ELECTRICAL ENGINEERING ELECTRIC SYSTEMS-GENERAL REQUIREMENTS Electric Power-Operated Boat Winches § 111.95-3 General requirements. (a) Each electrical...

21 CFR 900.11 - Requirements for certification.

Science.gov (United States)

2010-04-01

...) MAMMOGRAPHY QUALITY STANDARDS ACT MAMMOGRAPHY Quality Standards and Certification § 900.11 Requirements for... FDA, facilities are required to meet the quality standards in § 900.12 and to be accredited by an... requirements for reapplication for accreditation; (ii) Fully document its history as a previously provisionally...

48 CFR 9903.202-1 - General requirements.

Science.gov (United States)

2010-10-01

... ACCOUNTING STANDARDS CONTRACT COVERAGE CAS Program Requirements 9903.202-1 General requirements. (a) A... subcontract is of the type or value exempted by 9903.201-1 or (ii) In the most recently completed cost... of Germany. (3) United Kingdom. (f) Educational institutions—disclosure requirements. (1) Educational...

48 CFR 849.111-70 - Required review.

Science.gov (United States)

2010-10-01

... 48 Federal Acquisition Regulations System 5 2010-10-01 2010-10-01 false Required review. 849.111... MANAGEMENT TERMINATION OF CONTRACTS General Principles 849.111-70 Required review. (a) FAR 49.111 requires each agency to establish procedures, when necessary, for the administrative review of proposed...

37 CFR 1.143 - Reconsideration of requirement.

Science.gov (United States)

2010-07-01

... Inventions in One Application; Restriction § 1.143 Reconsideration of requirement. If the applicant disagrees... applicant must indicate a provisional election of one invention for prosecution, which invention shall be the one elected in the event the requirement becomes final The requirement for restriction will be...

7 CFR 1780.11 - Service area requirements.

Science.gov (United States)

2010-01-01

..., maintenance, debt service, and reserve requirements. Such guarantees from developers will meet the... 7 Agriculture 12 2010-01-01 2010-01-01 false Service area requirements. 1780.11 Section 1780.11... AGRICULTURE (CONTINUED) WATER AND WASTE LOANS AND GRANTS General Policies and Requirements § 1780.11 Service...

A review of tritium licensing requirements

International Nuclear Information System (INIS)

Meikle, A.B.

1982-12-01

Present Canadian regulations and anticipated changes to these regulations relevant to the utilization of tritium in fusion facilities and in commercial applications have been reviewed. It is concluded that there are no serious licensing obstacles, but there are a number of requirements which must be met. A license will be required from Atomic Energy Control Board if Ontario Hydro tritium is to be applied by other users. A license is required from the Federal Government to export or import tritium. A licensed container will be required for the storage and shipping of tritium. The containers being designed by AECL and Ontario Hydro and which are currently being tested will adequately store and ship all of the Ontario Hydro tritium but are unnecessarily large for the small quantities required by the commercial tritium users. Also, some users may prefer to receive tritium in gaseous form. An additional, smaller container should be considered. The licensing of overseas fusion facilities for the use of tritium is seen as a major undertaking offering opportunities to Canadian Fusion Fuels Technology Project to undertake health, safety and environmental analysis on behalf of these facilities

Household energy requirement and value patterns

International Nuclear Information System (INIS)

Vringer, Kees; Aalbers, Theo; Blok, Kornelis

2007-01-01

For an effective consumer energy policy, it is important to know why some households require more energy than others. The aim of the study described here was to examine whether there is a relationship between the total household energy requirement, on one hand, and value patterns, the motivation to save energy or the problem perception of climate change, on the other. To examine these relationships, we held a consumer survey among 2304 respondent households. We did not find significant differences in the energy requirement of groups of households with different value patterns, taking into account the differences in the socio-economic situation of households. Only for the 'motivation to save energy' we did find that the least motivated group requires 10 GJ more energy than the average and most motivated groups; this is about 4% of the total household energy requirement. This means that a self-regulating energy policy, solely based on the fact that a strategy of internalising environmental responsibility will not be effective in saving energy. There are indications that a social dilemma is one of the reasons why people's consumption patterns do not conform to their value patterns, problem perception or motivation to save energy

Security Requirements Management in Software Product Line Engineering

Science.gov (United States)

Mellado, Daniel; Fernández-Medina, Eduardo; Piattini, Mario

Security requirements engineering is both a central task and a critical success factor in product line development due to the complexity and extensive nature of product lines. However, most of the current product line practices in requirements engineering do not adequately address security requirements engineering. Therefore, in this chapter we will propose a security requirements engineering process (SREPPLine) driven by security standards and based on a security requirements decision model along with a security variability model to manage the variability of the artefacts related to security requirements. The aim of this approach is to deal with security requirements from the early stages of the product line development in a systematic way, in order to facilitate conformance with the most relevant security standards with regard to the management of security requirements, such as ISO/IEC 27001 and ISO/IEC 15408.

7 CFR 981.51 - Requirements for reserve.

Science.gov (United States)

2010-01-01

... 7 Agriculture 8 2010-01-01 2010-01-01 false Requirements for reserve. 981.51 Section 981.51... Regulating Handling Volume Regulation § 981.51 Requirements for reserve. Each handler may satisfy his reserve... include grade requirements for reserve almonds delivered to human consumption outlets. [41 FR 26853, June...

18 CFR 153.21 - Conformity with requirements.

Science.gov (United States)

2010-04-01

... 18 Conservation of Power and Water Resources 1 2010-04-01 2010-04-01 false Conformity with requirements. 153.21 Section 153.21 Conservation of Power and Water Resources FEDERAL ENERGY REGULATORY... Requirements § 153.21 Conformity with requirements. (a) General Rule. Applications under subparts B and C of...

Fusion Energy Sciences Network Requirements

Energy Technology Data Exchange (ETDEWEB)

Dart, Eli [ESNet, Berkeley, CA (United States); Tierney, Brian [ESNet, Berkeley, CA (United States)

2012-09-26

The Energy Sciences Network (ESnet) is the primary provider of network connectivity for the U.S. Department of Energy Office of Science, the single largest supporter of basic research in the physical sciences in the United States. In support of the Office of Science programs, ESnet regularly updates and refreshes its understanding of the networking requirements of the instruments, facilities, scientists, and science programs that it serves. This focus has helped ESnet to be a highly successful enabler of scientific discovery for over 25 years. In December 2011, ESnet and the Office of Fusion Energy Sciences (FES), of the DOE Office of Science (SC), organized a workshop to characterize the networking requirements of the programs funded by FES. The requirements identified at the workshop are summarized in the Findings section, and are described in more detail in the body of the report.

Quality Assurance Source Requirements Traceability Database

International Nuclear Information System (INIS)

MURTHY, R.; NAYDENOVA, A.; DEKLEVER, R.; BOONE, A.

2006-01-01

At the Yucca Mountain Project the Project Requirements Processing System assists in the management of relationships between regulatory and national/industry standards source criteria, and Quality Assurance Requirements and Description document (DOE/R W-0333P) requirements to create compliance matrices representing respective relationships. The matrices are submitted to the U.S. Nuclear Regulatory Commission to assist in the commission's review, interpretation, and concurrence with the Yucca Mountain Project QA program document. The tool is highly customized to meet the needs of the Office of Civilian Radioactive Waste Management Office of Quality Assurance

Smart gun technology requirements preliminary report

Energy Technology Data Exchange (ETDEWEB)

Weiss, D.R.; Brandt, D.J.; Tweet, K.D.

1995-05-01

Goal of the Smart Gun Technology project is to eliminate the capability of an unauthorized user from firing a law enforcement officer`s firearm by implementing user-recognizing-and-authorizing surety technologies. This project is funded by the National Institute of Justice. This document reports the projects first objective: to find and document the requirements for a user-recognizing-and-authorizing firearm technology that law enforcement officers will value. This report details the problem of firearm takeaways in law enforcement, the methodology used to develop the law enforcement officers` requirements, and the requirements themselves.

Time-varying Capital Requirements and Disclosure Rules

DEFF Research Database (Denmark)

Kragh, Jonas; Rangvid, Jesper

, implying that resilience in the banking system is also increased. The increase in capital ratios is partly due to a modest reduction in lending. Using a policy changes, we show that banks react stronger to changes in capital requirements when these are public. Our results further suggest that the impact......Unique and confidential Danish data allow us to identify how changes in disclosure requirements and bank-specific time-varying capital requirements affect banks' lending and capital accumu-lation decisions. We find that banks increase their capital ratios after capital requirements are increased...... of capital requirements differ for small and large banks. Large banks raise their capital ratios more, reduce lending less, and accumulate more new capital compared to small banks....

Preliminary waste acceptance requirements - Konrad repository project

International Nuclear Information System (INIS)

Brennecke, P.W.; Warnecke, E.H.

1991-01-01

In Germany, the planned Konrad repository is proposed for the disposal of all types of radioactive wastes whose thermal influence upon the host rock is negligible. The Bundesamt fuer Strahlenschutz has established Preliminary Waste Acceptance Requirements (as of April 1990) for this facility. The respective requirements were developed on the basis of the results of site-specific safety assessments. They include general requirements on the waste packages to be disposed of as well as more specific requirements on the waste forms, the packaging and the radionuclide inventory per waste package. In addition, the delivery of waste packages was regulated. An outline of the structure and the elements of the Preliminary Waste Acceptance Requirements of April 1990 is given including comments on their legal status. (Author)

49 CFR 229.206 - Design requirements.

Science.gov (United States)

2010-10-01

...-climber, emergency egress, emergency interior lighting, and interior configuration design requirements set... 49 Transportation 4 2010-10-01 2010-10-01 false Design requirements. 229.206 Section 229.206..., DEPARTMENT OF TRANSPORTATION RAILROAD LOCOMOTIVE SAFETY STANDARDS Locomotive Crashworthiness Design...

Tracing And Control Of Engineering Requirements

Science.gov (United States)

Turner, Philip R.; Stoller, Richard L.; Neville, Ted; Boyle, Karen A.

1991-01-01

TRACER (Tracing and Control of Engineering Requirements) is data-base/word-processing software system created to document and maintain order of both requirements and descriptions associated with engineering project. Implemented on IBM PC under PC-DOS. Written with CLIPPER.

Functional Requirements for an Electronic Work Package System

Energy Technology Data Exchange (ETDEWEB)

Oxstrand, Johanna H. [Idaho National Lab. (INL), Idaho Falls, ID (United States)

2016-12-01

This document provides a set of high level functional requirements for a generic electronic work package (eWP) system. The requirements have been identified by the U.S. nuclear industry as a part of the Nuclear Electronic Work Packages - Enterprise Requirements (NEWPER) initiative. The functional requirements are mainly applied to eWP system supporting Basic and Moderate types of smart documents, i.e., documents that have fields for recording input such as text, dates, numbers, and equipment status, and documents which incorporate additional functionalities such as form field data “type“ validation (e.g. date, text, number, and signature) of data entered and/or self-populate basic document information (usually from existing host application meta data) on the form when the user first opens it. All the requirements are categorized by the roles; Planner, Supervisor, Craft, Work Package Approval Reviewer, Operations, Scheduling/Work Control, and Supporting Functions. The categories Statistics, Records, Information Technology are also included used to group the requirements. All requirements are presented in Section 2 through Section 11. Examples of more detailed requirements are provided for the majority of high level requirements. These examples are meant as an inspiration to be used as each utility goes through the process of identifying their specific requirements. The report’s table of contents provides a summary of the high level requirements.

LHCb Online Networking Requirements

CERN Document Server

Jost, B

2003-01-01

This document describes the networking requirements of the LHCb online installation. It lists both quantitative aspects such as the number of required switch ports, as well as some qualitative features of the equipment, such as minimum buffer sizes in switches. The document comprises both the data acquisition network and the controls/general-purpose network. While the numbers represent our best current knowledge and are intended to give (in particular) network equipment manufacturers an overview of our needs, this document should not be confused with a market survey questionnaire or a formal tendering document. However the information contained in this document will be the input of any such document. A preliminary schedule for procurement and installation is also given.

School Library Media Certification Requirements: 1990 Update.

Science.gov (United States)

Perritt, Patsy H.

1990-01-01

Presents a compilation of school library media certification requirements taken from responses to a national survey. For each state, existing certificates or endorsements are listed, along with credit hours and/or experience required, whether it is an accredited or approved program, and required subject areas or competencies. A directory of…

24 CFR 232.610 - Certification of cost requirements.

Science.gov (United States)

2010-04-01

... 24 Housing and Urban Development 2 2010-04-01 2010-04-01 false Certification of cost requirements... ASSISTED LIVING FACILITIES Eligibility Requirements-Supplemental Loans To Finance Purchase and Installation of Fire Safety Equipment Cost Certification Requirements § 232.610 Certification of cost requirements...

Knowledge-based requirements analysis for automating software development

Science.gov (United States)

Markosian, Lawrence Z.

1988-01-01

We present a new software development paradigm that automates the derivation of implementations from requirements. In this paradigm, informally-stated requirements are expressed in a domain-specific requirements specification language. This language is machine-understable and requirements expressed in it are captured in a knowledge base. Once the requirements are captured, more detailed specifications and eventually implementations are derived by the system using transformational synthesis. A key characteristic of the process is that the required human intervention is in the form of providing problem- and domain-specific engineering knowledge, not in writing detailed implementations. We describe a prototype system that applies the paradigm in the realm of communication engineering: the prototype automatically generates implementations of buffers following analysis of the requirements on each buffer.

Responsibilities in the Usability Requirements Elicitation Process

Directory of Open Access Journals (Sweden)

Marianella Aveledo

2008-12-01

Full Text Available Like any other software system quality attribute, usability places requirements on software components. In particular, it has been demonstrated that certain usability features have a direct impact throughout the software process. This paper details an approach that looks at how to deal with certain usability features in the early software development stages. In particular, we consider usability features as functional usability requirements using patterns that have been termed usability patterns to elicit requirements. Additionally, we clearly establish the responsibilities of all the players at the usability requirements elicitation stage.

A Requirements Analysis Model Based on QFD

Institute of Scientific and Technical Information of China (English)

TANG Zhi-wei; Nelson K.H.Tang

2004-01-01

The enterprise resource planning (ERP) system has emerged to offer an integrated IT solution and more and more enterprises are increasing by adopting this system and regarding it as an important innovation. However, there is already evidence of high failure risks in ERP project implementation, one major reason is poor analysis of the requirements for system implementation. In this paper, the importance of requirements analysis for ERP project implementation is highlighted, and a requirements analysis model by applying quality function deployment (QFD) is presented, which will support to conduct requirements analysis for ERP project.

12 CFR 204.4 - Computation of required reserves.

Science.gov (United States)

2010-01-01

... RESERVE REQUIREMENTS OF DEPOSITORY INSTITUTIONS (REGULATION D) § 204.4 Computation of required reserves. (a) In determining the reserve requirement under this part, the amount of cash items in process of... reserves are computed by applying the reserve requirement ratios below to net transaction accounts...

The Sources and Methods of Engineering Design Requirement

DEFF Research Database (Denmark)

Li, Xuemeng; Zhang, Zhinan; Ahmed-Kristensen, Saeema

2014-01-01

to be defined in a new context. This paper focuses on understanding the design requirement sources at the requirement elicitation phase. It aims at proposing an improved design requirement source classification considering emerging markets and presenting current methods for eliciting requirement for each source...

Utility requirements for advanced LWR passive plants

International Nuclear Information System (INIS)

Yedidia, J.M.; Sugnet, W.R.

1992-01-01

LWR Passive Plants are becoming an increasingly attractive and prominent option for future electric generating capacity for U.S. utilities. Conceptual designs for ALWR Passive Plants are currently being developed by U.S. suppliers. EPRI-sponsored work beginning in 1985 developed preliminary conceptual designs for a passive BWR and PWR. DOE-sponsored work from 1986 to the present in conjunction with further EPRI-sponsored studies has continued this development to the point of mature conceptual designs. The success to date in developing the ALWR Passive Plant concepts has substantially increased utility interest. The EPRI ALWR Program has responded by augmenting its initial scope to develop a Utility Requirements Document for ALWR Passive Plants. These requirements will be largely based on the ALWR Utility Requirements Document for Evolutionary Plants, but with significant changes in areas related to the passive safety functions and system configurations. This work was begun in late 1988, and the thirteen-chapter Passive Plant Utility Requirements Document will be completed in 1990. This paper discusses the progress to date in developing the Passive Plant requirements, reviews the top-level requirements, and discusses key issues related to adaptation of the utility requirements to passive safety functions and system configurations. (orig.)

21 CFR 1302.03 - Symbol required; exceptions.

Science.gov (United States)

2010-04-01

... 21 Food and Drugs 9 2010-04-01 2010-04-01 false Symbol required; exceptions. 1302.03 Section 1302... REQUIREMENTS FOR CONTROLLED SUBSTANCES § 1302.03 Symbol required; exceptions. (a) Each commercial container of... § 1308.31 of this chapter) shall have printed on the label the symbol designating the schedule in which...

12 CFR 932.2 - Total capital requirement.

Science.gov (United States)

2010-01-01

... 12 Banks and Banking 7 2010-01-01 2010-01-01 false Total capital requirement. 932.2 Section 932.2 Banks and Banking FEDERAL HOUSING FINANCE BOARD FEDERAL HOME LOAN BANK RISK MANAGEMENT AND CAPITAL STANDARDS FEDERAL HOME LOAN BANK CAPITAL REQUIREMENTS § 932.2 Total capital requirement. (a) Each Bank shall...

29 CFR 18.1002 - Requirement of original.

Science.gov (United States)

2010-07-01

... 29 Labor 1 2010-07-01 2010-07-01 true Requirement of original. 18.1002 Section 18.1002 Labor... § 18.1002 Requirement of original. To prove the content of a writing, recording, or photograph, the original writing, recording, or photograph is required, except as otherwise provided in these rules, or by...

Integrating semi-formal and formal requirements

NARCIS (Netherlands)

Wieringa, Roelf J.; Olivé, Antoni; Dubois, Eric; Pastor, Joan Antoni; Huyts, Sander

1997-01-01

In this paper, we report on the integration of informal, semiformal and formal requirements specification techniques. We present a framework for requirements specification called TRADE, within which several well-known semiformal specification techniques are placed. TRADE is based on an analysis of

40 CFR 51.165 - Permit requirements.

Science.gov (United States)

2010-07-01

...; and (2) Shall include emissions associated with startups, shutdowns, and malfunctions; and, for an... data acquisition and availability requirements of this section, to sample, condition (if applicable... the equipment necessary to meet the data acquisition and availability requirements of this section, to...

40 CFR 233.31 - Coordination requirements.

Science.gov (United States)

2010-07-01

... 40 Protection of Environment 24 2010-07-01 2010-07-01 false Coordination requirements. 233.31 Section 233.31 Protection of Environment ENVIRONMENTAL PROTECTION AGENCY (CONTINUED) OCEAN DUMPING 404 STATE PROGRAM REGULATIONS Program Operation § 233.31 Coordination requirements. (a) If a proposed...

40 CFR 35.532 - Requirements summary.

Science.gov (United States)

2010-07-01

... 40 Protection of Environment 1 2010-07-01 2010-07-01 false Requirements summary. 35.532 Section 35.532 Protection of Environment ENVIRONMENTAL PROTECTION AGENCY GRANTS AND OTHER FEDERAL ASSISTANCE....532 Requirements summary. (a) Applicants and recipients of Performance Partnership Grants must meet...

40 CFR 35.132 - Requirements summary.

Science.gov (United States)

2010-07-01

... 40 Protection of Environment 1 2010-07-01 2010-07-01 false Requirements summary. 35.132 Section 35.132 Protection of Environment ENVIRONMENTAL PROTECTION AGENCY GRANTS AND OTHER FEDERAL ASSISTANCE... Requirements summary. Applicants and recipients of Performance Partnership Grants must meet: (a) The...

40 CFR 63.2252 - What are the requirements for process units that have no control or work practice requirements?

Science.gov (United States)

2010-07-01

... 40 Protection of Environment 12 2010-07-01 2010-07-01 true What are the requirements for process units that have no control or work practice requirements? 63.2252 Section 63.2252 Protection of... Pollutants: Plywood and Composite Wood Products General Compliance Requirements § 63.2252 What are the...

Energy Requirements in Critically Ill Patients

Science.gov (United States)

2018-01-01

During the management of critical illness, optimal nutritional support is an important key for achieving positive clinical outcomes. Compared to healthy people, critically ill patients have higher energy expenditure, thereby their energy requirements and risk of malnutrition being increased. Assessing individual nutritional requirement is essential for a successful nutritional support, including the adequate energy supply. Methods to assess energy requirements include indirect calorimetry (IC) which is considered as a reference method, and the predictive equations which are commonly used due to the difficulty of using IC in certain conditions. In this study, a literature review was conducted on the energy metabolic changes in critically ill patients, and the implications for the estimation of energy requirements in this population. In addition, the issue of optimal caloric goal during nutrition support is discussed, as well as the accuracy of selected resting energy expenditure predictive equations, commonly used in critically ill patients.

Energy Requirements in Critically Ill Patients.

Science.gov (United States)

Ndahimana, Didace; Kim, Eun-Kyung

2018-04-01

During the management of critical illness, optimal nutritional support is an important key for achieving positive clinical outcomes. Compared to healthy people, critically ill patients have higher energy expenditure, thereby their energy requirements and risk of malnutrition being increased. Assessing individual nutritional requirement is essential for a successful nutritional support, including the adequate energy supply. Methods to assess energy requirements include indirect calorimetry (IC) which is considered as a reference method, and the predictive equations which are commonly used due to the difficulty of using IC in certain conditions. In this study, a literature review was conducted on the energy metabolic changes in critically ill patients, and the implications for the estimation of energy requirements in this population. In addition, the issue of optimal caloric goal during nutrition support is discussed, as well as the accuracy of selected resting energy expenditure predictive equations, commonly used in critically ill patients.

Projected uranium requirements of developing countries

International Nuclear Information System (INIS)

Anon.

1990-01-01

The objective of this paper is to examine the uranium requirements of developing countries both in aggregate and individually. Although the cumulative uranium requirements of these countries are expected to account for less than eight percent of total requirements, the fact that many of these countries are expressing renewed interest in nuclear is, in itself, encouraging. The countries analyzed in this paper are Argentina, Brazil, Egypt, India, Israel, Mexico, Pakistan, South Africa, South Korea and Taiwan. For each country, the existing and planned nuclear capacity levels have been identified and capacity factors have been projected. For countries with no previous nuclear power, the world weighted average capacity factor for the specific reactor type is utilized. Other factors influencing nuclear power demand and operations of these developing countries will be discussed, and finally, uranium requirements based on a calculated optimal tails assay of .30 will be provided

Improving the requirements process in Axiomatic Design Theory

DEFF Research Database (Denmark)

Thompson, Mary Kathryn

2013-01-01

This paper introduces a model to integrate the traditional requirements process into Axiomatic Design Theory and proposes a method to structure the requirements process. The method includes a requirements classification system to ensure that all requirements information can be included...... in the Axiomatic Design process, a stakeholder classification system to reduce the chances of excluding one or more key stakeholders, and a table to visualize the mapping between the stakeholders and their requirements....

47 CFR 76.925 - Costs of franchise requirements.

Science.gov (United States)

2010-10-01

... 47 Telecommunication 4 2010-10-01 2010-10-01 false Costs of franchise requirements. 76.925 Section... MULTICHANNEL VIDEO AND CABLE TELEVISION SERVICE Cable Rate Regulation § 76.925 Costs of franchise requirements. (a) Franchise requirement costs may include cost increases required by the franchising authority in...

49 CFR 236.1033 - Communications and security requirements.

Science.gov (United States)

2010-10-01

... Train Control Systems § 236.1033 Communications and security requirements. (a) All wireless... 49 Transportation 4 2010-10-01 2010-10-01 false Communications and security requirements. 236.1033... exceeding the security strength required to protect the data as defined in the railroad's PTCSP and required...

24 CFR 700.175 - Other Federal requirements.

Science.gov (United States)

2010-04-01

... requirements described in 24 CFR part 44 (OMB Circular A-128). (b) Conflict of interest. In addition to the conflict of interest requirements in OMB Circular A-87 and 24 CFR part 85, no person who is an employee..., and the implementing regulations at 24 CFR part 108; and (3) Racial and ethnic collection requirements...

Nondestructive examination requirements for PWR vessel internals

International Nuclear Information System (INIS)

Spanner, J.

2015-01-01

This paper describes the requirements for the nondestructive examination of pressurized water reactor (PWR) vessel internals in accordance with the requirements of the EPRI Material Reliability Program (MRP) inspection standard for PWR internals (MRP-228) and the American Society of Mechanical Engineers Section XI In-service Inspection. The MRP vessel internals examinations have been performed at nuclear plants in the USA since 2009. The objective of the inspection standard is to provide the requirements for the nondestructive examination (NDE) methods implemented to support the inspection and evaluation of the internals. The inspection standard contains requirements specific to the inspection methodologies involved as well as requirements for qualification of the NDE procedures, equipment and personnel used to perform the vessel internals inspections. The qualification requirements for the NDE systems will be summarized. Six PWR plants in the USA have completed inspections of their internals using the Inspection and Evaluation Guideline (MRP-227) and the Inspection Standard (MRP-228). Examination results show few instances of service-induced degradation flaws, as expected. The few instances of degradation have mostly occurred in bolting

WFIRST: Update on the Coronagraph Science Requirements

Science.gov (United States)

Douglas, Ewan S.; Cahoy, Kerri; Carlton, Ashley; Macintosh, Bruce; Turnbull, Margaret; Kasdin, Jeremy; WFIRST Coronagraph Science Investigation Teams

2018-01-01

The WFIRST Coronagraph instrument (CGI) will enable direct imaging and low resolution spectroscopy of exoplanets in reflected light and imaging polarimetry of circumstellar disks. The CGI science investigation teams were tasked with developing a set of science requirements which advance our knowledge of exoplanet occurrence and atmospheric composition, as well as the composition and morphology of exozodiacal debris disks, cold Kuiper Belt analogs, and protoplanetary systems. We present the initial content, rationales, validation, and verification plans for the WFIRST CGI, informed by detailed and still-evolving instrument and observatory performance models. We also discuss our approach to the requirements development and management process, including the collection and organization of science inputs, open source approach to managing the requirements database, and the range of models used for requirements validation. These tools can be applied to requirements development processes for other astrophysical space missions, and may ease their management and maintenance. These WFIRST CGI science requirements allow the community to learn about and provide insights and feedback on the expected instrument performance and science return.

40 CFR 61.186 - Reporting requirements.

Science.gov (United States)

2010-07-01

...) NATIONAL EMISSION STANDARDS FOR HAZARDOUS AIR POLLUTANTS National Emission Standard for Inorganic Arsenic Emissions From Arsenic Trioxide and Metallic Arsenic Production Facilities § 61.186 Reporting requirements... at least 30 days prior notice of each reference opacity level determination required in § 61.183(a...

7 CFR 926.17 - Reporting requirements.

Science.gov (United States)

2010-01-01

... Federal cranberry marketing order (7 CFR Part 926) shall be required to submit four times annually, for... Agriculture Regulations of the Department of Agriculture (Continued) AGRICULTURAL MARKETING SERVICE (Marketing... RECORDKEEPING REQUIREMENTS APPLICABLE TO CRANBERRIES NOT SUBJECT TO THE CRANBERRY MARKETING ORDER § 926.17...

13 CFR 120.860 - Required objectives.

Science.gov (United States)

2010-01-01

... 13 Business Credit and Assistance 1 2010-01-01 2010-01-01 false Required objectives. 120.860 Section 120.860 Business Credit and Assistance SMALL BUSINESS ADMINISTRATION BUSINESS LOANS Development Company Loan Program (504) Project Economic Development Goals § 120.860 Required objectives. A Project...

40 CFR 141.90 - Reporting requirements.

Science.gov (United States)

2010-07-01

... facilities and organizations to which the system delivered public education materials during the period in....82(a). (3) For systems required to evaluate the effectiveness of corrosion control treatments under... activities have taken place. (f) Public education program reporting requirements. (1) Any water system that...

44 CFR 19.115 - Assurance required.

Science.gov (United States)

2010-10-01

... RECEIVING FEDERAL FINANCIAL ASSISTANCE Introduction § 19.115 Assurance required. (a) General. Either at the... 44 Emergency Management and Assistance 1 2010-10-01 2010-10-01 false Assurance required. 19.115 Section 19.115 Emergency Management and Assistance FEDERAL EMERGENCY MANAGEMENT AGENCY, DEPARTMENT OF...

Requirements engineering: foundation for software quality

NARCIS (Netherlands)

Daneva, Maia; Pastor, Oscar

2016-01-01

Welcome to the proceedings of the 22nd edition of REFSQ: the International Working Conference on Requirements Engineering – Foundation for Software Quality! Requirements engineering (RE) has been recognized as a critical factor that impacts the quality of software, systems, and services. Since the

7 CFR 1770.13 - Accounting requirements.

Science.gov (United States)

2010-01-01

... accrual basis of accounting. All transactions shall be recorded in the period in which they occur and... 7 Agriculture 12 2010-01-01 2010-01-01 false Accounting requirements. 1770.13 Section 1770.13... AGRICULTURE (CONTINUED) ACCOUNTING REQUIREMENTS FOR RUS TELECOMMUNICATIONS BORROWERS Uniform System of...

29 CFR 4010.3 - Filing requirement.

Science.gov (United States)

2010-07-01

... Relating to Labor (Continued) PENSION BENEFIT GUARANTY CORPORATION CERTAIN REPORTING AND DISCLOSURE REQUIREMENTS ANNUAL FINANCIAL AND ACTUARIAL INFORMATION REPORTING § 4010.3 Filing requirement. (a) General....10, all information specified in § 4010.6(a) with respect to all members of a controlled group and...

17 CFR 41.2 - Required records.

Science.gov (United States)

2010-04-01

... 17 Commodity and Securities Exchanges 1 2010-04-01 2010-04-01 false Required records. 41.2 Section 41.2 Commodity and Securities Exchanges COMMODITY FUTURES TRADING COMMISSION SECURITY FUTURES PRODUCTS General Provisions § 41.2 Required records. A designated contract market or registered derivatives...

15 CFR 971.418 - Diligence requirements.

Science.gov (United States)

2010-01-01

... 15 Commerce and Foreign Trade 3 2010-01-01 2010-01-01 false Diligence requirements. 971.418 Section 971.418 Commerce and Foreign Trade Regulations Relating to Commerce and Foreign Trade (Continued...: Terms, Conditions and Restrictions Terms, Conditions and Restrictions § 971.418 Diligence requirements...

49 CFR 601.45 - Required information.

Science.gov (United States)

2010-10-01

... 49 Transportation 7 2010-10-01 2010-10-01 false Required information. 601.45 Section 601.45 Transportation Other Regulations Relating to Transportation (Continued) FEDERAL TRANSIT ADMINISTRATION, DEPARTMENT OF TRANSPORTATION ORGANIZATION, FUNCTIONS, AND PROCEDURES Emergency Procedures for Public Transportation Systems § 601.45 Required...

40 CFR 191.13 - Containment requirements.

Science.gov (United States)

2010-07-01

... requirements. (a) Disposal systems for spent nuclear fuel or high-level or transuranic radioactive wastes shall... 40 Protection of Environment 24 2010-07-01 2010-07-01 false Containment requirements. 191.13 Section 191.13 Protection of Environment ENVIRONMENTAL PROTECTION AGENCY (CONTINUED) RADIATION PROTECTION...

42 CFR 3.106 - Security requirements.

Science.gov (United States)

2010-10-01

... ORGANIZATIONS AND PATIENT SAFETY WORK PRODUCT PSO Requirements and Agency Procedures § 3.106 Security requirements. (a) Application. A PSO must secure patient safety work product in conformance with the security... the confidentiality and security of patient safety work product. (2) Distinguishing patient safety...

40 CFR 267.147 - Liability requirements.

Science.gov (United States)

2010-07-01

... consideration of the guarantee. If the guarantor is a firm with a “substantial business relationship” with the... PERMIT Financial Requirements § 267.147 Liability requirements. (a) Coverage for sudden accidental... facilities, must demonstrate financial responsibility for bodily injury and property damage to third parties...

21 CFR 211.180 - General requirements.

Science.gov (United States)

2010-04-01

...: GENERAL CURRENT GOOD MANUFACTURING PRACTICE FOR FINISHED PHARMACEUTICALS Records and Reports § 211.180 General requirements. (a) Any production, control, or distribution record that is required to be... good manufacturing practices brought by the Food and Drug Administration. [43 FR 45077, Sept. 29, 1978...

36 CFR 801.7 - Information requirements.

Science.gov (United States)

2010-07-01

... 36 Parks, Forests, and Public Property 3 2010-07-01 2010-07-01 false Information requirements. 801.7 Section 801.7 Parks, Forests, and Public Property ADVISORY COUNCIL ON HISTORIC PRESERVATION HISTORIC PRESERVATION REQUIREMENTS OF THE URBAN DEVELOPMENT ACTION GRANT PROGRAM § 801.7 Information...

12 CFR 567.2 - Minimum regulatory capital requirement.

Science.gov (United States)

2010-01-01

... 12 Banks and Banking 5 2010-01-01 2010-01-01 false Minimum regulatory capital requirement. 567.2... Regulatory Capital Requirements § 567.2 Minimum regulatory capital requirement. (a) To meet its regulatory capital requirement a savings association must satisfy each of the following capital standards: (1) Risk...

EUROPEAN REQUIREMENTS REGARDING THE AUDIT REPORTING

Directory of Open Access Journals (Sweden)

Daniel Botez

2017-07-01

Full Text Available In recent years, reporting requirements on the statutory audit have been revised and amended to increase the audit report’s communication value. In addition to the stipulations of the International Auditing Standards Package on reporting, revised and enforceable on 15 December 2016, the European Union issued the Directive 2014/56 / EU amending Directive 2006/43 / EC on statutory audits of annual financial statements and consolidated financial statements and EU Regulation no. 537/2014 on specific requirements for the statutory audit of public interest entities, both of which were published on the same date, June 17, 2014 and with the same application deadline, June 17, 2016. These normative acts foresee increased requirements for the reporting procedure in the statutory audit. Thus, the Directive provides for additional content requirements for the audit report, and the regulation requires additional information in the report but also the issuance and provision of other reports by the auditor: report to the audit committee of the public interest entity; In some cases, a report to the public-interest entity's supervisory authority or to the auditor's supervisory authority, and a transparency report published annually on the auditor's website. Our article details this information with direct reference to the content of these European official documents.

An effective technique for the software requirements analysis of NPP safety-critical systems, based on software inspection, requirements traceability, and formal specification

International Nuclear Information System (INIS)

Koo, Seo Ryong; Seong, Poong Hyun; Yoo, Junbeom; Cha, Sung Deok; Yoo, Yeong Jae

2005-01-01

A thorough requirements analysis is indispensable for developing and implementing safety-critical software systems such as nuclear power plant (NPP) software systems because a single error in the requirements can generate serious software faults. However, it is very difficult to completely analyze system requirements. In this paper, an effective technique for the software requirements analysis is suggested. For requirements verification and validation (V and V) tasks, our technique uses software inspection, requirement traceability, and formal specification with structural decomposition. Software inspection and requirements traceability analysis are widely considered the most effective software V and V methods. Although formal methods are also considered an effective V and V activity, they are difficult to use properly in the nuclear fields as well as in other fields because of their mathematical nature. In this work, we propose an integrated environment (IE) approach for requirements, which is an integrated approach that enables easy inspection by combining requirement traceability and effective use of a formal method. The paper also introduces computer-aided tools for supporting IE approach for requirements. Called the nuclear software inspection support and requirements traceability (NuSISRT), the tool incorporates software inspection, requirement traceability, and formal specification capabilities. We designed the NuSISRT to partially automate software inspection and analysis of requirement traceability. In addition, for the formal specification and analysis, we used the formal requirements specification and analysis tool for nuclear engineering (NuSRS)

A closed-loop based framework for design requirement management

DEFF Research Database (Denmark)

Zhang, Zhinan; Li, Xuemeng; Liu, Zelin

2014-01-01

management from product lifecycle, and requirement and requirement management lifecycle views. This paper highlights the importance of requirement lifecycle management and aims at closing the requirement information loop in product lifecycle. Then, it addresses the requirement management in engineering...... design field with focusing on the dynamics nature and incomplete nature of requirements. Finally, a closed-loop based framework is proposed for requirement management in engineering design....

30 CFR 57.6132 - Magazine requirements.

Science.gov (United States)

2010-07-01

... 30 Mineral Resources 1 2010-07-01 2010-07-01 false Magazine requirements. 57.6132 Section 57.6132...-Surface Only § 57.6132 Magazine requirements. (a) Magazines shall be— (1) Structurally sound; (2... magazine; (6) Posted with the appropriate United States Department of Transportation placards or other...

30 CFR 56.6132 - Magazine requirements.

Science.gov (United States)

2010-07-01

... 30 Mineral Resources 1 2010-07-01 2010-07-01 false Magazine requirements. 56.6132 Section 56.6132....6132 Magazine requirements. (a) Magazines shall be— (1) Structurally sound; (2) Noncombustible or the... the inside; (5) Ventilated to control dampness and excessive heating within the magazine; (6) Posted...

32 CFR 298.5 - Information requirements.

Science.gov (United States)

2010-07-01

... 32 National Defense 2 2010-07-01 2010-07-01 false Information requirements. 298.5 Section 298.5... OF INFORMATION ACT PROGRAM DEFENSE INVESTIGATIVE SERVICE (DIS) FREEDOM OF INFORMATION ACT PROGRAM § 298.5 Information requirements. The DIS Office of Information and Public Affairs is responsible for...

An Online Graduate Requirements Engineering Course

Science.gov (United States)

Kilicay-Ergin, N.; Laplante, P. A.

2013-01-01

Requirements engineering is one of the fundamental knowledge areas in software and systems engineering graduate curricula. Recent changes in educational delivery and student demographics have created new challenges for requirements engineering education. In particular, there is an increasing demand for online education for working professionals.…

15 CFR 970.517 - Diligence requirements.

Science.gov (United States)

2010-01-01

... 15 Commerce and Foreign Trade 3 2010-01-01 2010-01-01 false Diligence requirements. 970.517 Section 970.517 Commerce and Foreign Trade Regulations Relating to Commerce and Foreign Trade (Continued..., Conditions and Restrictions Terms, Conditions, and Restrictions § 970.517 Diligence requirements. The terms...

5 CFR 330.208 - Qualification requirements.

Science.gov (United States)

2010-01-01

... perform the duties and responsibilities of the position. (c) The sex of an individual may not be... qualified for a position if he or she: (1) Meets OPM-established or approved qualification standards and requirements for the position, including any minimum educational requirements, and any selection placement...

7 CFR 97.8 - Specimen requirements.

Science.gov (United States)

2010-01-01

... 7 Agriculture 3 2010-01-01 2010-01-01 false Specimen requirements. 97.8 Section 97.8 Agriculture Regulations of the Department of Agriculture (Continued) AGRICULTURAL MARKETING SERVICE (Standards... required by the examiner to furnish representative specimens of the variety, or its flower, fruit, or seeds...

12 CFR 370.9 - Recordkeeping requirements.

Science.gov (United States)

2010-01-01

... 12 Banks and Banking 4 2010-01-01 2010-01-01 false Recordkeeping requirements. 370.9 Section 370.9 Banks and Banking FEDERAL DEPOSIT INSURANCE CORPORATION REGULATIONS AND STATEMENTS OF GENERAL POLICY TEMPORARY LIQUIDITY GUARANTEE PROGRAM § 370.9 Recordkeeping requirements. The FDIC will establish procedures...

1 CFR 10.10 - Publication required.

Science.gov (United States)

2010-01-01

... 1 General Provisions 1 2010-01-01 2010-01-01 false Publication required. 10.10 Section 10.10 General Provisions ADMINISTRATIVE COMMITTEE OF THE FEDERAL REGISTER SPECIAL EDITIONS OF THE FEDERAL REGISTER PRESIDENTIAL PAPERS Annual Publication § 10.10 Publication required. The Director of the Federal...

14 CFR 121.433 - Training required.

Science.gov (United States)

2010-01-01

... procedures set forth in a certificate holder's approved low-altitude windshear flight training program when... 14 Aeronautics and Space 3 2010-01-01 2010-01-01 false Training required. 121.433 Section 121.433..., FLAG, AND SUPPLEMENTAL OPERATIONS Crewmember Qualifications § 121.433 Training required. (a) Initial...

1 CFR 10.1 - Publication required.

Science.gov (United States)

2010-01-01

... 1 General Provisions 1 2010-01-01 2010-01-01 false Publication required. 10.1 Section 10.1 General Provisions ADMINISTRATIVE COMMITTEE OF THE FEDERAL REGISTER SPECIAL EDITIONS OF THE FEDERAL REGISTER PRESIDENTIAL PAPERS Regular Publication § 10.1 Publication required. The Director publishes a special edition...

49 CFR 1503.101 - TSA requirements.

Science.gov (United States)

2010-10-01

... 49 Transportation 9 2010-10-01 2010-10-01 false TSA requirements. 1503.101 Section 1503.101... Scope of Investigative and Enforcement Procedures § 1503.101 TSA requirements. (a) The investigative and enforcement procedures in this part apply to TSA's investigation and enforcement of violations of TSA...

15 CFR 806.3 - Reporting requirements.

Science.gov (United States)

2010-01-01

... be required from among others, U.S. persons which have direct investment abroad, U.S. persons in which foreign persons have direct investment, U.S. intermediaries, and U.S. persons which assist or... OF ECONOMIC ANALYSIS, DEPARTMENT OF COMMERCE DIRECT INVESTMENT SURVEYS § 806.3 Reporting requirements...

48 CFR 8.1102 - Presolicitation requirements.

Science.gov (United States)

2010-10-01

... ACQUISITION PLANNING REQUIRED SOURCES OF SUPPLIES AND SERVICES Leasing of Motor Vehicles 8.1102 Presolicitation requirements. (a) Except as specified in 8.1102(b), before preparing solicitations for leasing of... automobiles (sedans and station wagons) larger than Type IA, IB, or II (small, subcompact, or compact) are...

18 CFR 284.126 - Reporting requirements.

Science.gov (United States)

2010-04-01

... 18 Conservation of Power and Water Resources 1 2010-04-01 2010-04-01 false Reporting requirements. 284.126 Section 284.126 Conservation of Power and Water Resources FEDERAL ENERGY REGULATORY COMMISSION... AUTHORITIES Certain Transportation by Intrastate Pipelines § 284.126 Reporting requirements. (a) Notice of...

48 CFR 509.306 - Solicitation requirements.

Science.gov (United States)

2010-10-01

... 48 Federal Acquisition Regulations System 4 2010-10-01 2010-10-01 false Solicitation requirements... COMPETITION AND ACQUISITION PLANNING CONTRACTOR QUALIFICATIONS First Article Testing and Approval 509.306... contracting officer must address the requirements in FAR 9.306(d) and (f) through (j) in the solicitation's...

27 CFR 5.33 - Additional requirements.

Science.gov (United States)

2010-04-01

... 27 Alcohol, Tobacco Products and Firearms 1 2010-04-01 2010-04-01 false Additional requirements. 5.33 Section 5.33 Alcohol, Tobacco Products and Firearms ALCOHOL AND TOBACCO TAX AND TRADE BUREAU, DEPARTMENT OF THE TREASURY LIQUORS LABELING AND ADVERTISING OF DISTILLED SPIRITS Labeling Requirements for...

A fuzzy model for exploiting customer requirements

Directory of Open Access Journals (Sweden)

Zahra Javadirad

2016-09-01

Full Text Available Nowadays, Quality function deployment (QFD is one of the total quality management tools, where customers’ views and requirements are perceived and using various techniques improves the production requirements and operations. The QFD department, after identification and analysis of the competitors, takes customers’ feedbacks to meet the customers’ demands for the products compared with the competitors. In this study, a comprehensive model for assessing the importance of the customer requirements in the products or services for an organization is proposed. The proposed study uses linguistic variables, as a more comprehensive approach, to increase the precision of the expression evaluations. The importance of these requirements specifies the strengths and weaknesses of the organization in meeting the requirements relative to competitors. The results of these experiments show that the proposed method performs better than the other methods.

TWRSview system requirements specification

International Nuclear Information System (INIS)

Caldwell, J.A.; Lee, A.K.

1995-12-01

This document provides the system requirements specification for the TWRSview software system. The TWRSview software system is being developed to integrate electronic data supporting the development of the TWRS technical baseline

Standby battery requirements for telecommunications power

Energy Technology Data Exchange (ETDEWEB)

May, G.J. [The Focus Partnership, 126 Main Street, Swithland, Loughborough, Leics LE12 8TJ (United Kingdom)

2006-08-25

The requirements for standby power for telecommunications are changing as the network moves from conventional systems to Internet Protocol (IP) telephony. These new systems require higher power levels closer to the user but the level of availability and reliability cannot be compromised if the network is to provide service in the event of a failure of the public utility. Many parts of these new networks are ac rather than dc powered with UPS systems for back-up power. These generally have lower levels of reliability than dc systems and the network needs to be designed such that overall reliability is not reduced through appropriate levels of redundancy. Mobile networks have different power requirements. Where there is a high density of nodes, continuity of service can be reasonably assured with short autonomy times. Furthermore, there is generally no requirement that these networks are the provider of last resort and therefore, specifications for continuity of power are directed towards revenue protection and overall reliability targets. As a result of these changes, battery requirements for reserve power are evolving. Shorter autonomy times are specified for parts of the network although a large part will continue to need support for hours rather minutes. Operational temperatures are increasing and battery solutions that provide longer life in extreme conditions are becoming important. Different battery technologies will be discussed in the context of these requirements. Conventional large flooded lead/acid cells both with pasted and tubular plates are used in larger central office applications but the majority of requirements are met with valve-regulated lead/acid (VRLA) batteries. The different types of VRLA battery will be described and their suitability for various applications outlined. New developments in battery construction and battery materials have improved both performance and reliability in recent years. Alternative technologies are also being proposed

Warfarin Dosing Algorithms Underpredict Dose Requirements in Patients Requiring ≥7 mg Daily: A Systematic Review and Meta-analysis.

Science.gov (United States)

Saffian, S M; Duffull, S B; Wright, Dfb

2017-08-01

There is preliminary evidence to suggest that some published warfarin dosing algorithms produce biased maintenance dose predictions in patients who require higher than average doses. We conducted a meta-analysis of warfarin dosing algorithms to determine if there exists a systematic under- or overprediction of dose requirements for patients requiring ≥7 mg/day across published algorithms. Medline and Embase databases were searched up to September 2015. We quantified the proportion of over- and underpredicted doses in patients whose observed maintenance dose was ≥7 mg/day. The meta-analysis included 47 evaluations of 22 different warfarin dosing algorithms from 16 studies. The meta-analysis included data from 1,492 patients who required warfarin doses of ≥7 mg/day. All 22 algorithms were found to underpredict warfarin dosing requirements in patients who required ≥7 mg/day by an average of 2.3 mg/day with a pooled estimate of underpredicted doses of 92.3% (95% confidence interval 90.3-94.1, I 2 = 24%). © 2017 American Society for Clinical Pharmacology and Therapeutics.

Reserve Requirements and Monetary Management; An Introduction

OpenAIRE

International Monetary Fund

1993-01-01

Reserve requirements are widely used by central banks as a means to improve monetary control, an instrument for policy implementation, a source of revenue, and a safeguard of bank liquidity. The effectiveness of reserve requirements in fulfilling these functions is reviewed, and the detailed modalities of their use are examined. Reserve requirements in a sample of developing countries are described.

14 CFR 99.9 - Radio requirements.

Science.gov (United States)

2010-01-01

... 14 Aeronautics and Space 2 2010-01-01 2010-01-01 false Radio requirements. 99.9 Section 99.9 Aeronautics and Space FEDERAL AVIATION ADMINISTRATION, DEPARTMENT OF TRANSPORTATION (CONTINUED) AIR TRAFFIC AND GENERAL OPERATING RULES SECURITY CONTROL OF AIR TRAFFIC General § 99.9 Radio requirements. (a) A...

9 CFR 93.106 - Quarantine requirements.

Science.gov (United States)

2010-01-01

... Section 93.106 Animals and Animal Products ANIMAL AND PLANT HEALTH INSPECTION SERVICE, DEPARTMENT OF... other than ratites and hatching eggs of ratites. Each lot of pet birds, except as provided for in § 93.... (2) Physical plant requirements. The facility shall comply with the following requirements: (i...

Requirements elicitation for geo-information solutions

NARCIS (Netherlands)

Robbi Sluter, Claudia; van Elzakker, Corné P.J.M.; Ivanova, Ivana

2017-01-01

Geo-information solutions can achieve a higher level of quality if they are developed in accordance with a user-centred design that requires definition of the user requirements in the first step of solution construction. We treat a geo-information solution as a system designed to support human-based

27 CFR 4.38 - General requirements.

Science.gov (United States)

2010-04-01

... mandatory information required on labels by this part, except the alcoholic content statement, shall be in... OF THE TREASURY LIQUORS LABELING AND ADVERTISING OF WINE Labeling Requirements for Wine § 4.38... descriptive or explanatory information, the script, type, or printing of the mandatory information shall be of...

8 CFR 312.1 - Literacy requirements.

Science.gov (United States)

2010-01-01

... 8 Aliens and Nationality 1 2010-01-01 2010-01-01 false Literacy requirements. 312.1 Section 312.1... FOR NATURALIZATION § 312.1 Literacy requirements. (a) General. Except as otherwise provided in... determining English proficiency, as outlined in paragraph(c) of this section. (c) Literacy examination—(1...

40 CFR 59.105 - Reporting requirements.

Science.gov (United States)

2010-07-01

... 40 Protection of Environment 5 2010-07-01 2010-07-01 false Reporting requirements. 59.105 Section... Volatile Organic Compound Emission Standards for Automobile Refinish Coatings § 59.105 Reporting requirements. (a) Each regulated entity must submit an initial report no later than January 11, 1999 or within...

16 CFR 307.2 - Required warnings.

Science.gov (United States)

2010-01-01

... 16 Commercial Practices 1 2010-01-01 2010-01-01 false Required warnings. 307.2 Section 307.2 Commercial Practices FEDERAL TRADE COMMISSION REGULATIONS UNDER SPECIFIC ACTS OF CONGRESS REGULATIONS UNDER... Comprehensive Smokeless Tobacco Health Education Act of 1986 is the law that requires the enactment of these...

13 CFR 307.5 - Application requirements.

Science.gov (United States)

2010-01-01

... 13 Business Credit and Assistance 1 2010-01-01 2010-01-01 false Application requirements. 307.5 Section 307.5 Business Credit and Assistance ECONOMIC DEVELOPMENT ADMINISTRATION, DEPARTMENT OF COMMERCE... EDA) a CEDS, except that a CEDS is not required when applying for a Strategy Grant; and (2) Explain...

32 CFR 636.9 - Registration requirement.

Science.gov (United States)

2010-07-01

... 32 National Defense 4 2010-07-01 2010-07-01 true Registration requirement. 636.9 Section 636.9 National Defense Department of Defense (Continued) DEPARTMENT OF THE ARMY (CONTINUED) LAW ENFORCEMENT AND CRIMINAL INVESTIGATIONS MOTOR VEHICLE TRAFFIC SUPERVISION (SPECIFIC INSTALLATIONS) Fort Stewart, Georgia § 636.9 Registration requirement. In...

10 CFR 451.7 - Metering requirements.

Science.gov (United States)

2010-01-01

... 10 Energy 3 2010-01-01 2010-01-01 false Metering requirements. 451.7 Section 451.7 Energy DEPARTMENT OF ENERGY ENERGY CONSERVATION RENEWABLE ENERGY PRODUCTION INCENTIVES § 451.7 Metering requirements. The net electric energy generated and sold (kilowatt-hours) by the owner or operator of a qualified...

49 CFR 451.25 - Required information.

Science.gov (United States)

2010-10-01

... 49 Transportation 6 2010-10-01 2010-10-01 false Required information. 451.25 Section 451.25....25 Required information. (a) The safety approval number appearing on line 1 of the safety approval... safety approval plate on the freight container provided that all the information contained on the...

24 CFR 511.11 - Project requirements.

Science.gov (United States)

2010-04-01

... low income occupancy or rent projections, or restrictions on return on investment or other similar policies that prevent an owner, whether for-profit or non-profit, from maximizing return or setting rent... requirements and with the affirmative marketing requirements and procedures adopted under § 511.13, for the...

76 FR 3485 - Required Scale Tests

Science.gov (United States)

2011-01-20

...-AB10 Required Scale Tests AGENCY: Grain Inspection, Packers and Stockyards Administration, USDA. ACTION... their scales tested at least twice each calendar year at intervals of approximately 6 months. This final rule requires that regulated entities complete the first of the two scale tests between January 1 and...

76 FR 18348 - Required Scale Tests

Science.gov (United States)

2011-04-04

... RIN 0580-AB10 Required Scale Tests AGENCY: Grain Inspection, Packers and Stockyards Administration... published a document in the Federal Register on January 20, 2011 (76 FR 3485), defining required scale tests... the last sentence of paragraph (a) to read as follows: Sec. 201.72 Scales; testing of. (a...

1 CFR 9.1 - Publication required.

Science.gov (United States)

2010-01-01

... 1 General Provisions 1 2010-01-01 2010-01-01 false Publication required. 9.1 Section 9.1 General Provisions ADMINISTRATIVE COMMITTEE OF THE FEDERAL REGISTER SPECIAL EDITIONS OF THE FEDERAL REGISTER THE UNITED STATES GOVERNMENT MANUAL § 9.1 Publication required. The Director of the Federal Register shall...

21 CFR 610.40 - Test requirements.

Science.gov (United States)

2010-04-01

... appropriate: Donor Testing Status Label Tests negative Label as required under § 606.121 Tested negative... “DONOR UNTESTED” Tests negative Label as required under § 606.121 Reactive on current collection/reactive....121 of this chapter and with a label entitled “INTENDED RECIPIENT INFORMATION LABEL” containing the...

Problem-Oriented Requirements in Practice

DEFF Research Database (Denmark)

Lauesen, Søren

2018-01-01

[Context and motivation] Traditional requirements describe what the system shall do. This gives suppliers little freedom to use what they have already. In contrast, problem-oriented requirements describe the customer’s demands: what he wants to use the system for and which problems he wants...... to remove. The supplier specifies how his system will deal with these issues. The author developed the problem-oriented approach in 2007 on request from the Danish Government, and named it SL-07. [Question/problem] SL-07 has been used in many projects – usually with success. However, we had no detailed re......¬ports of the effects. [Princi¬pal ideas/results] This paper is a case study of SL-07 in acquisition of a complex case-management system. The author wrote the requirements and managed the supplier selection. Next, he was asked to run the entire acquisition project, although he was a novice project manager. Some...

Towards the Proper Integration of Extra-Functional Requirements

Directory of Open Access Journals (Sweden)

Elke Hochmuller

1999-05-01

Full Text Available In spite of the many achievements in software engineering, proper treatment of extra-functional requirements (also known as non-functional requirements within the software development process is still a challenge to our discipline. The application of functionality-biased software development methodologies can lead to major contradictions in the joint modelling of functional and extra-functional requirements. Based on a thorough discussion on the nature of extra-functional requirements as well as on open issues in coping with them, this paper emphasizes the role of extra-functional requirements in the software development process. Particularly, a framework supporting the explicit integration of extra functional requirements into a conventional phase-driven process model is proposed and outlined.

Toward the Decision Tree for Inferring Requirements Maturation Types

Science.gov (United States)

Nakatani, Takako; Kondo, Narihito; Shirogane, Junko; Kaiya, Haruhiko; Hori, Shozo; Katamine, Keiichi

Requirements are elicited step by step during the requirements engineering (RE) process. However, some types of requirements are elicited completely after the scheduled requirements elicitation process is finished. Such a situation is regarded as problematic situation. In our study, the difficulties of eliciting various kinds of requirements is observed by components. We refer to the components as observation targets (OTs) and introduce the word “Requirements maturation.” It means when and how requirements are elicited completely in the project. The requirements maturation is discussed on physical and logical OTs. OTs Viewed from a logical viewpoint are called logical OTs, e.g. quality requirements. The requirements of physical OTs, e.g., modules, components, subsystems, etc., includes functional and non-functional requirements. They are influenced by their requesters' environmental changes, as well as developers' technical changes. In order to infer the requirements maturation period of each OT, we need to know how much these factors influence the OTs' requirements maturation. According to the observation of actual past projects, we defined the PRINCE (Pre Requirements Intelligence Net Consideration and Evaluation) model. It aims to guide developers in their observation of the requirements maturation of OTs. We quantitatively analyzed the actual cases with their requirements elicitation process and extracted essential factors that influence the requirements maturation. The results of interviews of project managers are analyzed by WEKA, a data mining system, from which the decision tree was derived. This paper introduces the PRINCE model and the category of logical OTs to be observed. The decision tree that helps developers infer the maturation type of an OT is also described. We evaluate the tree through real projects and discuss its ability to infer the requirements maturation types.

Technical requirements document for the waste flow analysis

International Nuclear Information System (INIS)

Shropshire, D.E.

1996-05-01

Purpose of this Technical Requirements Document is to define the top level customer requirements for the Waste Flow Analysis task. These requirements, once agreed upon with DOE, will be used to flow down subsequent development requirements to the model specifications. This document is intended to be a ''living document'' which will be modified over the course of the execution of this work element. Initial concurrence with the technical functional requirements from Environmental Management (EM)-50 is needed before the work plan can be developed

Deep Borehole Field Test Requirements and Controlled Assumptions.

Energy Technology Data Exchange (ETDEWEB)

Hardin, Ernest [Sandia National Lab. (SNL-NM), Albuquerque, NM (United States)

2015-07-01

This document presents design requirements and controlled assumptions intended for use in the engineering development and testing of: 1) prototype packages for radioactive waste disposal in deep boreholes; 2) a waste package surface handling system; and 3) a subsurface system for emplacing and retrieving packages in deep boreholes. Engineering development and testing is being performed as part of the Deep Borehole Field Test (DBFT; SNL 2014a). This document presents parallel sets of requirements for a waste disposal system and for the DBFT, showing the close relationship. In addition to design, it will also inform planning for drilling, construction, and scientific characterization activities for the DBFT. The information presented here follows typical preparations for engineering design. It includes functional and operating requirements for handling and emplacement/retrieval equipment, waste package design and emplacement requirements, borehole construction requirements, sealing requirements, and performance criteria. Assumptions are included where they could impact engineering design. Design solutions are avoided in the requirements discussion. Deep Borehole Field Test Requirements and Controlled Assumptions July 21, 2015 iv ACKNOWLEDGEMENTS This set of requirements and assumptions has benefited greatly from reviews by Gordon Appel, Geoff Freeze, Kris Kuhlman, Bob MacKinnon, Steve Pye, David Sassani, Dave Sevougian, and Jiann Su.

BER Science Network Requirements

Energy Technology Data Exchange (ETDEWEB)

Alapaty, Kiran; Allen, Ben; Bell, Greg; Benton, David; Brettin, Tom; Canon, Shane; Dart, Eli; Cotter, Steve; Crivelli, Silvia; Carlson, Rich; Dattoria, Vince; Desai, Narayan; Egan, Richard; Tierney, Brian; Goodwin, Ken; Gregurick, Susan; Hicks, Susan; Johnston, Bill; de Jong, Bert; Kleese van Dam, Kerstin; Livny, Miron; Markowitz, Victor; McGraw, Jim; McCord, Raymond; Oehmen, Chris; Regimbal, Kevin; Shipman, Galen; Strand, Gary; Flick, Jeff; Turnbull, Susan; Williams, Dean; Zurawski, Jason

2010-11-01

The Energy Sciences Network (ESnet) is the primary provider of network connectivity for the US Department of Energy Office of Science, the single largest supporter of basic research in the physical sciences in the United States. In support of the Office of Science programs, ESnet regularly updates and refreshes its understanding of the networking requirements of the instruments, facilities, scientists, and science programs that it serves. This focus has helped ESnet to be a highly successful enabler of scientific discovery for over 20 years. In April 2010 ESnet and the Office of Biological and Environmental Research, of the DOE Office of Science, organized a workshop to characterize the networking requirements of the science programs funded by BER. The requirements identified at the workshop are summarized and described in more detail in the case studies and the Findings section. A number of common themes emerged from the case studies and workshop discussions. One is that BER science, like many other disciplines, is becoming more and more distributed and collaborative in nature. Another common theme is that data set sizes are exploding. Climate Science in particular is on the verge of needing to manage exabytes of data, and Genomics is on the verge of a huge paradigm shift in the number of sites with sequencers and the amount of sequencer data being generated.

LEGACY MANAGEMENT REQUIRES INFORMATION

International Nuclear Information System (INIS)

CONNELL, C.W.; HILDEBRAND, R.D.

2006-01-01

''Legacy Management Requires Information'' describes the goal(s) of the US Department of Energy's Office of Legacy Management (LM) relative to maintaining critical records and the way those goals are being addressed at Hanford. The paper discusses the current practices for document control, as well as the use of modern databases for both storing and accessing the data to support cleanup decisions. In addition to the information goals of LM, the Hanford Federal Facility Agreement and Consent Order, known as the ''Tri-Party Agreement'' (TPA) is one of the main drivers in documentation and data management. The TPA, which specifies discrete milestones for cleaning up the Hanford Site, is a legally binding agreement among the US Department of Energy (DOE), the Washington State Department of Ecology (Ecology), and the US Environmental Protection Agency (EPA). The TPA requires that DOE provide the lead regulatory agency with the results of analytical laboratory and non-laboratory tests/readings to help guide them in making decisions. The Agreement also calls for each signatory to preserve--for at least ten years after the Agreement has ended--all of the records in its or its contractors, possession related to sampling, analysis, investigations, and monitoring conducted. The tools used at Hanford to meet TPA requirements are also the tools that can satisfy the needs of LM

Margin Requirements and Equity Option Returns

DEFF Research Database (Denmark)

Hitzemann, Steffen; Hofmann, Michael; Uhrig-Homburg, Marliese

In equity option markets, traders face margin requirements both for the options themselves and for hedging-related positions in the underlying stock market. We show that these requirements carry a significant margin premium in the cross-section of equity option returns. The sign of the margin...... premium depends on demand pressure: If end-users are on the long side of the market, option returns decrease with margins, while they increase otherwise. Our results are statistically and economically significant and robust to different margin specifications and various control variables. We explain our...... findings by a model of funding-constrained derivatives dealers that require compensation for satisfying end-users’ option demand....

Margin Requirements and Equity Option Returns

DEFF Research Database (Denmark)

Hitzemann, Steffen; Hofmann, Michael; Uhrig-Homburg, Marliese

In equity option markets, traders face margin requirements both for the options themselves and for hedging-related positions in the underlying stock market. We show that these requirements carry a significant "margin premium" in the cross-section of equity option returns. The sign of the margin...... premium depends on demand pressure: If end-users are on the long side of the market, option returns decrease with margins, while they increase otherwise. Our results are statistically and economically significant and robust to different margin specifications and various control variables. We explain our...... findings by a model of funding-constrained derivatives dealers that require compensation for satisfying end-users’ option demand....

Nonfunctional requirements in systems analysis and design

CERN Document Server

Adams, Kevin MacG

2015-01-01

This book will help readers gain a solid understanding of non-functional requirements inherent in systems design endeavors. It contains essential information for those who design, use, and maintain complex engineered systems, including experienced designers, teachers of design, system stakeholders, and practicing engineers. Coverage approaches non-functional requirements in a novel way by presenting a framework of four systems concerns into which the 27 major non-functional requirements fall: sustainment, design, adaptation, and viability. Within this model, the text proceeds to define each non-functional requirement, to specify how each is treated as an element of the system design process, and to develop an associated metric for their evaluation. Systems are designed to meet specific functional needs. Because non-functional requirements are not directly related to tasks that satisfy these proposed needs, designers and stakeholders often fail to recognize the importance of such attributes as availability, su...

Instrumentation requirements for the ESF thermomechanical experiments

International Nuclear Information System (INIS)

Pott, J.; Brechtel, C.E.

1992-01-01

In situ thermomechanical experiments are planned as part of the Yucca Mountain Site Characterization Project that require instruments to measure stress and displacement at temperatures that exceed the typical specifications of existing geotechnical instruments. A high degree of instrument reliability will also be required to satisfy the objectives of the experiments, therefore a study was undertaken to identify areas where improvement in instrument performance was required. A preliminary list of instruments required for the experiments was developed, based on existing test planning and analysis. Projected temperature requirements were compared to specifications of existing instruments to identify instrumentation development needs. Different instrument technologies, not currently employed in geotechnical instrumentation, were reviewed to identify potential improvements of existing designs for the high temperature environment. Technologies with strong potentials to improve instrument performance with relatively high reliability include graphite fiber composite materials, fiber optics, and video imagery

40 CFR 63.1018 - Reporting requirements.

Science.gov (United States)

2010-07-01

...) National Emission Standards for Equipment Leaks-Control Level 1 § 63.1018 Reporting requirements. (a... § 63.1007(e)(2) and those pumps complying with the closed vent system provisions of § 63.1007(e)(3). (iv) Number of compressors subject to the requirements of § 63.1012, excluding those compressors...